MX2008005658A - Uses of anti-cd40 antibodies - Google Patents

Abstract

Methods for treating a human patient for an inflammatory or autoimmune disease that is associated with CD40-expressing cells are provided, where the human patient is heterozygous or homozygous for FcγRIIIa-158F (genotype V/F or F/F). Also provided are methods of inhibiting antibody production by B cells in a human patient who is heterozygous or homozygous for FcγRIIIa-158F (genotype V/F or F/F). The methods comprise administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody. Methods and kits for identifying a human patient with an inflammatory or autoimmune disease that is treatable with an anti-CD40 antibody and which is non-responsive or refractory to treatment with rituximab (Rituxan®), as well as methods and kits for selecting an antibody therapy for treatment of a human patienthaving an inflammatory or autoimmune disease that is non-responsive or refractory to treatment with rituximab (Rituxan®), are also provided. The methods of the present invention find use in treatment of inflammatory diseases and autoimmune diseases that are associated with CD40-expressing cells. These methods are particularly advantageous with respect to inflammatory diseases and autoimmune diseases that are associated with cells expressing both CD40 and CD20, as the methods enable the treatment of patients having an inflammatory or autoimmune disease that is non-responsive or refractory to therapy with other therapeutic agents such as anti-CD20 antibodies.

Description

USES OF ANTIBODIES ANTI-CD40 FIELD OF THE INVENTION This invention relates to new uses of anti-CD40 antibodies, in particular in the treatment of inflammatory diseases and autoimmune diseases that are associated with cells expressing CD40.

BACKGROUND OF THE INVENTION Many members of the family of tumor necrosis factor (TNF) ligands and their corresponding receptors regulate the growth of normal cells by inducing apoptosis or increasing cell survival and proliferation. It is this balance between apoptotic signals and survival and proliferation signals that maintains normal cellular homeostasis. At least 26 recipients of the TNF family and 18 ligands of the TNF family have been identified to date. The biologically active forms of both the receptors and the ligands are trimers of self-assembled proteins. Transmembrane and soluble forms of both the receptors and the ligands have been identified. Although the intracellular domains of the receptors do not share sequence homology, their extracellular domains comprise cysteine-rich repeat sequences of 40 amino acids. Their cytoplasmic tails emit signals by interacting with two main groups of intracellular proteins: factors associated with the TNF receptor (TRAFs) and proteins that contain domains of death (DD). The interaction between at least six human TRAFs and TRAF binding sites in the cytoplasmic tail of some of these receptors initiates several signaling pathways, including AKT (serine / threonine kinase referred to as protein kinase B or PKB), nuclear factor KB (NF-KB) and protein mitogen-activated kinases (MAPK). See, for example, the review by Younes and Kadin (2003) J Clin. Oncol. 18: 3526-3534. The CD40 member of receptors of the TNF family is a 50-55 kDa cell surface antigen present on the surface of B cells, dendritic cells, monocytes, macrophages, CD8 + T cells, endothelial cells and human epithelial and monocytic cells. The antigen CD40 is also expressed on activated T cells, activated platelets, smooth muscle vascular cells, eosinophils, synovial membranes in rheumatoid arthritis, dermal fibroblasts and other non-lymphoid cell types. Depending on the type of cell expressing CD40, ligation can induce intercellular adhesion, differentiation, activation and proliferation. For example, the binding of CD40 to its known ligand, CD40L (also referred to as CD154), stimulates the proliferation and differentiation of B cells into plasma cells, production of antibodies, isotype switching and generation of B cell memory. Differentiation of B cells, CD40 is expressed in pre B cells, but is lost with differentiation towards plasma cells. The CD40 ligand (CD40L), also known as CD154, is a 32-33 kDa transmembrane protein that also exists in two smaller soluble forms, biologically active, 18 kDa and 31 kDa, respectively (Graf ef al. (1995) Eur J. Immunol 25: 1749-1754; Mazzei ef al. (1995) J Biol. Chem. 270: 7025-7028; Pietravalle et al. (1996) J. Biol. Chem. 271: 5965-5967). CD40L is expressed on activated CD4 + helper T cells, but not at rest (Lañe ef al. (1992) Eur. J. Immunol. 22: 2573-2578; Spriggs ef al. (1992) J Exp. Med. 176: 1543-1550; and Roy ef al. (1993) J Immunol. 151: 1-14). Both CD40 and CD40L have been cloned and characterized (Stamenkovi et al. (1989) EMBO J. 8: 1403-1410; Armitage et al. (1992) Nature 357: 80-82; Lederman et al. (1992) J Exp. Med 175: 1091-1101; and Hollenbaugh et al. (1992) EMBO J. 11 4313-4321). See also U.S. Patent No. 5,945,513, which describes the human CD40L. Cells transfected with the CD40L gene and expressing the CD40L protein on their surface can activate the proliferation of B cells and, together with other stimulatory signals, can induce the production of antibodies (Armitage et al. (1992) supra).; and U.S. Patent No. 5,945,513). Patients with autoimmune disease have elevated serum levels of soluble CD40L (CD40Ls) that are not seen in healthy subjects. Overexpression of CD40L causes autoimmune diseases similar to systemic lupus erythematosus in rodent models (Higuchi et al (2002) J Immunol., 168: 9-12). In contrast, the absence of functional CD40L in activated T cells causes the hyperimmunoglobulinemia M syndrome associated with the X chromosome (Alien ef al (1993) Science 259: 990; Korthauer et al. (1993) Nature 361: 539). In addition, blocking the CD40 / CD40L interaction can prevent transplant rejection in non-human primate models. See, for example, Wee ef al. (1992) Transplantation 53: 501-7. The expression of CD40 in the APCs represents an important co-stimulatory role in the activation of these cells. For example, agonistic anti-CD40 monoclonal antibodies (mAbs) have been shown to mimic the effects of helper T cells on the activation of B cells. When presented in adherent cells expressing FcγRII, these antibodies induce B cell proliferation. (Banchereau et al (1989) Science 251: 70). Furthermore, agonistic anti-CD40 mAbs can replace the signal of helper T cells for the secretion of IgM, IgG, and IgE in the presence of IL-4 (Gasean et al (1991) J. Immunol. 147: 8) . Additionally, agonistic anti-CD40 mAbs can prevent programmed cell death (apoptosis) of B cells isolated from lymph nodes. These and other observations support the current theory that the interaction of CD40 and CD40L represents a fundamental role in the regulation of both humoral and cell-mediated immune responses. More recent studies have revealed a much wider role of the CD40 / CD40L interaction in various physiological and pathological processes. The signal transduction pathway of CD40 depends on the coordinated regulation of many intracellular factors. Like other members of the TNF receptor family, CD40 activates TRAFs, including TRAF-2, -3, -5 and -6, which over-regulate various signaling pathways after CD40 coupling with CD40L (either CD40L membrane-bound or soluble CD40L), including kinase regulated by extracellular signals (ERK), c-jun amino terminal kinase (JNK), p38 MAPK and NF-? B (see, for example, Younes and Carbone (1999) Int. J. Biol. Markers 14: 135-143; van Kooten and Banchereau (2000) J. Leukoc. Biol. 67: 2-17). It has been shown that CD40 signaling prevents cell death from apoptosis (Makus et al (2002) J. Immunol., 14: 973-982). Apoptotic signals are necessary to induce programmed cell death in a coordinated manner. Signs of cell death can include intrinsic stimuli from within the cell, such as reticule tension endoplasmic or extrinsic stimuli such as binding to FasL or TNFa receptors. The signaling pathway is complex, which involves activation of caspases such as Caspasa-3 and Caspasa-9, and poly (ADP ribose) polymerase (PARP). Over the course of the cascade, anti-apoptotic signaling proteins, such as mcl-1 and bcl-x, and members of the lAP protein family, such as the Chromosome-Associated Apoptosis Inhibitor (XIAP), are sub-regulated. (Budihardjo et al (1999) Annu. Rev. Cell Dev. Biol. 15: 269-290). For example, in dendritic cells, cell signaling by CD40 can block apoptosis signals transduced by FasL (Bjorck et al (1997) Intl. Immunol., 9: 365-372). In this way, the coupling of CD40 by CD40L, and the subsequent activation of CD40 signaling, are necessary steps for normal immune responses; however, destabilization in regulation of CD40 signaling can lead to disease. It has been shown that the CD40 signaling pathway is involved in the autoimmune disease (Ichikawa et al (2002) J Immunol. 169: 2781-2787 and Moore ef al (2002) J. Autoimmun. 19: 139-145) . Additionally, the CD40 / CD40L interaction represents an important role in inflammatory processes. For example, both CD40 and CD40L are overexpressed in human and experimental atherosclerosis lesions. The stimulation of CD40 induces the expression of matrix degradation enzymes and the expression of tissue factors in cell types associated with atheromas, such as endothelial cells, smooth muscle cells and macrophages. further, stimulation of CD40 induces the production of proinflammatory cytokines such as IL-5 IL-6 and IL-8, and adhesion molecules such as ICAM-I, E-selectin and VCAM. Inhibition of the CD40 / CD40L interaction prevents atherogenesis in animal models. In transplant models, the blocking of the CD40 / CD40L interaction prevents inflammation. It has been shown that the CD40 / CD40L junction acts synergistically with Alzheimer's amyloid-beta peptide to promote activation of the microglia cells, thereby leading to neurotoxicity. In patients with rheumatoid arthritis (RA), CD40 expression is increased in articular chondrocytes, thus, CD40 signaling probably contributes to the production of harmful matrix cytokines and matrix metalloproteinases. See, Gotoh ef al. (2004) J Rheumatol. 31: 1506-1512. In addition, it has been shown that the amplification of the synovial inflammatory response occurs through the activation of MAPKs and NF-? B through the ligation of CD40 in CD14 + synovial cells from RA patients (Harigai et al. (2004) Arthritis Rheum 50: 2167-2177). In an experimental model of RA, treatment with the anti-CD40L antibody prevented the induction of disease, joint inflammation and production of anti-collagen antibodies (Durie et al (1993) Science 261: 1328-1330). Finally, in clinical trials, it has been shown that completely removing CD20 + positive B cells from patients with RA by administering Rituxan® (generally indicated for B-cell lymphoma) improves symptoms (Shaw et al. (2003) Ann. Rheum, Dis. 62 (Suppl 2): 55-58). It has also been shown that blocking CD40 / CD40L interactions, during the presentation of antigens to T cells, induces tolerance of T cells. Therefore, blocking the CD40 / CD40L interaction prevents the initial activation of T cells as well as it induces long-term tolerance to re-exposure to the antigen. Human anti-CD40 monoclonal antibodies, and a number of uses thereof, are described in co-proprietary patent applications published as WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307, and WO 2005/044294. Those applications specifically describe an anti-CD40 human IgG1 monoclonal antibody, designated CHIR-12.12 therein, which was generated by immunization of transgenic mice carrying the human IgGi heavy chain locus and the human K light chain locus ( XenoMouse® technology, Abgenix, California). As shown by FACS analysis, CHIR-12.12 binds specifically to human CD40 and can prevent binding of the CD40 ligand (CD40L). The CHIR-12.12 can compete with the CD40L pre-bound to the cell surface CD40. The monoclonal antibody CHIR-12.12 is a strong antagonist and inhibits the proliferation mediated by CD40L in vitro of normal and malignant B cells. The monoclonal antibody CHIR-12.12 directly inhibits the survival and signaling pathways mediated by CD40L in normal human B lymphocytes. In vitro, CHIR-12.12 removes primary cancer cells from patients with NHL by ADCC. Dose-dependent tumor activity was observed in a model of human lymphoma by xenograft. He CHIR-12.12 is currently in Phase I trials for B-cell malignancies. CD20 is a cell surface antigen expressed early on in B cell differentiation and remains on the cell surface throughout the development of B cells. CD20 is involved in the activation of B cells, is expressed at very high levels in neoplastic B cells and is a clinically recognized therapeutic target (see, for example, Hboijberg et al (1995) Cancer Research 55: 2627). Antibodies that target CD20, such as rituximab (Rituxan®), have been approved by the US Food and Drug Administration. for the treatment of non-Hodgkin's lymphoma (see, for example, Boye et al. (2003) Ann. Oncol. 14: 520). It has been shown that Rituxan® is an effective treatment for low-grade, intermediate and high-grade non-Hodgkin's lymphoma (see for example, Maloney et al (1994) Blood 84: 2457-2466), McLaughlin et al. (1998) J Clin. Oncol 16: 2825-2833, Maloney ef al (1997) Blood 90: 2188-2195, Hainsworth et al. (2000) Blood 95: 3052-3056, Colombat ef al. (2001) Blood 97: 101-106, Coiffer ef al. (1998) Blood 92: 1927-1932), Foran ef al. (2000) J. Clin. Oncol. 18: 317-324, Anderson ef al. (1997) Biochem. Soc. Trans. 25: 705-708 or Vose ef al. (1999) Ann. Oncol. 10: 58a). Rituxan® also extracts normal B cells completely, which may play a role in inflammatory and autoimmune diseases. It is found in clinical tests for autoimmune diseases. Although the exact mechanism of action is not known, the evidence indicates that the effects against Rituxan® lymphomas are due in part to complement-mediated cytotoxicity (CMC), antibody-dependent cell-mediated cytotoxicity (ADCC), inhibition of cell proliferation and, finally, direct induction of apoptosis. Some patients, however, become resistant to treatment with Rituxan® (see Witzig et al (2002) J Clin Oncol 20: 3262, Grillo-Lopez et al (1998) J Clin. Oncol. 16: 2825 or Jazirehi ef al. (2003) Mol. Cancer Ther.2: 1183-1193). For example, some patients lose CD20 expression in malignant B cells after anti-CD20 antibody therapy (Davis et al (1999) Clin Cancer Res. 5: 611). Additionally, 30% to 50% of patients with low-grade NHL do not exhibit clinical response to this monoclonal antibody (Hainsworth et al (2000) Blood 95: 3052-3056; Colombat et al. (2001) Blood 97: 101-106). It has also been shown that the clinical activity of rituximab in the NHL correlates with the Fc? Rllla genotype of the patient. Patients with the V / V or V / F polymorphism of 158aa in Fc? Rllla are more sensitive to rituximab than those with F / F (for example, see Cartron et al. (2002) Blood 99 (3): 754- 758 or Dall'Ozzo et al. (2004) Cancer Res. 64: 4664-4669). For patients who develop resistance to this monoclonal antibody, or who have an inflammatory disease or autoimmune disease that is resistant to initial therapy with this antibody, alternative forms of therapeutic intervention are needed. In addition, Rituxan® extracts the normal B cells in patients completely. Therefore, it can be used to treat autoimmune and inflammatory diseases dependent on B cells. In this way, there is a continuing need for new therapeutic agents and new therapeutic strategies for inflammatory diseases and autoimmune diseases. In particular, there is a need for new therapeutic strategies for the treatment of patients resistant to treatment with anti-CD20 antibodies, such as rituximab (Rituxan®).

SUMMARY OF THE INVENTION Methods are provided for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, where the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). The methods comprise administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody. The invention also makes possible the use of a therapeutically or prophylactically effective amount of an anti-CD40 antibody in the manufacture of a medicament for the treatment of an inflammatory disease or autoimmune disease that is associated with cells expressing CD40 in a heterozygous human patient. or homozygous for Fc? Rllla-158F (genotype V / F or F / F). Also provided are methods for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F), which comprises administering to the human patient an effective amount of an anti-CD40 antibody. The invention also makes possible the use of an effective amount of an antibody anti-CD40 in the elaboration of a medicament for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). Methods and equipment are also provided to identify a human patient with an inflammatory disease or autoimmune disease, which can be treated with an anti-CD40 antibody, and which is resistant to treatment with rituximab (Rituxan®). In some embodiments, the methods comprise: a) identifying a human patient with an inflammatory disease or autoimmune disease that is associated with cells expressing CD40 and which is resistant to treatment with rituximab (Rituxan®); and b) determining the Fc? Rllla-158 genotype (V / V, V / F or F / F) of the human patient; wherein the inflammatory disease or autoimmune disease can be treated with an anti-CD40 antibody if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). The invention may further include the step of administering, to a human patient identified using this method, a therapeutically or prophylactically effective amount of an anti-CD40 antibody. The kits of the present invention, which make it possible to identify a human patient with an inflammatory disease or autoimmune disease that can be treated with an anti-CD40 antibody, comprise reagents to determine the Fc? Rllla-158 genotype of a human patient. The invention also provides methods and equipment for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease that is resistant to treatment with rituximab (Rituxan®). In some embodiments, the methods comprise: a) identifying a human patient who has an inflammatory or autoimmune disease that is associated with cells expressing CD40 and which is resistant to treatment with rituximab (Rituxan®); and b) determining the Fc? Rllla-158 genotype (V / V, V / F or F / F) of the human patient; wherein if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), an anti-CD40 antibody is selected for the treatment of inflammatory disease or autoimmune disease. The invention may further include the step of administering, to a human patient identified using this method, a therapeutically or prophylactically effective amount of an anti-CD40 antibody. The teams of the present invention, which make it possible to select an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease associated with cells expressing CD40, comprise reagents for determining an FcγRllla-158 genotype of the human patient. The present invention also provides methods for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, wherein the methods comprise administering to the human patient a slow internalizing antibody. In a similar embodiment, a therapeutically or prophylactically effective amount of an anti-CD40 antibody is administered to the human patient in such a manner that the anti-CD40 antibody is not significantly internalized by cells expressing CD40 after administration. In another such embodiment, a therapeutically or prophylactically effective amount of an anti-CD40 antibody is administered to the human patient in such a manner that the anti-CD40 antibody remains substantially and uniformly distributed on the surface of cells expressing CD40 after administration. In yet another such embodiment, an anti-CD40 antibody is administered to the human patient in such a manner that a therapeutically or prophylactically effective amount of the anti-CD40 antibody is present on the surface of cells expressing CD40 in the human patient after administration.

The anti-CD40 antibodies, for use according to the present invention, bind specifically to the CD40 antigen. In some embodiments, anti-CD40 antibodies, for use in the methods of the present invention, in particular monoclonal antibodies, exhibit a strong binding affinity for human FcγRllla-158V, a strong binding affinity for FcγRllla -158F human or a strong binding affinity for both Fc? Rllla-158V and human Fc? Rllla-158F. In some of these embodiments, the anti-CD40 antibodies can bind to either of the two (V or F) allotypes of 158 amino acids in Fc? Rllla in the natural cytolytic (NK) cells of the human patient, with binding characteristics that are adequate to induce a potent antibody-dependent cellular cytotoxicity (ADCC). Suitable anti-CD40 antibodies include, but are not limited to, anti-CD40 antibodies that do not have significant agonist activity, including, for example, anti-CD40 antibodies that are antagonists of CD40-CD40L signaling in cells expressing CD40. In some embodiments, the anti-CD40 antibody is selected from the group consisting of: a) the monoclonal antibody CHIR-12.12; b) the monoclonal antibody produced by the hybridoma cell line 12.12; c) a monoclonal antibody comprising an amino acid sequence selected from the group consisting of the sequence shown in SEQ ID NO: 2, the sequence shown in SEQ ID NO: 4, the sequence shown in SEQ ID NO: 5, the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 4, and the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 5; d) a monoclonal antibody having an amino acid sequence encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of the sequence shown in SEQ ID NO: 1, the sequence shown in SEQ ID NO: 3, and the sequences shown in SEQ ID NO: 1 and SEQ ID NO: 3; e) a monoclonal antibody that binds to an epitope capable of binding to the monoclonal antibody produced by the hybridoma cell line 12.12; f) a monoclonal antibody that binds to an epitope comprising residues 82-87 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; g) a monoclonal antibody that binds to an epitope comprising residues 82-89 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; h) a monoclonal antibody that competes with the monoclonal antibody CHIR-12.12 in a competitive binding assay; i) the monoclonal antibody of preceding article a) or a monoclonal antibody of any of preceding articles c) -h), wherein the antibody is produced recombinantly; and j) a monoclonal antibody that is an antigen-binding fragment of a monoclonal antibody of any of the preceding items a) -i), wherein the fragment retains the ability to specifically bind to the human CD40 antigen. The methods of the present invention find use in the treatment of inflammatory diseases or autoimmune diseases that are associated with cells expressing CD40. Examples include, but are not limited to, systemic lupus erythematosus (SLE), discoid lupus, lupus nephritis, sarcoidosis, inflammatory arthritis, including juvenile arthritis, rheumatoid arthritis, psoriatic arthritis, Reiter's syndrome, ankylosing spondylitis, and gout arthritis, rejection of an organ or tissue transplant, hyperacute, acute or chronic rejection and / or graft versus host disease, multiple sclerosis, hyperimmunoglobulinemia E syndrome, polyarteritis nodosa, primary biliary cirrhosis, bowel inflammation syndrome, Crohn's disease, celiac disease (gluten-sensitive enteropathy), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia , psoriasis, scleroderma, myasthenia gravis, autoimmune thrombocytopenic purpura, autoimmune thyroiditis, Grave's disease, Hasimoto thyroiditis, immune complex disease, chronic fatigue immune dysfunction syndrome (CFIDS), polymyositis and dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy , pemphigus vulgaris, pulmonary interstitial fibrosis, type I and type II diabetes mellitus, delayed type hypersensitivity types 1, 2, 3 and 4, allergy or allergic disorders, unwanted / unintended immune responses to therapeutic proteins, asthma, Churg-Strauss syndrome (allergic granulomatosis), atopic dermatitis, dermatitis by allergic and irritant contact, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis, idiopathic inflammatory myopathies, hemolytic disease, Alzheimer's disease and chronic inflammatory demyelinating polyneuropathy, pulmonary inflammation including, but not limited to, lung graft rejection, asthma, sarcoidosis, emphysema, cystic fibrosis, idiopathic pulmonary fibrosis, chronic bronchitis, allergic rhinitis and allergic lung diseases such as hypersensitivity pneumonitis, eosinophilic pneumonia, bronchiolitis obliterans due to bone marrow and / or lung transplantation or other causes, graft atherosclerosis / graft phlebosclerosis, pulmonary fibrosis resulting from collagen, vascular and auto immune diseases such as rheumatoid arthritis and lupus erythematosus, as well as inflammatory diseases or autoimmune diseases associated with cells expressing CD20. The methods of the invention are particularly favorable with respect to inflammatory diseases and autoimmune diseases which are associated with cells expressing both CD40 and CD20. In this form, the present invention enables the treatment of patients having an inflammatory or autoimmune disease that is insensitive or resistant to therapy with other therapeutic agents, including anti-CD20 antibodies for patients who are homozygous or heterozygous for Fc? Rllla -158F (genotype V / F or F / F).

BRIEF DESCRIPTION OF THE DRAWINGS Figures 1A-1F show the results of an antibody dependent cellular cytotoxicity analysis (ADCC) in six cell lines. 1A: A-ADCC-029 (Daudi lymphoma cell line); Anti-CD40 B-Abs eliminate Daudi cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-ED50 of 12.12: 4.07 pM; Rituxan: 66.93pM. 1B: A-ADCC-036 (Namalwa lymphoma cell line); B-DAC elimination activity of Anti-CD40 Abs on Namalwa cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-EC50 of 12.12: 1.01 pM; Rituxan: 189.72 pM; 1C: A-ADCC-028 (Multiple myeloma cell line ARH77); B-ADCC elimination activity of anti-CD40 Abs on ARH77 cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-ED50 of 12.12: 36.24 pM; Rituxan: 34.77 pM; 1D: A-ADCC-041 (IM-9 myeloma cell line); B-ADCC elimination activity on IM.9 cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-EC50 of 12.12: 5.92 pM; Rituxan: 61.02 pM; 1E: A-ADCC-056 (B-CLL cell line, EHEB); B-ADCC activity on EHEB cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-EC50 of 12.12: 5.56 pM; Rituxan: 76.43pM; 1F: A-ADCC-069 (B-ALL cell line, CCRF-SB); B-ADCC elimination activity on CCRF-SB cells with EC50 pM; C-Rituxan; D-% of specific lysis; E-Log [Abs] (μg / ml); F-EC50 of 12.12: 14.06 pM; Rituxan: 24.11pM. Figures 2A-2D show the results of an antibody dependent cellular cytotoxicity assay (ADCC) in cells from patients with CLL (n = 8). 2A-A-CHIR-12.12 mediates a better ADCC than rituximab against cells from patients with CLL; B-CLL # 4; C-CHIR12.12; D-rituximab; E-% specific lysis; F-Log of conc. of Abs [μg / ml]; G-CHIR12.12; H-Rituxan; l-CLL # 21. 2B-A-CHIR-12.12 mediates a better ADCC than rituximab against cells from patients with CLL; B-CLL # 22; C-% specific lysis; D-Log of conc. of Abs [μg / ml]; E-CHIR12.12; F-Rituximab; G-CLL # 27; 2C-A-CHIR-12.12 mediates an ADCC better than ntuximab against cells from patients with CLL; B-CLL # 32; C-% specific lysis; D-Log of conc. of Abs [μg / ml]; E-CHIR12.12; F-Rituximab; G-CLL # 30; 2D-A-CHIR-12.12 mediates a better ADCC than rituximab against cells from patients with CLL; B-CLL # 31; C-% specific lysis; D-Log of conc. of Abs [μg / ml]; E-CHIR12.12; F-Rituximab; G-CLL # 39. Figure 3 summarizes the results of an ADCC analysis in cells from patients with CLL (n = 9). A- comparative ADCC of CHIR-12.12 and rituximab against cells of patients with CLL (n = 9) by human NK cells from multiple donors; B-Conc of Abs (μg / ml); C- Fraction of lysis mediated by rituximab compared with CHIR-12.12; D-CHIR12.12 EC50) p; E-Rituximab EC50 (pM). Figure 4 shows the results of an ADCC analysis in cells of patients with CLL, using NK effector cells from two different donors. 4-A-Variation of NK donor in ADCC activities in the same object cells (CLL # 33); B-Donor # 1; C-% specific lysis; D-Log [Abs] (μg / ml); E-EC50 of 12.12: 49.53 pM; Rituxan: 473.28 pM; F-rituxan; G- EC50 of 12.12: 37.85 pM; Rituxan: 141.03 pM; H-Donor # 2 of NK. Figure 5 shows the results of the quantification of cell surface expression of CD40 and CD20 in cells from patients with CLL and normal B cells. A- Quantification of CD40 and CD20 molecules expressed in cells of patient CLL and normal B cells; B- CHIR-12.12; C-Patient with CLL #; D-Molecules CD40; E-Rituximab; F-Molecules CD20; G-Human B cells (n = 2). Figure 6 summarizes ADCC activity for cells with quantified cell surface expression of CD40 and CD20. A- Relative expression of CD40 and CD20 molecules in cells of relatives with CLL and ADCC activity; B- Maximum lysis%; C-Patient with CLL #; D-CHIR-12.12; E-Rituximab; F-Difference between CHIR-12.12 and Rituximab; G-Relationship of target molecules CD20 / CD40. Figure 7 is a bar graph showing levels of CHIR-12.12 bound to cell surface in the Daudi and ARH77 cell lines. A-Levels of CHIR-12.12 bound to cell surface (measured as fluorescence intensity) after 3 hours of incubation at 4 ° C and 37 ° C; B-MFI (geometric mean); C-hlgG1; D- CHIR-12.12; E-4 ° C Daudi; F-37 ° C Daudi; G- 4 ° C ARH77; H-37 ° C ARH77. Figure 8 shows the results of the investigation of the internalization of CHIR-12.12 and rituximab in cells of patients with CLL by FACS analysis. A-Internalization in percentage of CHIR-12.12 and rituximab in cells of CLL patients (n = 8): FACS; B-CHIR-12.12; C-rituximab; D-% internalization; E-CLL #; F-Average; G-% internalization = 100 *. { (GMF of Abs of test at 4 ° C-GMF of the Abs isotype at 4 ° C) - (GMF of Abs of test at 37 ° C-GMF of the Abs isotype at 37 ° C)} / (GMF of the test Abs at 4 ° C - GMF of the Abs isotype at 4 ° C); H-GMF: geometric mean fluorescence intensity; l - * negative% of internalization indicates higher binding of CHIR-12.12 at 37 ° C compared to 4 ° C. Figure 9 shows the results of the investigation of the internalization of CHIR-12.12 and rituximab in normal B cells by confocal microscopy of antibodies labeled with FITC. A-Inteization of CHIR-12.12 and rituximab in normal human B cells: confocal microscope; B-Rituximab-FITC; C-12.12-FITC; D-lgG1-FITC; Figure 10 shows the results of the investigation of the internalization of CHIR-12.12 and htuximab in cells of patients with CLL by confocal microscopy of antibodies labeled with Alexa488. A-Internalization of CHIR-12.12 and rituximab in patient cells with CLL # 33: confocal microscope; B-Rituximab-Alexa488; C-12.12-Alexa488; D-lgG1-Alexa488. Figure 11 summarizes the relationship between ADCC activity and internalization. A- Relationship between ADCC activity and internalization; B- Maximum lysis%; C-% internalization; D-Patient with CLL #; E-CHIR-12.12; F-Rituximab; G-Difference between CHIR-12.12 and Rituximab; H-CHIR-12.12; l-Rituximab. Figure 12 is a bar graph showing the maximum percentage of specific lysis of Daudi cells by CHIR-12.12 or rituximab by purified NK effector cells from donors with different Fc? Rllla genotypes. A- maximum% of specific lysis; B-NK cell genotype; C-Genotype FcR? Llla and ADCC (Daudi). Figure 13 is a bar graph showing the potency of ADCC (EC50) of CHIR-12.12 or rituximab in Daudi cells by purified NK effector cells from donors with different Fc? Rllla genotypes. A- Genotype FcR? Llla and ADCC (Daudi); B-NK cell genotype; C-ED50 (pM). Figure 14 summarizes the comparative ADCC of CHIR-12.12 and rituximab against cells from patients with CLL (n = 9) by human NK cells from human donors of genotypes multiple A-ADCC comparative of CHIR-12.12 and rituximab against cells of patients with CLL (n = 9) by NK cells of multiple donors with FcR genotype? Illa variant; B- Max. Lysis%; C-EC 50 (pM); D-Patient # with CLL; E-CHIR-12.12; F-Rituximab; G-Difference between CHIR-12.12 and Rituximab; H-CHIR-12.12; l-Rituximab; J-Rituximab vs CHIR-12.12 times; K-Genotype FcR? Llla from donor NK; L-Media DETAILED DESCRIPTION OF THE INVENTION The surprising finding has been made that anti-CD40 antibodies, such as CHIR-12.12, are capable of mediating a potent antibody-dependent cellular cytotoxicity (ADCC) of target cells expressing CD40 under conditions where other antibodies that mediate ADCC are less effective or relatively ineffective. Contrary to other antibodies, such as rituximab (Rituxan®), the anti-CD40 antibodies, used according to the invention, can bind to either of the two (158 or 158) allotypes (V or F) in Fc? Rllla in the cytolytic cells. natural (NK) of the human patient, with binding characteristics that are suitable to elicit a potent ADCC. This finding is unexpected and represents an advance in the ability to treat inflammatory diseases and autoimmune diseases in a whole cross-section of patients. Accordingly, anti-CD40 antibodies, such as CHIR-12.12, can be used in the treatment of inflammatory diseases and autoimmune diseases associated with cells expressing CD40 in human patients heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), in addition to human patients homozygous for Fc? Rllla158V (genotype and /). The invention thus provides a method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, wherein the human patient is heterozygous or homozygous for FcγRllla-158F (genotype V) / F or F / F), the method comprises administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody. The invention also provides the use of a therapeutically or prophylactically effective amount of an anti-cancer antibody.

CD40 in the preparation of a medicament for the treatment of an inflammatory disease or autoimmune disease that is associated with cells expressing CD40 in a human patient heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). As noted in the above, it has been shown that the clinical activity of rituximab in NHL correlates with the Fc? Rllla genotype of the patient. Patients with the F / F polymorphism of 158aa in Fc? Rllla are less sensitive to rituximab than those with V / V or V / F (for example, see Cartron et al. (2002) Blood 99 (3): 754- 758 or DalPOzzo ef al. Cancer Res. (2004) 64: 4664-4669). As noted in the above, Rituxan® is in clinical trials for autoimmune diseases. Accordingly, the present invention is favorable for the treatment of inflammatory diseases and autoimmune diseases that are not sensitive to treatment with an anti-CD20 antibody such as rituximab (Rituxan®). Moreover, such potent elimination of target cells without the need to use an antibody-toxin conjugate will result in a drug that is cheaper to make and has fewer side effects. Anti-CD40 antibodies, such as CHIR-12.12, can be used in methods to inhibit the production of antibodies by B cells in a human patient heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F) , in addition to human patients homozygous for Fc? Rllla-158V (genotype V / V). Thus, the invention provides a method for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F), which comprises administering to the patient human an effective amount of an anti-CD40 antibody, such as CHIR-12.12. The invention also provides the use of an effective amount of an anti-CD40 antibody in the manufacture of a medicament for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for FcγRllla-158F (genotype V) / F or F / F). It could not be expected, by a person skilled in the art, that the production of antibodies by B cells could be inhibited in a human patient heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). The present invention allows the treatment regimen selected for a Individual human patient is based on that Fc? Rllla-158 genotype of the patient when administering an anti-CD40 antibody that mediates ADCC. The invention provides a method for identifying a human patient with an inflammatory disease or autoimmune disease, treatable with an anti-CD40 antibody, and which is resistant to treatment with rituximab (Rituxan®), which comprises: a) identifying a human patient with an inflammatory disease or autoimmune disease that is associated with cells that express CD40 and which is resistant to treatment with rituximab (Rituxan®); and b) determining the Fc? Rllla-158 genotype (V / V, V / F or F / F) of the human patient; wherein the inflammatory disease or autoimmune disease is treatable with an anti-CD40 antibody if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). The invention may further include the step of administering, to a human patient identified using this method, a therapeutically or prophylactically effective amount of an anti-CD40 antibody. This method for identifying a human patient with an inflammatory disease or autoimmune disease, treatable with an anti-CD40 antibody, can be easily performed by a person skilled in the art using a suitable diagnostic equipment. The kit should comprise suitable reagents to determine an Fc? Rllla-158 genotype of the human patient. In this manner, the invention also provides a kit for identifying a human patient with an inflammatory disease or autoimmune disease treatable with an anti-CD40 antibody, comprising reagents for determining an FcγRllla-158 genotype of the human patient. The proper equipment is described in greater detail elsewhere in the present. The invention also provides a method for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease which is resistant to treatment with rituximab (Rituxan®), which comprises: a) identifying a human patient who has an inflammatory disease or autoimmune disease that is associated with cells that express CD40 and which is resistant to treatment with rituximab (Rituxan®); and b) determining the Fc? Rllla-158 genotype (V / V, V / F or F / F) of the human patient; wherein if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), an anti-CD40 antibody is selected for the treatment of inflammatory disease or autoimmune disease. In particular, an anti-CD40 antibody can be selected in preference to treatment with rituximab (Rituxan®). The invention may further include the step of administering, to a human patient identified using this method, a therapeutically or prophylactically effective amount of an anti-CD40 antibody. This method for selecting an antibody therapy, for the treatment of a human patient having an inflammatory disease or autoimmune disease, can be easily performed by a person skilled in the art using suitable diagnostic equipment. The kit should comprise suitable reagents to determine an Fc? Rllla-158 genotype of the human patient. In this manner, the invention also provides a kit for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease associated with cells expressing CD40, which comprises reagents to determine an Fc? Rllla genotype. 158 of the human patient. The surprising finding has also been made that anti-CD40 antibodies, such as CHIR-12.12, are not internalized significantly by cells expressing CD40 after administration. Instead, anti-CD40 antibodies, such as CHIR-12.12, are distributed substantially uniformly on the surface of cells expressing CD40 for a significant period of time after administration. This is in contrast to other antibodies, in particular anti-CD20 antibodies, such as rituximab (Rituxan®). The duration of binding to CD40 on the surface of cells expressing CD40, and the uniform distribution of the anti-CD40 antibody on the surface of cells expressing CD40, enables the anti-CD40 antibodies to mediate a potent antibody-dependent cellular cytotoxicity ( ADCC) of target cells expressing CD40, by binding to an FcR, such as Fc? Rllla in natural killer cells (NK). Thus, the invention provides a method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient a therapeutically or prophylactically effective amount of an anti-cancer antibody. CD40, such that the anti-CD40 antibody is not significantly internalized by cells expressing CD40 after administration. The invention also provides a method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody, such that the anti-CD40 antibody remains substantially uniformly distributed on the surface of cells expressing CD40 after administration. The invention also provides a method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient an anti-CD40 antibody, such that a therapeutic or Prophylactically effective anti-CD40 antibody is present on the surface of cells expressing CD40 in the human patient after administration. These aspects of the invention, in this way, involve administering a slow internalizing antibody to a patient. By "slow internalization antibody" is meant an antibody that remains disposed on the cell surface for a significant period of time. As the experienced person will realize, this property contrasts with properties estimated as favorable for many therapeutic applications that actually require internalization of the antibody-receptor complex in order for therapy to be effective. In this context, a significant period of time generally exceeds 3 hours, preferably 6 hours, more preferably 12 hours, more preferably 24 hours, 36 hours, 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, 168 hours or more.

Preferably, at least 5%, at least 10%, at least 20%, at least %, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more, of the antibody initially placed on the surface of a cell expressing CD40, remains disposed on the surface of the cell after the significant period of time mentioned above. The internalization of antibodies can be assessed by various tests. For example, cell lines such as the Daudi lymphoma cell line or the ARH77MM cell line can be used to evaluate the effect of binding of a candidate antibody on internalization. The cells are incubated with human IgG1 (control antibody) or candidate antibody on ice (with 0.1% sodium azide to block internalization) or at 37 ° C (without sodium azide) for a period of time, conveniently 3 hours. After washing with cold staining buffer (for example, PBS + 1% BSA + 0.1% sodium azide), the cells are stained, for example, with goat anti-human IgG-FITC for 30 minutes on ice . The degree of staining can then be assessed; in this example, the geometric mean fluorescent intensity (MFI) can be recorded, such as by FACS Calibur. Other suitable assays will be known to those skilled in the art (see, for example, http://www.abgenix.com/documents/SBS2003%20poster.pdf). In the experiments set forth in Examples 4 and 5 herein, no difference was observed in MFI between cells incubated with CH12.12 on ice in the presence of sodium azide or at 37 ° C in the absence of sodium azide (see Figures 7-10). These data show that CH12.12, with the binding to CD40, it does not internalize and continues to be exposed on the cell surface. A summary of standard techniques and procedures is given in the following, which may be used for the purpose of using the invention. It will be understood that this invention is not limited to the particular methodology, protocols, cell lines, vectors and reagents described. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and it is not intended that this terminology should limit the scope of the present invention. The scope of the invention is limited only by the terms of the appended claims. Standard abbreviations for nucleotides and amino acids are used in this specification.

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, recombinant DNA technology and immunology, which are within the experience of those working in the art. Such techniques are fully explained in the literature. Examples of texts particularly suitable for consultation include the following: Sambrook ef al. (1989) Molecular Cloning; A Laboratory Manual (2d ed.); D.N Glover, ed. (1985) DNA Cloning, Volumes I and II; M.J. Gait, ed. (1984) Oligonucleotide Synthesis; B.D. Hames & SJ. Higgins, eds. (1984) Nucleic Acid Hybridization; B.D. Hames & S.J. Higgins, eds. (1984) Transcription and Translation; R.l. Freshney, ed. (1986) Animal Cell Culture; Immobilized Cells and Enzymes (IRL Press, 1986); B. Perbal (1984) A Practical Guide to Molecular Cloning; the Methods in Enzymology series (Academic Press, Inc.), especially volumes 154 & 155; J.H. Miller and M.P. Calos, eds. (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Mayer and Walker, eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Scopes (1987) Protein Purification: Principles and Practice (2nd ed .: Springer Verlag, N.Y.); and D.M. Weir and C. C. Blackwell, eds. (1986) Handbook of Experimental Immunology, Volumes l-IV. The methods of the invention involve the use of anti-CD40 antibodies in the treatment of inflammatory diseases and autoimmune diseases associated with cells expressing CD40. By "CD40", "CD40 antigen" or "CD40 receptor" is intended the 50-55 kDa transmembrane glycoprotein of the family of tumor necrosis factor (TNF) receptors (see, for example, U.S. Patent Nos. 5,674,492 and 4,708,871; Stamenkovic et al. (1989) EMBO 8: 1403; Clark (1990) Tissue Antigens 36:33; Barclay et al. (1997) The Leucocyte Antigen Facts Book (2nd ed., Academic Press, San Diego)). Two isoforms of human CD40 have been identified, encoded by alternately spliced variants of the transcript of this gene. The first isoform (also known as the "long isoform" or "isoform 1") is expressed as a 277 amino acid precursor polypeptide (SEQ ID NO: 9, reported first as Access GenBank No. CAA43045, and identified as isoform 1 in Access GenBank No. NP_001241), codified by SEQ ID NO: 8 (see Accesses GenBank Nos. X60592 and NM_001250), which has a signal sequence represented by the first 19 residues . The second isoform (also known as the "short isoform" or "isoform 2") is expressed as a precursor polypeptide of 203 amino acids (SEQ ID NO: 7, Access GenBank No. NP 690593), encoded by SEQ ID NO: 6 (Access GenBank No. NM_152854), which also has a signal sequence represented by the first 19 residues. The precursor polypeptides of these two isoforms of human CD40 share in common their first 165 residues (ie, residues 1-165 of SEQ ID NO: 7 and SEQ ID NO: 9). The precursor polypeptide of the short isoform (shown in SEQ ID NO: 7) is encoded by a variant of the transcript (SEQ ID NO: 6) lacking a coding segment, which leads to a change in the translation framework; the resulting CD40 isoform contains a shorter and distinct C-terminus (residues 166-203 of SEQ ID NO: 7) than that contained in the long isoform of CD40 (C-terminus shown at residues 166-277 of SEQ ID NO. : 9). For purposes of the present invention, the term "CD40," or "CD40 antigen," "cell surface CD40 antigen," or "CD40 receptor" encompasses both the short and long isoforms of CD40. The CD40 antigen can be glycosylated completely or partially. As, it is observed elsewhere herein, CD40 is found on the surface of B cells, dendritic cells, monocytes, macrophages, CD8 + T cells, endothelial cells, human monocytic and epithelial cells., both normal and neoplastic, activated T cells, activated platelets, vascular smooth muscle cells, eosinophils, synovial membranes in rheumatoid arthritis, dermal fibroblasts and other non-lymphoid cell types. By "cells expressing CD40" herein, any normal or malignant cells expressing detectable levels of the CD40 antigen are intended. Preferably, cells expressing CD40 are cells expressing detectable levels of cell surface CD40 antigen. Methods for detecting CD40 expression in cells are well known in the art and include, but are not limited to, PCR techniques, immunohistochemistry, flow cytometry, Western blotting, ELISA and the like. These methods allow the detection of mRNA from CD40, CD40 antigen and CD40 cell surface antigen. Detection of cell surface CD40 expression can be performed as described in Example 3 herein, or by other suitable methods. By "CD40 ligand" or "CD40L" is intended primarily the transmembrane protein of 32-33 kDa which also exists in two smaller soluble forms biologically active, 18 kDa and 31 kDa, respectively (Graf et al (1995) Eur. J. Immunol., 25: 1749-1754, Mazzei et al. (1995) J Biol. Chem. 270: 7025-7028, Pietravalle ef al. (1996) J. Biol. Chem. 271: 5965-5967). Human CD40L is also known as CD154 or gp39. By "CD40 ligand" or "CD40L" is also meant any other peptide, polypeptide or protein that can bind to and activate one or more CD40 signaling pathways. In this manner, "CD40 ligands" include, but are not limited to, full length CD40 ligand proteins and variants and fragments thereof that retain sufficient activity to perform the function of binding to, and stimulating the CD40 signaling in cells expressing CD40. Modifications to a native CD40 ligand, e.g., human CD40L, include, but are not limited to, substitutions, deletions, truncations, extensions, fusion proteins, fragments, peptidomimetics, and the like. By "CD40 signaling" is meant any of the biological activities resulting from the interaction of cell surface CD40 with a CD40 ligand or other agonist, such as an agonist antibody. Examples of CD40 signaling are signals that lead to proliferation and survival of cells expressing CD40, and stimulation of one or more signaling pathways by CD40 within cells expressing CD40. A "signaling path" or "signal transduction path" by CD40 is intended to mean at least one biochemical reaction, or group of biochemical reactions, that results from the interaction of the CD40 receptor with a CD40 ligand, for example. , CD40L, and which generates a signal that, when transmitted through the signal path, leads to the activation of one or more molecules downstream in the signaling cascade. The signal transduction pathways involve a series of signal transduction molecules, which lead to the transmission of a signal from the cell surface CD40 receptor, from one side of the membrane to the other plasma of a cell, and through one or more in a series of signal transduction molecules, through the cytoplasm of the cell and, in some cases, into the nucleus of the cell. For the present invention, signal transduction pathways by CD40, including the AKT signaling pathway, which leads to the activation of AKT and, ultimately, to the activation of NF-? B by the signaling path of NF-? B; and the mitogen-activated protein kinase (MAPK) signaling pathways, including the MEK / ERK signaling pathway and the MEK / p38 signaling pathway, which lead to the activation of ERK and p38, respectively. As noted in the above, the present invention provides a method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, wherein the human patient is heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F), the method comprises administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody. By "human patient" is intended a human patient who is afflicted by, is at risk of developing or relapsing with, any inflammatory disease or autoimmune disease that is associated with cells expressing CD40. By "inflammatory disease or autoimmune disease associated with cells expressing CD40" is intended any inflammatory disease or autoimmune disease associated with cells expressing CD40. Inflammatory disease or autoimmune disease, associated with cells expressing CD40, may be an inflammatory disease or autoimmune disease associated with an undesirable level of CD40 signaling in cells expressing CD40, or the inflammatory disease or autoimmune disease may be associated only indirectly with cells expressing CD40. By "an inflammatory disease or autoimmune disease associated with an undesirable level of CD40 signaling" is intended an inflammatory disease or autoimmune disease whose development or progression is associated with an undesirable level of CD40 signaling. By "an undesirable level of CD40 signaling" is meant any physiologically undesirable level of CD40 signaling that can occur in cells that express CD40 in a human patient who has an inflammatory disease or autoimmune disease. Inflammatory diseases are characterized by inflammation and destruction of tissue, or a combination thereof. "Inflammatory disease" includes any inflammatory process mediated by the immune response where the initiating event or the target of the immune response involves the non-self antigen (s), including, for example, alloantigens, xenoantigens, viral antigens, bacterial antigens, unknown antigens or antigens. allergens As used herein, it is understood that the term "autoimmunity" generally encompasses inflammatory processes mediated by the immune response that involve "self" antigens. In autoimmune diseases, the antigen (s) itself activate the immune responses of the host. The present invention can be used in the treatment of inflammation associated with rejection of tissue transplants. "Rejection of transplants" or "rejection of grafts" refers to any immune response mounted by the host against a graft including, but not limited to, HLA antigens, blood group antigens and the like. The invention can also be used to treat graft versus host disease, such as that associated with bone marrow transplantation, for example. In such graft versus host disease, the bone marrow of the donor includes lymphocytes and cells that mature into lymphocytes. The donor lymphocytes recognize the recipient's antigens as non-self and mount an inflammatory immune response. Therefore, as used herein, "graft versus host disease" or "graft versus host reaction" refers to any T cell-mediated immune response in which the donor lymphocytes react with host antigens. Inflammatory diseases and autoimmune diseases that can be treated according to the methods of the invention include, but are not limited to, systemic lupus erythematosus (SLE), discoid lupus, lupus nephritis, sarcoidosis, inflammatory arthritis, including juvenile arthritis, rheumatoid arthritis, psoriatic arthritis, Reiter syndrome, ankylosing spondylitis and arthritis due to gout, rejection of an organ or tissue transplant, hyperacute, acute rejection or chronic and / or graft versus host disease, multiple sclerosis, hyperimmunoglobulinemia E syndrome, polyarteritis nodosa, primary biliary cirrhosis, bowel inflammation syndrome, Crohn's disease, celiac disease (gluten-sensitive enteropathy), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia , psoriasis, scleroderma, myasthenia gravis, autoimmune thrombocytopenic purpura, autoimmune thyroiditis, Grave's disease, Hasimoto thyroiditis, immune complex disease, chronic fatigue immune dysfunction syndrome (CFIDS), polymyositis and dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy , pemphigus vulgaris, pulmonary interstitial fibrosis, type I and type II diabetes mellitus, delayed type hypersensitivity types 1, 2, 3 and 4, allergy or allergic disorders, unwanted / unintended immune responses to therapeutic proteins (see for example, Patent Application North American No. US 2002/0119151 and Koren, ef al (2 002) Curr. Pharm. Biotechnol. 3: 349-60), asthma, Churg-Strauss syndrome (allergic granulomatosis), atopic dermatitis, allergic and irritant contact dermatitis, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis, idiopathic inflammatory myopathies, hemolytic disease, Alzheimer's disease , chronic inflammatory demyelinating polyneuropathy and the like. The methods of the invention are also useful in the treatment of pulmonary inflammation including, but not limited to, lung graft rejection, asthma, sarcoidosis, emphysema, cystic fibrosis, idiopathic pulmonary fibrosis, chronic bronchitis, allergic rhinitis and allergic lung diseases. such as hypersensitivity pneumonitis, eosinophilic pneumonia, bronchiolitis obliterans due to bone marrow and / or lung transplantation or other causes, graft atherosclerosis / phlebosclerosis, as well as pulmonary fibrosis resulting from collagen, vascular and autoimmune diseases such as rheumatoid arthritis and lupus erythematosus The inflammatory disease or autoimmune disease may be an inflammatory disease or autoimmune disease associated with cells expressing CD40. Examples of autoimmune antibody-dependent diseases include Rheumatoid Arthritis, Psoriasis, Systemic Lupus Erythematosus, Crohn's disease, Myasthenia gravis, Idiopathic thrombocytopenia purpura or Sjogren's syndrome. In addition, the extraction in its entirety of B cells and other cells that carry the CD40 they can limit the activation of T cells by emitting signals through the binding of the CD40 ligand. Therefore, the extraction of all B cells and other cells carrying CD40 can be used to treat autoimmune and inflammatory diseases mediated by T cells, such as Multiple Sclerosis, rejection of grafts, graft versus host disease, Alzheimer's disease or Diabetes. It can also be useful for bone marrow transplants. The present invention is particularly favorable with respect to inflammatory diseases and autoimmune diseases that are associated with cells expressing CD40. CHIR-12.12, described herein, can be used to treat patients having an inflammatory disease or autoimmune disease that is resistant to therapy with other therapeutic agents, including anti-CD20 antibodies, such as Rituxan®, as described with more detail elsewhere in the present. "Treatment" is defined herein as the application or administration of an anti-CD40 antibody to a subject, or the application or administration of an anti-CD40 antibody to a tissue or cell line isolated from a subject, wherein the subject has a autoimmune disease and / or inflammatory disease, a symptom associated with an autoimmune disease and / or inflammatory disease or a predisposition towards the development of an autoimmune disease and / or inflammatory disease, where the purpose is to cure, heal, calm, mitigate , alter, remedy, alleviate, improve or affect the autoimmune disease and / or inflammatory disease, any associated symptoms of the autoimmune disease and / or inflammatory disease, or the predisposition towards the development of the autoimmune disease and / or inflammatory disease . By "treatment" is also meant the application or administration of a pharmaceutical composition comprising an anti-CD40 antibody to a subject, or the application or administration of a pharmaceutical composition comprising an anti-CD40 antibody to a tissue or cell line isolated from a subject, wherein the subject has an autoimmune disease and / or inflammatory disease, a symptom associated with an autoimmune disease and / or inflammatory disease or a predisposition towards the development of an autoimmune disease and / or inflammatory disease, where the purpose is to heal, heal, soothe, mitigate, alter, remedy, alleviate, ameliorate or affect the autoimmune disease and / or inflammatory disease, any associated symptoms of autoimmune disease and / or inflammatory disease, or predisposition towards the development of autoimmune disease and / or inflammatory disease. By "anti-inflammatory activity" is intended a reduction or prevention of inflammation. Therapy with an anti-CD40 antibody, as defined elsewhere herein, elicits a physiological response that is beneficial with respect to the treatment of an autoimmune disease and / or inflammatory disease, where the disease involves cells expressing the antigen. CD40. It is recognized that the methods of the invention can be useful to prevent a phenotypic change in cells, such as proliferation, activation and the like. In the therapeutic methods of the present invention, at least one anti-CD40 antibody, as defined elsewhere herein, is used to promote a positive therapeutic response with respect to an inflammatory disease or autoimmune disease. By "positive therapeutic response" with respect to an inflammatory disease and / or autoimmune disease is intended an improvement in the disease, in association with the anti-inflammatory activity of the antibody, and / or an improvement in the symptoms associated with the disease. That is, an anti-proliferative effect can be observed, the prevention of subsequent proliferation of the cell expressing CD40, a reduction in the inflammatory response including, but not limited to, reduced secretion of inflammatory cytokines., adhesion molecules, proteases, immunoglobulins (in cases where the cell carrying the CD40 is a B cell), combinations thereof and the like, increased production of anti-inflammatory proteins, a reduction in the number of self-reactive cells, an increase in immune tolerance, inhibition of the survival of autoreactive cells and / or a decrease in one or more symptoms mediated by the stimulation of cells expressing CD40. Such positive therapeutic responses are not limited to the route of administration and may comprise administration to the donor, donor tissue (such as, for example, organ perfusion), the host, any combination thereof, and the like. The clinical response can be assessed using selection techniques such as magnetic resonance imaging (MRI) scanning, radiographic x-ray imaging, computed tomography (CT) scanning, flow cytometry or fluorescence activated cell sorter analysis (FACS), histology, ordinary pathology and blood chemistry including, but not limited to, detectable changes by ELISA, RIA, chromatography and the like. In addition to these positive therapeutic responses, the subject who undergoes anti-CD40 antibody therapy may experience the beneficial effect of an improvement in the symptoms associated with the disease. By "therapeutically or prophylactically effective dose" or "therapeutically or prophylactically effective amount" is meant an amount of anti-CD40 antibody that, when administered, causes a positive therapeutic response with respect to the treatment of a patient with an inflammatory disease or autoimmune disease. immune associated with cells expressing CD40. Suitable dosages are described in greater detail elsewhere herein. The method of treatment may comprise a single administration of a therapeutically effective dose or multiple administrations of a therapeutically effective dose of the anti-CD40 antibody, as described in greater detail elsewhere herein. The methods of the invention are particularly useful for treating inflammatory diseases or autoimmune diseases, including those listed above, which are resistant to one or more known therapies for inflammatory or auto immune diseases. Such therapies include, but are not limited to, surgery or surgical procedures (e.g., splenectomy, lymphadenectomy, thyroidectomy, plasmapheresis, leukophoresis, transplantation of cells, tissues or organs, intestinal procedures, organ perfusion and the like), radiation therapy, therapy such as steroid therapy and non-steroidal therapy, hormone therapy, cytokine therapy, therapy with dermatological agents (eg, topical agents used to treat skin conditions such as allergies, contact dermatitis and psoriasis), immunosuppressive therapy and other anti-inflammatory therapy -inflammatory with monoclonal antibodies, and the like, as described in greater detail elsewhere herein. By "resistant" it is intended that the inflammatory disease or particular autoimmune disease is resistant to, or insensitive to, a particular therapy. An inflammatory disease or autoimmune disease may be resistant to a particular therapy, either from the beginning of treatment with the particular therapy (ie, insensitive to initial exposure to therapy) or, as a result of developing resistance to therapy, either during the course of a first treatment period with the therapy or during a period of subsequent treatment therapy. In this manner, the present invention is useful for treating a human patient who is resistant to a therapy for inflammatory or auto immune diseases, when that human patient is either resistant, or insensitive, to that therapy. The methods of the present invention involve the use of anti-CD40 antibodies. The "antibodies" are usually heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each light chain is linked to a heavy chain by a covalent disulfide bond, while the number of disulfide bonds varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has disulfide bridges within the chain, spaced regularly. Each heavy chain has, at one end, a variable domain (VH) followed by a series of constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the variable domain of the light chain is aligned with the variable domain of the heavy chain. It is believed that particular amino acid residues form an interface between the variable domains of light and heavy chains. The term "variable" refers to the fact that certain portions of the variable domains differ widely in their sequence among the antibodies. The variable regions confer antigen binding specificity. Constant domains do not directly involve binding an antibody to an antigen, but exhibit various effector functions, such as Fe receptor binding (FcR), antibody involvement in antibody-dependent cellular toxicity, onset of complement-dependent cytotoxicity and degranulation of mast cells. The "light chains" of antibodies (immunoglobulins), of any vertebrate species, can be assigned to one of two clearly distinct types, called kappa (K) and lambda (?), Based on the amino acid sequences of their constant domains . Depending on the amino acid sequence of the constant domain of its "chains "immunoglobulins can be assigned to different classes." There are five main classes of human immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these can also be divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3 , lgG4, lgA1 and lgA2 The constant domains of heavy chains corresponding to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma and mu, respectively Subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known Different isotypes have different effector functions For example, the human IgG1 and IgG3 isotypes have ADCC (antibody-dependent cell-mediated cytotoxicity) activity IgG1 antibodies, in particular IgG1 human antibodies, are particularly useful in the methods of present invention "Human effector cells" are leukocytes that express one or more FcRs and perform effec functions Preferably, the cells express at least FcγRIII and carry out an antigen-dependent cell-mediated cytotoxicity (ADCC) effector function. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMC), natural killer cells (NK), monocytes, macrophages, eosinophils and neutrophils, with PBMCs and NK cells being preferred. Antibodies having ADCC activity are typically of the IgG1 or IgG3 isotype. Note that, in addition to isolating IgG1 and IgG3 antibodies, such antibodies that mediate ADCC can be made by designing a variable region from an antibody without ADCC, or variable region fragment for a constant region of the IgG1 or IgG3 isotypes. The terms "Fe receptor" or "FcR" are used to describe a receptor that binds to the Fe region of an antibody. The preferred FcR is a human FcR of native sequence. Moreover, a preferred FcR is one that binds to an IgG antibody (a gamma receptor) and includes recipients of the subclasses Fc? RI, Fc? RII and Fc? RIII, including allelic variants and alternatively spliced forms of these receptors . Fc? RII receptors include Fc? RIIA (an "activating receptor") and Fc? RIIB (an "inhibitory receptor"), which have similar amino acid sequences that differ mainly in the cytoplasmic domains thereof. The activating receptor Fc? RIIA contains an activation motive of tyrosine-based immunoreceptors (ITAM) in its cytoplasmic domain. The Fc? RIIB inhibitory receptor contains a motif of inhibition of tyrosine-based immunoreceptors (ITIM) in its cytoplasmic domain (see Daeron (1997) Annu., Rev. Immunol., 15: 203-234). The FcRs are reviewed in Ravetch and Kinet (1991) Annu. Rev. Immunol. 9: 457-492 (1991); Capel et al. (1994) Immunomethods 4: 25-34; and de Haas ef al. (1995) J Lab. Clin. Med. 126: 330-341. Other FcRs, including those that will be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al (1976) J. Immunol. 117: 587 and Kim et al. (1994) J Immunol. 249 (1994)). The term "antibody" is used herein in the broadest sense and covers fully assembled antibodies, fragments of antibodies which retain the ability to specifically bind to the CD40 antigen (eg, Fab, F (ab ') 2, Fv and other fragments), single chain antibodies, diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, humanized antibodies and the like), and recombinant peptides comprising the foregoing. The term "antibody" covers both polyclonal and monoclonal antibodies. As used herein, "anti-CD40 antibody" encompasses any antibody that specifically recognizes the CD40 antigen. In some embodiments, anti-CD40 antibodies, for use in the methods of the present invention, in particular the monoclonal anti-CD40 antibodies, exhibit a strong binding affinity to a single site for the CD40 antigen. Such monoclonal antibodies exhibit an affinity for CD40 (KD) of at least 10"5 M, at least 3 x 10" 5 M, preferably at least 10"6 M or at least up to 10" 7 M, more preferably at least 10'8 M or at least 10'12 M, when measured using a standard assay such as Biacore ™. Biacore analysis is known in the art and details are provided in the "BlApplications manual". The methods described in WO 01/27160 can be used to modulate the binding affinity. By "specifically recognizes" or "specifically binds to" it is intended that the anti-CD40 antibody does not bind to unrelated antigens, such as the CD20 antigen. In some embodiments, anti-CD40 antibodies, for use in the methods of present invention, in particular monoclonal antibodies, exhibit a strong binding affinity for human FcγRllla-158V. Preferably, an anti-CD40 antibody, for use in the methods of the invention, binds to human FcγRllla-158V with an affinity (KQ) of at least about 0.5 μM when measured using a standard assay such as Biacore ™ As described in Example 6 herein, the CHIR-12.12 antibody binds to human FcγRllla-158V with an affinity (KD) of 492 nM. In some embodiments, anti-CD40 antibodies, for use in the methods of the present invention, in particular monoclonal antibodies, exhibit a strong binding affinity for human FcγRllla-158F. Preferably, an anti-CD40 antibody, for use in the methods of the invention, binds to human FcγRllla-158F with an affinity (KQ) of at least about 12 μM when measured using a standard assay such as Biacore ™ Preferably, the anti-CD40 antibody, for use in the methods of the invention, binds to human FcγRllla-158F with an affinity (KD) of at least about 10 μM, at least about 8 μM, per at least about 6 μM, at least about 5 μM, at least about 4 μM or at least about 3 μM. As described in Example 6 herein, the CHIR-12.12 antibody binds to human FcγRllla-158F with an affinity (KD) of 2.8 μM. In some embodiments, anti-CD40 antibodies, for use in the methods of the present invention, in particular monoclonal antibodies, exhibit a strong binding affinity for both FcγRllla-158V and human FcγRllla-158F . Preferably, an anti-CD40 antibody, for use in the methods of the invention, binds to human FcγRllla-158V with an affinity (KD) of at least about 0 5 μM and binds to FcγRllla- Human 158F with an affinity (KD) of at least about 12 μM, when measured using a standard assay such as Biacore ™. The antibodies, for use in the methods of the present invention, can be produced using any suitable method of antibody production known to those of skill in the art. The anti-CD40 antibody, used in the methods of the present invention, can be a polyclonal antibody. In this way, polyclonal sera can be prepared by conventional methods. In general, a solution containing the antigen of interest (in this case, the CD40 antigen) is first used to immunize a suitable animal, preferably a mouse, rat, rabbit or goat. Rabbits or goats are preferred for the preparation of polyclonal sera due to the volume of serum that can be obtained, and the availability of antibodies labeled anti-rabbit and anti-goat. Sera from immunized animals can be selected for antibody reactivity against the initial antigen. Lymphocytes can be isolated from lymph nodes or spleen cells and can further be selected for B cells by screening for CD138 negative and CD19 positive cells. In one aspect, such B cell cultures (BCCs) can be fused to myeloma cells to generate hybridomas, as detailed herein. Polyclonal sera can also be prepared in a transgenic animal, preferably a mouse carrying gene fragments of human immunoglobulin. In a preferred embodiment, Sf9 cells expressing the protein of interest (in this case, the CD40 antigen), are used as the immunogen. The immunization can also be performed by mixing or emulsifying the solution containing antigens in saline, preferably in an adjuvant such as complete Freund's adjuvant, and injecting the mixture or emulsion in parenteral form (generally subcutaneously or intramuscularly). A dose of 50-200 μg / injection is typically sufficient. Immunization is usually reinforced 2-6 weeks later with one or more injections of the protein in saline, preferably using incomplete Freund's adjuvant. Alternatively, antibodies can be generated by in vitro immunization using methods known in the art which, for the purposes of this invention, are considered equivalent to immunization in vivo. Polyclonal antisera are obtained by bleeding the immunized animal in a glass or plastic container, incubating the blood at 25 ° C for one hour, followed by incubating at 4 ° C for 2-18 hours. The serum is recovered by centrifugation (eg, 1,000 x g for 10 minutes). Approximately 20-50 ml per bleed can be obtained from rabbits. The production of Sf 9 (Spodoptera frugiperda) cells is described in the patent North American No. 6,004,552, incorporated herein by reference. In the case of CD40, briefly, the sequences encoding human CD40 were recombined in a baculovirus using transfer vectors. The plasmids were co-transfected with wild type baculovirus DNA into Sf 9 cells. Sf 9 cells infected with recombinant baculovirus were identified and purified in clonal form. The anti-CD40 antibody, used in the methods of the present invention, can be a monoclonal antibody. The term "monoclonal antibody" (and "mAb"), as used herein, refers to an antibody obtained from a substantially homogeneous population of antibodies, ie, the individual antibodies comprising the population are identical except for possible mutations that occur naturally, which can occur in smaller amounts. The term is not limited as to the species of the antibody and does not require production of the antibody by any particular method. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different antigenic determinants (epitopes), each monoclonal antibody is directed against a single determinant (epitope) on the antigen. By "epitope" is meant the part of an antigenic molecule for which an antibody is produced and to which the antibody will bind. The epitopes may comprise linear amino acid residues (ie, the residues within the epitope are sequentially arranged one after the other linearly), non-linear amino acid residues (referred to herein as "non-linear epitopes", these epitopes do not are arranged in sequence) or amino acid residues, both linear and non-linear. An anti-CD40 monoclonal antibody, suitable for use in the methods of the present invention, will be capable of specifically binding to an epitope on the human CD40 antigen expressed on the surface of a human cell, i.e., an epitope that is exposed to the outside of the cell. The monoclonal antibodies for use in accordance with the present invention can be made by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or can be made by recombinant DNA methods (see, for example, US Patent No. 4,816,567). Monoclonal antibodies can also be isolated from of antibody phage libraries, generated using the techniques described, for example, in McCafferty et al. (1990) Nature 348: 552-554 (1990) and U.S. Patent No. 5,514,548. Clackson ef al. (1991) Nature 352: 624-628 and Marks ef al. (1991) J. Mol. Biol. 222: 581-597 describe the isolation of murine and human antibodies, respectively, using phage libraries. Subsequent publications describe the production of high affinity human antibodies (nM interval) by chain exchange (Marks et al (1992) Bio / Technology 10: 779-783), as well as combinatorial infection and in vivo recombination as a strategy to construct very large phage libraries (Waterhouse et al (1993) Nucleic Acids Res. 21: 2265-2266). In this way, these techniques are viable alternatives to the traditional techniques of monoclonal antibody hybridomas for the isolation of monoclonal antibodies. In the traditional method of Kohler ef al. (1975) Nature 256: 495-496, a mouse is typically immunized with a solution containing an antigen. The immunization can be performed by mixing or emulsifying the solution containing antigens in saline, preferably in an adjuvant such as Freund's complete adjuvant, and injecting the mixture or emulsion in parenteral form. Any method of immunization known in the art can be used to obtain the monoclonal antibodies of the invention. After immunization of the animal, the spleen (and optionally several large lymph nodes) is removed and dissociated into single cells. Spleen cells can be selected by applying a suspension of cells to a plate or well coated with the antigen of interest. B cells that express membrane-bound immunoglobulin, specific for the antigen, bind to the plate and do not remove with the rinse. The resulting B cells, or all of the dissociated cells of the spleen, are then induced to fuse with myeloma cells to form hybridomas, and are cultured in a selective medium. The resulting cells are plated by serial dilution and tested for the production of antibodies that specifically bind to the antigen of interest (and that do not bind to unrelated antigens). Selected hybridomas secreting monoclonal antibodies (mAbs) are then cultured either in vitro (for example, in tissue culture bottles or hollow fiber reactors), or in vivo (as ascites in mice). In another aspect, preferably, the B cell cultures can be further selected for reactivity against the initial antigen. Such selection includes enzyme-linked immunosorbent assay (ELISA) with the target protein / antigen, a competition assay with known antibodies that bind to the antigen of interest and in vitro binding to CHO cells, or others, transiently transfected, expressing the target antigen. Where the anti-CD40 antibodies for use in the methods of the invention are to be prepared using recombinant DNA methods, the DNA encoding the monoclonal antibodies is easily isolated and sequenced using conventional methods (eg, by using oligonucleotide probes that are capable of of binding specifically to genes encoding the heavy and light chains of murine antibodies). Hybridoma cells, described herein, serve as a preferred source of such DNA. Once isolated, the DNA can be placed in expression vectors, which are then transfected into host cells, such as E. coli cells, simian COS cells, Chinese hamster's ovary (CHO) cells or myeloma cells that another way they do not produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Review articles about recombinant expression in bacteria of the DNA encoding the antibody include Skerra ef al. (1993) Curr. Opinion in Immunol. 5: 256 and Phickthun (1992) Immunol. Revs. 130: 151. Alternatively, the antibody can be produced in a cell line such as a CHO cell line, as described in U.S. Patent Nos. 5,545,403; 5,545,405; and 5,998,144; incorporated herein by reference. Briefly, the cell line is transfected with vectors capable of expressing a light chain and a heavy chain, respectively. By transfecting the two proteins into separate vectors, chimeric antibodies can be produced. Another advantage is the correct glycosylation of the antibody. A "host cell," as used herein, refers to a microorganism or a eukaryotic cell or cell line cultured as a unicellular entity that can be used, or has been used, as a recipient for a recombinant vector or other transferred polynucleotides. , and include the progeny of the original cell that has been transfected. It is understood that the progeny of a single cell may not necessarily be completely identical in morphology or in complement of genomic or total DNA to the original parent, due to natural, accidental mutation or deliberate. In some embodiments, the anti-CD40 antibody, such as CHIR-12.12, is produced in CHO cells using the GS gene expression system (Lonza Biologies, Portsmouth, New Hampshire), which uses glutamine synthetase as a marker. See also U.S. Patent Nos. 5,122,464; 5,591, 639; 5,658,759; 5,770,359; 5,827,739; 5,879,936; ,891, 693; and 5,981, 216; whose contents are incorporated herein by reference in their entirety. Monoclonal antibodies to CD40 are known in the art. See, for example, sections devoted to B-cell antigen in McMichael, ed. (1987; 1989) Leukocyte Typing III and IV (Oxford University Press, New York); US Patents Nos. ,674,492; 5,874,082; 5,677,165; 6,056,959; WO 00/63395; International Publications Nos.

WO 02/28905 and WO 02/28904; Gordon ef al. (1988) J. Immunol. 140: 1425; Valley ef al. (1989) Eur.

J. Immunol. 19: 1463; Clark ef al. (1986) PNAS 83: 4494; Paulie ef al. (1989) J Immunol. 142: 590; Gordon ef al. (1987) Eur. J. Immunol. 17: 1535; Jabara ef al. (1990) J Exp. Med. 172: 1861; Zhang et al (1991) J Immunol. 146: 1836; Gas to ef. (1991) J Immunol. 147: 8; Banchereau ef al. (1991) Clin. Immunol. Spectrum 3: 8; and Banchereau ef al. (1991) Science 251: 70; whose entirety is incorporated herein by reference. As noted in the foregoing, the term "antibody", as used herein, encompasses chimeric antibodies. By "chimeric" antibodies we pretend antibodies that are derived more preferably using recombinant techniques of deoxyribonucleic acids and which comprise both human components (including immunologically "related", e.go, chimpanzees) and non-human. In this way, the constant region of the chimeric antibody of greater preference is substantially identical to the constant region of a natural human antibody; the variable region of the most preferred chimeric antibody is derived from a non-human source and has the desired antigenic specificity for the antigen of interest (CD40). The non-human source can be any vertebrate source that can be used to generate antibodies to the CD40 antigen. Such non-human sources include, but are not limited to, rodents (e.g., rabbit, rat, mouse, etc., see, e.g., Patent North American No. 4,816,567, incorporated herein by reference) and non-human primates (e.g., European Monkey, Mico, etc., see, for example, U.S. Patent Nos. 5,750,105 and 5,756,096, incorporated herein by reference). As noted in the foregoing, the term "antibody", as used herein, encompasses humanized antibodies. By "humanized" is meant forms of antibodies that contain a minimal sequence derived from non-human immunoglobulin sequences. In most cases, the humanized antibodies are human immunoglobulins (target antibody) in which the residues of a hypervariable region (also known as the complementarity determining region or CDR) of the recipient are replaced by residues from a hypervariable region of a species non-human (donor antibody) such as mouse, rat, rabbit or non-human primate having the specificity, affinity and desired capacity. The phrase "complementarity determining region" refers to amino acid sequences which together define the binding affinity and specificity of the native Fv region of a native immunoglobulin binding site. See, for example, Chothia ef al (1987) J Mol. Biol. 196: 901-917; Kabat ef al (1991) Dept. of Health and Human Services of the U., NIH Publication No. 91-3242). The phrase "constant region" refers to the portion of the antibody molecule that confers effector functions. In a previous work directed toward producing non-immunogenic antibodies for use in the therapy of human disease, the mouse constant regions were replaced by human constant regions. The constant regions of the subject humanized antibodies were derived from human immunoglobulins. However, these humanized antibodies can elicit an unwanted and potentially dangerous immune response in humans and there was a loss of affinity. Humanization can be performed following the method of Winter et al. (Jones et al. (1986) Nature 321: 522-525; Riechmann et al. (1988) Nature 332: 323-327; Verhoeyen et al. (1988) Science 239: 1534-1536), by replacing the CDRs or CDR sequences of rodent or mutant rodents with the corresponding sequences of a human antibody. See also U.S. Patent Nos. 5,225,539; 5,585,089; 5,693,761; 5,693,762; 5,859,205; incorporated herein by reference. In some cases, waste within the regions The scaffolds of one or more variable regions of human immunoglobulin are replaced by corresponding non-human residues (see, for example, U.S. Patent Nos. 5,585,089, 5,693,761, 5,693,762, and 6,180,370). Additionally, the humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine the antibody performance (e.g., to obtain the desired affinity). In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable regions correspond to those of a non-human immunoglobulin and all or substantially all framework regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of a constant region (Fe) of immunoglobulin, typically that of a human immunoglobulin. For additional details see Jones ef al. (1986) Nature 331: 522-525; Riechmann et al. (1988) Nature 332: 323-329; and Presta (1992) Curr. Op. Struct. Biol. 2: 593-596; incorporated herein by reference. Accordingly, such "humanized" antibodies can include antibodies wherein substantially less than an intact human variable domain has been replaced by the corresponding sequence of a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some framework residues are replaced by residues of analogous sites in rodent antibodies. See, for example, U.S. Patent Nos. 5,225,539; 5,585,089; 5,693,761; 5,693,762; 5,859,205. See also U.S. Patent No. 6,180,370, and International Publication No. WO 01/27160, wherein humanized antibodies and techniques for producing humanized antibodies having an improved affinity for a predetermined antigen are disclosed. Humanized anti-CD40 antibodies can also be produced using Human Engineering ™ technology (Xoma Ltd., Berkeley, California). Humanized anti-CD40 monoclonal antibodies include antibodies such as SGN-40 (Tai ef al. (2004) Cancer Res. 64: 2846-52; US Patent No. 6,838,261), which is the humanized form of the murine anti-CD40 antibody. SGN-14 (Francisco ef al. (2000) Cancer Res. 60: 3225-31), and the antibodies described in U.S. Patent Application Publication No. 2004/0120948; incorporated herein by reference in its entirety. The present invention can also be practiced using xenogeneic or modified antibodies, produced in a non-human mammalian host, more particularly a transgenic mouse, characterized by gene fragments of inactivated endogenous immunoglobulin (Ig). In such transgenic animals, the endogenous genes competent for the expression of light and heavy subunits of host immunoglobulins become non-functional and substituted with the gene fragments of analogous human immunoglobulins. These transgenic animals produce human antibodies in the substantial absence of subunits of light or heavy host immunoglobulins. See, for example, U.S. Patent Nos. 5,877,397 and 5,939,598; incorporated herein by reference. Thus, in some embodiments, fully human antibodies to CD40, for example, are obtained by immunizing transgenic mice. A similar mouse is obtained using the XenoMouse® technology (Abgenix, Fremont, California), and is described in US Patent Nos. 6,075,181, 6,091,001, and 6,114,598, the entirety of which is incorporated herein by reference. For example, to produce the CHIR-12.12 antibody, mice transgenic for the human IgG ^ heavy chain gene fragment and the human light chain fragment were immunized with Sf 9 cells expressing human CD40. Mice can also be transgenic for other isotypes. The fully human anti-CD40 antibodies, useful in the methods of the present invention, are characterized by binding properties similar to those exhibited by the monoclonal antibody CHIR-12.12. As noted in the foregoing, the term "antibody", as used herein, also encompasses fragments of antibodies that can bind to the antigen. "Antibody fragments" comprise a portion of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab, F (ab ') 2 and Fv fragments; diabodies; linear antibodies (Zapata ef al. (1995) Protein Eng. 10: 1057-1062); single chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies it produces two identical fragments of antigen binding, called "Fab" fragments, each with a unique antigen binding site, and a residual "Fe" fragment, whose name reflects its ability to easily crystallize. The pepsin treatment produces an F (ab ') 2 fragment that has two antigen combining sites and is still capable of binding the antigen. "Fv" is the minimum fragment of the antibody that contains a complete site of recognition and binding to the antigen. This region consists of a variable domain dimer of a heavy and a light chain in close non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the sce of the VH-VL dimer. Collectively, the six CDRs confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, albeit at a lower affinity than the entire binding site. The Fab fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. The Fab fragments differ from the Fab 'fragments by the addition of some residues at the carboxy terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab 'in which the cysteine residue (s) of the constant domains carry a free thiol group. Fab 'fragments are produced by reducing the disulphide bridge of the heavy chains of the F (ab') 2 fragments. Other chemical couplings of antibody fragments are also known. The anti-CD40 antibody fragments are suitable for use in the methods of the inventionas long as they retain the desired affinity of the full-length antibody. Thus, for example, a fragment of an anti-CD40 antibody will retain the ability to bind to the CD40 antigen. Such fragments are characterized by properties similar to those of the corresponding total length antibody. Thus, for example, a fragment of a full length antagonist anti-CD40 antibody will preferably be capable of specifically binding to a human CD40 antigen expressed on the surface of a human cell, and is free of significant agonist activity, but exhibits an antagonistic activity when it binds to a CD40 antigen in a human cell that expresses CD40. Such fragments are referred to herein as "antigen binding" fragments. The fragments of an anti-CD40 antibody, for use, in the methods of the invention, preferably will also retain the ability to bind to the relevant FcR or FcRs. Thus, for example, a fragment of an anti-CD40 antibody can retain the ability to bind to FcγRllla. Thus, for example, a fragment of a full-length anti-CD40 antibody may be capable of specifically binding to a cell surface CD40 antigen, and also capable of binding to FcγRllla in human effector cells, such as cytolytic cells. natural (NK). Such fragments are referred to herein as "FcR binding" fragments. Such fragments will generally include at least part of the constant domain of the heavy chain. Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived by proteolytic digestion of intact antibodies (see, for example, Morimoto, et al (1992) and Brennan et al (1985) Science 229: 81). However, these fragments can now be produced directly by recombinant host cells. For example, antibody fragments can be isolated from the phage libraries of antibodies discussed in the foregoing. Alternatively, Fab'-SH fragments can be recovered directly from E. coli and chemically coupled to form F (ab ') 2 fragments (Carter et al. (1992) Bio / Technology 10: 163-167). According to another methodology, F (ab ') 2 fragments can be isolated directly from culture of recombinant-looking cells. Other techniques for the production of antibody fragments will be apparent to the experienced practitioner. Suitable antigen-binding fragments of an antibody comprise a portion of a full-length antibody, generally the antigen-binding or variable region thereof. Examples of antibody fragments include, but are not limited to, Fab, F (ab ') 2, and Fv fragments and single chain antibody molecules. By "Fab" is intended a monovalent fragment of antigen binding of an immunoglobulin that is composed of the light chain and part of the heavy chain. By F (ab ') 2 a bivalent antigen-binding fragment of an immunoglobulin containing both light chains and part of both heavy chains. By fragments of "Sv single chain" or "sFv" antibodies, fragments comprising the VH and VL domains of an antibody are sought, wherein these domains are presented in a single chain of polypeptides. See, for example, U.S. Patent Nos. 4,946,778, 5,260,203, 5,455,030 and 5,856,456, incorporated herein by reference. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that allow the sFv to form the desired structure for antigen binding. For a review of sFv see Pluckthun (1994) in The Pharmacology of Monoclonal Antibodies, Vol. 113, ed. Rosenburg and Moore (Springer-Verlag, New York), pp. 269-315. The antigen binding fragments of the antagonist anti-CD40 antibodies described herein may also be conjugated to a cytotoxin to effect the removal of the target cancer cells, as described herein in the following. In some embodiments of the invention, the anti-CD40 antibody is an anti-CD40 antagonist antibody. When such antibodies bind to CD40 exposed on the surface of human cells, such as human B cells, they do not cause significant agonist activity. In some embodiments, its binding to CD40 exposed on the surface of human cells results in inhibition of the proliferation and differentiation of these human cells. Anti-CD40 antibodies suitable for use in the methods of the invention include those antibodies that can exhibit an antagonistic activity towards normal and malignant human cells expressing the CD40 antigen on cell surface. By "agonist activity" a substance is intended to function as an agonist. An agonist is combined with a receptor in a cell and initiates a reaction or activity that is similar or the same as that initiated by the natural ligand of the receptor. A CD40 agonist induces any or all, but is not limited to, the following responses: proliferation and / or differentiation of B cells; over-regulation of intercellular adhesion by such molecules as ICAM-1, E-selectin, VCAM, and the like; secretion of pro-inflammatory cytokines such as IL-1, IL-6, IL-8, IL-12, TNF, and the like; signal transduction through the CD40 receptor by routes such as TRAF (eg, TRAF2 and / or TRAF3), MAP kinases such as NIK (NF-kB induction kinase), l-kappa B kinases (IKK a / b ), transcription factor NF-kB, Ras and the MEK / ERK route, the route P13K / AKT, the P38 MAPK route, and the like; transduction of an anti-apoptotic signal by molecules such as XIAP, mcl-1, bcl-x, and the like; memory generation of B and / or T cells; production of B-cell antibodies; commutation of B cell isotypes, up-regulation of cell surface expression of MHC Class II and CD80 / 86, and the like. By "significant" agonist activity, an agonist activity of at least %, 35%, 40%, 45%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% greater than the agonist activity induced by a neutral substance or negative control as measured in an assay of a B cell response. Preferably, "significant" agonist activity is a agonist activity that is at least 2 times greater or at least 3 times greater than the agonist activity induced by a neutral substance or negative control as measured by an assay of a B cell response. Thus, for example, where the B cell response of interest is a B cell proliferation, "significant" agonist activity may be the induction of a level of B cell proliferation that is at least 2 times greater or at least 3 times higher than the level of B cell proliferation induced by a neutral or negative control substance. In one embodiment, a nonspecific immunoglobulin, for example lgG1, which does not bind to CD40, serves as the negative control. A substance "free of significant agonist activity" may exhibit an agonist activity of not more than about 25% greater than the agonist activity induced by a neutral or negative control substance, preferably not more than about 20% greater, 15% greater, 10% higher, 5% higher, 1% higher, 0.5% higher or even no more than about 0.1% greater than the agonist activity induced by a neutral or negative control substance as measured in a B cell response assay.

By "antagonistic activity" the substance is intended to function as an antagonist.

A CD40 antagonist prevents or reduces the induction of any of the responses induced by the binding of the CD40 receptor to an agonist ligand, particularly CD40L. The antagonist can reduce the induction of any one or more of the responses to the agonist binding by 5%, 10%, 15%, 20%, 30%, 35%, preferably 40%, 45%, 50%, 55 %, 60%, more preferably, 70%, 80%, 85%, and more preferably 90%, 95%, or 100%. The methods for measuring the binding specificity of the CD40 ligand and the antagonistic activity of an anti-CD40 therapeutic agent, by example, an anti-CD40 antibody, are known in the art and include, but are not limited to, standard competitive binding assays, assays to monitor the secretion of immunoglobulins by B cells, B-cell proliferation assays of the Banchereau type. Like-B, T-cell helper assays for antibody production, proliferation assays of B-cell co-stimulation, and assays for over-regulation of B-cell activation markers. See, for example, such assays described in Wo 00 / 75348 and U.S. Patent No. 6,087,329, incorporated herein by reference. See also WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307, and WO 2005/044294WO, the contents of each of which are incorporated in the present for reference in its entirety. Antagonist / non-agonist activity can be evaluated by assays showing that CHIR12.12 lacks agonist activity. Suitable assays are shown in the assays described in US 5677165 (Chiron Corporation). In one embodiment of the invention, the anti-CD40 antagonist antibody is free of significant agonist activity in a cellular response. In another embodiment of the invention, the anti-CD40 antagonist antibody is free of significant agonist activity in assays of more than one cellular response (e.g., proliferation and differentiation, or proliferation, differentiation, and, for B cells, production of antibodies). Of particular interest are anti-CD40 antagonist antibodies that are free of significant agonist activity as defined herein but that exhibit antagonistic activity when bound to the CD40 antigen in human B cells. In one embodiment of the invention, the anti-CD40 antagonist antibody is free of significant agonist activity in a B cell response. In another embodiment of the invention, the anti-CD40 antagonist antibody is free of significant agonist activity in assays of more than one B cell response (eg, proliferation and differentiation, or proliferation, differentiation and production of antibodies). Any of the assays known in the art can be used to determine whether an anti-CD40 antibody acts as an antagonist of one or more B cell responses. some embodiments, the anti-CD40 antibody acts as an antagonist of at least one B cell response selected from the group consisting of B cell proliferation, B cell differentiation, antibody production, intercellular adhesion, B cell memory generation , isotype switching, up-regulation of cell surface expression of MHC Class II and CD80 / 86, and secretion of pro-inflammatory cytokines such as IL-8, IL-12, and TNF. Of particular interest are antagonist anti-CD40 antibodies that are free of significant agonist activity with respect to the proliferation of B cells when they bind to the human CD40 antigen on the surface of a human B cell. In a similar embodiment, the anti-CD40 antibody is an antagonist of B cell proliferation induced by soluble or cell surface CD40L, as measured in a B-cell proliferation assay. Suitable cell proliferation assays B are known in the art. Suitable assays for B cell proliferation are also described in the following. In some embodiments, the anti-CD40 antagonist antibody stimulates proliferation at a level that is not more than about 25% greater than the B cell proliferation induced by a neutral or negative control substance, preferably not more than about 20% higher , 15%, 10% higher, 5% higher, 1% higher, 0.5% higher or even no more than approximately 0.1% higher than cell proliferation induced by a neutral or negative control substance. In other embodiments, the anti-CD40 antibody is a B cell proliferation antagonist that is induced by another anti-CD40 antibody, for example, the anti-CD40 S2C6 antibody, as measured by a proliferation of B cells, and the level of B cell proliferation stimulated by the other anti-CD40 antibody in the presence of the anti-CD40 antagonist antibody is not more than about 25% of the B cell proliferation induced by the other anti-CD40 antibody in the absence of the anti-CD40 antibody. -CD40 antagonist (ie, at least 75% inhibition), preferably not more than about 20%, 15%, 10%, 5%, 1%, 0.5%, or even no more than about 0.1% of the B cell proliferation induced by the other anti-CD40 antibody in the absence of the anti-CD40 antagonist antibody. In still other embodiments, the anti-CD40 antibody is an antagonist of the B cell proliferation that is induced by the EL4B5 cell line as measured in a B cell activation assay, and the level of B cell proliferation stimulated by the EL4B5 cell line in the presence of the anti-CD40 antagonist antibody is not more than about 25% of the B cell proliferation induced by this cell line in the absence of the anti-CD40 antagonist antibody (ie, at least 75% inhibition), preferably not more than about 20%, 15%, %, 5%, 1%, 0.5%, or even no more than about 0.1% of the B cell proliferation induced by this cell line in the absence of the anti-CD40 antagonist antibody. In yet other embodiments, the anti-CD40 antibody is an antagonist of the production of antibodies induced by human T cells by human B cells as measured in the assay of human B-helper cells for production of antibodies by B cells. In this form, the level of production of IgG antibodies, production of IgM antibodies or production of both IgG and IgM antibodies by B cells stimulated by T cells in the presence of the anti-CD40 antagonist antibody is not more than about 50% of the respective antibody production by B cells stimulated by T cells in the absence of the anti-CD40 antagonist antibody (ie, at least 75% inhibition), preferably not more than about 25%, 20%, 15% , 10%, 5%, 1%, 0.5%, or even no more than about 0.1% of the respective production of antibodies by B cells stimulated by T cells in the absence of the anti-C antibody D40 antagonist. Additional anti-CD40 antagonist antibodies include monoclonal antibodies referred to as 5D12, 3A8, and 3C6, which are secreted by a hybrid having the access numbers of ATCC HB 11339, HB 12024 and HB 11340, respectively. See, for example, U.S. Patent No. 6,315,998 incorporated herein by reference in its entirety. Anti-CD40 antagonist antibodies are known in the art. See, for example, the human anti-CD40 antibody produced by the hybridoma designated F4-465 described in US Patent Application Publication Nos. 20020142358 and 20030059427 incorporated herein by reference in its entirety. F4-465 was obtained from the HAC mouse (Kuroiwa et al. (2000) Nature Biotech 10: 1086 (2000)) and therefore expresses the light chain human lambda. See also WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307, and WO 2005/044294WO, the contents of each of which are incorporated in the present for reference in its entirety. In addition to antagonist activity, the anti-CD40 antibody for use in the methods of the present invention will preferably have another mechanism of action against a target cell. For example, the anti-CD40 antibody will preferably have an ADCC activity. Alternatively, the variable regions of the anti-CD40 antibody can be expressed in another isotype of antibody having ADCC activity. It is also possible to conjugate native forms, recombinant forms or antigen-binding fragments of the anti-CD40 antibodies to a cytotoxin, a therapeutic agent, or a radioactive metal ion or radioisotope, as also described elsewhere herein. As explained elsewhere herein, the surprising finding has been made that, contrary to other antibodies, anti-CD40 antibodies, such as CHIR-12.12, are capable of mediating potent antibody-dependent cellular cytotoxicity (ADCC). of target cells expressing CD40 by binding to either of the two (158 or F) allotypes (V or F) of Fc? Rllla in natural killer cells (NK) of a human patient. Accordingly, anti-CD40 antibodies, such as CHIR-12.12, can be used in the treatment of inflammatory diseases and autoimmune diseases associated with cells expressing CD40 in human patients heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), in addition to human patients homozygous for Fc? Rllla158V (genotype V / V). The present invention is especially favorable for the treatment of inflammatory diseases and autoimmune diseases that are not sensitive to treatment with rituximab (Rituxan®), since it has been shown that the clinical activity of rituximab in NHL correlates with the Fc genotype? Rllla of the patient. Thus, the anti-CD40 antibodies particularly preferred for use in the methods of the present invention are those which, in addition to antagonist activity, are capable of mediating ADCC of cells expressing CD40 by human effector cells, such as natural killer cells (NK cells) expressing Fc? Rllla. The best preferred are those anti-CD40 antibodies that are capable of binding to both FcγRllla-158F and FcγRllla-158V with a high affinity, as described elsewhere hereinbelow. Particularly preferred anti-CD40 antibodies are those described in WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307 and WO 2005/044294, the contents of each one of which are incorporated herein by reference in their entirety. Of particular interest for the present invention are the anti-CD40 antagonist antibodies which share the binding characteristics of the monoclonal antibody CHIR-12.12 described in WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005 / 044855, WO 2005/044307 and WO 2005/044294. Such antibodies include, but are not limited to, the following: a) the monoclonal antibody CHIR-12.12; b) the monoclonal antibody produced by the hybridoma cell line 12.12; c) a monoclonal antibody comprising an amino acid sequence selected from the group consisting of the sequence shown in SEQ ID NO: 2, the sequence shown in SEQ ID NO: 4, the sequence shown in SEQ ID NO: 5, the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 4, and the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 5; d) a monoclonal antibody having an amino acid sequence encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of the sequence shown in SEQ ID NO: 1, the sequence shown in SEQ ID NO: 3, and the sequences shown in SEQ ID NO: 1 and SEQ ID NO: 3; e) a monoclonal antibody that binds to an epitope capable of binding to the monoclonal antibody produced by the hybridoma cell line 12.12; f) a monoclonal antibody that binds to an epitope comprising residues 82-87 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; g) a monoclonal antibody that binds to an epitope comprising residues 82-89 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; h) a monoclonal antibody that competes with the monoclonal antibody CHIR-12.12 in a competitive binding assay; i) the monoclonal antibody of preceding article a) or a monoclonal antibody of any of preceding articles c) -h), wherein the antibody is produced recombinantly; and j) a monoclonal antibody that is an antigen-binding fragment of a monoclonal antibody of any of the preceding articles a) -), wherein the fragment retains the ability to specifically bind to the human CD40 antigen. The monoclonal antibody CHIR-12.12 is particularly preferred for use in the methods of the present invention. The monoclonal antibody CHIR-12.12 was described in detail in WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307 and WO 2005/044294. The CHIR-12.12 antibody is a fully human anti-CD40 monoclonal antibody of the lgG1 isotype produced from the hybridoma cell line 153.8E2.D10.D6.12.12 (referred to as cell line 12.12). The cell line was created using splenocytes from immunized xenotypic mice containing the human IgG heavy chain gene fragment and the human K chain fragment (XenoMouse® technology, Abgenix, Fremont, California). Spleen cells were fused with mouse myeloma SP2 / 0 cells (Sierra BioSource). The resulting hybridomas were sub-cloned several times to create the stable monoclonal cell line 12.12. Other antibodies suitable for use in the methods of the invention can be prepared in a similar manner using transgenic mice for gene fragments of human immunoglobulin, as described elsewhere herein. The monoclonal antibody CHIR-12.12 binds to soluble CD40 in ELISA-type assays, prevents binding of the CD40 ligand to cell surface CD40 and displaces the pre-bound CD40 ligand, as determined by flow cytometric assays. CHIR-5.9 antibodies and CHIR-12.12 compete with each other for binding to CD40 but not with 15B8, the anti-CD40 monoclonal antibody described in US Provisional Application Serial No. 60 / 237,556, entitled "Anti-CD40 Human Antibodies," filed on October 2, 2000, and PCT International Application No. PCT / USOI / 30857, also entitled "Anti-CD40 Human Antibodies" filed on October 2, 2001 (Attorney's File No. PP16092.003) and published as WO 2002/028904, which are incorporated herein by reference in their entirety. When tested in vitro for effects on the proliferation of B cells from normal human subjects, CHIR-12.12 acts as an anti-CD40 antagonist antibody. Additionally, CHIR-12.12 does not induce a strong proliferation of human lymphocytes from normal subjects. The antibody is capable of removing target cells that express CD40 by antibody-dependent cellular cytotoxicity (ADCC). The binding affinity of CHIR-12.12 for human CD40 is 5x10'10M, as determined by the Biacore ™ assay. The nucleotide and amino acid sequences of the variable regions of the CHIR-12.12 antibody are provided herein. More particularly, the amino acid sequences for the leader, variable and constant regions for the light chain and heavy chain for the CHIR-12.12 mAb are set forth in SEQ ID NO: 2 (complete sequence for the light chain of the CHIR-12.12 mAb) , SEQ ID NO: 4 (complete sequence for the heavy chain for mAb CHIR-12.12), and SEQ ID NO: 5 (complete sequence for a variant of the heavy chain for mAb CHIR-12.12 set forth in SEQ ID NO: 4, wherein the variant comprises a serine substitution for the alanine residue at position 153 of SEQ ID NO: 4). The nucleotide sequences encoding the light chain and heavy chain for mAb CHIR-12.12 are set forth in SEQ ID NO: 1 (coding sequence for the light chain for mAb CHIR-12.12) and SEQ ID NO: 3 (coding sequence for the heavy chain for mAb CHIR-12.12). Hybridomas expressing the CHIR-12.12 antibody have been deposited with the ATCC with a patent deposited designation of PTA-5543. Anti-CD40 antibodies for use in the methods of the present invention include antibodies that differ from the monoclonal antibody CHIR-12.12 but retain the CDRs, and antibodies with one or more of the amino acid additions, deletions or substitutions. Anti-CD40 antibodies for use in the methods of the present invention can also be de-immunized antibodies, particularly antagonistic anti-CD40 antibodies. deimmunized, which can be produced as described, for example, in International Publications Nos. WO 98/52976 and WO 0034317; incorporated herein by reference. In this form, the residues within the anti-CD40 antagonist antibodies of the invention are modified so that the antibodies become no or less immunogenic for humans while retaining their antagonistic activity towards human cells expressing CD40, where such activity is measured by the trials observed elsewhere in the present. Also included within the scope of the present invention are fusion proteins comprising an antibody of interest, for example, an anti-CD40 antagonist antibody or an anti-CD40L antagonist antibody, or a fragment thereof, whose fusion proteins they can be synthesized or expressed from corresponding polynucleotide vectors, as is known in the art. Such fusion proteins are described with reference to conjugation of antibodies as observed elsewhere herein. Any known antibody having the binding specificity of interest may have sequence variations produced using the methods described, for example, in Patent Publications Nos. EP 0983303 A1, WO 00/34317, and WO 98/52976, incorporated herein by reference. present for reference. For example, it has been shown that sequences within the CDR can cause an antibody to bind to MHC Class II and activate an undesired response of helper T cells. A conservative substitution may allow the antibody to retain its binding activity, while at the same time losing its ability to activate an undesired T cell response. Any such conservative or non-conservative substitutions may be made using methods recognized in the art, such as those observed. elsewhere herein, and the resulting antibodies can also be used in the methods of the present invention. The antibodies of the variants can be routinely tested for the particular activity, for example, antagonist activity, affinity and specificity using the methods described herein. For example, variants in amino acid sequences of an anti-CD40 antagonist antibody, for example, the monoclonal antibody CHIR-12.12, can be prepared by mutations in the cloned sequence of DNA encoding the antibody of interest. Methods for mutagenesis and alterations in nucleotide sequences are well known in the art. See, for example, Walker and Gaastra, eds. (1983) Techniques in Molecular Biology (MacMillan Publishing Company, New York); Kunkel (1985) Proc. Nati Acad. Sci. USA 82: 488-492; Kunkel ef al. (1987) Methods Enzymol. 154: 367-382; Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (Cold Spring Harbor, New York); U.S. Patent No. 4,873,192; and the references cited therein; incorporated herein by reference. A guide for appropriate substitutions of amino acids, which do not affect the biological activity of the polypeptide of interest, can be found in Dayhoff's model ef el. (1978) in > 4f / as of Protein Sequence and Structure (Nat. Biomed. Res. Found., Washington, D.C.), incorporated herein by reference. Conservative substitutions, such as exchanging one amino acid with another having similar properties, may be preferred. Examples of conservative substitutions include, but are not limited to, GlyoAla, Val < = > lleoLeu, AspoGlu, Lys = Arg, Asn = Gln and Phe «rp = > Tyr. To construct variants of an antibody of interest, for example, a polypeptide of the antagonist anti-CD40 antibody of interest, modifications are made in such a way that the variants continue to possess the desired activity, i.e., similar binding affinity and, in In the case of anti-CD40 antagonist antibodies, they are able to bind specifically to a human CD40 antigen expressed on the surface of a human cell, and being free of significant agonist activity but exhibiting antagonistic activity when they bind to the CD40 antigen in a cell expressing human CD40. Obviously, any mutations made in the DNA encoding the variant polypeptide should not place the sequence outside the reading frame and preferably will not create complementary regions that could produce a secondary structure of mRNA. See EP Patent Publication No. 75,444. In addition, the constant region of an antibody, for example, an anti-CD40 antagonist antibody, can be mutated to alter the effector function in a number of ways. For example, see U.S. Patent No. 6,737,056B1 and U.S. Patent Application Publication No. 2004/0132101 A1, which describe Fe mutations that optimize antibody binding to Fe receptors. Preferably, the variants of an antibody of reference, for example, an antibody anti-CD40 antagonist, have amino acid sequences that have at least 70% or 75% sequence identity, preferably at least 80% or 85% sequence identity, most preferably at least 90%, 91% 92%, 93%, 94% or 95% sequence identity with the amino acid sequence for the reference antibody, for example, an antagonist anti-CD40 antibody molecule, for example, the monoclonal antibody CHIR-12.12 described in present, or to a shorter portion of the reference antibody molecule. Most preferably, the molecules share at least 96%, 97%, 98% or 99% sequence identity. For purposes of the present invention, the percent identity of sequences is determined using the Smith-Waterman homology search algorithm, using a matching space search with an open penalty of 12 spaces and a space extension penalty of 2, matrix BLOSUM of 62. The Smith-Waterman algorithm of homology search is taught in Smith and Waterman (1981) Adv. Appl. Math. 2: 482-489. A variant, for example, may differ from the reference antibody, for example, an anti-CD40 antagonist antibody, in only 1 to 15 amino acid residues, only 1 to 10 amino acid residues, such as 6-10, only 5, only 4, 3, 2 or even 1 amino acid residue. With respect to the optimal alignment of two amino acid sequences, the contiguous segment of the amino acid sequence of the variant may have additional amino acid residues or deleted amino acid residues with respect to the reference amino acid sequence. The contiguous segment used for comparison to the reference amino acid sequence will include at least 20 contiguous amino acid residues, and may be 30, 40, 50 or more amino acid residues. Corrections can be made for sequence identity associated with conservative substitutions of residues or spaces (see the Smith-Waterman algorithm for homology search). The precise chemical structure of a polypeptide capable of specifically binding to CD40 and retaining the antagonist activity, particularly when binding to the CD40 antigen in target cells, depends on a number of factors. Since ionizable amino and carboxyl groups are present in the molecule, a particular polypeptide can be obtained as an acid or basic salt, or in a neutral form. All similar preparations which retain their biological activity when placed under suitable environmental conditions, they are included in the definition of anti-CD40 antagonist antibodies as used herein. In addition, the primary amino acid sequence of the polypeptide can be increased by derivatization using sugar portions (glycosylation) or by other complementary molecules such as lipids, phosphate, acetyl groups and the like. It can also be increased by conjugation with saccharides. Certain aspects of such an increase are achieved through post-translational processing systems of the producing host; other similar modifications can be introduced in vitro. In any case, such modifications are included in the definition of an anti-CD40 antibody used herein, as long as the antagonistic properties of the anti-CD40 antibody are not destroyed. It is expected that such modifications may affect the activity quantitatively or qualitatively, either by increasing or decreasing the activity of the polypeptide, in the various assays. In addition, individual amino acid residues in the chain can be modified by oxidation, reduction or other derivation, and the polypeptide can be cleaved to obtain fragments that retain activity. Similar alterations, which do not destroy the antagonist activity, do not remove the polypeptide sequence from the definition of anti-CD40 antibodies of interest as used herein. The technique provides substantial guidance in the preparation and use of polypeptide variants. To prepare the anti-CD40 antibody variants, one skilled in the art can readily determine which modifications to the native protein, nucleotide sequence or amino acids will result in a variant that is suitable for use as a therapeutically active component of a pharmaceutical composition used in the methods of the present invention. The anti-CD40 antibody, for use in the methods of the invention, preferably possesses at least one of the following biological activities in vitro and / or in vivo: inhibition of immunoglobulin secretion by normal human peripheral B cells stimulated by cells T; inhibition of survival and / or proliferation of normal human peripheral B cells stimulated by cells expressing CD40L or by soluble CD40 ligand (sCD40L); inhibition of survival and / or proliferation of normal human peripheral B cells stimulated by Jurkat T cells; inhibition of intracellular anti-apoptotic signals of "survival" in any cell stimulated by sCD40L or solid phase CD40L; e, inhibition of signal transduction by CD40 in any cell with ligation with sCD40L or solid phase CD40L, elimination induction, anergy and / or tolerance of CD40-bearing target cells or cells carrying known ligands for CD40 including, but not limited to T cells and B cells, induction of expansion or activation of CD4 + CD25 + regulatory T cells (see, for example, rejection of alloantigen-specific donor tissues by interference CD40-CD40L, van Maurik et al. (2002) J Immunol 169: 5401-5404), cytotoxicity by any mechanism (including, but not limited to, antibody-dependent cell-mediated cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), sub-regulation of proliferation, and / or apoptosis in target cells), modulation of cytokine secretion and / or expression of cell surface molecules in target cells, and combinations thereof. Assays for such biological activities can be performed as described herein and in the provisional applications entitled "Monoclonal Antibodies Anti-CD40 Antagonists and Methods for Their Use" presented on November 4, 2003, November 26, 2003 and April 27 of 2004, and assigned US Patent Applications Nos. 60 / 517,337 (Attorney's File No. PP20107.001 (035784/258442)), 60 / 525,579 (Attorney's File No. PP20107.002 (035784/271525)), and 60 / 565,710 (File of the Representative No. PP20107.003 (035784/277214)), respectively; and International Patent Application No. PCT / US2004 / 037152 (Attorney's File No. PP20107.004 (035784/282916)), published as WO 2005/044854, also entitled "Anti-CD40 Monoclonal Antibodies Antagonists and Methods for Their Use" presented on November 4, 2004; the contents of each of which are incorporated herein for reference in their entirety. See also the assays described in Schultze et al. (1998) Proc. Nati Acad. Sci. USA 92: 8200-8204; Dentón ef al. (1998) Pediatr. Transplant. 2: 6-15; Evans et al. (2000) J Immunol. 164: 688-697; Noelle (1998) Agents Actions Suppl. 49: 17-22; Lederman et al. (1996) Curr. Opin. Hematol. 3: 77-86; Coligan ef al. (1991) Current Protocols in Immunology 13:12; Kwekkeboom ef al. (1993) Immunology 79: 439-444; and U.S. Patent Nos. 5,674,492 and 5,847,082; incorporated in the present for reference. A representative assay for detecting anti-CD40 antibodies specific for the epitopes of the CD40 antigen identified herein is a "competitive binding assay." Competitive binding assays are serological assays in which unknown compounds are detected and quantified for their ability to inhibit the binding of a known ligand labeled to its specific antibody. This is also referred to as a competitive inhibition assay. In a representative competitive binding assay, the tagged CD40 polypeptide is precipitated by candidate antibodies in a sample, for example, in combination with monoclonal antibodies raised against one or more epitopes of the monoclonal antibodies of the invention. Anti-CD40 antibodies that specifically react with an epitope of interest can be identified by selecting a series of antibodies prepared against a CD40 protein or fragment of the protein comprising the particular epitope of the CD40 protein of interest. For example, for human CD40, epitopes of interest include epitopes comprising linear and / or non-linear amino acid residues of the short isoform of human CD40 (see GenBank Accession No. NP 690593) set forth in SEQ ID NO: 10, encoded by the set sequence SEQ ID NO: 9; see also GenBank Access No. NM 152854), or of the long isoform of human CD40 (see Accesses GenBank Nos. CAA43045 and NP 001241, set forth in SEQ ID NO: 12, encoded by the sequence set forth in SEQ ID NO: 11; see Access GenBank Nos. X60592 and NM_001250). Alternatively, competitive binding assays with suitable antagonist anti-CD40 antibodies, previously identified, can be used to select monoclonal antibodies comparable with previously identified antibodies. The antibodies used in such immunoassays may be labeled or unlabeled. Non-labeled antibodies can be used in agglutination; labeled antibodies can be employed in a wide variety of assays, employing a wide variety of labels. Detection of the formation of an antibody-antigen complex, between an anti-CD40 antibody and an epitope of interest, can be facilitated by binding a detectable substance to the antibody. Suitable detection means include the use of labels such as radionuclides, enzymes, coenzymes, fluorescents, chemiluminescers, chromogens, substrates or co-factors of enzymes, enzyme inhibitors, complexes of prosthetic groups, free radicals, particles, dyes and the like. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material is luminol; examples of bioluminescent materials include luciferase, luciferin and aequorin; and examples of suitable radioactive material include 125 I, 131 I, 35 S or 3 H. Such labeled reagents can be used in a variety of well-known assays, such as radioimmunoassays, immunoassays with enzymes, for example, ELISA, fluorescent immunoassays and the like. See, for example, U.S. Patent Nos. 3,766,162; 3,791, 932; 3,817,837; and 4,233,402. It is also possible to design an antibody to have an increased activity of ADCC. In particular, the carboxy-terminal half of the CH2 domain is critical for ADCC mediated through the FcRIII receptor. Since CH2 and hinge regions have an important role in effector functions, a series of multi-domain antibodies containing extra CH2 and / or hinge regions can be created and investigated for any changes in effector potency (see Greenwood, J. , Gorman, SD, Routledge, EG, Lloyd, LS. &Waldmann, H., Ther Immunol, 1994 Oct; l (5): 247-55). An alternative methodology may be to design extra domains in parallel, for example, through the creation of dimers when designing a cysteine in the H chain of a chimeric Ig (see Shopes B. (1992) J. Immunol. 1992 1; 148 ( 9): 2918-22). Additionally, changes can be designed to increase the activity of ADCC by introducing mutations in the Fe region (see, for example, US 6,737,056 B1), expressing cells in fucosyl transferase-deficient cell lines (see, for example, US2003 / 0115614) or other changes to the glycosylation of the antibody (see, for example, US 6,602,684). The present invention is especially favorable for the treatment of inflammatory diseases and autoimmune diseases that are associated with cells expressing CD20, particularly in patients resistant to Rituxan®, more particularly in those who are heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F). As used herein, "anti-CD20 antibody" encompasses any antibody that specifically recognizes the cell surface antigen CD20, including polyclonal antibodies, monoclonal antibodies, single chain antibodies and fragments thereof such as Fab, F (ab ' ) 2, Fv and other fragments that retain the antigen-binding function of the parent anti-CD20 antibody. Of particular interest, in relation to the methods of the present invention, are the anti-CD20 antibodies or antigen-binding fragments thereof, which have the binding properties exhibited by the monoclonal antibody IDEC-C2B8 (Biogen IDEC Inc., Cambridge, MA). In some embodiments, the anti-CD40 antibodies used in the methods of the invention exhibit a more potent therapeutic activity than the chimeric anti-CD20 monoclonal antibody IDEC-C2B8, where the therapeutic activity is assayed with equivalent amounts of these antibodies in a model appropriate experimental IDEC-C2B8 (IDEC Pharmaceuticals Corp., San Diego, California, commercially available under the trademark Rituxan®, also referred to as rituximab) is a chimeric anti-CD20 monoclonal antibody that contains human IgG1 and kappa constant regions with murine variable regions isolated from a murine anti-CD20 monoclonal antibody, IDEC-2B8 (Reff et al. (1994) Blood 83: 435-445). Rituximab is approved for the treatment of non-Hodgkin's lymphoma (NHL) of B-cells, low-grade or follicular, recidivist, and is found in clinical trials for autoimmune diseases. The discovery of antibodies with superior therapeutic activity, compared to rituximab, can dramatically improve therapy methods for inflammatory diseases and autoimmune diseases. There are adequate models to test the activity in systemic lupus erythematosus (SLE), multiple sclerosis, inflammation and atherosclerosis, transplantation and Alzheimer's disease, as described in the following. For example, to test the efficacy in human systemic lupus erythematosus (SLE) in which peripheral blood mononuclear cells (PMBCs) from patients with SLE are grafted onto SCID mice. See, for example, the model described in Duchosal et al. (1990) J Exp. Med. 172: 985-8. After the transfer of the PBMCs from patients with SLE in SCID mice, it is determined whether the treatment influences or does not influence the response of T lymphocytes to the production of auto-antigens and auto-antibodies, and manifestations of disease such as glomerulonephritis. Experimental autoimmune encephalitis (EAE) in marmosets is a model for human multiple sclerosis. See, for example, the model described in Raine et al. (1999) Ann. Neurol. 46: 144-60 and Hart ef al. (2004) Lancet Ne? Rol. 3: 588-97. Antibodies can be tested in vitro for their ability to inhibit CD40L-induced production of matrix degradation enzymes, expression of tissue factors, proinflammatory cytokines and upregulation of adhesion molecules. Subsequent studies prove the ability of antibodies to show anti-inflammatory activities in vivo using transgenic mice expressing human CD40 and / or CD20 molecules. See, for example, the model described in Yasui (2002) Int. Immunol. 14: 319-29. Antibodies can be tested for their ability to prevent rejection of transplants in non-human primate models. Recipients of rhesus kidney allografts are treated with antibody to demonstrate the effect on graft acceptance with or without additional immunosuppressive drugs such as cyclosporin, FK506, rapamycin, corticosteroids, CTLA4-lg and anti-Stimulator antibody of B lymphocytes, and the like . See, for example, the model described in Wee ef al. (1992) Transplantation 53: 501-7. For Alzheimer's disease, antibodies can be tested first in vitro for their ability to block the activation of microglia cells. In vivo efficacy studies with the antibodies can be conducted in double transgenic mice expressing human CD40 and / or CD20 and overproducing amyloid-beta peptide. See, for example, the model described in Tan et al. (2002) Nat. Neurosci. 5: 1288-93. By "equivalent amount" of the anti-CD40 antibody of the invention and Rituxan®, it is intended that the same dose or less in mg be administered on a per-weight basis. Thus, where the anti-CD40 antibody is dosed at 0.01 mg / kg of body weight of the mouse used in the model, Rituxan® is also dosed at 0.01 mg / kg of mouse body weight. By way of Similar, where the anti-CD40 antibody is dosed at 0.1, 1 or 10 mg / kg of body weight of the mouse used in the model, the Rituxan® is also dosed at 0.1, 1 or 10 mg / kg, respectively, of body weight mouse. Another difference in the effectiveness of the antibody is to measure in vitro the concentration of antibody necessary to obtain the maximum lysis of target cells in vitro, in the presence of NK cells. For example, the anti-CD40 antibodies of the invention reach a maximum lysis of Daudi cells at an EC50 of less than Vi, and preferably VA, and more preferably, 1/10 of the concentration of Rituxan®. This type of measurement is also described in the Examples herein. Anti-CD40 antibodies that benefit from having significantly greater efficacy than equivalent amounts of Rituxan® in the assays described in the foregoing may include: a) the monoclonal antibody CHIR-12.12; b) the monoclonal antibody produced by the hybridoma cell line 12.12; c) a monoclonal antibody comprising an amino acid sequence selected from the group consisting of the sequence shown in SEQ ID NO: 2, the sequence shown in SEQ ID NO: 4, the sequence shown in SEQ ID NO: 5, the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 4, and the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 5; d) a monoclonal antibody having an amino acid sequence encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of the sequence shown in SEQ ID NO: 1, the sequence shown in SEQ ID NO: 3, and the sequences shown in SEQ ID NO: 1 and SEQ ID NO: 3; e) a monoclonal antibody that binds to an epitope capable of binding to the monoclonal antibody produced by the hybridoma cell line 12.12; f) a monoclonal antibody that binds to an epitope cosing residues 82-87 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; g) an antibody monoclonal that binds to an epitope cosing residues 82-89 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; h) a monoclonal antibody that competes with the monoclonal antibody CHIR-12.12 in a competitive binding assay; i) the monoclonal antibody of preceding article a) or a monoclonal antibody of any of preceding articles c) -h), wherein the antibody is produced recombinantly; and j) a monoclonal antibody that is an antigen-binding fragment of a monoclonal antibody of any of the preceding articles a) -i), wherein the fragment retains the ability to specifically bind to the human CD40 antigen. The present invention provides a method for identifying a human patient with an inflammatory disease or autoimmune disease, treatable with an anti-CD40 antibody, comprising: a) identifying a human patient with an inflammatory disease or autoimmune disease that is associated with cells expressing CD40; and b) determining the Fc? Rllla-158 genotype (V / V, V / F or F / F) of the human patient; wherein the inflammatory disease or autoimmune disease is treatable with an anti-CD40 antibody if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F). The inflammatory disease or autoimmune disease may be resistant to treatment with rituximab (Rituxan®). Once a human patient with an inflammatory disease or autoimmune disease treatable with an anti-CD40 antibody has been identified, that human patient can then be treated with an anti-CD40 antibody. In this manner, the method may include the additional step of (c) administering to a human patient identified as heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F) a therapeutically or prophylactically effective amount of an antibody anti-CD40. This method for identifying a human patient with an inflammatory disease or autoimmune disease, treatable with an anti-CD40 antibody, can be easily performed by a person skilled in the art using a suitable diagnostic equipment. The team it must comprise suitable reagents to determine a Fc? Rllla-158 genotype of the human patient. In this manner, the invention also provides a kit for identifying a human patient with an inflammatory disease or autoimmune disease treatable with an anti-CD40 antibody, comprising reagents for determining an FcγRllla-158 genotype of the human patient. The proper equipment is described in greater detail elsewhere in the present. The invention also provides a method for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease, comprising: (a) identifying a human patient who has an inflammatory disease or autoimmune disease that is associates with cells that express CD40; and (b) determining the Fc? Rllla-158 (VA /, V / F or F / F) genotype of the human patient; wherein if the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), an anti-CD40 antibody is selected for the treatment of inflammatory disease or autoimmune disease. The inflammatory disease or autoimmune disease may be resistant to treatment with rituximab (Rituxan®). Once a therapy with anti-CD40 antibodies for the treatment of a human patient having an inflammatory disease or autoimmune disease has been selected, that human patient can then be treated with an anti-CD40 antibody. In this manner, the method may include the additional step of (c) administering to a human patient identified as heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F) a therapeutically or prophylactically effective amount of an antibody anti-CD40. This method for selecting an antibody therapy, for the treatment of a human patient having an inflammatory disease or autoimmune disease, can also be easily performed by a person skilled in the art using suitable diagnostic equipment. The kit should comprise suitable reagents to determine an Fc? Rllla-158 genotype of the human patient. In this way, the invention also provides a kit for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease associated with cells that express CD40, which comprises reagents to determine an Fc? Rllla-158 genotype of the human patient. By "treatable with an anti-CD40 antibody" it is intended that the human patient (i.e., an individual with an inflammatory disease or autoimmune disease), when treated with the anti-CD40 antibody, may benefit from a "positive therapeutic response". "(as defined elsewhere herein) with respect to the inflammatory disease or autoimmune disease for which the treatment is sought. Any method for determining the Fc? Rllla-158 genotype of a human patient is contemplated using a biological sample obtained from the human patient. For example, the invention provides a kit for its use to determine the genotype Fc? Rllla-158 of a human patient, which includes a DNA microarray comprising at least one probe of 10 or more nucleotides in length and of a sequence suitable for determining Fc? Rllla-158 genotype of a human patient. The labeled RNA or DNA hybridizes to complement probes in the array and is then detected by laser scanning. Hybridization intensities for each probe in the array are determined and converted to a quantitative value representing relative levels of gene expression. The selection of sequences and lengths of probes can be easily performed by the experienced person. The nucleotide sequence of the human gene and mRNA encoding the Fc? Rllla-158 F and V allotypes is known. In this way, the experienced person can select the probe or probes that, under the appropriate experimental conditions, allow a determination of the genotype. Fc? Rllla-158 of the target sequences. Techniques for the synthesis of these arrays using mechanical synthesis methods are described, for example, in U.S. Patent No. 5,384,261, incorporated herein by reference in its entirety. Although a planar surface of the array is preferred, the array can be fabricated on a surface of virtually any conformation or even a multiplicity of surfaces. The arrays may be peptides or nucleic acids in beads, gels, polymeric surfaces, fibers such as optical fibers, glass or any other suitable substrate, see U.S. Patents Nos. 5,770,358, 5,789,162, 5,708,153, 6,040,193 and 5,800,992, each of which is hereby incorporated in its entirety for all purposes. Arrangements can be packaged in a way that allows diagnosis or other manipulation of an all-inclusive device. See, for example, U.S. Patent Nos. 5,856,174 and 5,922,591; incorporated herein by reference. For example, the invention also provides a kit for use in determining the FcγRllla-158 genotype of a human patient, comprising oligonucleotides suitable for use as primers in a polymerase-catalyzed amplification of the region of the gene or mRNA encoding the 158 amino acids of Fc? Rllla. The selection of sequences and lengths of primers can be easily performed by the experienced person. The nucleotide sequence of the human gene and mRNA encoding the Fc? Rllla-158 F and V allotypes is known. In this way, the experienced person can select primers which, under the appropriate experimental conditions, will allow the amplification of the region of the gene or mRNA that encodes the 158 amino acids of Fc? Rllla. The amplified sequence can then be sequenced using known methods to determine the Fc? Rllla-158 genotype of the patient. Another method to determine the Fc? Rllla158 genotype of a human patient is to use a nucleic acid-based method that detects the fragmentation of DNA that is characteristic of the Fc? Rllla-158 genotype of the human patient. When resolved using agarose gel electrophoresis, the DNA of each Fc? Rllla-158 genotype has a characteristic pattern. In this way, the invention also provides a kit for use in determining the Fc? Rllla-158 genotype of a human patient, comprising one or more suitable restriction enzymes for determining the Fc? Rllla-158 genotype of a human patient. Suitable restriction enzymes are known in the art (for example, see Koene ef al Blood, 1997, Vol 90, No. 3, p 1109-1114). The kits of the invention may also include instructions which indicate how to use the equipment to determine the Fc? Rllla-158 genotype of a human patient. The kit may also comprise, for example, a buffering agent, a preservative or a protein stabilizing agent. Each component of the equipment is usually enclosed within an individual container, and all the various containers are contained in a single package together with instructions which indicate how to use the equipment to determine the Fc? Rllla-158 genotype of a human patient. The invention provides the use of anti-CD40 antibodies in the manufacture of medicaments for treating an inflammatory disease or autoimmune disease associated with cells expressing CD40, as described elsewhere herein. The anti-CD40 antibodies of this invention are administered at a concentration that is therapeutically effective to prevent or treat an inflammatory disease or autoimmune disease associated with cells expressing CD40. To achieve this goal, antibodies can be formulated using a variety of acceptable carriers and / or excipients known in the art. The anti-CD40 antibody can be administered by a parenteral route of administration. Typically, the antibodies are administered by injection, either intravenously or subcutaneously. Methods for achieving this administration are known to those of ordinary skill in the art. Intravenous administration is preferably by infusion over a period of about half an hour to 1 hour to about 10 hours (less than 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 hours). Subsequent infusions can be administered for a period of from less than about 1 to about 6 hours, including, for example, about 1 to about 4 hours, about 1 to about 3 hours or about 1 to about 2 hours or less than an hour. Alternatively, an anti-CD40 antibody such as CHIR-12.12 can be administered subcutaneously. A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Solutions or suspensions used for parenteral application may include the following components: a sterile diluent, such as water for injection, saline; antibacterial agents, such as benzyl alcohol or methyl parabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates or phosphates and tonicity adjusting agents such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be included in ampules, disposable syringes or multi-dose vials made of glass or plastic. Anti-CD40 antibodies are typically provided by a standard technique within a pharmaceutically acceptable buffer, eg, sterile saline, sterile buffered water, combinations of the foregoing, etc. Methods for preparing agents that can be administered parenterally are described in Remington's Pharmaceutical Sciences (18th ed .; Mack Publishing Company, Eaton, Pennsylvania, 1990), incorporated herein by reference. See also, for example, WO 98/56418, which discloses stabilized pharmaceutical formulations of antibodies, suitable for use in the methods of the present invention. The amount of at least one anti-CD40 antibody to be administered is easily determined by one of ordinary skill in the art without unfounded experimentation. Factors influencing the mode of administration and the respective amount of at least one anti-CD40 antibody include, but are not limited to, the severity of the disease, the history of the disease and age, height, weight, health and physical condition of the individual who undergoes therapy. Similarly, the amount of anti-CD40 antibody to be administered will depend on the mode of administration and whether the subject will be subjected to a single dose or multiple doses of this agent against tumors. Generally, a higher dosage of the anti-CD40 antibody is preferred with the increasing weight of the subject being subjected to therapy. The dose of anti-CD40 antibody to be administered is in the range of about 0.003 mg / kg to about 50 mg / kg, from about 0.1 mg / kg to about 40 mg / kg, from about 0.01 mg / kg to about 30 mg / kg, from about 0.1 mg / kg to about 30 mg / kg, from about 0.5 mg / kg to about 30 mg / kg, gives about 1 mg / kg to about 30 mg / kg, from about 3 mg / kg to about 30 mg / kg, from about 3 mg / kg to about 25 mg / kg, from about 3 mg / kg to about 20 mg / kg, from about 5 mg / kg to about 15 mg / kg or from about 7 mg / kg to about 12 mg / kg. In this way, for example, the dose can be 0.3 mg / kg, 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 5 mg / kg, 7 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg or 50 mg / kg, or other similar doses that fall within the range of about 0.3 mg / kg to about 50 mg / kg. The treatment of a subject with a therapeutically effective amount of an antibody may include a single treatment or, preferably, may include a series of treatments. Thus, in another embodiment of the invention, the method comprises administering multiple doses of the anti-CD40 antibody. The method may comprise the administration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or more therapeutically effective doses of a pharmaceutical composition comprising a anti-CD40 antibody. The frequency and duration of multiple dose administration of the pharmaceutical compositions comprising the anti-CD40 antibody can be readily determined by one skilled in the art without unfounded experimentation. The same therapeutically effective dose of an anti-CD40 antibody can be administered during the course of a treatment period. Alternatively, different therapeutically effective doses of an anti-CD40 antibody can be used during the course of a treatment period. In a preferred example, a subject is treated with the anti-CD40 antibody in the range of between about 0.1 to 20 mg / kg body weight, once a week for between about 1 to 10 weeks, preferably between about 2 to 8 weeks. weeks, more preferably between approximately 3 to 7 weeks and even more preferably for approximately 4, 5 or 6 weeks. The treatment can be presented twice a year or annually to prevent recidivism or with the indication of recidivism. It will also be appreciated that the effective dosage of the antibody used for the treatment may be increased or decreased during the course of a particular treatment. Changes in dosage can result and become apparent from the results of diagnostic assays as described herein. Thus, in one embodiment, the dosage regimen includes a first administration of a therapeutically effective dose of at least one anti-CD40 antibody on days 1, 8, 15 and 22 of the treatment period. In another modality, the dosage regimen includes a dosage regimen having a first administration of a therapeutically effective dose of at least one anti-CD40 antibody daily, or on days 1, 3, 5 and 7 of a week in a treatment period; a dosage regimen including a first administration of a therapeutically effective dose of at least one anti-CD40 antibody on days 1 and 3-4 of a week in a treatment period; and a preferred dosage regimen including a first administration of a therapeutically effective dose of at least one anti-CD40 antibody on day 1 of a week in a treatment period. The treatment period can include 1 week, 2 weeks, 3 weeks, one month, 2 months, 3 months, 6 months or a year. The treatment periods can be subsequent or separated from each other for a week, 2 weeks, a month, 3 months, 6 months or a year. In other embodiments, the therapeutically effective initial dose of an anti-CD40 antibody, as defined elsewhere herein, may be in the lower dosage range (i.e., about 0.003 mg / kg to about 20 mg / kg) with subsequent doses that fall within the highest dosage range (i.e., from about 20 mg / kg to about 50 mg / kg). In alternative embodiments, the therapeutically effective initial dose of an anti-CD40 antibody, as defined elsewhere herein, may be in the upper dosage range (i.e., about 20 mg / kg to about 50 mg / kg) with subsequent doses that fall within the lower dosage range (ie, 0.003 mg / kg to approximately 20 mg / kg). Thus, in some embodiments of the invention, anti-CD40 antibody therapy can be initiated by administering an "attack dose" of the antibody to the subject in need of therapy. By "attack dose" is meant an initial dose of the anti-CD40 antibody that is administered to the subject, wherein the dose of the administered antibody falls within the highest dosage range (i.e., from about 20 mg / kg to about 50 mg / kg). The "attack dose" can be administered as a single administration, for example, a single infusion where the antibody is administered IV, or as multiple administrations, for example, multiple infusions where the antibody is administered IV, as long as the "dose of "full" attack is administered within a period of approximately 24 hours. After administration of the "attack dose," the subject is then administered one or more additional therapeutically effective doses of the anti-CD40 antibody. Subsequent therapeutically effective doses may be administered, for example, according to a dosing schedule weekly or once every two weeks, once every three weeks or once every four weeks. In such embodiments, subsequent therapeutically effective doses generally fall within the lower dosage range (ie, 0.003 mg / kg to about 20 mg / kg). Alternatively, in some embodiments, after the "attack dose", the subsequent therapeutically effective doses of the anti-CD40 antibody are administered according to a "maintenance schedule," wherein the therapeutically effective dose of the antibody is administered a once a month, once every 6 weeks, once every two months, once every 10 weeks, once every three months, once every 14 weeks, once every four months, once every 18 weeks, once every five months, once every 22 weeks, once every six months, once every 7 months, once every 8 months, once every 9 months, once every 10 months, once every 11 months, or once every 12 months. In such embodiments, the therapeutically effective doses of the anti-CD40 antibody fall within the lower dosage range (ie, 0.003 mg / kg to about 20 mg / kg), particularly when the subsequent doses are administered at more frequent intervals, for example , once every two weeks to once a month, or within the highest dosage range (i.e., from about 20 mg / kg to about 50 mg / kg), particularly when the subsequent doses are administered at less frequent intervals, for example, where subsequent doses are administered approximately one month to approximately 12 months apart. The anti-CD40 antibodies, present in the pharmaceutical compositions described herein, for use in the methods of the invention, can be native or obtained by recombinant techniques, and can be from any source, including mammalian sources such as, for example, example, mouse, rat, rabbit, primate, pig and human. Preferably, such polypeptides are derived from a human source and, most preferably, are recombinant human proteins of hybridoma cell lines. Pharmaceutical compositions useful in the methods of the invention may comprise biologically active variants of the anti-CD40 antagonist antibodies of the invention, as described elsewhere herein. Any pharmaceutical composition comprising an anti-CD40 antibody, which has the binding properties described herein as the therapeutically active component, can be used in the methods of the invention. In this manner, liquid, lyophilized or spray-dried compositions, comprising one or more of the anti-CD40 antibodies, can be prepared as an aqueous or nonaqueous solution or suspension for subsequent administration to a subject according to the methods of the invention. Each of these compositions will comprise at least one anti-CD40 antibody as a therapeutic or prophylactically active component. By "therapeutically or prophylactically active component" it is intended that the anti-CD40 antibody is specifically incorporated into the composition to give rise to a desired therapeutic or prophylactic response in consideration of the treatment, prevention or diagnosis of a disease or condition within a subject when The pharmaceutical composition is administered to that subject. Preferably, the pharmaceutical compositions comprise stabilizing agents, appropriate consistency agents, or both, to minimize problems associated with loss of protein stability and biological activity in the course of preparation and storage. Formulation agents can be added to pharmaceutical compositions comprising an anti-CD40 antibody of the invention. These formulation agents can include, but are not limited to, oils, polymers, vitamins, carbohydrates, amino acids, salts, buffers, albumin, surfactants or consistency agents. Preferably, the carbohydrates include sugar or sugar alcohols such as mono, di or polysaccharides, or water-soluble glycans. The saccharides or glucans can include fructose, glucose, mannose, sorbose, xylose, maltose, sucrose, dextran, pullulana, dextrin, a and β-cyclodextrin, soluble starch, hydroxyethylstarch and carboxymethylcellulose, or mixtures thereof. "Sugar alcohol" is defined as a C4 to C8 hydrocarbon that has a hydroxyl group and includes galactitol, inositol, mannitol, xylitol, sorbitol, glycerol and arabitol. These sugars or sugar alcohols can be used individually or in combination. The sugar or sugar alcohol concentration is between 1.0% and 7% w / v, more preferably between 2.0% and 6.0% w / v. Preferably, the amino acids include levogy (L) forms of carnitine, arginine and betaine; however, other amino acids can be added. Preferred polymers include polyvinylpyrrolidone (PVP) with an average molecular weight between 2,000 and 3,000, or polyethylene glycol (PEG) with an average molecular weight between 3,000 and 5,000. The surfactants that can be added to the formulation are shown in EP Nos. 270,799 and 268,110. Additionally, the antibodies can be chemically modified by covalent conjugation to a polymer to increase their half-life in circulation, for example. Preferred polymers, and methods for binding them to peptides, are shown in U.S. Patent Nos. 4,766,106; 4,179,337; 4,495,285; and 4,609,546; which are incorporated by this for reference in their totalities. Preferred polymers are polyoxyethylated polyols and polyethylene glycol (PEG). PEG is soluble in water at room temperature and has the general formula: R (0-CH 2 -CH 2) n O-R where R can be hydrogen, or a protecting group such as an alkyl or alkanol group. Preferably, the protecting group has between 1 and 8 carbons, more preferably it is methyl. The symbol n is a positive whole number, preferably between 1 and 1, 000, more preferably between 2 and 500. The PEG has a preferred average molecular weight between 1,000 and 40,000, more preferably between 2,000 and 20,000, more preferred between 3,000 and 12,000. Preferably, the PEG has at least one hydroxy group, more preferably it is a terminal hydroxy group. It is this hydroxy group which is preferably activated to react with a free amino group in the inhibitor. However, it will be understood that the type and amount of the reactive groups can be varied to achieve a covalently conjugated PEG / antibody of the present invention. Water-soluble polyoxyethylated polyols are also useful in the present invention.

These include polyoxyethylated sorbitol, polyoxyethylated glucose, polyoxyethylated glycerol (POG) and the like. POG is preferred. One reason is because the glycerol backbone of the polyoxyethylated glycerol is the same main chain that occurs naturally, for example, in animals and humans in mono, di, triglycerides. Therefore, this branching might not necessarily be observed as a foreign agent in the body. POG has a preferred molecular weight in the same range as PEG. The structure for the POG is shown in Knauf et al. (1988) J Bio. Chem. 263: 15064-15070, and a discussion of POG / IL-2 conjugates is found in U.S. Patent No. 4,766,106, which are hereby incorporated by reference in their totals. Another system of drug delivery to increase the circulatory half-life is the liposome. Methods for preparing liposome delivery systems are discussed in Gabizon et al. (1982) Cancer Research 42: 4734; Cafiso (1981) Biochem Biophys Acta 649: 129; and Szoka (1980) Ann. Rev. Biophys. Eng. 9: 467. Other drug delivery systems are known in the art and are described, for example, in Poznansky et al. (1980) Drug Delivery Systems (R.L. Juliano, ed., Oxford, N. Y.) pp. 253-315; Poznansky (1984) Pharm Revs 36: 277. Formulation agents that must be incorporated into a pharmaceutical composition must allow stability or anti-CD40 antibody. That is, the anti-CD40 antibody must retain its physical and / or chemical stability and have the desired biological activity, i.e., one or more of the antagonist activities defined herein, including, but not limited to, inhibition. of secretion of immunoglobulins by normal human peripheral B cells stimulated by T cells; inhibition of survival and / or proliferation of normal human peripheral B cells stimulated by Jurkat T cells; inhibition of survival and / or proliferation of normal human peripheral B cells stimulated by cells expressing CD40L or by soluble CD40 ligand (sCD40L); inhibition of intracellular anti-apoptotic signals of "survival" in any cell stimulated by sCD40L or solid phase CD40L; inhibition of signal transduction by CD40 in any cell with ligation with sCD40L or CD40L in solid phase; and inhibition of the proliferation of human malignant B cells as observed elsewhere herein. Methods for monitoring protein stability are well known in the art. See, for example, Jones (1993) Adv. Drug Delivery Rev. 10: 29-90; Lee, ed. (1991) Peptide and Protein Drug Delivery (Marcel Dekker, Inc., New York, New York); and stability tests described herein in the following. Generally, protein stability is measured at a chosen temperature for a specified period of time. In preferred embodiments, a stable pharmaceutical formulation of antibodies enables stability of the anti-CD40 antibody when stored at room temperature (approximately 25 ° C) for at least 1 month, at least 3 months, or at least 6 months , and / or is stable at approximately 2-8 ° C for at least 6 months, at least 9 months, at least 12 months, at least 18 months, at least 24 months. A protein, such as an antibody, when formulated in a pharmaceutical composition, is considered to retain its physical stability at a given point in time if it does not show visual signs (i.e., discoloration or loss of clarity) or measurable signs (eg, example, using size exclusion chromatography (SEC) or UV light scattering) of precipitation, aggregation and / or denaturation in that pharmaceutical composition. With respect to chemical stability, a protein such as an antibody, when formulated into a pharmaceutical composition, is considered to retain its chemical stability at a given point in time if the chemical stability measurements are indicative of the protein (i.e. , antibody) retains the biological activity of interest in that pharmaceutical composition. Methods for monitoring changes in chemical stability are well known in the art and include, but are not limited to, methods for detecting chemically altered forms of the protein such as those resulting from truncation, using, for example, SDS-PAGE, SEC , and / or mass spectrometry by laser desorption-ionization / matrix-assisted flight time; and degradation associated with changes in molecular charge (e.g., associated with deamidation), using, for example, ion exchange chromatography. See, for example, the methods described herein in the following. An anti-CD40 antibody, when formulated in a pharmaceutical composition, is considered to retain a desired biological activity at a given point in time if the desired biological activity at that time is within about 30%, preferably within about 20. % of the desired biological activity exhibited at the time the pharmaceutical composition was prepared, as determined in a suitable assay for the desired biological activity. Assays for measuring the desired biological activity of the anti-CD40 antibodies can be performed as described in the Examples herein. See also the assays described in Schultze et al. (1998) Proc. Nati Acad. Sci. USA 92: 8200-8204; Dentón ef al. (1998) Pediatr. Transplant. 2: 6-15; Evans ef al. (2000) J Immunol. 164: 688-697; Noelle (1998) Agents Actions Suppl. 49: 17-22; Lederman et al. (1996) Curr. Opin. Hematol. 3: 77-86; Coligan ef al. (1991) Current Protocols in Immunology 13:12; Kwekkeboom ef al. (1993) Immunology 79: 439-444; and U.S. Patent Nos. 5,674,492 and 5,847,082; incorporated herein by reference. In some embodiments of the invention, the anti-CD40 antibody is formulated in a liquid pharmaceutical formulation. The anti-CD40 antibody can be prepared using any method known in the art, including those methods described herein in the foregoing. In one embodiment, the anti-CD40 antibody is produced recombinantly in a CHO cell line. Where the anti-CD40 antibody is to be stored prior to its formulation, it can be frozen, for example, at < -20 ° C, and then thaw at room temperature for later formulation. The liquid pharmaceutical formulation comprises a therapeutically effective amount of the anti-CD40 antibody. The amount of antibody of the same, present in the formulation, takes into consideration the route of administration and desired volume of dose. In this form, the liquid pharmaceutical composition comprises the anti-CD40 antibody at a concentration of about 0.1 mg / ml to about 50.0 mg / ml, about 0.5 mg / ml to about 40.0 mg / ml, about 1.0 mg / ml to about 30.0 mg / ml, about 5.0 mg / ml to about 25.0 mg / ml, about 5.0 mg / ml to about 20.0 mg / ml or about 15.0 mg / ml to approximately 25.0 mg / ml. In some embodiments, the liquid pharmaceutical composition comprises the anti-CD40 antibody at a concentration of about 0.1 mg / ml to about 5.0 mg / ml, about 5.0 mg / ml to about 10.0 mg / ml, about 10.0 mg / ml to about 15.0. mg, approximately 15.0 mg / ml a about 20.0 mg / ml, about 20.0 mg / ml to about 25.0 mg / ml, about 25.0 mg / ml to about 30.0 mg / ml, about 30.0 mg / ml to about 35.0 mg / ml, about 35.0 mg / ml to about 40.0 mg / ml, approximately 40.0 mg / ml to approximately 45.0 mg / ml or approximately 45.0 mg / ml to approximately 50.0 mg / ml. In other embodiments, the liquid pharmaceutical composition comprises the anti-CD40 antibody at a concentration of about 15.0 mg / ml, about 16.0 mg / ml, about 17.0 mg / ml, about 18.0 mg / ml, about 19.0 mg / ml, about 20.0 mg / ml, approximately 21.0 mg / ml, approximately 22.0 mg / ml, approximately 23.0 mg / ml, approximately 24.0 mg / ml or approximately 25.0 mg / ml. The liquid pharmaceutical composition comprises the anti-CD40 antibody and a buffer which maintains the pH of the formulation in the range from about pH 5.0 to about pH 7.0, including about pH 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, and other similar values within the range of about pH 5.0 to about pH 7.0. In some embodiments, the buffer maintains the pH of the formulation in the range of from about pH 5.0 to about pH 6.5, about pH 5.0 to about pH 6.0, about pH 5.0 to about pH 5.5, about pH 5.5 to about 7.0, about pH 5.5 at about pH 6.5 or about pH 5.5 at about pH 6.0. Any suitable buffer that maintains the pH of the liquid formulation of anti-CD40 antibodies in the range of from about pH 5.0 to about pH 7.0 can be used in the formulation, as long as the desired physical-chemical stability and biological activity of the antibody is retained as observed in the present in the previous. Suitable buffers include, but are not limited to, conventional acids and salts thereof, where the counter-ion may be, for example, sodium, potassium, ammonium, calcium or magnesium. Examples of conventional acids and salts thereof which can be used to buffer the liquid pharmaceutical formulation include, but are not limited to, succinic acid or succinate buffers, citric acid or citrate, acetic acid or acetate, tartaric acid or tartarate, phosphoric acid or phosphate, acid Gluconic or gluconate, glutamic acid or glutamate, aspartic acid or aspartate, maleic acid or maleate and malic acid or malate. The concentration of the buffer within the formulation can be from about 1 mM to about 50 mM, including about 1 mM, 2 mM, 5 mM, 8 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM , 40 mM, 45 mM, 50 mM, or other similar values within the range of about 1 mM to about 50 mM. In some embodiments, the concentration of the buffer within the formulation is from about 5 mM to about 15 mM, including about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM , 14 mM, 15 mM, or other similar values within the range of about 5 mM to about 15 mM. In some embodiments of the invention, the liquid pharmaceutical formulation comprises a therapeutically effective amount of the anti-CD40 antibody and succinate buffer or citrate buffer at a concentration that maintains the pH of the formulation in the range of about pH 5.0 to about pH 7.0 , preferably about pH 5.0 to about pH 6.5. By "succinate buffer" or "citrate buffer" is meant a buffer comprising a salt of succinic acid or a salt of citric acid, respectively. In a preferred embodiment, the counter-ion of succinate or citrate is the sodium cation and, thus, the buffer is sodium succinate or sodium citrate, respectively. However, any cation is expected to be effective. Other possible succinate or citrate cations include, but are not limited to, potassium, ammonium, calcium and magnesium. As noted above, the concentration of the succinate or citrate buffer within the formulation can be from about 1 mM to about 50 mM, including about 1 mM, 2 mM, 5 mM, 8 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, or other similar values within the range of about 1 mM to about 50 mM. In some embodiments, the concentration of the buffer within the formulation is from about 5 mM to about 15 mM, including about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM , 14 mM or approximately 15 mM. In other embodiments, the liquid pharmaceutical formulation comprises the anti-CD40 antibody at a concentration of about 0.1 mg / ml to about 50.0 mg / ml, or about 5.0 mg / ml at about 25.0 mg / ml, and succinate or citrate buffer, for example, sodium succinate or sodium citrate buffer, at a concentration of about 1 mM to about 20 mM, about 5 mM to about 15 mM, preferably about 10 mM. Where it is desirable for the liquid pharmaceutical formulation to be almost isotonic, the liquid pharmaceutical formulation comprising the anti-CD40 antibody and a buffer may further comprise an amount of an isotonicity agent sufficient to render the formulation almost isotonic. By "quasi isotonic" it is intended that the aqueous formulation have an osmolarity of about 240 mmol / kg to about 360 mmol / kg, preferably about 240 to about 340 mmol / kg, more preferably about 250 to about 330 mmol / kg, even more preferably about 260 to about 320 mmol / kg, even more preferably about 270 to about 310 mmol / kg. Methods for determining the isotonicity of a solution are known to those skilled in the art. See, for example, Setnikar ef al. (1959) J Am. Pharm. Assoc. 48: 628. Those skilled in the art are familiar with a variety of pharmaceutically acceptable solutes, useful for providing isotonicity in pharmaceutical compositions. The isotonicity agent can be any reagent capable of adjusting the osmotic pressure of the liquid pharmaceutical formulation of the present invention to a value almost equal to that of a body fluid. It is desirable to use a physiologically acceptable isotonicity agent. In this manner, the liquid pharmaceutical formulation comprising a therapeutically effective amount of the anti-CD40 antibody and a buffer may further comprise components that can be used to provide isotonicity, for example, sodium chloride; amino acids such as alanine, valine and glycine; sugars and sugar alcohols (polyols), including, but not limited to, glucose, dextrose, fructose, sucrose, maltose, mannitol, trehalose, glycerol, sorbitol and xylitol; acetic acid, other organic acids or their salts, and relatively minor amounts of citrates or phosphates. The ordinary experienced person may know of additional agents that are suitable to provide an optimum tonicity of the formulation liquid In some preferred embodiments, the liquid pharmaceutical formulation comprising an anti-CD40 antibody and a buffer further comprises sodium chloride as the isotonicity agent. The concentration of sodium chloride in the formulation will depend on the contribution of other components to the tonicity. In some embodiments, the concentration of sodium chloride is about 50 mM to about 300 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, about 50 mM to about 175 mM, about 50 mM to about 150 mM , about 75 mM to about 175 mM, about 75 mM to about 150 mM, about 100 mM to about 175 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 125 mM to about 175 mM, approximately 125 mM to approximately 150 mM, approximately 130 mM to approximately 170 mM, approximately 130 mM to approximately 160 mM, approximately 135 mM to approximately 155 mM, approximately 140 mM to approximately 155 mM or approximately 145 mM to approximately 155 mM. In a similar embodiment, the concentration of sodium chloride is approximately 150 mM. In other similar embodiments, the sodium chloride concentration is about 150 mM, the buffer is sodium succinate buffer or sodium citrate at a concentration of about 5 mM to about 15 mM, the liquid pharmaceutical formulation comprises a therapeutically effective amount of the anti-CD40 antibody and the formulation has a pH of about pH 5.0 at about pH 7.0, about pH 5.0 to about pH 6.0 or about pH 5.5 to about pH 6.5. In other embodiments, the liquid pharmaceutical formulation comprises the anti-CD40 antibody at a concentration of about 0.1 mg / ml to about 50.0 mg / ml or about 5.0 mg / ml to about 25.0 mg / ml, about 150 mM sodium chloride and succinate of sodium or sodium citrate approximately 10 mM, at a pH of approximately pH 5.5.

The degradation of proteins due to freeze-thawing or mechanical cutting, in the course of the processing of the liquid pharmaceutical formulations of the present invention, can be inhibited by the incorporation of surfactants in the formulation in order to decrease the interfacial surface tension. solution-air. Thus, in some embodiments, the liquid pharmaceutical formulation comprises a therapeutically effective amount of the anti-CD40 antibody, a buffer and further comprises a surfactant. In other embodiments, the liquid pharmaceutical formulation comprises an anti-CD40 antibody, a buffer, an isotonicity agent and further comprises a surfactant. Typical surfactants employed are nonionic surfactants, including polyoxyethylene sorbitol esters such as polysorbate 80 (Tween 80) and polysorbate 20 (Tween 20); polyoxypropylene-polyoxyethylene esters such as Pluronic F68; polyoxyethylene alcohols such as Brij 35; simethicone; polyethylene glycol such as PEG400; lysophosphatidylcholine; and polyoxyethylene-p-t-octylphenol such as Triton X-100. The classical stabilization of pharmaceuticals by surfactants or emulsifiers is described, for example, in Levine et al. (1991) J Parenteral Sci. Technol. 45 (3): 160--165, incorporated herein by reference. A preferred surfactant, employed in the practice of the present invention, is polysorbate 80. Where a surfactant is included, typically it is added in an amount from about 0.001% to about 1.0% (w / v), about 0.001% to about 0.5% , about 0.001% to about 0.4%, about 0.001% to about 0.3%, about 0.001% to about 0.2%, about 0.005% to about 0.5%, about 0.005% to about 0.2%, about 0.01% to about 0.5%, about 0.01% to about 0.2%, about 0.03% to about 0.5%, about 0.03% to about 0.3%, about 0.05% to about 0.5% or about 0.05% to about 0.2%. Thus, in some embodiments, the liquid pharmaceutical formulation comprises a therapeutically effective amount of the anti-CD40 antibody, the buffer is sodium succinate or sodium citrate buffer at a concentration of about 1 mM at about 50 mM, about 5 mM to about 25 mM or about 5 mM to about 15 mM; the formulation has a pH of about pH 5.0 to about pH 7.0, about pH 5.0 to about pH 6.0 or about pH 5.5 to about pH 6.5; and the formulation further comprises a surfactant, for example, polysorbate 80, in an amount from about 0.001% to about 1.0% or about 0.001% to about 0.5%. Such formulations may optionally comprise an isotonicity agent, such as sodium chloride, at a concentration of about 50 mM to about 300 mM, about 50 mM to about 200 mM or about 50 mM to about 150 mM. In other embodiments, the liquid pharmaceutical formulation comprises the anti-CD40 antibody at a concentration of about 0.1 mg / ml to about 50.0 mg / ml or about 5.0 mg / ml to about 25.0 mg / ml, including about 20.0 mg / ml; sodium chloride approximately 50 mM to approximately 200 mM, including sodium chloride approximately 150 mM; sodium succinate or sodium citrate at about 5 mM to about 20 mM, including sodium succinate or about 10 mM sodium citrate; sodium chloride at a concentration of about 50 mM to about 200 mM, including about 150 mM; and optionally a surfactant, for example, polysorbate 80, in an amount from about 0.001% to about 1.0%, including from about 0.001% to about 0.5%; wherein the liquid pharmaceutical formulation has a pH of from about pH 5.0 to about pH 7.0, about pH 5.0 to about pH 6.0, about pH 5.0 to about pH 5.5, about pH 5.5 to about pH 6.5 or about pH 5.5 to about pH 6.0. The liquid pharmaceutical formulation can be essentially free from any preservatives and other carriers, excipients or stabilizers noted herein. Alternatively, the formulation may include one or more preservatives, e.g., antibacterial agents, carriers, excipients or pharmaceutically stabilizers. acceptable methods described herein in the above to the condition that they do not adversely affect the physicochemical stability of the anti-CD40 antibody. Examples of acceptable carriers, excipients and stabilizers include, but are not limited to, additional buffering agents, cosolvents, surfactants, antioxidants including ascorbic acid and methionine, chelating agents such as EDTA, metal complexes (eg, Zn-protein complexes) and biodegradable polymers such as polyesters. A thorough discussion of the formulation and selection of pharmaceutically acceptable carriers, stabilizers and physiological solutions can be found in Remington's Pharmaceutical Sciences (18th ed .; Mack Publishing Company, Eaton, Pennsylvania, 1990), incorporated herein by reference. "Carriers", as used herein, include pharmaceutically acceptable carriers, excipients or stabilizers that are non-toxic to the cell or mammal that is exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous solution of buffered pH. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate, succinate and other organic acids; antioxidants including ascorbic acid; low molecular weight polypeptides (less than about 10 residues); proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN, polyethylene glycol (PEG) and Pluronics. The administration "in combination with" one or more additional therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order. After the liquid pharmaceutical formulation, or other pharmaceutical composition described herein, is prepared, it can be lyophilized to prevent its degradation. Methods for lyophilizing liquid compositions are known to those of ordinary skill in the art. Just before use, the composition can be reconstituted with a sterile diluent (Ringer's solution, distilled water or sterile saline, for example) which may include additional ingredients Upon reconstitution, the composition is preferably administered to the subjects using those methods that are known to those skilled in the art. In some embodiments, anti-CD40 antibodies may be administered in combination with at least one other known therapy for inflammatory and / or autoimmune diseases. Such therapies include, but are not limited to, surgery or surgical procedures (e.g., splenectomy, lymphadenectomy, thyroidectomy, plasmapheresis, leukophoresis, transplantation of cells, tissues or organs, intestinal procedures, organ perfusion and the like), radiation therapy, therapy such as steroid therapy and non-steroidal therapy, hormone therapy, cytokine therapy, therapy with dermatological agents (eg, topical agents used to treat skin conditions such as allergies, contact dermatitis and psoriasis), immunosuppressive therapy and other anti-inflammatory therapy -inflammatory with monoclonal antibodies, and the like. Where the methods of the present invention comprise combined therapeutic regimens, these therapies can be given simultaneously, i.e., the anti-CD40 antagonist antibody, or antigen-binding fragment thereof, is administered concurrently or within the same timeframe as the other therapy (ie, the therapies are continued concurrently, but the anti-CD40 antibody, or antigen-binding fragment thereof), it is not administered precisely at the same time as the other therapy). Alternatively, the anti-CD40 antagonist antibody of the present invention, or antigen-binding fragment thereof, may also be administered prior to, or subsequent to, the other therapy. The sequential administration of the different therapies can be carried out regardless of whether the treated subject responds to the first course of therapy to reduce the possibility of remission or recidivism. In this form, anti-CD40 antibodies are administered in combination with at least one other therapy, including, but not limited to, surgery, organ perfusion, radiation therapy, steroid therapy, non-steroidal therapy, antibiotic therapy, therapy of antifungals, hormonal therapy, cytokine therapy, therapy with dermatological agents (for example, topical agents used to treat skin conditions such as allergies, contact dermatitis and psoriasis), immunosuppressive therapy, other anti-inflammatory therapy with monoclonal antibodies, combinations of the same and similar. In this way, where Combination therapies comprise the administration of an anti-CD40 antibody in combination with the administration of another therapeutic agent, such as with steroids as an example, the methods of the invention encompass co-administration, using separate formulations or as a single pharmaceutical formulation, and consecutive administration in any order. Examples of immunosuppressive drugs that can be administered in combination with anti-CD40 antibodies include, but are not limited to, methotrexate, cyclophosphamide, mizoribine, chlorambucil, cyclosporin, such as, for example, cyclosporin aerosol (see, Patent Application Publication). North American No. US20020006901, incorporated herein by reference in its entirety), tacrolimus (FK506, ProGraf ™), mycophenolate-mofetil and azathioprine (6-mercaptopurine), sirolimus (rapamycin), deoxyspergualin, leflunomide and its malononitriloamide analogues; and immunosuppressant proteins, including, Examples of suitable anti-inflammatory agents that can be administered in combination with anti-CD40 antibodies include, but are not limited to, corticosteroids such as, for example, clobetasol, halobetasol, hydrocortisone, triamcinolone, betamethasone, fluocinol. , fluocinonide, prednisone, prednisolone, methylprednisolone; non-steroidal anti-inflammatory drugs (NSAIDs) such as, for example, sulfasalazine, medicines containing mesalamine (known as 5-ASA agents), celecoxib, diclofenac, etodolac, fenprofen, flurbiprofen, ibuprofen, ketoprofen, meclofamate, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, rofecoxib, salicylates, sulindac and tolmetin; Enbrel ® (a soluble TNF receptor), anti-inflammatory antibodies such as adalimumab (HUMIRA®, a TNF-α antagonist) and infliximab (Remicade®, a TNF-α antagonist) and the like. Rejection of transplants and graft versus host disease can be hyperacute (humoral), acute (mediated by T cells) or chronic (unknown etiology) or a combination thereof. In this manner, the anti-CD40 antibodies of the invention are used in some embodiments to prevent and / or alleviate rejection and / or symptoms associated with rejection of hyperacute, acute and / or chronic transplants of any tissue, including, but not limited to, liver, kidney, pancreas, pancreatic islet cells, small intestine, lung, heart, corneas, skin, blood vessels, bone, heterologous or autologous bone marrow and the like.

Graft tissues can be obtained from any donor and transplanted into any recipient host and, thus, the transplant procedure can comprise transplanting animal tissue to humans (e.g., xenografts), transplanting tissue from one human to another human (e.g. allografts), and / or transplant tissue from one part of a human body to another (for example, autografts). The treatment with the antibodies of the invention can also reduce the sequelae of transplantation, such as fever, anorexia, hemodynamic abnormalities, leukopenia, infiltration of leukocytes from the transplanted organ / tissue, as well as opportunistic infections. In embodiments where the anti-CD40 antibodies of the invention are used to treat rejection of grafts, rheumatoid arthritis or multiple sclerosis, the antibodies can be used in combination with suitable immunosuppressive drugs as described herein. In this manner, the invention provides the use of a therapeutically or prophylactically effective amount of an anti-CD40 antibody in the manufacture of a medicament for the treatment of an inflammatory disease or autoimmune disease that is associated with cells expressing CD40 in a patient human heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), wherein the drug is coordinated with treatment with at least one other therapy. By "coordinated" it is intended that the medicament comprising the anti-CD40 antibody should be used either before, during or after treatment of the subject with at least one other therapy. The invention also makes possible the use of an anti-CD40 antibody in the manufacture of a medicament for treating a human patient for an inflammatory disease or autoimmune disease which is associated with cells expressing CD40, wherein the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F) and has been pretreated with at least one other therapeutic agent. By "pretreated" or "pretreatment" it is intended that the subject has received one or more other therapies (i.e., has been treated with at least one other therapy) before receiving the medicament comprising the anti-CD40 antibody. "Pretreated" or "pretreatment" includes subjects who have been treated with at least one other therapy within 2 years, within 18 months, within 1 year, within 6 months, within 2 months, within 6 weeks, within 1 month, within 4 weeks , within 3 weeks, within 2 weeks, within 1 week, within 6 days, within 5 days, within 4 days, within 3 days, within 2 days or even within 1 day before the start of treatment with the medicament comprising the anti-CD40 antibody. It is not necessary for the subject to respond well to pretreatment with previous therapy, or previous therapies. In this manner, the subject receiving the medicament comprising the anti-CD40 antagonist antibody may have responded, or may have failed to respond (ie, the disease was resistant), to pretreatment with the previous therapy, or to one or more of the previous therapies, where the pretreatment comprised multiple therapies. Examples of other therapies for which a subject may have received pretreatment prior to receiving the medicament comprising the anti-CD40 antibody include, but are not limited to, the therapies for the inflammatory or auto immune diseases described elsewhere herein. "Treatment", in the context of the coordinated use of a medicament described herein with one or more other therapies, is defined herein as the application or administration of the medicament or other therapy to a subject, or the application or administration of the medicament or other therapy to a tissue or cell line isolated from a subject, where the subject has a disease, a symptom of a disease or a predisposition towards a disease, where the purpose is to cure, heal, soothe, mitigate, alter, remedy , alleviate, improve or affect the disease, the symptoms of the disease, or the predisposition towards the disease.

In some embodiments, combination therapy provides a synergistic improvement in therapeutic efficacy with respect to individual therapeutic agents when administered alone. The term "synergy" is used to describe a combined effect of two or more active agents, which is greater than the sum of the individual effects of each respective active agent. In this way, where the combined effect of two or more agents results in "synergistic inhibition" of an activity or process, it is intended that the inhibition of the activity or process be greater than the sum of the inhibitory effects of each respective active agent. The term "synergistic therapeutic effect" refers to a therapeutic effect observed with a combination of two or more therapies, where the therapeutic effect (as measured by any of a series of parameters) is greater than the sum of the individual therapeutic effects observed with the respective individual therapies. Various aspects and embodiments of the present invention will now be described in more detail by way of example only. It will be appreciated that the modification of the detail can be made without departing from the scope of the invention.

EXPERIMENTS The anti-CD40 antibody, used in the examples in the following, is CHIR-12.12. The production, sequencing and characterization of the CHIR-12.12 antibody is described in detail in the international patent applications published as WO 2005/044854, WO 2005/044304, WO 2005/044305, WO 2005/044306, WO 2005/044855, WO 2005/044307 and WO 2005/044294. The hybridoma line 153.8E2.D10.D6.12.12 (CMCC # 12056) expressing the CHIR-12.12 antibody has been deposited with the American Type Culture Collection [ATCC; 10801 University BIvd., Manassas, Virginia 20110-2209 (USA)] under Patent Deposit Number PTA-5543. A series of the following examples are based on the binding of CHIR-12.12 to cancer cell lines and cells of cancer patients. However, all of the following examples are directly relevant to the use of CHIR-12.12 to treat inflammatory diseases and / or autoimmune diseases, since they illustrate the characteristics of the CHIR-12.12 antibody that are equally relevant for the treatment of inflammatory diseases and / or autoimmune diseases using anti-CD40 antibodies.

Example 1: Analysis of ADCC in cell lines CHIR-12.12 and rituximab were compared for their relative ADCC activity against a variety of malignant B cell lines expressing both CD40 and CD20 antigens, including lymphoma cell lines (Daudi, Namalwa) , multiple myeloma cell lines (ARH77, IM-9), a B-ALL cell line (CCRF-SB) and a B-CLL cell line (EHEB). The efficacy and potency of ADCC, measured as maximum percentage lysis and ED50, respectively, they were compared for CHIR-12.12 and rituximab. The results of these experiments are shown in Figures 1A-1F. For all target cell lines, CHIR-12.12 was a mediator of ADCC more potent and effective than rituximab. In the six cell lines tested, the number of CD20 molecules on cell surface per cell was 2.6 to 30.8 times higher than CD40. These data show that, despite showing fewer CD40 molecules than CD20, malignant B cell lines are lysed more effectively by CHIR-12.12 than by rituximab.

Example 2: Analysis of ADCC in cells from patients with CLL The relative ADCC activity of CHIR-12.12 and rituximab against cells of Primary CLL ex vivo of 8 patients. CHIR-12.12 exhibited a higher ADCC than rituximab against CLL of all patients (see Figure 2A-D and Figure 3). The results are summarized in Figure 3. CHIR-12.12 is more potent than rituximab.

Experimental Design for Antibody-Dependent Cell Cytotoxicity (ADCC) Target Cylinders: Patient CLL, 5000 / well. Effector cells: purified normal human NK cells, 50,000 / well. ratio E: 0 ratio: 10. Abs concentration: 0.00001, 0.0001, 0.001, 0.01, 0.1, 1 and 10 μg / ml. Incubation time: 4 hours. Culture medium: RPMI (without phenol red) + 10% FBS + 1% P / S. Growing device: 96-well round bottom plate. Reading: Release of calcein AM measured by Arbitrary Fluorescent Units (AFU) with 485nm excitation / 535 nm emission. Calculation:% specific lysis = 100 x (AFU test - spontaneous release 1 AFU) / (maximum release 2 AFU - spontaneous AFU). Negative control: Calcein released by the target cells in the absence of antibody or NK cells. Positive control: Calcein released by the target cells with lysis by detergent (1% NP40). The results illustrated in Figures 2 and 3 show that CHIR-12.12 mediates a greater ADCC than rituximab against cells from patients with CLL. The magnitude of the difference in ADCC may depend on either the target cells or the NK donor cells, but observed against all patient samples. When CLL cells from a single patient were tested with two different NK donors, CHIR-12.12 measured a higher ADCC than rituximab for both NK donor cells, although the magnitude of the differential ADCC was not identical (see Figure 4). ). The mechanistic basis for this higher ADCC may include the relative levels of expression of the target antigens (CD20 and CD40), the extent of internalization of the antibody and the affinity of the antibody for the Fc? Illa receptor in NK cells. Therefore, the influence of these factors on the ADCC activity of CHIR-12.12 and rituximab was investigated.

Example 3: Quantitation of CD40 and CD20 molecules on cell surface The quantitative density of CD20 and CD40 density in CLL cells (Example 3) and the degree of internalization of antibodies (Example 4) were investigated as potential reasons for the difference in activity of ADCC described in the above. The higher activity of ADCC and efficacy of CHIR-12.12 was not dependent on a higher density of CD40 molecules on the cell surface, since there were 1.3 to 14 times higher numbers of CD20 than CD40 molecules on the cell surface (see Figure 5). and Figure 6).

Methods Cells were preincubated with human IgG1 at 1 mg / ml in staining buffer (PBS contains 1% BSA, 0.1% Na Azide) to block nonspecific binding sites. They were incubated for 30 minutes at 4 ° C (on ice). Then, the isotype control of human IgG1 conjugated with FITC, CHIR-12.12 conjugated with FITC or rituximab conjugated with FITC was added at 100, 10, 1, 0.1 μg / ml, and the cells were incubated for 30 minutes at 4 ° C (in ice). The cells were washed with staining buffer (PBS + 1% FBS + 0.1% Sodium Azide), and analyzed by FACS Calibur. The geometric mean fluorescence intensity was measured by FACS. The Equivalent Soluble Fluorchrome Molecules (MESF) were then calculated based on the standard curve established by the calibrated FITC beads.

Example 4: CH-12.12 does not induce internalization with CD40 binding in cell lines Daudi, a lymphoma cell line, and ARH77, a MM cell line, were used to evaluate the effect of CHIR-12.12 binding on internalization. The cells were incubated with human IgG1 (control antibody) or CHIR-12.12 at 1 μg / mL on ice (with 0.1% sodium azide to block internalization) or at 37 ° C (without sodium azide) for 3 hours. After washing with cold staining buffer (PBS + 1% BSA + 0.1% sodium azide), the cells were stained with goat anti-human IgG-FITC for 30 minutes on ice. The geometric mean fluorescent intensity (MFI) was recorded by FACS Calibur. No difference was observed in MFI between cells incubated with CHIR-12.12 on ice in the presence of sodium azide or at 37 ° C in the absence of sodium azide (Figure 7). These data show that CHIR-12.12, with the binding to CD40 does not internalize and continues to be exposed on the cell surface.

Example 5: Internalization of CHIR-12.12 and Rituximab after joining cells of patients with CLL: FACS and confocal microscope FACS methodology The cells were incubated with hulgGI, CHIR-12.12 or rituximab at 10 μg / ml for 3 hrs at 40 ° C (with 0.1% Na azide) or 37 ° C (without Na azide). The cells were washed with staining buffer (PBS + 1% FBS + 0.1% Na Azide), FITC-Goat-anti-human IgG was then added, and then the cells were incubated for 30 minutes at 40 ° C and they were analyzed by FACS Calibur Confocal Microscope Methodology The cells were incubated with CHIR-12.12, rituximab and IgG1 conjugated with Alexa 488 or FITC at 10 μg / ml, for 3 hrs at 40 ° C (with 0.1% Na azide) or 37 ° C (without azide of Na). The cells were then washed and fixed with 2% formaldehyde, 5 min at RT. The cells were then washed and placed on slides coated with poly-L-lysine, mounted and sealed, and then analyzed by confocal imaging.

Results The results of these experiments are illustrated in Figure 8 (FACS) and Figures 9 and 10 (confocal microscope). The results of these experiments are summarized in Figure 11. These internalization studies of antibodies, using primary CLL cells, conducted by flow cytometry and confocal microscopy, show that, with binding to CD40 at 37 ° C, the CHIR- 12.12 remains evenly distributed on the cell surface, even after 3 hours. In contrast, after binding at 37 ° C, rituximab is redistributed in coatings and internalized. These data suggest that the potent ADCC activity of CHIR-12.12 may be related to its ability to expose itself uniformly on the surface of target cells, allowing for optimal interaction with effector cells. These results suggest that CHIR-12.12 may be effective in mediating a potent ADCC against CLL cells in vivo.

Example 6: Biacore analysis of binding to Fc? Rllla by Rituxan® and CHIR-12.12 The affinities of the alleles aa158F and aa158V of Fc? Rllla by CHIR-12.12 and rituximab were compared by standard Biacore® analysis. CHIR-12.12 bound to the aa158F allele with a 4.6 fold higher affinity when compared to rituximab (KD of 2.8 μM versus 13 μM, respectively). The results of these experiments are summarized in the following table: Example 7: The effect of the FcyRIIIa polymorphism on ADCC by NK effector cells The antibody dependent cellular cytotoxicity (ADCC) is a mechanism of action principal for many monoclonal antibodies marketed and investigated. Rituximab (Rituxan®), marketed for the treatment of follicular and non-Hodgkin's lymphoma (NHL) and active in other B-cell malignancies, is considered to have ADCC as one of its primary mechanisms of action. Notably, it has been shown that the clinical activity of rituximab in NHL correlates with the Fc? Rllla genotype. Patients with the V / V or V / F polymorphism of 158aa in Fc? Rllla are more sensitive to rituximab than those with F / F (for example, see Cartron ef al. (2002) Blood 99 (3): 754-758 or Dall'Ozzo ef al. (2004) Cancer Res. 64: 4664-4669). In these experiments, purified NK effector cells from multiple human donors expressing various FcγRllla aa158 polymorphisms were evaluated using the Daudi human lymphoma cell line as the target cells (see Figures 12 and 13). As illustrated by those figures, CHIR-12.12 induced a potent ADCC with the NK cells of the three genotypes. The ED50s of CHIR-12.12 for the lysis of the Daudi cell line were 4, 2 and 0.4 pM for F / F, V / F and V / V, respectively (Figure 13). The ED50s of rituximab for lysis of the Daudi cell line were 53, 21 and 9 pM for F / F, V / F and V / V, respectively (Figure 13). Purified NK effector cells from multiple human donors expressing various Fc? Rllla aa158 polymorphisms were also evaluated using the cells of CLL patients as the target cells (see Figure 14). It was found that CHIR-12.12 is a more potent mediator of ADCC than rituximab against all cells tested in patients with CLL (Figure 14). These data suggest that CHIR-12.12 is a more potent ADCC mediator than rituximab, even with NK cells of genotype aa158 V / F or F / F. These findings are surprising since it could have been expected that CHIR-12.12 could be significantly less potent in ADCC assays using NK cells with the F / F or V / F polymorphism of 158aa in Fc? Rllla than those with V / V . Again, it has been shown that the clinical activity of rituximab in NHL correlates with the Fc? Rllla genotype. Patients with the V / V or V / F polymorphism of 158aa in Fc? Rllla are more sensitive to rituximab than those with F / F. Rituximab is also an IgG1 monoclonal antibody that binds to an antigen expressed on the surface of B cells, and thus it could have been expected that the CHIR-12.12 could show the same preference for the Fc? Rllla-158 V polymorphism. Instead, it was found that CHIR-12.12 induces a potent ADCC with the NK cells of the three genotypes. Many modifications and other embodiments of the inventions set forth herein will come to mind to one skilled in the art to which these inventions pertain, having the benefit of the teachings presented in the foregoing descriptions and associated drawings. Therefore, it should be understood that the inventions should not be limited to the specific embodiments described and that the modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only, and not for purposes of limitation. All publications and patent applications cited herein are incorporated by reference in their entirety as if each publication or individual patent application was specifically and individually indicated as incorporated for reference.

Claims (53)

1. CLAIMS 1. A method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, wherein the human patient is heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F), the method comprises administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody. The method according to claim 1, characterized in that the inflammatory disease or autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), discoid lupus, lupus nephritis, sarcoidosis, inflammatory arthritis, including juvenile arthritis, rheumatoid arthritis, psoriatic arthritis, Reiter's syndrome, ankylosing spondylitis and gout arthritis, rejection of an organ or tissue transplant, hyperacute, acute or chronic rejection and / or graft versus host disease, multiple sclerosis, hyperinmunoglobulinemia syndrome E, polyarteritis nodosa , primary biliary cirrhosis, bowel inflammation syndrome, Crohn's disease, celiac disease (gluten-sensitive enteropathy), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia, psoriasis, scleroderma, myasthenia gravis, autoimmune thrombocytopenic purpura, auto thyroiditis immune, Grave's disease, Hasimoto thyroiditis, disease d by immune complexes, chronic fatigue immune dysfunction syndrome (CFIDS), polymyositis and dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy, pemphigus vulgaris, pulmonary interstitial fibrosis, type I and type II diabetes mellitus, delayed type hypersensitivity types 1, 2, 3 and 4, allergy or allergic disorders, unwanted / unintended immune responses to therapeutic proteins, asthma, Churg-Strauss syndrome (allergic granulomatosis), atopic dermatitis, allergic and irritant contact dermatitis, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis , idiopathic inflammatory myopathies, hemolytic disease, Alzheimer's disease, and chronic inflammatory demyelinating polyneuropathy, pulmonary inflammation including, but not limited to, lung graft rejection, asthma, sarcoidosis, emphysema, cystic fibrosis, idiopathic pulmonary fibrosis, chronic bronchitis, rhinitis allergic and allergic lung diseases such such as hypersensitivity pneumonitis, eosinophilic pneumonia, bronchiolitis obliterans due to bone marrow and / or lung transplantation
2. other causes, atherosclerosis by grafts / phlebosclerosis by grafts, pulmonary fibrosis resulting from collagen, vascular and auto immune diseases such as rheumatoid arthritis and lupus erythematosus.
3. 3. The method according to claim 1, characterized in that the inflammatory disease or autoimmune disease is an inflammatory disease or autoimmune disease associated with cells expressing CD20.
4. 4. The method according to claim 3, characterized in that the inflammatory disease or autoimmune disease is rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Crohn's disease, myasthenia gravis, idiopathic thrombocytopenic purpura or Sjogren's syndrome.
5. 5. The method according to claim 3, characterized in that the inflammatory disease or autoimmune disease is multiple sclerosis, rejection of grafts, graft versus host disease, Alzheimer's disease or diabetes.
6. 6. The method according to claim 3, characterized in that the inflammatory disease or autoimmune disease is an inflammatory disease or autoimmune disease that is associated with cells expressing both CD40 and CD20.
7. The method according to claim 6, characterized in that the inflammatory disease or autoimmune disease is rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Crohn's disease, myasthenia gravis, idiopathic thrombocytopenic purpura or Sjogren's syndrome.
8. 8. The method according to claim 6, characterized in that the inflammatory disease or autoimmune disease is multiple sclerosis, rejection of grafts, graft versus host disease, Alzheimer's disease or diabetes.
9. 9. The method according to any of claims 1-8, characterized in that the anti-CD40 antibody is administered by a parenteral route of administration.
10. The method according to claim 9, characterized in that the anti-CD40 antibody is administered intravenously or subcutaneously.
11. 11. A use of a therapeutically or prophylactically effective amount of an antibody
12. anti-CD40 in the manufacture of a medicament for the treatment of an inflammatory disease or autoimmune disease that is associated with cells expressing CD40 in a human patient heterozygous or homozygous for Fc? Rllla-158F (genotype V / F or F / F ). The use according to claim 11, characterized in that the inflammatory disease or autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), discoid lupus, lupus nephritis, sarcoidosis, inflammatory arthritis, including juvenile arthritis, rheumatoid arthritis, psoriatic arthritis, Reiter's syndrome, ankylosing spondylitis and gout arthritis, rejection of an organ or tissue transplant, hyperacute, acute or chronic rejection and / or graft versus host disease, multiple sclerosis, hyperinmunoglobulinemia syndrome E, polyarteritis nodosa , primary biliary cirrhosis, bowel inflammation syndrome, Crohn's disease, celiac disease (gluten-sensitive enteropathy), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia, psoriasis, scleroderma, myasthenia gravis, autoimmune thrombocytopenic purpura, auto thyroiditis Immune, Grave's disease, Hasimoto thyroiditis, disease for immune complexes, chronic fatigue immune dysfunction syndrome (CFIDS), polymyositis and dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy, pemphigus vulgaris, pulmonary interstitial fibrosis, type I and type II diabetes mellitus, delayed type hypersensitivity types 1, 2, 3 and 4, allergy or allergic disorders, unwanted / unintended immune responses to therapeutic proteins, asthma, Churg-Strauss syndrome (allergic granulomatosis), atopic dermatitis, allergic and irritant contact dermatitis, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis, idiopathic inflammatory myopathies, hemolytic disease, Alzheimer's disease, and chronic inflammatory demyelinating polyneuropathy, lung inflammation including, but not limited to, lung graft rejection, asthma, sarcoidosis, emphysema, cystic fibrosis, idiopathic pulmonary fibrosis, chronic bronchitis, allergic rhinitis and allergic lung diseases such as or hypersensitivity pneumonitis, eosinophilic pneumonia, bronchiolitis obliterans due to bone marrow and / or lung transplantation or other causes, graft atherosclerosis / phlebosclerosis by grafts, as well as pulmonary fibrosis resulting from collagen, vascular and autoimmune diseases such as rheumatoid arthritis and lupus erythematosus.
13. 13. The use according to claim 11, characterized in that the inflammatory disease or autoimmune disease is an inflammatory disease or autoimmune disease associated with cells expressing CD20.
14. 14. The use according to claim 13, characterized in that the inflammatory disease or autoimmune disease is rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Crohn's disease, myasthenia gravis, idiopathic thrombocytopenic purpura or Sjogren's syndrome.
15. 15. The use according to claim 13, multiple sclerosis, graft rejection, graft versus host disease, Alzheimer's disease or diabetes.
16. 16. The use according to claim 13, characterized in that the inflammatory disease or autoimmune disease is an inflammatory disease or autoimmune disease that is associated with cells expressing both CD40 and CD20.
17. 17. The use according to claim 16, characterized in that the inflammatory disease or autoimmune disease is rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Crohn's disease, myasthenia gravis, idiopathic thrombocytopenic purpura or Sjogren's syndrome.
18. 18. The use according to claim 16, multiple sclerosis, graft rejection, graft versus host disease, Alzheimer's disease or diabetes.
19. 19. Use according to any of claims 11-18, characterized in that the medicament is formulated for administration by a parenteral administration route.
20. 20. The use according to claim 19, characterized in that the medicament is formulated for administration in intravenous or subcutaneous form.
21. 21. A method for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for FcγRllla-158F (genotype V / F or F / F), which comprises administering to the human patient an effective amount of an anti-CD40 antibody. The method according to claim 21, characterized in that the human patient has an inflammatory disease or autoimmune disease that is associated with cells
22. that express CD40.
23. 23. Use of an effective amount of an anti-CD40 antibody in the manufacture of a medicament for inhibiting the production of antibodies by B cells in a human patient heterozygous or homozygous for FcγRllla-158F (genotype V / F) or F / F).
24. 24. The method or use according to any of the preceding claims, characterized in that the anti-CD40 antibody is a human monoclonal antibody.
25. 25. The method or use according to claim 24, characterized in that the human anti-CD40 monoclonal antibody comprises a heavy chain constant region of human Ig1.
26. 26. The method or use according to claims 1-25, characterized in that human IgG1 comprises the amino acid sequence listed in SEQ ID NO: 4 or SEQ ID NO: 5.
27. 27. The method or use according to any of the preceding claims, characterized in that the anti-CD40 antibody is free of significant agonist activity.
28. The method or use according to any of the preceding claims, characterized in that the anti-CD40 antibody is an antagonist of CD40-CD40L signaling in cells expressing CD40.
29. 29. The method or use according to any of the preceding claims, characterized in that the anti-CD40 antibody is selected from the group consisting of: a) the monoclonal antibody CHIR-12.12; b) the monoclonal antibody produced by the hybridoma cell line 12.12; c) a monoclonal antibody comprising an amino acid sequence selected from the group consisting of the sequence shown in SEQ ID NO: 2, the sequence shown in SEQ ID NO: 4, the sequence shown in SEQ ID NO: 5, the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 4, and the sequences shown in SEQ ID NO: 2 and SEQ ID NO: 5; d) a monoclonal antibody having an amino acid sequence encoded by a nucleic acid molecule comprising a nucleotide sequence selected from the group

    which consists of the sequence shown in SEQ ID NO: 1, the sequence shown in SEQ ID NO: 3, and the sequences shown in SEQ ID NO: 1 and SEQ ID NO: 3; e) a monoclonal antibody that binds to an epitope capable of binding to the monoclonal antibody produced by the hybridoma cell line 12.12; f) a monoclonal antibody that binds to an epitope comprising residues 82-87 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; g) a monoclonal antibody that binds to an epitope comprising residues 82-89 of the human CD40 sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9; h) a monoclonal antibody that competes with the monoclonal antibody CHIR-12.12 in a competitive binding assay; i) the monoclonal antibody of preceding article a) or a monoclonal antibody of any of preceding articles c) -h), wherein the antibody is produced recombinantly; and j) a monoclonal antibody that is an antigen-binding fragment of a monoclonal antibody of any of the preceding articles a) -i), wherein the fragment retains the ability to specifically bind to the human CD40 antigen.
30. 30. The method or use according to claim 29, characterized in that the anti-CD40 antibody is the monoclonal antibody CHIR-12.12.
31. The method or use according to claim 29, characterized in that the antigen-binding fragment is selected from the group consisting of a Fab fragment, an F (ab ') 2 fragment, an Fv fragment and a chain Fv fragment. only.
32. 32. A method for identifying a human patient with an inflammatory disease or autoimmune disease, treatable with an anti-CD40 antibody, and which is resistant to treatment with rituximab (Rituxan®), which comprises: a) identifying a human patient with an inflammatory disease or autoimmune disease that is associated with cells that express CD40; and b) determining the Fc? Rllla158 genotype (V / V, V / F or F / F) of the human patient; where the inflammatory disease or autoimmune disease is treatable with a

    anti-CD40 antibody if the human patient is heterozygous or homozygous for Fc? RIMa-158F (genotype V / F or F / F).
33. 33. A method for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease which is resistant to treatment with rituximab (Rituxan®), which comprises: a) identifying a human patient who have an inflammatory disease or autoimmune disease that is associated with cells that express CD40 and which is resistant to treatment with rituximab (Rituxan®); and b) determining the Fc? Rllla158 genotype (V / V, V / F or F / F) of the human patient; where if the human patient is heterozygous or homozygous for Fc? Rllla-158F

    (genotype V / F or F / F), an anti-CD40 antibody is selected for the treatment of inflammatory disease or autoimmune disease.
34. 34. A method or use according to any preceding claim, characterized in that the human patient is resistant to a therapy for an inflammatory or autoimmune disease.
35. 35. The method according to claim 34, characterized in that the human patient resists therapy with an anti-CD20 monoclonal antibody.
36. 36. The method according to claim 35, characterized in that the human patient is resistant to therapy with an anti-CD20 monoclonal antibody.
37. 37. The method according to claim 35, characterized in that the human patient is not sensitive to therapy with an anti-CD20 monoclonal antibody.
38. 38. The method according to any of claims 35-37, characterized in that the anti-CD20 monoclonal antibody is rituximab (Rituxin®).
39. 39. A kit for identifying a human patient with an inflammatory disease or autoimmune disease treatable with an anti-CD40 antibody, comprising reagents to determine an Fc? Rllla-158 genotype of the human patient.
40. 40. A kit for selecting an antibody therapy for the treatment of a human patient having an inflammatory disease or autoimmune disease associated with

    CD40 expressing cells, which comprises reagents to determine a Fc? Rllla-158 genotype of the human patient.
41. 41. The equipment according to claim 39 or claim 40, characterized in that it includes a DNA microarray comprising at least one probe of 10 or more nucleotides in length and of a suitable sequence to determine the genotype Fc? Rllla-158 of a human patient.
42. 42. The equipment according to claim 39 or claim 40, characterized in that it comprises oligonucleotides suitable for use as primers is a polymerase-catalyzed amplification of the genomic region encoding the 158 amino acids of Fc? Rllla.
43. 43. The equipment according to claim 39 or claim 40, characterized in that it comprises one or more suitable restriction enzymes for determining the Fc? Rllla-158 genotype of a human patient.
44. 44. A method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody, in such a manner that the anti-CD40 antibody is not significantly internalized by cells expressing CD40 after administration.
45. 45. A method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient a therapeutically or prophylactically effective amount of an anti-CD40 antibody, in such a manner that the anti-CD40 antibody remains substantially uniformly distributed on the surface of cells expressing CD40 after administration.
46. 46. A method for treating a human patient for an inflammatory disease or autoimmune disease that is associated with cells expressing CD40, the method comprising administering to the human patient an anti-CD40 antibody, such that a therapeutically or prophylactically effective amount of the anti-CD40 antibody is present on the surface of cells expressing CD40 in the human patient after administration.
47. 47. The method or use according to any of claims 1-31 or 44-46, characterized in that the method or use results in antibody-dependent cellular cytotoxicity (ADCC) of cells expressing CD40 by natural cytolytic (NK) cells expressing Fc Rllla of a human patient.
48. 48. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody is more potent than rituximab (Rituxin®) in an antibody dependent cellular cytotoxicity assay (ADCC) , wherein the assay comprises incubating cells expressing CD40 and cells expressing CD20 with isolated human natural killer (NK) cells in the presence of the relevant antibody.
49. 49. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody is more potent than rituximab (Rituxin®) in a model of systemic lupus erythematosus (SLE), sclerosis multiple, inflammation and atherosclerosis, transplant or Alzheimer's disease.
50. 50. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody binds to human CD40 with an affinity (KD) of at least about 10"6 M to a at least about 10"12 M.
51. 51. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody binds to the human FcγRllla-158V with an affinity (KD). ) of at least about 0.5 μM.
52. 52. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody binds to the human Fc? Rllla-158F with an affinity

    (KD) of at least about 12 μM.
53. 53. The method or use according to any of claims 1-31 or 44-46, characterized in that the anti-CD40 antibody binds to human FcγRllla-158V with an affinity (KD) of at least about 0.5 μM. , and binds to human FcγRllla-158F with an affinity (KD) of at least about 12 μM.