SI9520118A - Compositions and treatment for multiple sclerosis - Google Patents

Abstract

The present invention provides isolated peptides and peptide combinations derived from myelin autoantigens, such as MBP, MOG, PLP and MAG, suitable for the treatment of multiple sclerosis, including prophylactic and therapeutic compositions and methods for the prevention or treatment of multiple sclerosis. Preferred compositions of the invention comprise at least one isolated purified peptide which is free of all other polypeptides or contaminants, the peptide comprising an amino acid sequence, a myelin autoantigen having T cell activity. The therapeutic composition of the invention is capable of down-regulating the autoantigen-specific immune response to the myelin autoantigen in a population of people suffering from or susceptible to multiple sclerosis by reducing, eliminating or preventing disease symptoms and / or preventing or slowing the onset or progression of disease or progression . Additionally, the compositions and methods of the present invention when administered in the advanced stage of the disease, eradicate ongoing paralysis or other signs of the disease when administered during the acute phase of the disease, or prevent the disease from returning during remission.ŕ

Description

Multiple sclerosis preparations and treatment

Related applications

This application is a partial continuation application of USSN 08 / 328,224, filed October 25, 1994. This application is also a partial continuation application of USSN 08 / 300,811, filed September 1, 1994, which is a partial continuation application of USSN 08 / 116,824, filed September 3, 1993 This application is also a partial continuation application of USSN 08 / 241,246, filed May 10, 1994. The above applications are fully incorporated herein by reference.

BACKGROUND OF THE INVENTION

Autoimmune diseases are a major human health problem and are relatively poorly understood. As no microbial or viral agent is clearly directly responsible, the prevention, treatment and diagnosis of such diseases must be based on the etiology of the disease. This always involves a complex series of reactions of endogenous metabolic intermediates, structural components, cells, etc. However, with regard to the nature of the autoimmune condition, there is an opinion that at least one autoantigen must be involved in the generation of a sequence of events that results in a sympatome. Autoimmune demyelinating diseases, such as multiple sclerosis, are no exception.

MS is usually expressed in the form of recurrent seizures, which are reflected in lesions within the central nervous system (CNS = Central Nervous System). Attacks seem to be randomly repeated, relented, and repeated over many years. The incidence of flares is highest during the first 3 to 4 years of the disease, but the first attack, which may have been so mild as to have escaped medical attention and can hardly be remembered, may not be repeated for 10 to 20 years. The extent of recovery after an episodic event varies greatly between patients. Remission can be complete, especially after an early onset, but is often incomplete, and when one attack follows another, a gradual downward progression with an increasing permanent deficit follows. The clinical picture of MS is determined by locating foci of demyelination within the CNS. Classical features include impaired vision, nystagamus, dysarthria, decreased vibration perception and decreased sense of position, ataxia and intentional tremor, weakness or paralysis of one or more limbs, spasticity and bladder problems. Criteria for the diagnosis of clinically definitive MS should include a reliable history of at least 2 episodes of neurological deficit and an objective clinical sign of the lesion at more than one site within the CNS. No effective treatment is known for MS.Current therapeutic efforts are directed toward improving the acute event and preventing the relapse or progression of the disease (Harrison's Principles of Internal Medicine, 12th Edition, Vol. 2, pp. 2038-2043; McGravv Hill, 1991).

The commonly used animal model for human multiple sclerosis is Experimental Allergic Encephalomyelitis (EAE), a central nervous system demyelinating disease that can be induced in susceptible mouse strains by immunization with myelin basic protein (MBP), proteolipid protein (PLP), myelin protein oligod MOG), or with synthetic peptides based on the sequences of these myelin-related peptides. MBP is one of the predicted autoantigens in multiple sclerosis (MS) and has been epitopically mapped both in human (Ota et al., Nature, 346: 183-187 (1990)) and in the murine system (Zamvil et al. Nature 324: 258-260 (1986). Peptides thought to comprise at least one T cell epitope of MBP (myelin basic protein = myelin basic protein) have been identified in ΛΥΟ 93/21222, EP 0 304 279, WO 91/15225, Ota et al. Letters to Nature, 346: 183-187 (1990), Wucherpfennig et al., J. Exp. Med., 170: 279-290 (1994). Other MBP peptides containing the T cell epitope have been identified in the applications, which are incorporated herein by reference in their entirety, in the U.S.S.N. No. 08 / 328,224, filed Oct. 25, 1994, and in U.S.S.N. No. 08 / 241,246, filed May 10, 1994. MOG peptides having T cell activity have been identified in USSN 08 / 300,811, which is incorporated herein by reference.

Proteolipid protein (PLP) and myelin-associated glycoprotein (MAG) have also been implicated as possible autoantigens in multiple sclerosis. Studies describing the pathogenesis of the autoimmune response to PLP in multiple sclerosis have been described in Trotter et al., J. Neuroimmunol., 33: 55-62 (1991); PLP T cell epitopes have been described in Pelfrey et al., J. Neuroimmunol., 46: 33-42 (1993). Studies describing MAG as a potential autoantigen in multiple sclerosis are described in Johnson et al., J. Neuroimmunol., 13: 99-108 (1986).

Experimental autoimmune encephalomyelitis (EAE) is an autoimmune disease mediated by T CD4 + cells that is similar to multiple sclerosis in some of its clinical and histological features and serves as an experimental model for this and other autoimmune diseases. EAE is an inflammatory disease of the central nervous system that results in paralysis and other neurological abnormalities. It is typically induced by purified myelin proteins and peptides. The EAE model is widely used in investigating the mechanisms of autoimmunity and in exploring possible therapists for autoimmune disease.

As early as 1965, it was observed that EAE could be treated by administering MBP in a non-encephalitogenic form, presumably by inducing immunologic immunity or tolerance. (Alvord, EC, et al., Ann. NY Acad Sci., 1965,122: 333; Levine, S.E., et al. Science, 1968, 161: 1155, Bernard CCA, 1977 Ciin. Exp. Immunol., 1977, 29: 100). This early observation has been expanded over the past few years by numerous studies showing that neonatal administration of MBP or MBP peptides or administration to adults prevents EAE (Bernard, CCA, 1977 Ciin. Exp. Immunol ;, 1977, 29: 100, Clayton, JP, et al., J. Exp Med 1989, 169: 1681, Smilek, DE, et al., Nat Nat, Acad. Sci., 1991, 88: 9633, Guar, A. ., et al., Science, 1992, 258: 1491, Metzler, B. et al., Int. Immunol, 1993, 5: 1159, Miller, A., et al., J. Neuroimmunol, 1993, 46:73 , Critchfield, J. M. et al., Science, 1994, 263: 1139; Miller, A., et al., Proc. Nat'l, Acad, Sci., USA, 1992, 89: 421). These studies have suggested different routes of administration that include subcutaneous, intraperitoneal, intranasal, intravenous, and oral administration. The induction of immunological unresponsiveness in adult animals with intravenous peptides has been demonstrated in a variety of antigen systems. For many of the reasons described herein, there are limits to the clinical utility of oral, enteral, or aerosol administration of autoantigens, such as the inability to characterize the active component of a therapeutic composition once introduced into the stomach due to subsequent enzymatic degradation in the stomach. Thus, using these methods, it can be difficult to achieve predictable and reproducible therapeutic effects, not to mention the potential for adverse side effects, as a result of further physical recovery of the drug, which may also be unpredictable.

In most of these cases, the mechanism of disease prevention or induction of tolerance has been attributed to clonal anergy (Gaur et al., Supra), peripheral deletion (Critchfield, et al., Supra), or other forms of antigen-specific tolerance. However, TGF-) 3-mediated bystander suppression seems to be an additional mechanism by which orally administered MBPs and MBP peptides can inhibit EAE (Miller et al., Supra in Nat. 1 Acad. Sci. ). MBP peptides, as well as substituted MBP peptide analogs, were investigated as alternative therapeutics for EAE in PU, B10.PL and (PLJ x SJL) F1 mice. MBP Acl-11 is an immunodominant encephalitogenic peptide for each of these strains and binds readily to Aa ^u A / 3 ^u (Wraith, DC, et al., Supra). MBP Acl-11 was thoroughly studied by substitution analysis and its requirements for identification of T cells and binding of the major histocompatibility complex (MHC) were well established. The side chains of residues 3 and 6 mainly contribute to the identification of T cells, while the side chains of residues 4 and 5 mainly contribute to MHC binding. The binding of MBP Acl-11 to Aa ^u A / 3 ^u can be greatly enhanced by a variety of amino acid substitutions at residue 4, including alanine (Acl-11 [4A]) and tyrosine (Acl11 [4Y]) (Wraith, DC et al., above Fairchild, PJ et al., Int. Immunol, 1993, 5: 1151). The substitutions of residue 4, especially tyrosine, appear to improve the stability of peptide-MHC complexes formed by these peptides. Acl-11 [4A] and Acl-11 [4Y] retain the T cell receptor (TCR) contact residues required for antigenicity and are more potent than MBP Acl-11 in stimulating MBP-specific T cells in vitro. Acl-11 [4Y] is also encephalitogenic in vivo (Acl-11 [4Aj is poorly encephalitogenic. For unknown reasons). Both Acl-11 [4A] and Acl11 [4Y] are known to prevent EAE and are thought to act by antigen-specific mechanisms (Smilek, et al., Supra, Wraith, DC et al., Supra).

In previous studies, peptides or peptide analogues of MBP administered in an incomplete adjuvant just before the onset of the disease prevented the subsequent development of EAE (Smilek et al., Supra, Gaur et al., Supra). In a separate study, because they used lymphocytes from an MBP-specific TCR transgenic mouse, adoptively tolerated EAE was prevented by early and aggressive administration of intravenous MBP before the onset of clinical signs (Critchfield, et al., Supra). These studies suggested that it was possible to prevent EAE by injecting MBP peptides after encephalitogenic T cells were activated, but did not address the question of whether pending paralysis could be eliminated (and probably active CNS inflammation) system) or whether it could prevent recurrent illnesses following remission. In addition, in some of the experiments previously performed, effective administration of extremely high doses of MBP or MBP peptides was required for effective treatment. The inventors here were the first to describe the elimination of paralysis, as well as the prevention of recurrent disease following remission, in their previous case USSN 08 / 328,224, filed October 25, 1994, which continues here. Since the inventors of the present invention have submitted a prior subject, others working in the field have been surprised to find that the injection of certain peptide analogs derived from MBP in EAE could eliminate the ongoing paralysis (Karin et al. J. Exp. Med. 180: 2227-2237 (Dec. 1994)).

The present invention overcomes the disadvantages described above and provides novel peptides, compositions and methods for the treatment of multiple sclerosis using compositions comprising at least one peptide having a sequence of amino acid residues containing T cell activity of MBP. Further, the present invention is directed to a still unresolved problem, namely that, in order to be generally applicable, the treatment of multiple sclerosis using peptides or peptide analogs must be effective even when given late in the course of the disease or in advanced stages of disease or during remissions, or during disease recurrences while an adverse immune response takes place.

It is therefore an object of the present invention to provide peptides and peptide combinations that are suitable as a therapist for multiple sclerosis, including the prevention of the onset of disease. Another object of the present invention is to provide prophylactically and therapeutically effective dosage regimens and routes of administration of identified proteins, peptides and peptide analogs for the effective treatment of MS. It is another object of the present invention to provide a treatment that successfully cures late-stage MS, prevents disease recurrence, stops disease, and / or suppresses MS progression.

Summary of the Invention

The present invention provides isolated peptides and peptide combinations derived from myelin autoantigens, such as MBP, MOG, PLP and MAG, suitable for the treatment of multiple sclerosis, including prophylactic and therapeutic compositions and methods for preventing or treating multiple sclerosis. Preferred compositions of the invention comprise at least one isolated purified peptide substantially free of all other polypeptides or contaminants, the peptide comprising an amino acid sequence of a myelin autoantigen having T cell activity. The therapeutic composition of the invention is capable of down-regulating an autoantigen-specific immune response to a myelin autoantigen in a population of people with multiple sclerosis or susceptible to multiple sclerosis by reducing, eliminating or preventing and / or preventing or delaying the onset of disease symptoms progression of disease symptoms.

In addition, the compositions and methods of the present invention, when administered in the advanced stage of the disease, eradicate ongoing paralysis or other signs of the disease when administered during the acute phase of the disease or prevent the disease from returning during remission.

Brief description of the drawings

FIG. 1 shows the full length of the amino acid sequence of human MBP, and also shows the numbering of the amino acid residues referred to herein.

FIG. 2 shows the amino acid sequence of the preferred peptides derived from MBP.

FIG. 3 shows the amino acid sequence of the overlapping peptides as well as the longer peptides used in Example 1.

FIG. 4a is a graphical representation of the percentage of the total MBP response for each peptide shown in FIG. 3. MBP reactivity was calculated on the basis of the percentage of MBP positive microtiter groups in each group of individual patients who gave a positive result also for one of the MBP peptides. Overall MBP positive microtiter cultures ranged from 89-184 in each group of 19-43 patients tested for each peptide.

FIG. 4b is a branch view of the percentage of MBP responders that recognize each peptide shown in FIG. 3. Patients were considered MBP responders if at least one of their microtiter cultures had a positive result for MBP reactivity. In each group of 19-43 patients tested for each peptide, there were 1231 MBP response patients. The peptide reactivity was calculated based on the percentage of MBP response patients in each group with at least one microtiter culture that gave a positive result for one of the MBP peptides.

FIG. 5a is a graph of median clinical outcomes over the period 0-40 days after disease induction in mice treated with i.p. injections of MBP Acl-11 alone (SEQ ID NO: 65), compared to control.

FIG. 5b is a graph of median clinical outcomes for the period 0-40 days after disease induction in mice treated with i.v. injections of MBP Acl-11 (SEQ ID NO: 65) combined with MBP 31-47 (SEO ID NO: 68) compared to control.

FIG. 5c is a graph of mean clinical outcomes in the range of 0-40 days after disease induction in mice treated with i.v. injections of OVA 323-339 (SEQ ID NO: 69) compared to control.

SL 6a is a graph of median clinical results over a period of 0 to at least 30 days after induction of disease in specimens treated with either 250 nmolar injection of Acl11 [4Y] (SEQ ID NO: 67) or Acl-11 (SEO ID NO : 65), compared to control.

FIG. 6b is a graph of median clinical results over a period of 0 to at least 30 days after disease induction in mice treated with 2.5 nmol Acl-11 [4Y] (SEQ ID NO: 67) compared to the control.

FIG. 6c is a graph of mean clinical results over a period of 0 to at least 30 days after disease induction in mice treated with 2.5 nmol Acl-11 (SEQ ID NO: 65) compared to control.

FIG. 7 is a bar graph showing the mean histologic results of peptide-treated mice versus control-treated mice, the lower result being the reduced number and severity of CNS inflammatory infiltrates.

FIG. 8 is a bar graph showing the capacity of spleen cells contacted in vivo at 250 nmol Acl11 [4Y] for specific time intervals (1-10 hours) to activate in vitro hybridoma cell line (1934 hybrid) derived from MBP-specific T cell clone.

FIG. 9 is a graph showing a comparison of treatment of mice with 250 nmol Acl-11 [4Y] (SEQ ID NO: 67) against control, PBS and OVA 323-337 (SEQ ID NO: 70).

FIG. 10 is a graph showing a comparison of the treatment with 25 nmol Acl-11 [4YJ (SEQ ID NO: 67) initiated during remission.

FIG. Ila is a graph showing the in vitro lymph node proliferation (as indicated by the incorporation of 3H-thymidine) from mice previously obtained i.v. treatment with 250 nmol MBP Acl-11 (SEQ ID NO: 65) compared to controls.

SL 1 lb is a bar graph showing the relative production of IL-2 lymph node in response to MBP Acl-11 (SEQ ID NO: 65) in mice pretreated with

Acl-11 [4Yj (SEQ ID NO: 67), compared to control.

FIG. 12a is a graphical description of an experiment showing the effects of IFN- / 3 on EAE in two groups of 10 (SJL x TLP) Fj adult mouse females, in which EAE was induced with guinea pig MBP in complete adjuvant plus pertussis toxin on day 0 and to which either the incomplete Freund's adjuvant (control) was administered or treated intraperitoneally (ip) with 2000 IFN- / 3 units at 9, 12, 16, and 20 days (indicated on the x axis by arrows), the Y axis represents the mean clinical result (MCS = mean clinical score) for each group, 0 = no EAE clinical signs, 1 = weak, unresponsive tail, 2 = partial hind limb paralysis, 3 = complete hind limb paralysis, 4 = partial to full limb paralysis, and 5 = dying.

FIG. 12b is a graphical description of an experiment showing the effects of the MBP Acl-11 peptide (SEQ ID NO: 65) in two groups of 10 (SJL x TLP) Fj adult female mice in which EAE was induced using a guinea pig MBP in complete adjuvant plus pertussis toxin on day 0 and given either PBS (control) or treated with 250 nmol Acl-11 (SEQ ID NO: 65) intravenously at days 10, 13, 17 and 21 (indicated by arrows on the x axis) The Y axis represents the mean mean clinical score for each group as described in FIG. 12a.

FIG. 12c is a graphical description of an experiment showing the effects of the MBP Acl-11 peptide (SEQ ID NO: 65) administered in combination with IFN- / 3 on EAE in two groups of 10 (SJL x TLP) Fj adult mice females to whom EAE was induced in complete adjuvant plus pertussis toxin on day 0 and given either PBS (control) or treated intravenously with 250 nmol Acl-11 (SEQ ID NO: 65) 10, 13, 17 and 21 days (indicated by blank arrows on the x axis) and which were treated intraperitoneally (ip) with 2000 IFN- / 3 units on 9, 12, 16, and 20 days (indicated by full arrows on the x axis), Y axis showing the mean clinical result , as discussed in FIG. 12a.

FIG. 13 is a graphical description of an experiment showing the effects of different doses of IFN- / 3 (10,000 units or 2000 units) on induced EAE in two groups of 10 (SJL x TLP) Fj adult mouse females, which were EAE induced using MBP guinea pig plus pertussis toxin on day 0 and given either PBS (control) or treated with 10,000 units or. 2000 units of IFN- / 3 intraperitoneally (i.p.) For 9, 13, 16 days (indicated by solid arrows on the x axis), and the Y axis indicates the mean clinical result as discussed in FIG. 12a.

FIG. 14 shows various peptides derived from MBP that may be suitable in the compositions and methods of the invention.

FIG. 15 is a graphical representation of the positivity index (y axis) (average S.I./MBP response patient) multiplied by the percentage of individuals responding to each peptide (x-axis) derived from the same experimental data shown in FIG. 4a and 4b, and which are described in Example 1.

FIG. 16 is a graphical representation of the mean clinical outcome relative to the initiation of therapy using Acl-11 [4Yj (SEQ ID NO: 67) for each individual (day 1 = onset of clinical signs for each individual mouse). Mice were treated with PBS (full squares), 250 nmol Acl-11 [4Y] i.v., (SEQ ID NO: 67) empty triangles, or 250 nmol Acl-11 [4Y] s.c. on day 1, 3, 6, and 9 of the disease. (SEQ ID NO: 67) (blank circles). These data indicate that Acl-11 [4Y] (SEQ ID NO: 67) suppresses EAE when administered after paralysis.

FIG. 17 is a graphical representation of the mean clinical result when Acl-11 [4Y] is given before symptoms of the disease appear. Data are plotted against disease induced by gp brain homogenate (gpSCH) at day 0 and days after therapy. Survival mice groups were treated for 8, 10, 13, and 17 days with 25 nmol Acl-11 [4Y] (SEQ ID NO: 67) i.v. (empty triangles), 25 nmol Acl-11 [4Y] (SEQ ID NO: 67) s.c. (full circles), or we didn't process them (empty squares). These data indicate that Acl-11 [4Y] (SEQ ID NO: 67) prevents EAE from occurring.

S1.18 is a graphical representation of the mean clinical result relative to the initiation of therapy using Acl-11 [4Yj (SEQ ID NO: 67) administered intravenously (empty triangles) or subcutaneously (solid circles). Blank squares show control.

FIG. 19 is a graphical representation of the mean clinical outcome with respect to the day of disease induced using 75 µg gpMBP (solid circles), 100 µg huMOG (empty triangles), and combinations of 75 µg gpMBP and 100 µg huMOG (solid squares). As shown, the disease induced by both MOG and MBP is much more serious than the disease induced by each.

FIG. 20 is a graphical representation of the mean clinical score relative to days of therapy using Acl-11 [4Y] (SEQ ID NO: 67) (blank squares) versus PBS controls (full squares). The disease was induced as in Example 10 using huMOG + gpMBP. These data indicate that Acl-11 [4Y] (SEQ ID NO: 67) significantly decreased the median clinical outcome compared to controls.

Detailed description of the invention

The patent and scientific literature referred to herein provides knowledge that is available to experts. The U.S. cited here cited patents, PCT publications and other publications are incorporated by reference.

The present invention provides isolated peptides and combinations thereof derived from myelin autoantigens useful for the treatment of multiple sclerosis, as well as therapeutic compositions and methods for the treatment of multiple sclerosis. As used herein, the term treatment of multiple sclerosis includes: the prophylactic treatment of those mammals susceptible to MS; treatment of MS onset; and treatment of all advanced stages of MS, including returning-relapsing MS, chronic progressive MS, primary progressive MS, and benign MS. The therapeutic compositions of the invention comprise at least one purified peptide substantially free of all other proteins or contaminants and comprising a defined sequence of amino acid residues of myelin antigen having T cell-stimulating activity, wherein the peptide may also be an isolated peptide. As used herein, the term isolated refers to a peptide that is free of all other polypeptides, contaminants, starting reagents or other materials, and which is not conjugated to any other molecule.

According to the present invention, the peptide refers to a specific sequence of amino acid residues that is smaller than the amino acids of the native protein antigen. The peptide of the invention preferably comprises at least about 7 amino acid residues in length and preferably at least about 12-40 amino acid residues and more preferably at least 13-30 amino acid residues, and, when derived from a protein antigen, contains fewer amino acids than the whole protein has antigen and preferably not more than about 75% of the amino acid residues of the whole protein antigen. The peptides used in the invention have T cell activity. A peptide having T cell activity may have any one or more of the following characteristics: a) the ability to elicit a T cell response, such as stimulation (ie, proliferation or lymphokine secretion): b) the ability to cause T cell unresponsiveness or reduced T cell responsiveness of the relevant T cell subpopulations so that they do not participate in the stimulation of the immune response to the causative autoantigen (eg via anergy, tolerance or apoptosis); c) the ability to modify the lymphokine secretion profile in comparison to autoantigen exposure occurring in nature; d) the ability to induce the suppression of T suppressor cells; and e) the ability to cause down-regulation of autoimmune disease symptoms by any mechanism; or f) is derived from an bystander an11 tigen and has the ability to elicit suppressor T cells at the site of a myelin autoimmune attack, resulting in downregulation of immune responses at the scene of myelin autoimmune attack.

Peptides comprising at least one T cell epitope have T cell activity and are capable of eliciting a T cell response, such as stimulation of T cells (ie, T cell proliferation or lymphokine secretion) and / or capable of down-regulating the autoantigen-specific response of T cells it may result in autoantigen-specific T cell responsiveness, or which reduce the level of autoantigen-specific T cell responsiveness. The T cell epitope is the basic element or smallest unit recognized by the T cell receptor, the epitope comprising amino acids essential for receptor recognition. We think that T. cell epitopes are involved in the initiation and duration of an immune response to an antigen or autoantigen. These T cell epitopes are thought to trigger early immune response events at the T helper cell level by binding to the corresponding HLA molecule on the surface of the antigen-presenting cell and by stimulating a significant subpopulation of T. cells. These events lead to proliferation of T cells, lymphokine secretion. local inflammatory reactions, enhancement of additional immune cells at the site, and activation of the B cell cascade leading to antibody production. In the case of autoimmune disease, the antibodies being produced are autoantibody auto-antibodies such as MBP, which results in clinical symptoms of autoimmune disease.

Peptides. .which have defined amino acid compositions and that comprise T cell epitopes, can be identified for any myelin autoantigen, including MBP. One method involves splitting the protein antigen into non-overlapping or overlapping peptides of the desired lengths and synthesizing, purifying and testing these peptides using any number of assays (i.e., T cell proliferation assay, lymphokine secretion assay, and T cell unresponsiveness study) to determine whether peptides comprise at least one T cell epitope of MBP. In another method, we use an algorithm to predict those peptides that are likely to comprise T cell epitopes, and then synthesize, purify, and test peptides predicted by the algorithm in T cell assays or in vivo studies to determine whether such predicted peptides induce T cell proliferation or lymphokine secretion or T cell unresponsiveness and are therefore likely to contain T cell epitopes. As discussed in many of the documents cited above, human T cell activity can be tested by culturing T cells obtained from an autoantigen-sensitive individual such as MBP with an antigen derived peptide and determining whether proliferation occurs of T cells in response to the peptide as measured, e.g. with cellular uptake of tritiated thymidine. Stimulation indices for T cell responses to peptides can be calculated as maximal beats per minute (CPM = counts per minute, beats per minute) in response to the peptide divided by control CPMs. The stimulation index (S.I.) of T cells, which is equal to or twice and preferably three times the background level, is considered positive. Positive results are used in the analysis of the potential therapeutic efficacy of peptides, as discussed hereinafter in Example 1. Preferred peptides useful in the present invention comprise at least one T cell epitope and preferably at least two or more T cell epitopes.

One aigontem for predicting peptides having T cell stimulating activity is described in Rothbard, lst Forum and Virology, Annals of the Pasteur Institute, p. 518-526 (December, 1986), Rothbard and Taylor, Embo, 7: 93-100 (1988), and EP 0 304 279. These documents describe the determination of a general pattern (algorithm) for peptide binding to MHC class II, its statistical significance and correlating the sample with known T cell epitopes, as well as its successful use in predicting previously unidentified T cell epitopes of various protein antigens and autoantigens. As stated in the documents mentioned above, the general pattern for a peptide known to bind well to MHC class II appears to contain a linear pattern consisting of a charged amino acid residue or glycine followed by two iTiarotobiia residues. After determining whether the peptide is in accordance with the general pattern, the peptide can then be tested for T cell reactivity. Other algorithms used to predict T cell epitopes of previously undefined proteins include the algorithm described by Margalit et al., J. Immunol. 138: 2213-2229 (1987) based on an amphipathic helix model.

In addition, peptides may be identified that contain latent T cell epitopes and are also useful in the methods and compositions of the invention. Covert T cell epitopes are those determinants in a protein antigen that are not normally detected in the immune system due to the formation and introduction of a native protein antigen to the corresponding MHC molecule. However, a peptide containing a hidden T cell epitope is capable of causing T cells to become inactive and when the subject is initiated with the peptide, the T cells obtained from the subject will proliferate in vitro in response to a peptide or protein antigen from which is a peptide derived. Peptides comprising at least one covert T cell epitope derived from a protein antigen are referred to herein as covert peptides. To confirm the presence of covert T cell epitopes, we can use T cell proliferation assay, as is known in the art, where antigen-primed T cells are cultured in vitro in the presence of each peptide individually to demonstrate peptide-reactive T cell lines. A peptide is considered to contain at least one disguised T cell epitope if a given peptide can be shown to have a T cell line and if T cells are able to proliferate after challenge with the peptide and protein antigen from which the peptide is derived.

In addition, the peptide used according to the process of the invention need not be derived from an amino acid sequence of a myelin antigen such as MBP. According to the process of the invention, any peptide containing a defined sequence of amino acid residues capable of down-regulating an antigen-specific immune response to a myelin autoantigen can be used. For example, peptides containing a defined amino acid sequence not based on the amino acid sequence of the myelin antigen, which are capable of down-regulating the antigen-specific immune response, e.g. the peptide mimics the T cell epitope of the myelin autoantigen and down-regulates the immune response to the myelin autoantigen or causes down-regulation of the immune response for another reason such as this. that it is derived from an adjacent (bystander) .antigen. Without limitation to any theory, we think that neighboring antigens, which are also tissue-specific (but not targeted by the immune or autoimmune attack), may have the ability to elicit suppressor T cells at the site of the immune attack, which can then result in downregulation of immune responses on the site of an immune attack (eg, affected own tissue in case of autoimmune disease, or nasal mucosa, skin and lungs in case of allergy). Neighboring antigens include, but are not limited to, portions of the antigen or autoantigen that are not themselves targeted by the immune attack and which have suppressive activity at the site of the immune attack. As used herein, they include the terms myelin antigen or myelin autoantigen adjacent antigens, which may have suppressive activity at the site of myelin autoimmune attack.

In addition, any compound that mimics a peptide capable of down-regulating an antigen-specific immune response to a myelin autoantigen (e.g., peptidomimetic) can be used in the invention. Such a compound may not necessarily consist entirely of subunits that are coupled to peptide bonds, but may be coupled to other compounds (such as thiolester bonds, reduced bond analogues, amide bond isosterases), provided that a peptide compound mimics a peptide capable of down-regulating an antigen-specific immune response to the antigen in question, as demonstrated by effective therapeutic / prophylactic treatment of symptoms. Peptidomimetics may be based on any of the peptides of the invention, but instead of one or more normal peptide bonds, they may include e.g.

peptide bond analogs (e.g. N-methylamide bonds NH C _a2 [-CO-NH3-] C _a] ) and reduced bond analogs (NH-C _a2 [-CH ₂ -NH-] C a _l )).

Once the peptides containing the T cell epitope have been identified once, it is also possible to modify the structure of such peptides for use in the present invention for such purposes as enhancing solubility (especially desired if the composition is to be injected), promoting therapeutic or preventive efficacy (see discussion of peptide analogs having accelerated MHC binding, below), or stability (e.g., ex vivo shelf life and resistance to proteolytic degradation in vivo, or to facilitate peptide synthesis. Modified peptide or peptide analogue in which the amino acid was sequence altered from the native protein sequence from which it is derived or from the unmodified peptide from which the modified peptide is to be derived. Can be produced by amino acid substitution, deletion or addition by modifying immunogenicity, improving peptide solubility or facilitating peptide synthesis (eg automated peptide synthesis).

The peptide can be modified e.g. by at least retaining, if not already improving, the ability to downregulate the autoimmune response in MS (e.g., by inducing T cell unresponsiveness or by decreased T cell responsiveness) and still retain the ability to bind MHC proteins. In this case, the critical binding residues for the T cell receptor can be determined using known techniques (e.g., substitution of each residue and determination of the presence or absence of T cell reactivity). Those residues that have been shown to be an esadial T cell receptor interaction can be modified by replacing an essential amino acid with another amino acid, preferably with a similar amino acid residue (conservative substitution), the presence of which has been shown to increase, decrease but not eliminate or not. affected T cell activities. In addition, those amino acid residues that are not essential for T cell receptor interaction can be modified by replacing them with another amino acid, the inclusion of which may increase, decrease but not eliminate, or affect T cell activity, but does not eliminate binding to significant MHC.

In addition, the peptides of the invention can be modified by replacing an amino acid that has been shown to be essential for interacting with a protein-MHC complex with another amino acid, preferably with a similar amino acid residue (conservative substitution), whose presence appears to increase, decrease, but does not eliminate or affect T cell activity. In addition, amino acid residues that are not essential for interaction with the MHC protein complex but which still bind the MHC protein complex can be modified by replacing them with another amino acid, the inclusion of which may increase, impair, or decrease but not eliminate T cell reactivity. Preferred amino acid substitutions of non-essential amino acids include, but are not limited to, substitutions with alanine, glutamic acid or methylamino acid. In one example, WO 94/06828 describes substituted peptides in which substantially any amino acid residue may be substituted by a conservative amino acid, or by a non-naturally occurring amino acid or by alanine, a substituted peptide still capable of down-regulating antigen-specific immune response. In another example, Karin et al., J. Exp. Med. 180: 2227-2237 (1994) describe studies using analogues of the immunodominant rat MBP epitope having the amino acid sequence 87-99 (SEQ ID NO: 43) as shown in FIG. 14. The analogs were from a series of 13 substituted peptides based on sequence 87-99 (SEQ ID NO: 43) that differed from the original peptide 87-99 in a single alanine substitution at each position along peptides 87-99 (SEQ ID NO: 43). These studies were designed to identify putative sites where the immunodominant peptide reacts with MHC and TCR in Lewis rats, as well as to explain modified peptides that have improved desired characteristics for potential therapeutic use. These studies showed that the substitution of lysine (K) at position 91 of peptide 87-99 (SEQ ID NO: 43) with alanine (A) (see 87-99 [K> A] (SEQ ID NO: 3b) in FIG. .14) Prevented and abolished EAE in Lewis rats. Based on this information, one would expect alanine (A) substitution with lysine (K) for the only lysine (K) present in peptides containing sequence 87-99 (SEQ ID NO: 43) within its amino acid sequences , ie MBP-2 (SEQ ID NO: 5), MBP2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEO ID NO: 9), MBP-2.5 (SEQ ID NO: 10) and MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2, as a result also increased T cell activity of the substituted peptides. E.g. MBP-2.1 (SEQ ID NO: 6) having lysine (K) substituted at analogue (A) at position 10 of MBP-2.1 peptide (SEQ ID NO: 6) (i.e. DENPVVHFFANIVTPRTPPPSOGK) (SEQ ID NO: 71) increased T ceic activity, which would result in increased therapeutic property compared to the parent MBP-2.1 (SEQ ID NO: 6) peptide.

To enhance stability and / or reactivity, peptides can also be modified to include one or more polymorphisms in the amino acid sequence of a protein antigen resulting from a natural allelic variation. For the production of a modified protein or peptide within the scope of the present invention, D-amino acids, non-natural amino acids and non-amino acid analogs can be added or substituted.

Further, for the production of a protein or peptide conjugated to PEG, the peptides of the present invention can be modified using the polyethylene glycol (PEG) method of A. Sehon et al. (Wie et al. Above). In addition, PEG can be added during the chemical synthesis of a protein or peptide of the invention. Modifications of peptides or portions thereof may also include reduction / alkylation (Tarr in: Methods of Protein Microcharacterization, J.E. Silver ed. Humana Press, Clifton, NJ, pp. 155-194 (1986)); acylation (Tarr. above), chemical coupling to a suitable vehicle (Mishell and Shiigi, eds., Selected Methods in Cellular Immunology, WH Freeman, San Francisco, CA a (1980); US patent 4,939,239; or mild formalin treatment (Marsh International Archives of Allergy and Applied Immunology, 41: 199-215 (1971).

In order to facilitate purification and potentially increase the solubility of the proteins or peptides of the invention, it is possible to add reporter group (s) to the peptide framework. For example, for the purification of a peptide on immobilized metal ion affinity chromatography, (Hochuli, E. et al., Bio / Technology, 6: 1321-1325 (1988)) may be peptkiudxadam ".- pialihistidine. In addition, if desired, specific endoprotease cleavage sites can be introduced between the reporter group and the amino acid sequences of the peptide in order to facilitate peptide isolation without irrelevant sequences. The proper formation of T cell epitope antigen within the peptide can be greatly assisted by the standard recombinant or synthetic design of standard protease sensitive sites between regions, with each site containing at least one T cell epitope. For example, during recombinant peptide construction, charged amino acid pairs such as KK or RR may be introduced between regions within the peptide. The resulting peptide can be made sensitive to cleavage by cathepsin and / or other trypsin-like enzymes to form portions of a peptide containing one or more T cell epitopes.

Another example of peptide modification is the substitution of cysteine residues, preferably with serine, threonine, leucine or glutamic acid, in order to minimize dimerization via disulfide bonds. In addition, for use in buffered aqueous solutions such as pharmaceutically acceptable carriers or diluents, peptides can be modified to increase the solubility of the peptide by adding functional groups, terminal portions of the peptide, or by not incorporating hydrophobic regions into the peptides. To increase solubility, e.g. To the carboxyterminal terminus or amino terminus of the peptide or to both are added charged amino acids or charged amino acid pairs or triplets. Examples of charged amino acids include arginine (R). lysine (K), histidine (H), glutamic acid (E) and aspartic acid (D). In order to facilitate peptide synthesis, such as automated peptide synthesis, it may be desirable to remove or replace amino acids that may impede peptide synthesis or which may make it more expensive. For example, if the N-terminal or C-terminal amino acid of a peptide is capable of cyclization, or if it can be degraded or during or after peptide synthesis, such amino acids can be removed or replaced, or alternatively, one or more additional amino acids may be added. to block the less desired amino or carboxy-terminal amino acids. Such added amino acids may either be derived from the native protein sequence or may be a non-native amino acid residue. To increase peptide T cell activity as defined above, additional amino acids can be added either to the amino-terminal end of the peptide, to the carboxyterminal end of the peptide, or to both. Such additional amino acids may be derived from the native protein sequence or may be non-native amino acid residues.

The peptide compositions administered according to the present invention preferably contain a sufficient percentage of T cell epitopes of myelin autoantigen (i.e. at least about 20%, more preferably about 30%, more preferably about 40%, and more preferably about 60% or more) with total T cell reactivity to myelin autoantigen in a population of autoantigen-responsive individuals who have multiple sclerosis (e.g., at least 10 individuals and more preferably at least 20 individuals) and are included in the composition such that it has a therapeutic regimen for administration of the composition an individual with MS of the present invention as a result of down-regulation of the autoimmune response of MS. Several analytical schemes can be used to determine whether a peptide (preferably a peptide, therapeutic candidate) or combination of candidate peptides is likely to contain a sufficient percentage of the total T cell reactivity of myelin autoantigen to down-regulate the autoimmune response of MS in a significant percentage of the MS population. .

According to one analytical scheme (using MBP as an example), peptides containing a T cell epitope are classified by the number of MBP microtiter culture lines responsive to epitope-containing peptides and by the number of patients with MS undergoing them. they respond. Because MBP-specific T cell frequency in PBL in patients with MS may be very low, it is often not possible to test all MBP peptides in each individual patient with MS. For this reason, the following method is suitable for determining the most useful therapeutic peptides and is further described in Example 1. PBL is isolated from blood and introduced into 96-well microtiter plates. PBLs were purified from fresh peripheral blood samples (approximately 75 cm ³ ) of patients with definitive MS using a Ficoll density gradient. Microtiter cultures were introduced with 2 χ 10 ⁵ PBL per well and K) Mg / ml MBP of purified human brain in the culture medium.

RPMI 1640 is supplemented with 5% human AB serum, penicillin-streptomycin and L-glutamine. Start at 6-7 days of culture supplemented with IL2 (20 units / ml) and IL4 (5 units / ml). After 11-13 days, the microtiter cultures were washed, resuspended in fresh medium and divided into 12 fresh microtiter wells. Autologous frozen PBLs were added as antigen presenting cells in an amount of 5 χ 10 ⁴ PBL per well. To the 12 replication wells from each microtiter culture, antigens were selected in duplicate. Medium is always used as a negative control and purified human recombinant MBP of 10 ^ g / ml is used as a positive control. Each patient is also tested for reactivity to a maximum of 4 MBP peptides, each at a concentration of 10 μΜ. After 48 hours of analysis, contact

27.75 kBq ³ H-thymidine and were collected after 6-16 hours of contact. Cultures were evaluated as positive for each peptide according to the following criteria: stimulation index greater than 3.0, change in CPM greater than or equal to 500, and standard error of the mean less than change in CPM. In addition, for analytical purposes, cultures are evaluated as peptide-positive only if they respond to both MBP and peptide and if they do not respond to more than one non-overlapping peptide. Groups of about 10 - 50 patients are preferably tested with each of the MBP peptides, and each patient group is tested with up to 4 peptides. The peptides are then classified according to the following criteria: 1) the percentage of MBP positive microtiter cultures in each patient group (total MBP reactivity) that give a positive result also for one of the MBP peptides; 2) the percentage of MBP response individuals in each group with at least one microtiter culture that gave a positive result for one of the MBP peptides, with the MBP response individual defined as a patient with at least one microtiter culture that gave a positive result for MBP. Individual candidate peptides are then selected only if 1) they contain at least 5%, more preferably at least 10% and most preferably at least 20% of the total MBP reactivity and 2) the reactivity found for them is at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50%, and most preferably at 60% MBP response individuals. If desired, a positive index for the classification of peptides can also be considered a positive index for a given peptide. The positive index represents both the strength of the T cell response to the peptide (SI) and the frequency of the T cell response to the peptide in a population of individuals responding to the myelin autoantigen. For example, as shown in FIG. 15, the positivity index 141-165 (MBP-4) (SEQ ID NO: 14) is about 2500, which was calculated using the data described here in Example 1. Mean SI peptide MBP-4 (SEQ ID NO: 14) ) to the MBP of the responding patient is multiplied by the percentage of individuals responding to MBP-4 (SEQ ID NO: 14) in the population of patients responding to the MBP.

Highly purified peptides, essentially free of all other polypeptides and contaminants having a defined sequence of amino acid residues comprising at least one T cell epitope used in the therapeutic compositions of the invention, can be produced synthetically by chemical synthesis using standard techniques. Various methods of chemical synthesis of peptides are known in the art, such as solid-phase synthesis on commercially available peptide synthesizers, which is fully or semi-automated. The synthetically produced peptides can then be purified, preferably to homogeneity, more typically up to at least 90%, more preferably up to at least 95% and even more preferably up to at least 97% purity, using any number of techniques known in the protein purification literature, essentially free of all other polypeptides and contaminants.

The synthetically produced peptides of the invention, comprising up to about 45 amino acid residues in length and most preferably up to about 30 amino acid residues, are particularly desirable since length increases may result in difficulty in peptide synthesis. Larger length peptides can be produced by recombinant DNA techniques as explained below.

Peptides useful in the methods of the present invention can also be produced using recombinant DNA techniques in a host cell transformed with a nucleic acid sequence encoding for such a peptide. When produced by recombinant techniques, we cultivate host cells that have been transformed with a nucleic acid encoding the desired peptide into cell-appropriate medium, and isolated peptides can be purified from cell culture medium, host cells, or both using techniques known in the art. for the purification of peptides and proteins, including ion exchange chromatography, ultrafiltration, electrophoresis, or immune purification of "antibodies specific for the desired peptide. For use in accordance with the procedures described above for synthetically produced peptides, recombinantly produced peptides can be isolated and purified, preferably to homogeneity, so that they are substantially free of cellular material, other polypeptides or culture medium.

In certain limited circumstances, peptides can also be produced by chemical or enzymatic cleavage of a highly purified full-length protein or a natural protein that has been previously identified by sites of chemical metabolism or enzymatic cleavage and whose resulting secretion can be reproduced. Peptides having defined amino acid sequences can be well purified and isolated essentially without any other polypeptides or contaminants present in the enzymatic or chemical secretion by any of the processes described above for highly purified and isolated synthetically or recombinantly produced peptides.

In addition, the peptides of the present invention can be used in conjugates as described e.g. in the U.S. 5,130,297 (Sharma et al.), wherein the therapeutic agents have been prepared using the formula X-MHC-peptide or MHC-peptide-X, in which X represents a functional moiety selected from the toxin and the labeling group; MHC is an effective part of the MHC glycoprotein, said glycoprotein being dissociated from the cell surface on which it is normally located; and the peptide represents any of the peptides listed herein, more specifically MBP or MOG, and more specifically represents the peptides shown in FIG. 2 and 14.

The preferred peptides of the invention further comprise at least one T-cell epitope of the protein? Polnt ΐ1ο1ζίη6, more specifically MBP or MOG. Peptides may contain tandem repeats of each epitope and / or more than one epitope.

According to the methods described herein for the identification of peptides comprising the T cell activity of MBP (see discussion above as well as example 1 below), the preferred peptides derived from MBP that are candidates for therapeutic use comprise the following peptides: MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15) ), all as shown in FIG. 2, or any part thereof, or any modification thereof. These peptides were tested for T cell activity as described in Example 1, and were shown to encompass a implanted Tscellular epiiop, as evidenced by the MBP ratio of positive rmcriteric cultures, which also gave a positive result for one of the MBP peptides (Fig. 4a) and the detectable response for each of the preferred peptides in a significant percentage of MBP of the patients being tested (Fig. 4b). MBP-4 (141-165) (SEQ ID NO: 14) is the most reactive of the four peptides, attributable to 21% of the overall MBP response and detectable in 64% of the MBP response patients being tested. Among the four peptides, MBP-4 (SEQ ID NO: 14) showed surprisingly much more reactivity than the combined reactivities of MBP 141-160 (SEQ ID NO: 28) and MBP 151-170 (SEQ ID NO: 29). as shown in FIG. 3. The present inventors were the first to identify this immunodominant peptide, which appears to comprise multiple T cell epitopes. This new peptide is particularly preferred for therapeutic use.

In addition, we think MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), and MBP-3.1 (SEQ ID NO: 13) as shown in FIG. 2, suitable candidate peptides for therapeutic use. These peptides are modified versions of MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5) or. MBP-3 (SEQ ID NO: 12), and are expected to have similar T cell activity to that of their respective parent peptides. These peptides were modified by amino acid deletion, addition, or both, according to peptide modification techniques as described above, mainly to facilitate peptide synthesis.

Other peptides that have been shown to be immunodominant (i.e. having T cell MBP activity) or derived from peptides known to have T cell MBP activity have been identified by the inventors or others working in the field (see, e.g., USSN 08 / 328,224 filed October 25, 1994; USSN 08 / 241,246 filed

May 10, 1994; WO 93/21222; EP 0 304 279, WO 91/15225; Ota et al. Letters to Nature, 346: 183-187 (1990); Wucherpfennig et al., J. Exp. Med. 170: 279-290 (1994); Martin et al., J. Immunol. (1990) 145: 540-548); Karin et al., J. Exp. Med. 180: 22272237 (1994). Such peptides may also be suitable for therapeutic use in the compositions and methods of the invention, especially when combined with the preferred candidate peptides described above. Such peptides include, but are not limited to, all or part of the following peptides having residue numbers corresponding to the amino acid residues of the human MBP protein shown in FIG. 1, and having the individual amino acid sequences shown in FIG. 14: 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32),

82-96 (SEQ ID NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100P> Yj (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37),

83- 101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 84-100 (SEQ ID NO: 40), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 88-100 (SEQ ID NO) : 44), 88-99 (SEQ ID NO: 45), 111-135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141- 160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153-170 (SEQ ID NO: 53), and more preferably comprise the following peptides: 13-25 (SEQ ID NO: 31), 87-99 (SEQ ID

NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 821 () () [] OOP> Y] (SEQ ID NO: 46) , all as shown in FIG. 14. Preferred portions of these peptides or preferred modifications should preferably have similar or greater T cell activity in an equal or greater percentage of patients to be tested and / or have similar or greater therapeutic efficacy in the methods of the invention, such as that which has the parent peptide from which the modified peptide is derived.

One aspect of the present invention provides therapeutic compositions comprising at least one peptide derived from a myelin antigen having T cell activity, or a combination of peptides derived from a myelin antigen, each peptide having T cell activity, and a pharmaceutically acceptable carrier or diluent. Preferred therapeutic compositions comprise a sufficient percentage of the T cell activity of the myelin autoantigen such that when administered to a patient with MS in a therapeutic regimen, preferably in a non-immunogenic form, they are capable of down-regulating a specific immune response to the myelin autoantigen in a population of such antigen-specific responses. As used herein, it includes, but is not limited to, down-regulation, prevention of the onset of disease symptoms, reduction of disease symptoms of multiple sclerosis caused by an antigen-specific immune response to MBP or other myelin autoantigen, more specifically including reduction, nullification, non-progression or mitigation simpotomov. Non-progression may be characterized, but not limited, by (a) shorter periods of active disease or exacerbation. (b) _ less severe symptoms or inability, (c) delay in disease progression, with the underlying state of health not going down as quickly (d), increasing or prolonging the time between periods of active disease or exacerbation (eg longer periods of remission), ( e) with fewer disease recurrences or exacerbations and / or (f) slowing or stopping the progression of the extent of MRL-detected lesions Other determining tools used by experts in the art are Expanded Disability Status Scale (EDSS) and Neurological evaluation systems Rating Scale. As used herein, the advanced stage is any point beyond the pure clinical signs of overt disease, whether the disease is returning-relapsing MS, chronic progressive MS, benign MS or primary progressive MS. In addition to this definition, the advanced stage may include acute phase (s), remission (s) and exacerbation (s). As used herein, the term acute phase may mean ongoing seizure, acute illness, active disease and exacerbation, and may be used interchangeably. As used herein, these terms generally refer to a condition in which a person with the disease (whether diagnosed or not) exhibits active symptoms or signs that are commonly understood by one of skill in the art in conjunction with a specific immune response characteristic of multiple sclerosis. It is understood that disease recurrence represents an acute phase following remission. The term exacerbation, when used in an appropriate context, can also be interpreted as meaning new and worsening symptoms or signs. Symptoms are those indications of a disease that causes the patient to complain. The signs are those indications that are noticed or measured by the diagnostician. Of course, unless otherwise stated, the terms symptoms and signs may be used interchangeably.

The therapeutic compositions of the invention preferably comprise at least one T cell epitope comprising a peptide or a modified peptide or peptide analog and a pharmaceutically acceptable carrier or diluent. Such compositions may preferably comprise an MBP peptide selected from the following peptide group: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9) ), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 ( SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), more preferably the MBP peptide is selected from MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), and more preferably, the MBP peptide MBP-4 (SEQ ID NO: 14).

The compositions of the invention may comprise at least two peptides (e.g., a physical mixture of at least two peptides), each peptide having T cell activity and preferably comprising at least one T cell epitope of a myelin autoantigen such as MBP. Such compositions may be administered in the form of a therapeutic composition with a pharmaceutically acceptable carrier or diluent. A therapeutically effective amount of one or more such compositions may be administered to an individual suffering from MS simultaneously or sequentially. Preferred compositions comprise at least one peptide, and preferably at least two peptides selected from the following group of peptides: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4 ), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7) .MBP 2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ) ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO:

13), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15) all as shown in FIG. 2 and more preferably selected from the following peptide group: MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2.1 (SEO ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15) and most preferably selected from the following peptide group: MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEO ID NO: 14), and MBP-5 (SEQ ID NO: 15). In addition, the compositions of the invention may further comprise peptides derived from MBP having residue numbers corresponding to the amino acid residues of the human MBP protein shown in FIG. 1 and having individual amino acid sequences as shown in FIG. 14: 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEO ID NO: 21), 82-92 (SEO ID NO: 32), 82-96 ( SEO ID NO: 33), 82-97 (SEO ID NO: 34j, 82-98 (SEO ID NO: 35),

82-100 (SEQ ID NO: 30), 82-100 [100P> Y] (SEQ ID NO: 46), 83-100 (SEO ID NO: 37), 83-101 (SEQ ID NO: 38), 84 -97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO. : 43), 87-99 [91K> A] (SEQ ID NO: 36), 88-100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111-135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141-160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153-170 (SEQ ID NO: 53) and more preferably comprise the following peptides: 13-25 (SEQ ID NO: 31), 87- 99 (SEQ ID NO: 43), 8799 [91K> Aj (SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 82-100 [100P> Y] (SEQ ID NO: 46). Preferably, the compositions of the invention comprise at least two peptides, wherein at least one peptide is MBP-4 (SEQ ID NO: 14) and at least one peptide is selected from the following group of peptides: MBP-1 (SEQ ID NO: 2), MBP- 1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEO ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13) and MBP-5 (SEQ ID NO: 15), all as shown in FIG. 2 and further may comprise at least one of the following MBP derived peptides as shown in FIG. 14: 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32), 82-96 ( SEO ID NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEO ID NO: 35), 82-100 (SEO ID NO: 30), 82-100 [100P> Y] (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83-101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 88 -100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111-135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO. : 49), 141-160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153- 170 (SEQ ID NO: 53) and more preferably comprise at least one of the following peptides: 13-25 (SEQ ID NO: 31), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] ( SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 82-100 [100P> Υ] (SEQ ID NO: 46).

Preferred compositions of the invention comprise the following peptides:

MBP-1 (SEQ ID NO: 2), MBP-2 (SEO ID NO: 5), MBP-3 (SEQ ID NO: 12) and MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-3 (SEQ ID NO: 12),

MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15):

MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEO ID NO: 5), MBP-4 (SEO ID NO: 14) and

MBP-5 (SEQ ID NO: 15);

MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEO ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-3 (SEQ ID NO: 12); MBP-1 (SEQ ID NO: 2) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11);

MBP-1.1 (SEQ ID NO: 3) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5). MBP-2.1 (SEO ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10) or MBP-2.6 (SEQ ID NO: 11);

MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11);

MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-1 (SEQ ID NO: 2) or MBP-1.1 (SEQ ID NO: 3);

MBP-1.1 (SEQ ID NO: 3), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO; 30), 82-100 [100P> Y ] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3) and MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO) : 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10) or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2;

MBP-1.1 (SEO ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO : 30), 82-100 [100P> Y] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43), and 87-99 [91K> A] (SEQ ID NO: 36), all as are shown in FIG. 14;

MBP-1.1 (SEO ID NO: 3), MBP-5 (SEO ID NO: 15), MBP-4 (SEO ID NO: 14) and a peptide selected from the group consisting of MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all of which are shown in FIG. 2;

MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO. : 30), 82-100 [100P> Y] (SEO ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14;

MBP-1.1 (SEQ ID NO: 3), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of:

MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG.

The present invention also describes a treatment regimen comprising administering combinations of therapeutically effective peptides as a single treatment event. Such peptide combinations can be administered simultaneously or sequentially, as therapeutic compositions comprising only one peptide or comprising multiple peptides. Such a treatment regimen does not necessarily have to be a physical mixture of more than one peptide, but comprises a combination of peptides administered simultaneously or sequentially as a single treatment event. Preferred peptide combinations (in the form of one or more compositions, each comprising one or more peptides) that can be administered simultaneously or sequentially as a single treatment event include the following peptide combinations:

MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-3 (SEQ ID NO: 12) and MBP-4 (SEO ID NO: 14) and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEO ID NO: 6), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and

MBP-5 (SEQ ID NO: 15);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEO ID NO: 14);

MBP-1 (SEO ID NO: 2), MBP-2.1 (SEO ID NO: 6) and MBP-4 (SEO ID NO: 14);

MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5) and MBP-4 (SEO ID NO: 14);

MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-3 (SEQ ID NO: 12); MBP-1 (SEQ ID NO: 2) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2-6 (SEQ ID NO: 11);

MBP-1.1 (SEQ ID NO: 3) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11);

MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11);

MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-1 (SEQ ID NO: 2) or MBP-1.1 (SEQ ID NO: 3);

MBP-1.1 (SEQ ID NO: 3), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO: 30), 82-100 [100P> Y ] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14;

MBP-1.1 (SEQ ID NO: 3) and MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO) : 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10) or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2;

MBP-1.1 (SEO ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO : 30), 82-100 [100P> Yj (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> Aj (SEQ ID NO: 36), all shown in FIG. . 14;

MBP-1.1 (SEO ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all as are shown in FIG. 2;

MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO. : 30), 82-100 [100P> Y] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14;

MBP-1.1 (SEQ ID NO: 3), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEO ID

NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10) or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2.

In addition, preferred compositions and preferred combinations of MBP peptides that can be administered simultaneously and / or sequentially may include any of the above compositions and combinations, and may further comprise at least one T cell epitope-containing peptide derived from myelin oligodentrocyte protein (MOG ), another protein thought to be one of the autoantigens involved in multiple sclerosis (see Lebar, et al., J. Immunol. (1976) 116: 1439-1446; Lebar et al., J.Exp. Immunol (1986) 66: 423-443; Linington and Lassman, J. Neuroimmunol. / 1987) 17: 61-69; Lassman et al., Acta Neuorpathol. (Barry) (1988) 75: 566-576; and Sun et al., J. Immunol. (1991) 146: 1490-1495). Peptides that may comprise T cell epitopes derived from MOG are described in USSN 08 / 116,824 filed September 3, 1993 and USSN 08 / 300,811 filed September 1, 1994 (incorporated herein by reference) and which are expected to be effective in the treatment multiple sclerosis when prepared and / or administered in conjunction with the compositions described above, and combinations of T cell epitope-containing peptides of MBP of the present invention are:

Human MOG 1-13 (SEQ ID NO: 54) Human MOG 103-115 (SEQ ID NO: 55) Human MOG 1-121 (SEQ ID NO: 56)

G Q F R V I G P R H P I R

H S Y Q E E A A M E L K. V

GQFRVIGPRHPIRALVGDEV

ELPCRTSPGKNATGMEVGWY

RPPFSRVVHLYRNGKDQDGD

QAPEYRGRTELLKDAIGEGK

VTLRIRNVRFSDEGGFTCFF

RDHSYQEEAAMELKVEDPFYW

Additional MOG peptides containing the epitope were identified by the inventors using an experimental analysis similar to that described above and in Example 1 (data not shown). Such peptides include:

HumaniMOG 1-20 (SEQ ID NO: 57) HumaniMOG 11-30 (SEQ ID NO: 58) HumaniMOG 21-40 (SEQ ID NO: 59) HumaniMOG 31-50 (SEQ ID NO: 60) HumaniMOG 141-160 ( SEQID: 61) HumaniMOG 151-170 (SEQID: 62) HumaniMOG I6I-I80 (SEQ ID NO: 63) HumaniMOG 199-218 (SEQID: 64)

GQFRVIGPRHPIRALVGDEV;

PIRALVGDEVELPCR1SPGK;

ELPCRISPGKNATGMEVGWY;

NATGMEVGWYRPPFSRVVHL;

TVGLVFLCLQYRLRGKLRAE;

YRLRGKLRAEIENLHRTFDP;

IENLHRTFDPHFLRVPC WKI; and ynwlhrrlagqfleelrnpf.

The present invention further contemplates modifications or analogues (previously discussed) of MOG top peptides containing a T cell epitope that retain similar or greater T cell activity to that of the parent peptide from which the modification or analog is derived.

Compositions comprising the T cell epitope-containing peptides of MOG and MBP, their modifications, their analogs or peptidomimetics based on them, and combinations of such peptides which can be administered simultaneously or sequentially, are thought to have the advantage of maximizing the downregulation -regulatory effect on both MBP and MOG-specific T cells participating in the autoimmune response in MS. In this way, we can target a set of T cells that may participate in the autoimmune response or MBP or MOG, resulting in clinical manifestations of MS (demyelination) in order to regulate them, thereby accelerating the therapeutic effect of the compounds and compositions. according to the invention. Similarly, T cell epitope-containing peptides derived from other myelin antigens thought to be autoantigens in MS (eg, proteolipid protein (PLP) and myelin-associated glycoprotein (GI) may also be suitable in the compositions and methods of the invention. MAG)).

The therapeutic / prophylactic treatment regimen of the present invention (resulting in the elimination, prevention or delay of onset of disease symptoms caused by the causative autoantigen, or resulting in the reduction, non-progression or mitigation of the symptoms caused by the causative autoantigen, ie, down-regulation autoantigen-specific, immune response) comprises administering a therapeutic composition of the invention in a non-immunogenic form comprising at least one isolated peptide derived from an autoantigen responsible for the condition being treated (e.g., MBP, MOG, PLP, MAG ). Although we do not intend to confine ourselves to any theory, we think that administering a therapeutic composition of the invention may: a) cause T cell non-responsiveness of the corresponding T cell subpopulations by becoming unresponsive to the causative antigen and not participating in stimulating the immune response after exposure to the causative agent. protein antigen (eg via anergy or apoptosis); b) modify the lymphokine secretion profile compared to the exposure to the causative autoantigen; c) causing the T cell subpopulations normally involved in the response to the antigen to be withdrawn from the normal exposure sites against the sites of administration (this redistribution of the T cell subpopulations may enhance or decrease the ability of the individual's immune system to stimulate the normal immune response at the site normal exposures to the causative antigen resulting in the reduction or elimination of symptoms); d) induce the induction of suppressor cells T or

e) induce the suppression of T suppressor cells via the bystander antigen.

The highly purified and isolated peptides produced as described above can be formulated into therapeutic compositions of the invention suitable for human therapy. If the therapeutic composition of the invention is to be administered by injection (e.g., subcutaneous injection, intravenous injection), then it is preferable that the highly purified peptide is soluble in aqueous solution at a pharmaceutically acceptable pH (i.e., a pH range of about 4-9) the composition is fluid and easily injectable. The composition preferably also includes a pharmaceutically acceptable carrier. As used herein, the pharmaceutically acceptable carrier includes any and all excipients, solvents, dispersion media, coatings, antibacterial and antifungal agents, toxic agents, buffers, absorption retarding or promoting agents, surfactants and micelles, lipids, liposomes and liquid complexing agents, stabilizing agents and the like. The use of such media and agents for the pharmaceutically active substance is known in the art. It is intended to use them in therapeutic compositions, except to the extent that any conventional medium or agent is incompatible with the active compound. Additional active compounds may also be included in the compositions.

As described above, the therapeutic compositions of the invention suitable for injection are preferably sterile aqueous suspensions prepared by combining the active compound (i.e., one or more highly purified and isolated peptides as described above) in the required amount in a suitable amount. solvent, with one ingredient or combination of ingredients listed above and below as required, followed by filter sterilization. Preferred pharmaceutically acceptable carriers include at least one excipient, such as sterile water, sodium phosphate, mannitol, sorbitol or sodium chloride, or any combination thereof. Other pharmaceutically acceptable carriers that may be suitable include solvents or a dispersion medium containing e.g. water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol and the like), suitable mixtures thereof and vegetable oils. Appropriate fluid can be maintained e.g. using a coating such as lecithin in the case of dispersions by maintaining the required particle size and using surfactants. The prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, e.g. parabens, chlorobutanol, phenol, ascorbic acid, tymerozole and the like. Prolonged absorption of injectable compositions can be achieved by incorporating an absorbent-retaining agent into the composition, e.g. aluminum monostearate and gelatin.

Preferred therapeutic compositions of the invention must be sterile, stable under conditions of manufacture, storage, distribution and use, and must be protected against the contamination effects of microorganisms such as bacteria and fungi. A preferred method of manufacturing a therapeutic composition that maintains the integrity of the composition (i.e., prevents contamination, prolongs storage, etc.) is to prepare a formulation of the peptide and pharmaceutically acceptable carrier (s) such that the composition can be in the form of a lyophilized powder, which is reconstituted immediately before use in a pharmaceutically acceptable carrier such as sterile water. In the case of sterile powders for the preparation of sterile injection solutions, vacuum drying, lyophilization or spin drying processes which give the powder of the active ingredient plus any desired ingredient from its pre-sterile filtered solution are preferred.

As described above, the therapeutic composition of the invention may comprise more than one isolated peptide. A therapeutic composition comprising a multipeptide formulation suitable for pharmaceutical administration to humans may be desirable for the administration of several inactive peptides. The multipeptide formulation includes at least two or more isolated peptides having a defined amino acid sequence and is capable of down-regulating the antigen-specific immune response. Any of the previously described compositions comprising at least two peptides may be suitable as a multipeptide formulation. As described previously, highly desirable peptides that are suitable for use in the multipeptide formulation include at least two of the following peptides:

MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEO ID NO: 10), MBP -2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO) : 15). In preparing the multipeptide formulation, special care must be taken to maintain the solubility and stability of all the peptides in the formulation in aqueous solution at a physiologically acceptable pH. This requires the selection of one or more pharmaceutically acceptable solvents and excipients that are compatible with all peptides in the multipeptide formulation. Suitable excipients include e.g. sterile water, sodium phosphate, mannitol, or both, sodium phosphate and mannitol. Additionally, if necessary, care should be taken in the multipeptide formulation to prevent dimerization of the peptides.

Administration of the therapeutic compositions as described above in a non-immunogenic form can be carried out in a person using known methods, at doses and at periods effective to cause down-regulation of MBP of an antigen-specific immune response of a person being treated for MS . The downregulation of an antigen-specific immune response to an antigen associated with a human condition can be clinically determined, where possible, or subjectively determined (ie, the patient feels if some or all of the associated symptoms have been alleviated) with the condition we are treating).

The effective amounts of the therapeutic compositions of the invention may vary depending on factors such as the degree of sensitivity of the individual to the antigen, age, sex and weight of the individual, and the ability of the peptide to induce down-regulation of the antigen-specific immune response in the individual. The therapeutic composition of the invention may be administered in a non-immunogenic form in a suitable manner, such as by injection (subcutaneously, intravenously, etc.), by oral administration, sublingually, by inhalation, by transdermal administration, by rectal administration, or by any combination of routes of administration, which are said to promote therapeutic efficacy, or any other route of administration known in the art to administer therapeutic agents. It may be desirable to administer a therapeutically effective amount of one or more therapeutic compositions of the invention to the individual simultaneously or sequentially. Each of such compositions for simultaneous or sequential administration may comprise only one peptide, or may comprise a multipeptide formulation as described above.

For administration of a peptide or peptide composition other than parenteral administration, it may be necessary to coat the peptide with a material to prevent its inactivation, or to administer the material simultaneously with the peptide. For example, the peptide composition may be administered simultaneously with enzyme inhibitors or in liposomes. Enzyme inhibitors include pancreatic trypsin inhibitor, diisopropylfluorophosphate (DEP) and trazilol. Liposomes include water-in-oil-in-water CGF emulsions as well as conventional liposomes (Strejan et al., (1984) J. Neuroimmunol. 7:27). When the peptide is suitably protected as described above, it can be administered orally, for example, by an inert diluent or by an assimilation-capable edible carrier. The peptide composition and other ingredients may also be included in hard or soft gelatin capsules, compressed into tablets or incorporated directly into an individual's diet. For oral therapeutic administration, the active compound may be combined with excipients and used in the form of digestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wraps and the like. Such compositions and preparations must contain at least 1% by weight of active compound. The percentage of the compositions and preparations may, of course, vary and may be appropriate between about 5 to 80% by weight of the unit. The amount of active compound in such therapeutically useful compositions is such that a suitable dose is obtained. Preferred compositions or preparations of the invention are prepared in such a way that the oral dosage unit contains between about 10 µg to about 200 mg of the active compound. Tablets, lozenges, pills, capsules and the like may also contain the following: a binder such as gumitragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as maize starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose or saccharin or a flavoring agent such as peppermint, green oil or cherry aroma. When the unit dosage form is a capsule, it may contain a liquid carrier in addition to the materials of the above type. No different materials may be present as coatings, or for any other modification of the physical form of the dosage unit. For example, tablets, pills or capsules may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparaben as a preservative, colorant and flavor, such as cherry or orange flavor. Of course, any material used in the preparation of any unit dosage form must be pharmaceutically pure and substantially non-toxic in the amounts used. In addition, the active compound may be included in sustained release formulations and formulations.

For injection (subcutaneous IV, IM, intraperitoneal) one or more therapeutic compositions of the invention can be administered, preferably about 1 / wt-3 mg and more preferably from about 20 ^ gl, 5 mg and more preferably about 50 / wt-750 / wg and more preferably about 75 / wg to about 750 gg of each active component (peptide) per dosage unit. Depending on the regimen as described below, doses of 1500 / g or more may be used. In order to facilitate administration and uniformity of dosage, it is particularly advantageous to formulate the parenteral compositions in a unit dosage form. The unit dosage form as used herein refers to physical discrete units that are suitable as unitary doses for the people to be treated, each unit containing a predetermined amount of the active compound calculated to produce the desired therapeutic effect in in association with the desired pharmaceutical carrier. The specification for new unit dosage forms of the invention is dictated and directly dependent on (a) the unique properties of the active compound and the particular therapeutic effect to be achieved and (b) the limitations present in the preparation of such active compound for human treatment.

The dosage regimen can be adjusted to provide an optimal prophylactic or therapeutic response. For example, multiple divided doses may be administered over days, weeks, months, or years, or with each subsequent injection, the dose may be increased or decreased proportionally, as indicated by the requirements of the therapeutic situation. In a preferred therapeutic regimen, subcutaneous injections of therapeutic compositions are administered once daily during the acute phase and once every other day during remission during the lifetime of the individual suffering from the disease. Alternatives would include weekly, monthly or other periodic injections. The dose may remain constant for each injection or may be increased or decreased with each subsequent injection. A continuous, lifelong treatment program may be most desirable. Alternatively, a boost injection may be given at intervals of about three months to about 1 year after the initial treatment period, and may include only one injection, but may include another series of injections similar to those of the original treatment.

Due to the highly individual response of the immune system to individuals with or with impaired MS, primary progressive MS, benign MS or chronic progressive MS, careful and different dosing regimens need to be developed. First, we need to evaluate the intake of the patient in question. We perform a complete test, in which, among other things, we look for impaired vision, nystagamus, dysarthria, impaired vigilance percussion, and impaired sense of position, ataxia and intent tremor, weakness or paralysis of one or more limbs, spasticity and bladder problems. Experts will use accepted tools such as EDSS, the Neurological Ranking Scale, and other similarly known tools in the art, as well as previously discussed disease status indicators, to determine the baseline from which any changes can be measured, including the progression of disability, but preferred down-regulation. Although there is no typical acute or impaired phase of MS, certain patterns have emerged that will guide an experienced physician. As mentioned above, the incidence of inflammation or acute conditions is highest during the first 3 to 4 years of the disease, but the first attack is not necessarily followed by another observed attack of 10 to 20 years (although it may show accumulation of lesions that can only be detected by MRI , a different clinical picture). During typical episodic events, symptoms worsen over a period of several days, up to 2 to 3 weeks, and then subside. Recovery is usually quick, over the course of a few weeks, though it can sometimes extend to several months.

MS is a chronic disease and continuous therapy is required. Continuous, long-term treatment may be the most effective treatment to stop disease progression and incapacity. The dosing regimen may be appropriate every other day, at least once a week or once a month. Meaningful modifications are within the skill of the expert. Intervention at the onset of an attack is considered an important component of effective treatment.

The severity of the attack should be evaluated for the patient in the acute stage. The initial injection can be given at the beginning of the acute stage. It would obviously be useful to start treatment as early as possible in the acute phase. If desired, this treatment can be combined with concomitant treatment with / 3-interferon, steroids, and / or other therapies, especially those designed to reduce inflammation. The patient is observed daily after the first treatment, preferably by injection. It is suggested that additional treatments be given daily or at least every third day during the acute phase. As with any medication, the treating physician must modify the dose according to clinical changes that indicate a need for modification. Any such discretion is within the scope of the experts using the proposed dosage program and quantities as guidance. A dosage range of about 75 to about 750 / g per dose gives the GP a great deal of freedom (and not an absurd amount). The descriptions include, but are not limited to, daily treatments during the acute phase followed by the course of remission treatment listed above. This does not in any way exclude the possibility of more or less frequent dosing if the treating physician determines that intervention is required.

The present invention describes that the advanced stage of multiple sclerosis can be treated according to the compositions and methods of the invention. Compositions comprising at least one peptide having T cell activity and derived from myelin antigen are suitable for the treatment of advanced stage MS. The compositions of MBP peptides, MOG peptides and combinations thereof described above are suitable for the treatment of advanced stage MS. As described above, these peptides include, but are not limited to:

MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP -2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), MBP-5 (SEQ ID NO) : 15), 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32), 82- 96 (SEQ ID NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100P> Yj ( SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83-101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40) , 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91 K> Aj (SEQ ID NO: 36), 88-100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111, -135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141-160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52), and 153-170 (SEQ ID NO: 53) and i n preferably MBP-1.1 (SEQ ID NO: 3),

MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), MBP-5 (SEQ ID NO: 15), 13-25 (SEQ ID NO: 31), 87-99 (SEQ ID NO: 43), 87-99 [91 K> AJ (SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 82-100 [100P> Y] (SEQ ID NO: 46} as well as peptides derived from human MOG:

Human MOG 1-13 (SEQ ID NO: 54) Human MOG 103-115 (SEQ ID NO: 55) Human MOG 1 -121 (SEQ ID NO: 56)

Human MOG 1-20 (SEQ ID NO: 57) Human MOG 11-30 (SEQ ID NO: 58) Human MOG 21-40 (SEQ ID NO: 59) Human MOG 31-50 (SEQ ID NO: 60) Human MOG 141-160 (SEQ ID NO: 61) Human MOG 151-170 (SEQ ID NO: 62) Human MOG 161-180 (SEQ ID NO: 63) Human MOG 199-218 (SEQ ID NO: 64)

GQFR VIGPRHPIR

HSYQEEAAMELKV

GQFRVIGPRHPIRALVGDEV

ELPCRTSPGKNATGMEVGWY

RPPFSRVVHLYRNGKDQDGD

Q A P EYRGRTELLKDAIGEGK

V T L R I R N V R F S D E G G F TC F F

RDHSYQEEAAMELKVEDPFYW

GQFRVIGPRHPIRALVGDEV;

PIRALVGDEVELPCRISPGK;

ELPCR1SPGKNATGMEVGWY;

NATGMEVGWYRPPFSRVVHL;

TVGLVFLCLQYRLRGKLRAE;

YRLRGKLRAEIENLHRTFDP;

IENLHRTFDPHFLRVPCVVKI; and

YNWLHRRLAGQFLEELRNPF.

The inventors here were the first to come to the surprising discovery that an intervention in the middle of an ongoing attack actually improves the condition of the person being treated according to the process of the invention. At a minimum, such intervention does not impair the person's condition. Furthermore, the applicants show that the intervention during remission down-regulates the immune response and that, as a result of any such intervention, the condition is not activated. Thus, the intervention can not only lead to improvement, but appears to be completely safe when used in the context of the process of the present invention. Using the EAE model described above, the Applicants illustrate that repeated intravenous administration of the immunodominant peptide MBP Acl-11 (SEQ ID NO: 65) successfully cures EAE induced by whole MBP protein. Successful treatment (T cell reactivity indicator) is shown with a native peptide (an unsubstituted peptide derived from the native sequence according to the procedures described herein for identification of a T cell epitope containing peptides derived from a protein autoantigen), suitably modified peptides however, it can be identified by comparing the binding affinities of the modified peptide with the native peptide. A modified peptide having a binding affinity lower than the native peptide would be a suitable candidate peptide for the therapeutic composition of the present invention. Assays for determining binding affinities can be obtained from the references in USSN 08 / 300,811 filed September 1, 1994, which are incorporated herein by reference. For example, we have found that Acl-11 (SEQ ID NO: 65) therapy is effective. Acl-11 therapy (SEQ ID NO: 65) was then compared to Acl-11 therapy [4Y] (SEQ ID NO: 67), an Acl-11 analog (SEQ ID NO: 65) that binds to A ^U A ^U with higher affinity and greater stability (Wraith, DC et al., Above and Fairchild. PJ et al. Above). Interestingly, the Applicants found that Acl-11 [4Y] (SEQ ID NO: 67) was effective at 100 times lower dose than Acl-11 (SEQ ID NO: 65) and that its therapeutic effect lasted longer, indicating that the selected modified peptides would also be suitable in the therapeutic composition. Applicants further show that the particular tested peptide Acl-11 [4Y] (SEQ ID NO: 67) in stable mice forms stable peptide-MHC complexes in vivo. This suggests that it is very likely that the modified peptides forming stable peptide-MHC complexes in vivo are suitable peptides for human treatment, since their potency can be greatly increased by conservative amino acid substitutions (see also Karin et al., J. Exp. Med. 180: 2227-2237 (1994). Applicants also found that the peptide analogue MBP Acl11 [4Y] (SEQ ID NO: 67) eliminated running paralysis when administered during the acute phase of EAE and prevented disease recurrence when administered during remission. This suggests that modified peptides and analogs of selected peptides can suppress active symptoms and prevent disease recurrence in people suffering from MS.

In another model of the course of human disease, applicants have initiated intravenous peptide therapy after initiation of EAE and during the acute phase. Quite surprisingly, this therapy nullified the disease in progress when given during the acute phase. This finding is the first of its kind. Past experiments made by others were initiated with peptide therapy when most mice in the treatment group had not yet shown signs of EAE, prior to immunization or at the latest near onset of the disease. For example, in Gaur et al. above, a group of 13 mice were exposed to the disease-causing MBP regimen. When only one of the 13 mice showed minimal signs of disease, i.e. one mouse had clinical result 1 (see Example 2 for discussion of clinical evaluation in mice), seven mice were intraperitoneally injected with a mixture of MBP peptides. The other six mice, one of them an animal showing signs of disease, were not further treated. In this case, Gaur et al. for mice without clinical symptoms of the disease, unable to draw any meaningful conclusion about their data. The presence of low clinical score 1 in one mouse cannot be interpreted as indicative of active disease in any of the other mice. Gaur et al. therefore, they describe the treatment prior to the onset of the disease and most specifically, the clinical signs of the disease. In contrast, in order to be useful in the clinical setting of treating people with MS, therapy should also be effective if started long after the onset of the disease process. As a model of appropriate human therapy, the applicants were administered to the mice peptide therapy with 250 nmol Acl-11 [4Y] (SEQ ID NO: 67), which was started after the onset of the acute phase of EAE. FIG.

shows the effect of this therapy. We started therapy in each mouse individually after showing initial signs of EAE. Mice were characterized as having developed EAEs on the Acl-11 [4Y] (SEQ ID NO: 67) peptide therapeutic group and on each of the three control groups (no treatment, PBS or A ^u A ^u- binding peptide OVA 323-337 ) (SEO ID NO: 70). Figure 9 shows that treatment of EAE in progress with Acl-11 [4Y] (SEQ ID NO: 67) leads to a remarkable improvement in disease outcomes, which is evident within 48 hours of the initial injection of the peptide. In these treated mice, EAE is almost completely eliminated by the fourth day after onset and the effect of the 18-day course of peptide therapy is prolonged.

In yet another surprising development, applicants have discovered that intravenous peptide therapy initiated during the EAE remission phase prevents disease recurrence. Using EAE, as a model for human disease, as mentioned above, in (PLJ x SJL) F1 mice, a relapse of EAE disease develops after immunization with MBP (Fritz, RB et al., J. Immunol, 1983, 130: 1024) . EAE usually develops between 9 and 16 days after immunization and in this model the acute phase of the disease lasts from about 3 to 20 days. Depending on the severity of the disease in the experiment, 20% or more of the mice may die or die during the current phase. Most survivors in the acute phase then enter a short remission phase, expressing mild or no clinical signs of the disease for several days. In some experiments, all mice enter the remission phase at about the same time (illustrated in Figure 5), but in most cases the length of the acute phase of the disease is more variable. The remission phase is followed by the first disease return phase, which is generally equal in severity to the acute disease phase. Subsequently surviving mice develop residual chronic paralysis, which persists until the end of the experiment.

In the mouse model, the Acl-11 [4Y] peptide (SEQ ID NO: 67) is an extremely effective therapy when administered during the EAE remission phase. To investigate the capacity of Acl-11 [4Y} (SEO ID NO: 67) to prevent EAE recurrence, mice were pursued over the course of their initial paralysis events, which ranged from 3 to 19 days. The surviving mice individually entered the remission period (defined as a reduction in clinical score to 1 or 0 for at least two days), were alternatively grouped into two groups and started with intravenous therapy with 25 nmol Acl-11 [4Y] (SEQ ID NO : 67) but with control PBS. Mice were treated individually, starting on their second day of remission. Figure 10 shows that intravenous treatment with Acl-11 [4Y] (SEQ ID NO: 67), initiated during the EAE remission phase, greatly reduced the incidence and severity of disease recurrence, and indicates that intravenous peptide therapy may be effective even when we start with it late in the course of the disease.

The mechanism of intravenous peptide therapy has not yet been fully elucidated, although it has been previously suggested in many other antigen systems that intravenous administration of the antigen peptide prior to immunization induces antigen-specific immune system responses. As a further indication of the viability of the therapeutics administered to patients with MS, applicants I.V. administered an encephalitogenic peptide or peptide analog and found to have reduced in vitro lymph node proliferation and IL2 production. Mice were injected intravenously with 250 nmol of MBP Acl-11 (SEQ ID NO: 65) 10 and 5 days before immunization with the MBP protein. Figure Ila shows the proliferative response of the lymph node in these treated mice measured 10 days after immunization. Pre-intravenous treatment with the immunodominant peptide MBP Acl-11 (SEQ ID NO: 65) without adjuvant prior to immunization, reduces the subsequent in vitro proliferative T cell response to MBP Acl-11 (SEQ ID NO: 65). Similar results were obtained when mice were pretreated with intravenous injections of the peptide analogue MBP, Acl-11 [4Y] (SEQ ID NO: 67) (data not shown). In addition, the production of lymph node IL2 in response to MBP Acl-11 (SEQ ID NO: 65) was prevented by intravenous treatment with Acl-11 [4Y] (SEQ ID NO: 67) (Figure Ilb). The results confirm that pretreatment with MBP Acl-11 (SEQ ID NO: 65) or Acl-11 [4Yj (SEQ ID NO: 67) induces T cell responsiveness to MBP Acl-11 (SEO'ID NO: 65).

Applicants have therefore investigated the efficacy of intravenous peptide therapy and MBP peptide analogues in the treatment of EAE disease recurrence, and have made a number of unique observations. First and probably most important, the treatment of an autoimmune disease with a single autoantigen peptide may also be effective when treatment is initiated after the onset of disease signs. The specific disease signs used to evaluate the mice are addressed in the examples here. Median clinical outcomes of grade 2 ah higher were defined as advanced stage EAE. The results show that intravenous peptides of MBP interfere with the encephalitogenic activity of autoaggressively displaced cells even after the blood brain barrier has been removed and lymphocytic infiltration of the central nervous system has occurred. We have never observed peptides at least temporarily activate, exacerbate, or cause exacerbation of an ongoing disease. The EAE that is treated after the onset of the acute phase is eliminated within days of initiation of therapy, although the initially treated mice were severely affected. Histological examination confirmed that the clinical response to peptide therapy was correlated with the observed reduction in brain and spinal cord lymphocyte infiltrates. As discussed here, so was I.V. administration of MBP Acl-11 [4Υ] (SEO ID NO: 67) effectively even when started as late as 30 days after MBP immunization, during the EAE remission phase. Thus, initiation of treatment during the apparently calm phase of the disease did not lead to exacerbations again, and completely prevented all but the mildest relapses. This result is particularly interesting because there is no evidence that recognition of a plurality of MBP epitopes occurs late in the immune response to MBP, and it has been suggested that these epitopes may contribute to the disease (Lehmann, PV, et al., Nature, 1992, 358: 155). The present invention proposes that administration of the peptide very early in the course of the disease could prevent the development of a late immune response to such epitopes. Of course, it is notable that a single peptide analogue of MBP effectively prevented disease recurrence when given after resolution of a serious acute phase EAE, when the immune response to MBP and probably to other myelin antigens may have already progressed.

In the embodiment of the present invention, the choice of an effective therapist is preferred and is described hereinafter. In addition, the inventors of the invention have suggested that one criterion for the best candidate peptides could be that they are those with T cell reactivity and with a higher MHC binding affinity than the native peptide forming stable MHC class II complexes in vivo . In particular, applicants have studied the peptide analogue of MBP, which forms stable complexes with MHC class II in vivo, which is more effective in treating EAE than Acl-11 (SEQ ID NO: 65). It has been shown previously that the peptide analogue of MBP Acl-11 [4Y] (SEQ ID NO: 67) binds to A ^U A ^U with higher affinity and that the complexes formed in vitro during Acl-11 [4Y] (SEQ ID NO: 67) and A ^U A ^U more stable than those formed between Acl-11 (SEQ ID NO: 65) and A ^U A ^U. Acl-11 [4Y] (SEQ ID NO: 67) has also been shown to prevent EAE when high doses are inhaled before immunization (Metzler, B. et al., Supra). The present invention describes that a modified peptide analogue of MBP Acl-11 [4Υ] (SEQ ID NO: 67) as a therapeutic agent is at least 100 times more effective than MBP Acl-11 (SEQ ID NO: 65) and requires a less frequent schedule dosing. This increased efficacy results in the therapist being both more practical and more effective. Applicants have observed that intravenous administration of Acl-11 [4Yj (SEQ ID NO: 67) (but not Acl-11 (SEQ ID NO: 65)) results in the formation of stable peptide-MHC complexes in vivo. An improved efficiency of Acl-11 [4Yj (SEQ ID NO: 67) is thought to be associated with the formation of stable peptide-MHC complexes. These complexes can be detected on spleen cells using MBP Acl-11 (SEQ ID NO: 65) -specific hybridoma for up to 10 hours after injection. We also think that the formation of stable peptide-MHC complexes may be one of the features of such an effective therapeutic peptide. Surprisingly, the in vivo therapeutic effect of Acl-11 [4Y] (SEO ID NO: 67) was present long after the peptide-MHC complexes on the spleen cells could no longer be detected (see Figure 6a and Figure 9). It is possible that peptide-MHC complexes form antigen-specific functional therapeutic units that express a long-lasting effect on T encephalitogenic cells, which persists even after the in vivo complexes can no longer be detected. It is unknown whether such complexes express their efficacy peripherally or in the central nervous system. When they function in the central nervous system, complexes can form there, or cells that carry complexes can migrate across the blood brain barrier.

The present invention further provides methods for treating multiple sclerosis comprising administering a therapeutically effective amount of at least one MBP peptide having T cell activity in a treatment regimen comprising a therapeutically effective amount of IFN-1S.

As a result of the work described herein, the combination of at least one peptide having T cell activity derived from myelin antigens (e.g. MBP) and IFN- / 3 when administered in a therapeutic regimen has a synergistic effect (Fig. 12c). which surprisingly reduces the clinical symptoms of EAE in mice to a much greater extent than the effect of each on the mitigation of EAE symptoms when administered individually (Fig. 12a-b) and which is greater than would be expected for the primarily additive effect of peptide + IFN- / 3.

Because EAE serves as a murine model of human MS and is induced by various myelin antigens (e.g., PLP, MBP, MOG), we expect to see a similar effect in humans. Therefore, the present invention provides a method of treating individuals who have multiple sclerosis or are susceptible to developing multiple sclerosis, comprising administering an effective amount of a composition of the invention comprising at least one peptide having T cell activity derived from a myelin antigen, preferably in a non-immunogenic form, in a therapeutic regimen including the administration of IFN- / 3. Preferred compositions include those compositions of the invention previously described comprising MBP peptides as well as MBP peptides coupled to MOG peptides in conjunction with IFN- / 3 simultaneously or sequentially administered.

The administration of a composition of the invention comprising at least one peptide having T cell myelin antigen activity in a therapeutic regimen involving the administration of IFN-β can be carried out using known dose procedures and for periods that effectively reduce, eliminate or prevent symptoms associated with multiple sclerosis.

The effective amounts of both peptide and IFN-β, when administered together in the therapeutic regimen, vary according to the factors discussed above. The active compounds (eg peptide MBP or its composition and IFN-β) can be administered appropriately, such as by injection (subcutaneously, intravenously, etc.), by oral administration, by inhalation, by transdermal administration or by rectal administration as described above. .

By injection, e.g. preferably about 1 / xg-3 mg and more preferably about 20-500 μ-g peptide derived from myelin antigen per dosage unit. Preferably, an injection of 100-10000 IFN-β units can be administered by injection. The dosage regimen of these two compounds can be adjusted to provide an optimal therapeutic response. For example, IFN-β and the composition of the invention may be administered simultaneously, or may preferably be administered at least 6 hours apart, preferably at least 12 hours apart, or more preferably at least 24 hours apart. The therapeutic regimen of administration of both the antigenic peptide and IFN-β may be continued for several days or weeks and may be shortened or prolonged as dictated by the urgency of the therapeutic situation as discussed above.

The present invention also provides a novel composition comprising a physical mixture of a peptide having a T cell activity derived from a myelin antigen such as MBP or MOG and IFN-β in a pharmaceutically acceptable carrier or diluent. This composition can be used as part of a therapeutic regimen for treating or preventing multiple sclerosis in an individual.

Other autoimmune diseases, such as type I diabetes and rheumatoid arthritis, are generally accepted as being the result of T cell-mediated antigen-specific autoantigen responses. Various approaches for the treatment of T-cell-mediated autoimmune diseases include administration of whole autoantigens or peptide-derived peptides containing T cell epitopes to the patient in order to down-regulate T-cell mediated response responsible for the deleterious symptoms of autoimmune disease.

In addition to myelin autoantigens that play a role in MS, it has been found to cause disease symptoms in other autoimmune diseases such as diabetes, Graves' disease, myastenia gravis, Good Pasture's syndrome, psoriasis, thyroiditis and rheumatoid arthritis, many antigens (eg autoantigens such as isoline; rh factor; acetylcholine receptors; thyroid cell receptors, core membrane proteins; thyroid proteins; ICA-69 (PM-1); glutamic acid decarboxylases (64K or 65K); proteolipid, collagen (type II), thermal protein shock and carboxypeptidase H). We think that for the treatment of autoimmune diseases such as rheumatoid arthritis, diabetes, myasthenia gravis, Graves disease, Good Pasture syndrome, psoriasis and thyroiditis, where the antigen responsible for the disease is a protein autoantigen, compositions and methods similar to those can be used described here.

Peptides comprising defined sequences of amino acid residues having T cell activity and preferably containing at least one T cell epitope and / or which induce T cell unresponsiveness or reduced T cell responsiveness have been identified and isolated for many of the above-mentioned autogens. E.g. peptides thought to be capable of down-regulating an antigen-specific response to soluble type II collagen, a protein antigen thought to be an autoantigen in rheumatoid arthritis, have been identified in WO 94/07520. WO 92/06704 describes methods for identifying insulin peptides thought to contain T cell epitopes and which may be useful in compositions and methods analogous to those of the present invention for the treatment of diabetes I. In addition, the corresponding peptides, containing the T cell epitope are identified for an antigen or autoantigen that is assumed to be responsible for any of the above-mentioned diseases, by any of the methods described above and in the cases for the identification of T cell epitope-containing peptides of a protein autoantigen. Once peptides containing such a T cell epitope have been identified for the target autoantigen, such peptides can be used in compositions and methods analogous to those described herein for the treatment of multiple sclerosis.

The following invention is illustrated by the following non-limiting examples.

EXAMPLE 1

Study of the Multiple Sclerosis Immune Response in the Human Population to Myelin Basic Protein and Peptides and Selection of MBP Peptides Suitable for Therapeutic Use

Synthesis of peptides

The peptides were synthesized using standard Fmoc / tBoc chemistry and purified by reverse phase HPLC. Figure 3 shows the MBP peptides used in these studies. Peptide names or amino acid residues are consistent throughout.

Protocol: Analysis of human PBL for reactivity with MBP and MBP peptides:

PBLs were purified from fresh peripheral blood samples (approximately 75 cm ³ ) of 222 patients with definitive MS using a Ficoll density gradient. Microtiter cultures were filed with 2 χ 10 ⁵ PBL per well and 10 Mg / ml of purified human cerebellum MBP in RPMI 1640 culture medium supplemented with 5% human AB serum, penicillin-streptomycin and L-glutamine. Beginning on day 6-7, the cultures were supplemented with IL2 (20 units / ml) and IL4 (5 units / ml). After 11-13 days, the microtiter cultures were washed, resuspended in fresh medium and divided into 12 fresh microtiter wells. Autologous frozen PBLs, with 5 χ 10 ⁴ PBL per well, were added as cells representing anitgen.

Antigen screening was added in duplicate to 12 replication wells from each microtiter culture. Medium was always used as a negative control, and purified human recombinant MBP with 10 Mg / ml was used as a positive control. Each patient was also tested for reactivity with up to 4 MBP peptides, each at a concentration of 10 mM. After 48 hours, the assays were contacted with 27.75 kBq ³ Htimidine and collected after 6-16 hours of contact.

Cultures were evaluated as positive for each of the peptides according to the following criteria: stimulation index greater than 3.0, change in CPM greater than or equal to 500, and standard error of the mean less than change in CPM. In addition, for the purposes of culture analysis, we evaluated peptide-positive only if they responded to both MBP and peptide and if they did not respond to more than one non-overlapping peptide. Reactivity with each of the peptides was tested using at least 19 and at most 43 patients.

Results:

An average of 6% microtiter cultures gave a positive result on MBP reactivity in each patient (range 0-37 MBP positive cultures per patient). At least one microtiter culture gave a positive result for MBP reactivity in 77% of patients, and these individuals were considered MBP responders. MBP response status did not correlate with gender, MS (VP vs. KP) category, HLA-DR type, age, or whether or not the patient was taking betaserone.

The MBP peptides tested included a panel of 16 20-mer (Fig. 3) overlapping with 10 amino acids and covering the entire sequence of human MBP (18.5 kD isoform). We also tested two additional longer peptides (MBP sequences (MBP 83-105 (SEQ ID NO: 5) and MBP 141-165 (SEQ ID NO: 14), Figure 3). Then, based on the proportion of MBP positive microtiter cultures that also gave a positive result for one of the MBP peptides, calculating the reactivity of the MBP peptides, Each of the four peptides contributed at least 10% of the total MBP reactivity (MBP 11-30 (SEQ ID NO: 2), MBP 81-100 (SEQ ID NO: 23), MBP 83-105 (SEQ ID NO: 5) and MBP 141165 (SEQ ID NO: 14). In addition, reactivity to each of these peptides was detected in at least one-third of individual MBP-responsive MS patients in whom further, 77% of MBP-responsive patients showed reactivity to both MBP 83-105 (SEQ ID NO: 5) and MBP 141-165 (SEQ ID NO: 14), or both peptides.

Among the four peptides, MBP 141-165 (SEQ ID NO: 14) is the most reactive, contributing 21% of the total MBP response and can be determined in 64% of MBP response patients tested. MBP 141-165 (SEQ ID NO: 14) appears to include T-cell epitopes from both MBP 141-160 (SEQ ID NO: 28) and MBP 151-170 (SEQ ID NO: 29), and it is surprising showed remarkably more reactivity than both 20-mer peptides. MBP 81-100 (SEQ ID NO: 23) and MBP 83-105 (SEQ ID NO: 5) are very similar in sequence, and are likely to include similar, if not identical, T cell epitopes. They correspond to the region once thought to be related to the HLA-DR2 haplotype. Our results indicate that both DR2 and non-DR2 patients with MS have good reactivity to these peptides. MBP 11-30 (SEQ ID NO: 2) is a region of peptide reactivity not previously thought to be commonly recognized in MS patients.

Each of the other MBP peptides contributed less than 5% of the total MBP reactivity, and was detected in 20% or less of the MBP response patients tested (see Figures 4a and 4b). The highest reactivity among this subset of peptides was determined by MBP 111-130 (SEQ ID NO: 12), which contributed 4.5% of MBP reactivity and was found in 19% of MBP responders tested.

Selection of MBP peptide preference

Priority MBP peptides were selected based on two criteria:

1. Number of MS patients responding to candidate peptides (at least 75% of the tested patients respond to a protein antigen recognized by at least one of the peptides).

2. Magnitude of T cell response to candidate peptide in patients with MS (response to candidate peptides equals 40% of total response to antigen).

MBP 83-105 (MBP-2 (SEQ ID NO: 5)) and MBP 141-165 (MBP-4 (SEQ ID NO: 14)) meet the first criterion (77% of MBP responders recognize one or both). See Table 1 below. These two peptides together contribute 32% of MBP reactivity (see Table 2 below).

TABLE 1

Responsiveness for MBP-2 (SEQ ID NO: 5) and MBP-4 (SEO ID NO: 14): Analysis by individual% of individuals responding to MBP-2 MBP-4

MBP-2 and / or MBP-4 among MBP responders

11/31 [35.5%] 20/30 [64.5%] 24/31 [77.4%]

TABLE 2

Response to MBP-2 (SEQ ID NO: 5) and MBP-4 (SEQ ID NO: 14): Analysis by T Cell Lines

% MBP of the positive of lines MBP positive and MBP-2 MBP positive and 141-165 MBP positive and MBP-2 or MBP-4 during MBP 20/184 39/184 [21.1%] 59/184 [32,1%] of the positive [10,95%] of lines

Addition of MBP 11-30 (MBP-1 (SEQ ID NO: 2)) thus fulfills the second criterion, since the three peptides together contribute 42% of MBP reactivity. MBP 111-130 (MBP-3 (SEQ ID NO: 12)) would also be suitable as a preferred peptide for therapeutic use, especially in combination with MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5) and MBP-4 (SEQ ID NO: 14). MBP 81-100 (SEQ ID NO: 23) seems equivalent to MBP 83-105 (SEQ ID NO: 5) and can also be used in conjunction with other peptides selected as preferred.

EXAMPLE 2

Administration of peptides to mammals for the treatment of EAE as a model for multiple sclerosis

Synthesis of peptides

Peptides were prepared by automated peptide synthesis (ABI 430A, Applied Biosciences. Foster City, CA) using standard 9-fluorenylmethoxycarbonyl chemistry. The peptides were purified by high-performance liquid chromatography and the amino acid composition confirmed. The peptide sequences selected were MBP Acl-11 (SEQ ID NO: 65) ASQKRPSQRHG; MBP Ac 1-11 [4Α] (SEQ ID NO: 66) ASQARPSQRHG; MBP Acl-11 [4Υ] (SEQ ID NO: 67) ASQYRPSQRHG; MBP 31-47 (SEQ ID NO: 68) RHRDTGILDSIGRFFSG; This 323-339 (SEQ ID NO: 69) ISQAVHAAHAEINEAGR;

in This 323-337 (SEQ ID NO: 70) ISQAVHAAHAEINEA.

MBP Cleanup

From the guinea pig pig brain (Keystone Biologicals, Cleveland, OH), using a method modification according to Smith, M.E. J. Neurochem., 1969,16: 83, prepared by MBP. In summary, MBP was extracted from the isolated myelin membranes, extracted with potassium citrate, acid extracted and lyophilized using chloroform and methanol. SDS-PAGE analysis of this material revealed a major band at an expected molecular weight of 18.5 Kd.

EAE Induction, Evaluation and Peptide Therapy

EAEs were induced in (PLJ x SJL) Fl mice obtained from Jackson Laboratory (Bar Harbor, ME). When the mice reached 8-14 weeks of age, EAE was induced by immunization with 50-100 MBP emulsified in complete Freund's adjuvant (Gibco Laboratories, Grand Island, NY) supplemented with an additional 400 μg per mouse M. tuberculosis H37Ra (Difco Laboratories , Detroit, MI). Immunization and 48 hours later, 200 ng of pertussis toxin were given intravenously (JRH Biosciences, Lenexa, - 'J <Sj.'Na'based clinical signs, mice were evaluated according to the following scale: 1, tail paralysis; 2, partial hind limb paralysis ; 3, complete hind limb paralysis; 4, anterior limb paralysis; 5, dying or dead. Fifth-degree mice were euthanized. After death, mice were excluded from subsequent mid-clinical outcome calculations. Peptides were administered intravenously as described in the individual cases below. .

Suspensions of inguinal and paraaortic lymph nodes from mice immunized as described above for EAE induction were prepared for lymph node proliferation and IL2 production assays (pertussis toxin was not included for proliferation experiments). Lymphocytes were cultured in 5 x 10 ⁵ round bottom microtitre plates per well with MBP and RPMI 1640 peptides supplemented with 0.5% fresh normal mouse serum, penicillin-streptomycin, L-glutamine, and 5 χ IO ' ⁵ M

2-mercaptoethanol. Proliferation assay cultures were contacted for ³ h with ³ H-thymidine, followed by 72 h incubation. For the IL2 assay, culture supernatants were collected after 24 hours and examined for their ability to support the growth of IL2-dependent HT2 cell line. To differentiate IL2 production from IL4 production, HT2 bioassays were performed in the presence of 10% supernatant of the monoclonal antibody ll.B.ll culture (O'Hara, J., et al., Nature, 1985, 315: 333), which inhibits IL4-induced HT2 cell proliferation.

The results are shown in Figure 9, which shows that Acl-11 [4Y] (SEO ID NO: 67) nullifies EAE when administered after paralysis. EAE was induced by MBP in groups of 14 mice. We started therapy in each individual mouse after developing clinical signs of EAE. Data are plotted against the start of therapy for each mouse individually (day 1 is the onset of clinical signs in each individual mouse). Mice were treated intravenously for days 1, 4, 11, and 18 with PBS (empty circles), 250 nmol Acl-11 [4Υ] (SEQ ID NO: 67) (full squares), or 250 nmoli Ov 323-337 (SEO ID NO: 70) (full circles). We did not process additional control groups (blank boxes). Small crosses indicate single mice that were killed or killed at EAE grade 5. MMS for the untreated control group is 4.6 and mortality is 64%. The MMS for the PBS treated group is 4.4 and the mortality rate is 35%. The MMS for the group treated with Ova 323-337 (SEQ ID NO: 70) is 4.2 and the mortality rate is 21%. The MMS for the group treated with Acl-11 [4Y] (SEQ ID NO: 67) is 3.2 and the mortality rate is 7%.

Figure 10 shows that Acl-11 [4Y] (SEQ ID NO: 67) prevents EAE recurrence when administered during the remission phase of the disease. EAE is induced by MBP. Therapy was started in each mouse individually on the second day after remission, following the initial episodic event of EAE. Data are plotted against the start of therapy in each mouse individually (day 1 = second day of remission). Groups of 6-7 surviving mice 1, 4, 7, and 18 days after initiation of remission were treated intravenously with PBS (empty circles) or 25 nmol Acl11 [4 Υ] (SEQ ID NO: 67) (full squares). Small crosses indicate single mice that died or were killed at EAE stage 5. Initial episodic events before remission lasted from 3 to 18 days (averaging 9 days for both groups). MMS during the initial event was similar for both groups of mice (3.0 for the PBS treated group and 3.9 for the Acl-11 [4Y] treated group (SEQ ID NO: 67). For the PBS treated group, post-treatment MMS was 4.1 and the mortality rate was 33%. For the peptide treated group, post-treatment MMS is 0.3 and mortality is 0%.

Figure 11 shows that intravenous administration of Acl-11 (SEQ ID NO: 65) or Acl-11 [4Y] (SEQ ID NO: 67) induces T cell unresponsiveness.

a. Groups of 3-4 mice were pretreated intravenously with PBS (full circles) or 250 nmol Acl11 (SEQ ID NO: 65) (full triangles) ten and five days before immunization with 150 / xg MBP. On the ninth day, we performed lymph node proliferation tests with MBP Acl-11 (SEO ID NO: 65). The controls were as follows: PBS pretreatment PPD 108277 / medium 1855 and Acl-11 (SEO ID

NO: 65) PPD pretreatment 88499 / medium 1335.

b. Groups of 4 mice were pretreated intravenously with either PBS or 250 nmol Acl-11 [4Y] ten and five days before immunization with 150 µgx MBP (SEQ ID NO: 67). On day 10, we performed IL2 lymph node production assays with 150 μΜ Acl-11 (SEQ ID NO: 65) or 17 μΜ Acl-11 [4Y] (SEQ ID NO: 67).

EXAMPLE 3

Daianie peptides in advanced stage

Following immunization with MBP, a return of EAE develops in (PLJxSJL) Fl mice (Fritz, RB, et al., J. Immunol., 1983, 130: 1024), making this model of EAE ideal for exploring therapeutic intervention. Although MBP Acl-11 (SEQ ID NO: 65) is an immunodominant encephalitogen in (PLJ x SJL) F1 mice, the subdominant MBP 31-47 epitope in this strain is also encephalitogenic (SEQ ID NO: 68). To determine whether one or both encephalitogen peptides are required for the treatment of MBP-induced EAE, the steps of Example 2 were followed, except that the groups of mice in the advanced stage of the disease were treated with intravenous injections of MBP Acl-11 (SEQ ID NO: 65) and MBP 31-47 (SEQ ID NO: 68) together, or with Acl-11 alone (SEQ ID NO: 65). Groups of control mice were treated with intravenous injection of PBS or the control peptide Ova 323-339 (SEQ ID NO: 69), an A ^U A ^U binding peptide that is not related to MBP and which is non-immunogenic in (PLJxSJL) Fl mice.

Figure 5 shows that repeated intravenous injections of MBP Acl-11 alone (SEQ ID NO: 65), (Figure 5a) or in combination with MBP 31-37 (SEQ ID NO: 68) (Figure 5b) reduce the incidence, severity and mortality EAE. Treatment with both peptides seems to be somehow more effective than treatment with MBP Acl-11 alone (SEQ ID NO: 65), which is consistent with the observations of others as mentioned above. Mice were treated with six intravenous peptide injections during the acute phase and the first disease return phase. Mice were then monitored for up to 125 days during the chronic, debilitating phase, with no evidence of late disease in the MBP-treated groups (data not shown). Control peptide Ovate 323-339 (SEQ ID NO: 69) had no effect on the disease when administered via the intravenous route (Figure 5c). In a separate experiment, intravenous administration of MBP 31-47 alone (SEQ ID NO: 68) was not effective in the treatment of EAE (data not shown). Figures 5 (a), (b), and (c) show the efficiency of intravenous treatment of mice with MBE-induced EAE. In groups of 11-16 mice, EAE was induced with MBP as given above. Mice were treated by intravenous injection of 250 nmol of each peptide starting at 9,12,15, 21, 29 and 37 days. Small crosses were used in all the drawings to indicate single mice that were killed or killed in EAE grade 5. The control group was treated with PBS (full circles). The results indicate that the immunodominant encephalitogenic peptide MBP is both necessary and sufficient for the treatment of MBP-induced EAE in (PLJxSJL) Fl mice, as evidenced by a decrease in the severity and mortality of EAE in treated mice.

EXAMPLE 4

Multiple I.V. injections of Acl-11 (SEQ ID NO: 65)

Following the preparation steps of Example 2, multiple injections of Acl-11 (SEQ ID NO: 65) were prepared. One injection of peptide on day 9 delayed the disease by two days and reduced mortality from 100% to 63%, but all of the treated mice eventually developed severe EAE (data not shown). Three peptide injections during the acute phase of EAE initially treated the disease effectively, but during the disease return phase, the treated group of mice developed a late onset of disease (data not shown). In summary, the results indicate that multiple MBP peptide intravenous injections between acute and first EAE return phase are required for MBP-induced disease in (PLJ x SJL) F1 mice. Thus, repeated injections of Acl-11 (SEQ ID NO: 65) are required for prolonged reduction of disease severity. Figure 6 shows that Acl11 [4Y] (SEQ ID NO: 67) treats EAE at a lower dose than Acl-11 (SEQ ID NO: 65) and has a longer lasting effect on the disease. In groups of 8-10 mice, EAE was induced with MBP. Figure 6a shows mice treated with I.V. 12 and 15 days with PBS (phosphate buffered saline) (empty circles), with 250 nmol Acl-11 (SEQ ID NO: 65) (solid squares) or with 250 nmol Acl-11 [4Yj (SEQ ID NO: 67) ( blank triangles). The MMS for PBS-treated mice is 4.4 and the mortality rate is 60%. The MMS for Acl-11 treated mice (SEQ ID NO: 65) is 3.9 and the mortality rate is 37.5%. The MMS for Acl-11 [4Y] treated mice (SEQ ID NO: 67) is 2.1 and the mortality rate is 0%. Figure 6b shows mice treated with I.V. On days 12, 15, 18, 21, 24 and 27 either with PBS (empty circles) or 2.5 nmol Acl-11 (SEQ ID NO: 65) (full circles). The MMS for PBS treated mice is 2.9 and the mortality rate is 20%. The MMS for peptide treated mice is 4.3 and the mortality rate is 50%.

EXAMPLE 5

Treatment with peptide analogue MBP forming stable complexes with MHC class II in vivo

Because peptide-MHC complexes are functional therapeutic units that treat EAE, applicants have a theory that Acl-11 [4Y] (SEQ ID NO: 67) would be more effective in treating MBP-induced active EAE than Acl-11 (SEQ ID NO: 65). In order to test this hypothesis, the steps of Example 2 were performed, except that we changed the schedule of injections and dosages. Figure 6a shows that only two Acl-U [4Y] injections (SEQ ID NO: 67) produce a longer effect early in the course of the disease than two Acl-11 injections (SEQ ID NO: 65). In addition, Figure 6b shows that repeated intravenous injections of Acl-11 [4Y] (SEQ ID NO: 67) are very effective in treating EAE at a dose of 2.5 nmol (Figure 6c). We then performed histological tests. Histological sections from EAE brain and spinal cord were prepared and stained with hematoxylin and eosin (CVD, Inc., West Sacramento, CA). Sections were evaluated with an aperture observer with respect to inflammatory infiltrates on a scale of 1 to 4. The efficacy of Acl11 [4Y] (SEQ ID NO: 67) was confirmed by histological examination of the brain and spinal cord sections of peptide-treated mice and mice. treated with PBS, showing a markedly reduced number and severity of inflammatory central nervous system infiltrates in peptide treated mice (Fig. 7). Figure 7 illustrates that inflammatory infiltrates are reduced in mice treated with Acl-11 [4Y] (SEQ ID NO: 67). The improved performance of Acl-11 [4Υ] (SEQ ID NO: 67) is evident.

In the example above, the improved efficacy of Acl-11 [4Y] (SEQ ID NO: 67) was apparent. Applicants have a theory that stable peptide-MHC complexes are formed in vivo between A ^U A ^U and Acl-11 [4Y] (SEQ ID NO: 67), which were injected intravenously, thereby producing the observed improved efficacy. These in vivo-formed complexes can be detected using hybridoma 1934, which was derived from an encephalitogenically MBP-specific T cell clone. The steps in Example 2 above were followed, except that the number of injections and volumes were adjusted as described below. The T cell hybridoma 1934 is specific for MBP Acl-11 (SEQ ID NO: 65), following the procedure described in Wraith, DC, et al., Whole 1989, 59: 247. In summary, hybridoma cells at 5 χ 10 ⁴ per well were incubated in flat-bottom microtiter plates with peptides or peptide analogues of MBP. 5 χ 10 ⁵ (PU x SJL) F1 spleen cells irradiated with 3000 rad were added as antigen presenting cells. The medium was the same as for lymph node proliferation assays except that the serum supplement, instead of normal mouse serum, was 10% fetal calf serum. After 24 hours, the supernatants were collected and their ability to support HT2 cell growth was investigated. Then, at different times after injection of 250 nmol Acl-11 [4Y] (SEQ ID NO: 67), spleens were removed from the injected mice and splenocytes were used as cells presenting antigen for Acl-11-specific hybrids 1934. No additional cultures were added to the cultures peptides. Figure 8 shows that spleen cells that were contacted (pulsed) in vivo with Acl-11 [4Y] (SEQ ID NO: 67) were highly effective in activating hybridoma 1934. The capacity to activate hybridoma was a maximum of 2 hours after the injection and then 4 to 6 hours after the injection began to decrease. Activation was at least 10 hours after Acl-11 [4Y] injection (SEQ ID NO: 67) but was no longer present after 20 hours (data not shown). As can be seen from Figure 8, the experiments with Acl-11 (SEQ ID NO: 65) did not show activation of the hybridoma with crow cells from the injected mice even 1 hour after injection (data not shown). As shown in more detail in Figure 8, in vivo peptide-MHC complexes can be detected in mice injected with Acl-11 [4Y] (SEQ ID NO: 67). In groups of two mice, 250 nmol Acl-11 [4Y] (SEQ ID NO: 67) was injected intravenously at various time points before spleen removal (1-10 h). Splenocytes from injected mice were examined for their capacity to activate 1934 hybridoma cells to produce IL2. The results are expressed as HT2 proliferation. The results support the hypothesis that the formation of stable peptide-MHC complexes in vivo contributes to the efficacy of the therapeutic peptide analog of MBP.

EXAMPLE 6

Synthesis of the mouse peptide MBP Ac-1-11 fSEQ ID NO: 651

The mouse peptide MBP, Acl-11 (SEQ ID NO: 65) was synthesized using standard Fmoc / tBoc synthesis and purified by HPLC. The amino acid sequence for the peptide Acl-11 (SEQ ID NO: 65) is as follows:

Induction of EAE

In 6- to 8-week-old female (SJLxPL) Fl mice (Jackson Labs, Bar Harbor, ME), EAE was induced by immunization of mice with 100 μ-g purified MBP guinea pig in CFA (GIBCO Lab., Grand Island, NY), which contains 400 pg of H37RA strain M. tuberculosis (DIFCO Lab., Detroit, MI) subcutaneously into the tail base. On the day of immunization and also 48 hours later, they were given twice intravenously (i.v.) 200 ng pertussis toxin (JHL BIOSCIENCE, Lenexa, Kansas). Mice were monitored daily for disease symptoms and the severity of the disease was evaluated according to the following scale 0 = no clinical signs of EAE, 1 = weak, unresponsive tail, 2 = partial hind limb palsy, 3 = complete hind limb palsy, and 4 = partial to full anterior palsy limbs, and 5 = dying. Data are expressed as the mean of the disease severity scores on a daily basis, including all animals in the group. Mice were followed for 26 days. When the mouse died for EAE, the result 5 was included in the calculations for all subsequent days.

Effect of IFN-ff on EAE in a titration experiment to determine the effects of different doses of IFN- / 3 on EAE (Figure 13). On days 9, 3, and 16 after EAE induction, one group of mice was treated intraperitoneally with PBS (control), one group of mice was treated on day 9, 13, and 16 with 10,000 IFN- / 3 units (empty circles) and one group of mice 9, 13 and day 16 with 2000 IFN- / 3 (full circles). As shown in Figure 13, the symptoms of EAE at each time point for both IFN- / 3 doses are similar, indicating that a lower dose would be appropriate for IFN-jS trials and that this is the preferred dose since chances of toxicity due to higher IFN-β dose, less likely. In the remaining experiments shown in Figures 12a - 12c, 2000 IFN- / 3 units were used as this dose showed an improvement in clinical outcome.

As shown in Figure 12a, on day 9,12, 16 and 20, the control group of mice was treated with PBS and the second group of mice with 2000 IFN- / 3 units i.p. As shown in Figure 12a, the IFN-β treated group had only slightly less severe symptoms than the control mice over time.

Effect of mouse peptide MBP Acl-11 (SEQ ID NO: 65) on EAE

The effect of the mouse peptide MBP Acl-11 (SEO ID NO: 65) was determined and the results are shown in Figure 12b. 10, 13, 17 and 21 days after EAE induction, one group of mice was treated i.p. with PBS (control), and the second group of mice were treated i.v. for 10, 13, 17, and 21 days. with 250 nmol of Acl-11 peptide (SEQ ID NO: 65). Mice were observed as described above. As shown in Figure 12b, mice treated with Acl-11 (SEQ ID NO: 65) had less severe symptoms than those in the control group.

The effect of treatment with the combination of the peptide Acl-11 (SEQ ID NO: 65) and IFN-ff on

EAE

The effects of treatment with the combination of the peptide Acl-11 (SEQ ID NO: 65) and IFN-β are shown in Figure 12c. After EAE induction, one group of mice was treated with i.p. with PBS (control), and one group of mice was treated i.v. for 10, 13, 17, and 21 days. with 250 nmol of peptide Acl-11 (SEQ ID NO: 65) (empty arrows) and 9, 12, 16 and 20 on i.p. with 2000 IFN-β units. As shown in Figure 12c, the group of mice treated with the combination of peptide and IFN-β showed a remarkable reduction in the severity of symptoms compared to the control group as well as compared to IFN-β or peptide alone treatment, as shown in Figures 12a and 12b, indicating the synergistic effect of the combination. Therefore, a treatment regimen that incorporates a combination of peptide and IFN-β provides an increased effect on reducing the severity of EAE symptoms.

EXAMPLE 7

Administration of Peptides to Mammals for the Treatment of EAE Induced by Spinal Cord Homogenate (SCH = Spinal Cord Homogenate) as a Model for Multiple Sclerosis

Synthesis of peptides

Peptides were prepared by automated peptide synthesis (ABI 430A, Applied Biosciences, Foster City, CA) using standard 9-fluorenylmethoxycarbonyl chemistry. The peptides were purified by high-performance liquid chromatography and the amino acid composition confirmed. The peptide sequences selected were: MBP Acl-11 (SEQ ID NO: 65) ASQKRPSQRHG; MBP Acl-11 [4Α] (SEQ ID NO: 66) ASQARPSQRHG; MBP Acl-11 [4Υ] (SEQ ID NO: 67) ASQYRPSQRHG; MBP 31-47 (SEQ ID NO: 68) RHRDTGILDSIGRFFSG; This 323-339 (SEQ ID NO: 69) ISQAVHAAHAEINEAGR;

in This 323-337 (SEQ ID NO: 70) ISQAVHAAHAEINEA.

MBP Cleanup

Using a modification of the method according to Smith, M.E., J. Neurochem., 1969, 16:83, we prepared an SCH from the brain of a guinea pig (Keystone Biologicals, Cleveland, OH). The SCH protein mixture was lyophilized and used according to dry weight.

EAE Induction, Evaluation and Peptide Therapy

EAE was induced in (PLJ x SJL) F1 mice obtained from Jackson Laboratory (Bar Harbor, ME). When the mice reached 8-14 weeks of age, EAE was induced by immunization with 500-1000 / g SCH, emulsified in complete Freund's adjuvant (Gibco Laboratories, Grand Island, NY) supplemented with an additional 400 / g per mouse M. tuberculosis H37Ra ( Difco Laboratories, Detroit, MI). Immunization and 48 hours later, 200 ng of pertussis toxin (JRH Biosciences, Lenexa, KS) were administered intravenously. Based on clinical signs, mice were evaluated according to the following scale:

1, tail paralysis; 2, partial paralysis of hind limb; 3, complete paralysis of hind limb; 4, anterior limb paralysis; 5, dying or dead. Mice with a value of 5 were euthanized. After death, mice were excluded from subsequent calculations of the mean clinical outcome. Peptides were administered intravenously as described in the individual cases below.

The results are shown in Figure 16, which shows that Acl-11 [4Y] (SEQ ID NO: 67) prevents EAE when given after paralysis. EAE was induced by SCH in groups of 14 mice each. We started therapy in each mouse individually after developing clinical signs of EAE. Data are plotted against initiation of therapy in each individual (day 1 = onset of clinical signs for each individual mouse. Mice were treated for days 1, 3, 6, and 9 with PBS (solid squares), 250 nmol Acl-11 [4Y] (SEQ ID NO: 67) iv (blank triangles), or with 250 nmol Acl-11 [4Y] (SEQ ID NO: 67) (blank circles) A small asterisk indicates individual mice that are dead or killed at level 5 EAE .

Figure 17 shows that Acl-11 [4Y] (SEQ ID NO: 67) prevents the onset of EAE when given before the onset of symptoms. EAE was induced by SCH. Therapy was started in each mouse individually 8 days after induction of disease with gpSCH. Data are plotted against the start of therapy in each mouse individually. Groups of surviving mice were treated i.v. on day 8,10,13 and day 17. with 25 nmol Acl-11 [4Y] (SEQ ID NO: 67) (empty triangles) or s.c. with 25 nmol Acl-11 [4Y] (SEQ ID NO: 67) (full circles). We did not process one group (empty squares).

The results shown in Figures 16 and 17 show that any route of administration is effective in preventing the onset of symptoms.

EXAMPLE 8

We followed the same procedure as in Example 2, except that we did not perform lymph node proliferation analysis and IL2 production. EAEs were induced on day 0 using gpMBP, after which 8, 10, 13 and 17 days were given Acl11 [4Υ] to three groups of mice (SEQ ID NO: 67). The results for intravenous administration (empty triangles), subcutaneous injection (full circles) and control (empty circles) are presented graphically in Figure 18. Pre-treatment (eg pre-treatment with symptom formation) with Acl-11 [4Y] (SEQ ID NO : 67) can prevent EAE-induced gpMBP disease.

EXAMPLE 9 cDNA encoding a human MOG protein

In order to obtain human DNA encoding MOG protein, in the initial experiment, the human cDNA library was subjected to polymerase chain reaction (PCR) using 3 'and 5' primers formed from the MOG rat encoding sequence published by Gardinier et al. (above). The human MOG sequences could not be obtained in this way, presumably because of insufficient homology at the 5 'and / or 3' ends of the human and rat sequences.

Because of this, we formed four rat internal oligonucleotides. Two of them were homologous to the upstream gene (cf. 94-111 and 166-183 (SEQ ID NO: 74) of base 1, starting with ATG), and two were homologous to the downstream gene (cf. 538-555 ( SEQ ID NO: 75) and 685-702). A combination of primers 166-183 (SEO ID NO: 74) and 538-555 (SEQ ID NO: 75) was successful in performing fragment amplification with an expected size of approximately 400 bp from the human brain cDNA library. The sequence of these primers was:

a) 166-183: CAGAATCCGGGAAGAATGCCACGGGC_ (SEQ ID NO:

74); and

b) 538-555: CAGCGGCCGCACGGAGTTTTCCTCTCAG (SEQ ID

NO: 75).

The EcoRI site is present in primer 166-183 (SEQ ID NO: 74); The note site is present in primer 538-555 (SEQ ID NO: 75).

The 400 bp PCR product was cloned into the expression vector pVL1393 by degradation of pVL1393 (Pharmingen CA) with EcoRI and Noti, by digestion of the amplified product with the same enzymes and ligation of the resulting fragments. The insert was verified by degradation of several clones derived from ligated plasmids with EcoRI and Noti and by sequencing the resulting 400 bp fragment of human MOG. Based on the 738 bp rat open reading frame, the resulting insert is said to have no 184 bp 5 'sequence and 201 bp 3' sequence.

From 400 bp inclusions, we made two primers from positions 346-363 of the top and bottom chains as follows:

5'-CAGAATTCTCAGGTTCTCAGATGAAGGA-3 '(SEQ ID NO: 76); and

5′-AAGCGGCCGCTATCCTTCATCTGAGAACCT-3 '(SEQ ID NO:

77).

wherein the EcoRI site is present in the first chain and the Noti site is present in the second chain. The underlined regions correspond to the MOG sequence.

The human MOG 346-363 top and bottom primers (SEQ ID NOS: 76 and 77) were used in combination with the above 5 'or. 3 'rat primers that we amplified the 5' and 3 'missing ends of the gene from the same human brain cDNA library as we used previously. We obtained a PCR product corresponding to the 3 'end of the gene, but did not obtain the corresponding 5' end.

t

The resulting 3 'fragment had an expected size of 400 bp and this fragment was cloned into pVL1393 and sequenced.

To obtain the 5 'portion of the gene, a gtlO human brain medulla library purchased from Clontech, which was previously amplified and had a titer of 8x10 ¹⁰ pfu / ml, was selected according to the protocol described by the manufacturer. The library was planted on 12 large plates of 30,000 plates / plate and the plates were mounted on nitrocellulose filters (2 replication filters / plate). Then, 12 filters lifted from 12 different plates were hybridized to a ³² P labeled sample corresponding to the originally cloned inner 400 bp fragment of human MOG (positions 184-534). We received 22 strong positives. 7a, each positive was removed from the original plates and incubated overnight with dilution buffer to elute the phage from the agar. The tube was then centrifuged and the supernatant transferred.

From each single pool, DNA was amplified using or gt 10 anterior primer with the Sstll site:

5'-CTTTTGAGCAAGTTCAGCCTGGTTAAG-3 '(SEQ ID NO: 78) or by gtlO reverse primer with Xhol site:

5-. ACCTCGAGGAGGTGGCTTATGAGTATTTCTTCCAGGGTA-3 '(SEQ ID NO: 79) as well as an internal primer of the human or lower chain MOG: 5-GGTGCGGGAAAGGTGACTCTCAGAGATCCGGAAT-3' (SEQ ID NO: 80) or

5'-ATTCCGGATCCTGAGAGTCACCTTTCCCGCACC-3 '(SEQ ID NO: 81).

The last two primers (SEQ ID NOS: 80 and 81) include the BamH1 site (underlined in sequences), which is naturally present in the human MOG sequence.

The primers were used in four different combinations: 1) anterior upper / inner MOG below; 2) reverse bottom / inner MOG below; 3) internal MOG top / reverse bottom; and 4) internal MOG above / front above.

The first two combinations provided the 5 'end of the gene (up to the BamHl site) and the last two 3' end of the gene. Both portions 5 'and 3' include non-translated regions. Which of the two members of each combination actually resulted in the desired fragment depends on the orientation of the cDNAs cloned into gtlO.

The size of the fragments obtained varied from one pole to another. The five largest 5 'fragments or 3' fragments were subcloned into Sstll and BamHl or BamHl and Xhol sites of the SK polylinker. Three clones from each pool were then sequenced to eliminate the presence of PCR errors. This provided the complete sequence of the gene coding region as well as the 174 bp 5 'untranslated sequence.

The human MOG gene encodes a 248 amino acid preprotein that has 87% homology to 246 amino acids in rat protein. The mature protein contains 218 amino acids. The mature protein starts at the glycine residue and is derived from the 248 amino acid preprotein by cleavage from a pre-sequence that extends from the MET start codon to the alanine residue just before the glycine in position 1.

EXAMPLE 9A

Expression of truncated human MOG in SF-9 insect and E.coli cells

SF-9 expression

The transfer vector PVL1393 containing human MOG truncated cDNA encoding amino acids 1-121 of human MOG (first 121 amino acids of SEQ ID NO: 48) was cotransfected into SF-9 cells together with Baculovirus DNA linearized by Baculogold (Pharmingen, San Diego, CA). A culture supernatant containing recombinant viruses was collected after 4 days. The recombinant virus was purified to be plaque free and subjected to 3x amplification to obtain a high titre viral stock. AF-9 cells were then infected with viral stock at MOI 2.0. The supernatant from infected cells was collected 48 h after infection and applied to a NiNTA agarose column. The recombinant MOG protein was eluted under non-denaturing conditions using 250 mM imidazole, dialyzed against 5% propionic acid and H ₂ O, and then lyophilized. Protein concentration was assessed by BCA. Purified MOG protein was visualized on a 12.5% polyacrylamide gel stained with Coomassie blue.

EXAMPLE 10

The truncated human MOG (huMOG) was prepared as described in Examples 9 and 9a. MBPs (Guinea pigs) were prepared as described. The procedures of Example 2 were followed, except that EAE was induced in three separate groups of mice using 75 pg gpMBP and 100 pg huMOG (empty triangles) and a combination of 75 pg gpMBP and 100 pg huMOG (full squares). The graph in Figure 19 shows the course of the disease for each of these cases. The disease induced by both MOG and MBP is much more serious than the disease induced by each. The results show that both MOG and MBP contribute to the disease.

EXAMPLE 11

Using huMOG + gpMBP (day 0 = disease induction), disease was induced as in example 10. Before the onset of symptoms, mice were treated with 250 nmol Acl-11 [4Y] (SEQ ID NO: 67) and control (PBS) 6, 8 , 10, 13, 17, 22, and 27 days (arrows indicate processing). As shown in the graph in Figure 20, treatment with Acl-11 [4Y] (SEQ ID NO: 67) (blank spaces) significantly reduced the mean clinical result compared to controls (solid squares).

Equivalents

Experts will recognize or. will be able to find out, through the use of more than one routine experimentation, many equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of the present invention and to be covered by the claims which follow.

For

Immulogic Pharmaceutical Corporation:

LIST OF SEQUENCES (1) GENERAL INFORMATION:

(i) APPLICANT: Smilek, Dawn;

Samson, Michael F.;

Gefter, Malcolm, Hsu, Di-Hwei;

Shi, Jia-Dong;

Paliard, Xavier, Devaux, Brigitte;

Rothbard, Jonathan; and Franzen, Henry M.

(ii) TITLE OF INVENTION: Compositions and Treatment for Multiple Sclerosis (iii) NUMBER OF SEQUENCES: 81 (iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: LAHIVE & COCKFIELD (B) STREET: 60 State Street, Suite 510 (C) CITY: Boston (D) STATE: Massachusetts (E) COUNTRY: USA (F) ZIP: 02109-1875 (v) COMPUTER READABLE FORM:

(A) MEDIUM :Ε: Floppy Disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTNARE: Patent Release # 1.0, Version # 1.25 (vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: PCT / US95 / 13682 (B) FILING DATE: 25-October-1995 (vii) PRIOR APPLICATION DATE:

(A) APPLICATION NUMBER: US 08 / 404,228 (B) FILING DATE: 15-March-1995 (A) APPLICATION NUMBER: US 08 / 328,224 (B) FILING DATE: 25-October-1994 (A) APPLICATION NUMBER: US 08 / 300,811 (B) FILING DATE: 1-September-1994 (A) APPLICATION NUMBER: US 08 / 241,246 (B) FILING DATE: 10-May-1994 (A) APPLICATION NUMBER: US 08 / 116,824 (B) FILING DATE: 3-September-1993 (villas) ATTORNEY / AGENT INFORMATION:

(A) NAME: Amy E. Mandragouras (B) REGISTRATION NUMBER: Reg. No. 36,207 (C) REFERENCES / DOCKET NUMBER: 094.2 PCT (IMI-052C2PC) (ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: (617) 227-7400 (B) TELEFAX: (617) 227-5941 (2) INFORMATION FOR SEQ ID NO: 1:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 170 amino acids (B) ΤΥΡΕ: amino acids (D) TOPGLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 1:

Ala Sir Gin Lys Arg Pro Sir Gin Arg His Gly Sir Lys Tyr Leu Ala 1 5 10 15 Thr Ala Sir Thr Met Asp His Ala Arg His Gly Phe Leu Pro Arg His 20 25 30 Arg Asp Thr Gly lle Leu Asp Sir lle Gly Arg Phe Phe Gly Gly Asp 35 40 45 Arg Gly Ala Pro Lys Arg Gly Sir Gly Lys Asp Sir His His Pro Ala 50 55 60 Arg Thr Ala His Tyr Gly Sir Leu Pro Gin Lys Sir His Gly Arg Thr 65 70 75 80 Gin Asp Glu Asn Pro Val Val His Phe Phe Lys Asn lle Val Thr Pro 85 90 95 Arg Thr Pro Pro Pro Sir Gin Gly Lys Gly Arg Gly Leu Sir Leu Sir 100 105 110 Arg Phe Sir Trp Gly Ala Glu Gly Gin Arg Pro Gly Phe Gly Tyr Gly 115 120 125 Gly Arg Ala Sir Asp Tyr Lys Sir Ala His Lys Gly Phe Lys Gly Val 130 135 140 Asp Ala Gin Gly Thr Leu Sir Lys lle Phe Lys Leu Gly Gly Arg Asp 145 150 155 160 Sir Arg Sir Gly Sir Pro Met Ala Arg Arg

165 170 (2) INFORMATION FOR SEQ ID NO: 2:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 2:

Gly 1 Sir Lys Tyr Leu Ala Thr Ala 5 Sir Thr 10 Met Asp His Ala Arg 15 His Gly Phe Leu Pro 20 INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 amino acids (B) ΤΥΡΕ: amino acid (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: peptides (V) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 3: Gly Sir Lys Tyr Leu Ala Thr Ala Sir Thr Met Asp His Ala Arg His

10 15

Gly Phe Leu (2) INFORMATION FOR SEQ ID NO: 4:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 21 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 4:

Gly Ser Lys Tyr Leu Ala Thr Ala Ser Thr Met Asp His Ala Arg His 15 10 15

Gly Phe Leu Pro Arg 20 (2) INFORMATION FOR SEQ ID NO: 5:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 23 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT internalΕ: internal

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: : 5: Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr 1 5 10 15

Pro Pro Pro Gin Gly Lys 20 (2) INFORMATION FOR SEQ ID NO: 6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 24 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 15

Thr Pro Pro Pro Gin Gly Lys 20 (2) INFORMATION FOR SEQ ID NO: 7:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 23 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 15

Thr Pro Pro Gin Gly (2) INFORMATION FOR SEQ ID NO: 8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8:

Thr Gin Asp Glu Asn Pro Val His Phe Phe Lys Asn Ile Val Thr 15 10 15

Pro Arg Thr (2) INFORMATION FOR SEQ ID NO: 9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 21 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 15

Thr Pro Pro Pro Ser 20 (2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (Xi) SECUENCE DESCRIPTION: SEQ ID NO: 10:

Thr Gin Asp Glu Asn Pro Val His Phe Phe Lys Asn Ile Val Thr 15 10 15

Pro Arg Thr Pro Pro Pro Gin Gly 20 25 (2) INFORMATION FOR SEQ ID NO: 11:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 23 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

Thr Gin Asp Glu Asn Pro Val His Phe Phe Lys Asn Ile Val Thr 15 10 15

Pro Arg Thr Pro Pro Pro Pro 20 (2) INFORMATION FOR SEQ ID NO: 12:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 12:

Leu Ser Arg Phe Ser Trp Gly Ala Glu Gly Gin Arg Pro Gly Phe Gly 15 10 15

Tyr Gly Gly Arg 20 (2) INFORMATION FOR SEQ ID NO: 13:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13:

Leu Ser Arg Phe Ser Trp Gly Ala Glu Gly Gin Arg Pro Gly Phe Gly 15 10 15

Tyr Gly Gly (2) INFORMATION FOR SEQ ID NO-.14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) :Ε: amino acids (D) TOPOLOGV: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14:

Phe Lys Gly Val Asp Ala Gin Gly Thr Leu Ser Lys Ile Phe Lys Leu 15 10 15

Gly Gly Arg Ser Ser Arg Ser Gly Ser 20 25 (2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

Pro Ser Gin Gly Lys Gly Arg Gly Leu Ser Leu Ser Arg Phe Ser Trp 15 10 15

Gly Ala Glu Gly Gin Arg Pro Gly Phe 20 25 (2) INFORMATION FOR SEQ ID NO: 16:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (Β) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:

Ala Ser Gin Lys Arg Pro Ala Gin Arg His Gly Ser Lys Tyr Leu Ala 15 10 15

Thr Ala Ser Thr 20 (2) INFORMATION FOR SEQ ID NO: 17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (XX) SEQUENCE DESCRIPTION: SEQ ID NO: 17:

Met Asp His Ala Arg His Gly Phe Leu Pro Arg His Arg Asp Thr Gly 15 10 15

Ile Leu Asp Ser 20 (2) INFORMATION FOR SEQ ID NO: 18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18:

Arg His Arg Asp Thr Gly Ile Leu Asp Ser Ile Gly Arg Phe Phe Gly 15 10 15

Gly Asp Arg Gly 20 (2) INFORMATION FOR SEQ ID NO: 19:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:

Ile Gly Arg Phe Phe Gly Gly Asp Arg Gly Ala Pro Lys Arg Gly Ser 15 10 15

Gly Lys Asp Ser 20 (2) INFORMATION FOR SEQ ID NO: 20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:

Aly Pro Lys Arg Gly Ser Gly Lys Asp Ser His His Pro Ala Arg Thr 15 10 15

Ala His Tyr Gly 20 (2) INFORMATION FOR SEQ ID NO: 21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21:

His His Pro Ala Arg Thr Ala His Tyr Gly Ser Leu Pro Gin Lys Ser 15 10 15

His Gly Arg Thr 20 (2) INFORMATION FOR SEQ ID NO: 22:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22:

Ser Leu Pro Gin Lys Ser His Gly Arg Thr Gin Asp Glu Asn Pro Val 15 10 15

Val His Phe Phe 20 (2) INFORMATION FOR SEQ ID NO: 23:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acid (D) TOPOLOGY : linear (ii) MOLECULE ΤΥΡΕ : peptides (v) FRAGMENT ΤΥΡΕ : internal

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: : 23: Gin Asp Glu Asn Pro Val Val His Phe Phe Lys Asn lle Val Thr Pro 1 5 10 15

Arg Thr Pro Pro 20 (2) INFORMATION FOR SEQ ID NO: 24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24:

Lys Asn Ile Val Thr Pro Arg Thr Pro Pro Pro Ser Gin Gly Lys Gly 15 10 15

Arg Gly Leu Ser 20 (2) INFORMATION FOR SEQ ID NO: 25:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGV: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25:

Pro Ser Gin Gly Lys Gly Arg Gly Leu Ser Leu Ser Arg Phe Ser Trp 15 10 15

Gly Ala Glu Gly 20 (2) INFORMATION FOR SEQ ID NO: 26:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26:

Gly Arg Pro Gly Phe Gly Tyr Gly Gly Arg Ala Ser Asp Tyr Lys Ser 15 10 15

Ala His Lys Gly 20 (2) INFORMATION FOR SEQ ID NO: 27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (Β) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27:

Ala Ser Asp Tyr Lys Ser Ala His Lys Gly Phe Lys Gly Val Asp Ala 15 10 15

Gin Gly Thr Leu 20 (2) INFORMATION FOR SEQ ID NO: 28:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28:

Phe Lys Gly Val Asp Ala Gin Gly Thr Leu Ser Lys Ile Phe Lys Leu 15 10 15

Gly Gly Arg Asp 20 (2) INFORMATION FOR SEQ ID NO29:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 29:

Ser Lys Ile Phe Lys Leu Gly Gly Arg Asp Ser Arg Ser Gly Ser Pro 15 10 15

Met Ala Arg Arg 20 (2) INFORMATION FOR SEQ ID NO: 30:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn lle Val Thr Pro Arg 15 10 15

Thr Pro Pro (2) INFORMATION FOR SEQ ID NO: 31:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 31:

Lys Tyr Leu Ala Thr Ala Ser Thr Met Asp Ala Arg 15 10 (2)

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 32:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn 1 5 10 (2) INFORMATION FOR SEQ ID NO: 33:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 15 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 33:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro 15 10 15 (2) INFORMATION FOR SEQ ID NO: 34:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 16 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 15 (2) INFORMATION FOR SEQ ID NO: 35:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 17 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT internalΕ: internal

Thr (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 35:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg

10 15

2) INFORMATION FOR SEQ ID NO: 36:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 36:

Val His Phe Phe Ala Asn Ile Val Thr Pro Arg Thr Pro 15 10 (2) INFORMATION FOR SEQ ID NO: 37:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 18 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 37:

Glu Asn Pro Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr 15 10 15

Pro Pro (2) INFORMATION FOR SEQ ID NO: 38:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (Xi) SEOUENCE DESCRIPTION: SEQ ID NO: 38:

Glu Asn Pro Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr 15 10 15

Pro Pro Pro

2 (2) INFORMATION FOR SEQ ID NO: 39:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 14 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 39:

Asn Pro Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 (2) INFORMATION FOR SEQ ID NO: 40:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 17 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 40:

Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro 15 10 15

Pro (2) INFORMATION FOR SEQ ID NO: 41:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 16 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (Xi) SEOUENCE DESCRIPTION: SEQ ID NO: 41:

Pro Val Val Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro Pro (2) INFORMATION FOR SEQ ID NO: 42:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42:

Val Val His Phe Phe Lys Asn Ile Val-Thr Pro Arg Thr Pro Pro Pro 15 10 15

Ser Gin Gly Lys 20 (2) INFORMATION FOR SEQ ID NO: 43:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43:

Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro 15 10 (2) INFORMATION FOR SEQ ID NO: 44:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44:

His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro Pro 15 10 (2) INFORMATION FOR SEQ ID NO: 45:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 12 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45:

His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro 15 10 (2) INFORMATION FOR SEQ ID NO: 46:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 46:

Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr Pro Arg 15 10 15

Thr Pro Tyr (2) INFORMATION FOR SEQ ID NO: 47:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47:

Leu Ser Arg Phe Ser Trp Gly Ala Glu Gly Gin Arg Pro Gly Phe Gly

10 15

Tyr Gly Gly Arg Ala Ser Asp Tyr Lys 20 25 (2) INFORMATION FOR SEQ ID NO: 48:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 48:

Arg Pro Gly Phe Gly Tyr Gly Gly Arg Ala Ser Asp Tyr Lys Ser Ala 15 10 15

His Lys Gly (2) INFORMATION FOR SEQ ID NO: 49:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 32 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 49:

Lys Gly Phe Lys Gly Val Asp Ala Gin Gly Thr Leu Ser Lys Ile Phe 15 10 15

Lys Leu Gly Gly Arg Asp Ser Arg Ser Gly Ser Pro Met Ala Arg Arg 20 25 30 (2) INFORMATION FOR SEQ ID NO: 50:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50:

Lys Gly Val Asp Ala Gin Gly Thr Leu Ser Lys Ile Phe Lys Leu Gly 15 10 15

Gly Arg Ser Ser Arg Ser Gly Ser Pro 20 25 (2) INFORMATION FOR SEQ ID NO: 51:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 27 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51:

Lys Gly Val Asp Ala Gin Gly Thr Leu Ser Lys Ile Phe Lys Leu Gly 15 10 15

Gly Arg Asp Ser Arg Ser Gly Ser Pro Met Ala 20 25 (2) INFORMATION FOR SEQ ID NO: 52:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 15 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 52:

Ala Gin Gly Thr Leu Ser Lys Ile Phe Lys Leu Gly Gly Arg Asp 15 10 15 (2) INFORMATION FOR SEQ ID NO: 53:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 18 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53:

lle Phe Lys Leu Gly Gly Arg Asp Ser Arg Ser Gly Ser Pro Met Ala 15 10 15

Arg Arg (2) INFORMATION FOR SEQ ID NO: 54:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54:

Gly Gin Phe Arg Val lle Gly Pro Arg His Pro lle Arg 15 10 (2) INFORMATION FOR SEQ ID NO: 55:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 13 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 55:

His Ser Tyr Gin Glu Glu Ala Ala Met Glu Leu Lys Val 15 10 (2) INFORMATION FOR SEQ ID NO: 56:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 121 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 56:

Gly 1 Gin Phe Arg Val 5 Ile Gly Pro Arg His 10 Pro Ile Arg Ala Leu Val 15 Gly Asp Glu Val Glu Leu Pro Cys Arg Thr Sir Pro Gly Lys Asn Ala 20 25 30 Thr Gly Met Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val 35 40 45 His Leu Tyr Arg Asn Gly Lys Asp Gin Asp Gly Asp Gin Ala Pro Glu 50 55 60 Tyr Arg Gly Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys 65 70 75 80 Val Thr Leu Arg Ile Arg Asn Val Arg Phe Sir Asp Glu Gly Gly Phe 85 90 95 Thr Cys Phe Phe Arg Asp His Ser Tyr Gin Glu Glu Ala Ala Met Glu 100 105 110 Leu Lys Val Glu Asp Pro Phe Tyr Trp 115 120 INFORMATION FOR SEQ ID NO: 57: (i) SEOUENCE CHARACTERISTICS: (A) LENGTH : 20 amino acids (B) ΤΥΡΕ: amino acid (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ :; peptides (v) FRAGMENT ΤΥΡΕ: internal

(xi) SEOUENCE DESCRIPTION: SEQ ID NO: ; 57: Gly Gin Phe Arg Val Ile Gly Pro Arg His Pro Ile Arg Ala Leu Val 1 5 10 15

Gly Asp Glu Val 20 (2) 58:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOG '/: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 58:

Pro lle Arg Ala Leu Val Gly Asp Glu Val Glu Leu Pro Cys Arg lle 1 5 10 15

Ser Pro Gly Lys 20 (2) INFORMATION FOR SEQ ID NO: 59:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 59:

Glu Leu Pro Cys Arg lle Ser Pro Gly Lys Asn Ala Thr Gly Met Glu 15 10 15

Val Gly Trp Tyr 20 (2) INFORMATION FOR SEQ ID NO: 60:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 60:

Asn Ala Thr Gly Met Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg

10 15

Val Val His Leu 20 (2) INFORMATION FOR SEQ ID NO: 61:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 61:

Thr Val Gly Leu Val Phe Leu Cys Leu Gin Tyr Arg Leu Arg Gly Lys 15 10 15

Leu Arg Ala Glu 20 (2) INFORMATION FOR SEQ ID NO: 62

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) ΤΥΡΕ: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO-.62:

Tyr Arg Leu Arg Gly Lys Leu Arg Ala Glu Ile Glu Asn Leu His Arg 15 10 15

Thr Phe Asp Pro 20 (2) INFORMATION FOR SEQ ID NO: 63:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 63:

Ile Glu Asn Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro

10 15

Cys Trp Lys Ile 20

2) INFORMATION FOR SEQ ID NO: 64:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 64:

Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gin Phe Leu Glu Glu Leu 15 10 15

Arg Asn Pro Phe 20 (2) INFORMATION FOR SEQ ID NO: 65:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65:

Ala Ser Gin His Gly Pro 10 G (2) INFORMATION FOR SEQ ID NO: 66:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66:

Ala Ser Gin Ala Arg Pro Ser Gin Arg His Gly 15 10 (2) INFORMATION FOR SEQ ID NO: 67:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67:

Ala Ser Gin Tyr Arg Pro Ser Gin Arg His Gly 15 10 (2) INFORMATION FOR SEQ ID NO: 68:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 17 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 68:

Arg His Arg Asp Thr Gly lle Leu Asp Ser lle Gly Arg Phe Phe Ser 15 10 15

Gly (2) INFORMATION FOR SEQ ID NO: 69:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 17 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEO ID NO: 69:

Ile Ser Gin Ala Val His Ala Ala His Ala Glu Ile Asn Glu Ala Gly 15 10 15

Arg (2) INFORMATION FOR SEQ ID NO: 70:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 15 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 70:

Ile Ser Gin Ala Val His Ala Ala His Ala Glu Ile Asn Glu Ala 15 10 15 (2) INFORMATION FOR SEQ ID NO: 71:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 24 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 71:

Asp Glu Asn Pro Val Val His Phe Phe Ala Asn Ile Val Thr Pro Arg 15 10 15

Thr Pro Pro Pro Gin Gly Lys 20 (2) INFORMATION FOR SEQ ID NO: 72:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 12 amino acids (B) :Ε: amino acids (D) TOPOLOGY: linear (ii) MOLECULE peptΕ: peptides (v) FRAGMENT ΤΥΡΕ: internal (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 72:

His Phe Phe Ala Asn Ile Val Thr Pro Arg Thr Pro 15 10 (2) INFORMATION FOR SEQ ID NO: 73:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 18 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 73:

GAAAGGCTCC ACACACCG 18 (2) INFORMATION FOR SEQ ID NO: 74:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 26 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 74:

CAGAATCCGG GAAGAATGCC ACGGGC 26 (2) INFORMATION FOR SEQ ID NO: 75:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 28 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEO ID NO: 75:

CAGCGGCCGC ACGGAGTTTT CCTCTCAG 28 (2) INFORMATION FOR SEO ID NO: 76:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 28 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 76:

CAGAATTCTC AGGTTCTCAG ATGAAGGA 28 (2) INFORMATION FOR SEQ ID NO: 77:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 30 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 77:

AAGCGGCCGC TATCCTTCAT CTGAGAACCT 30 (2) INFORMATION FOR SEQ ID NO: 78:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 27 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: CDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 78:

CTTTTGAGCA AGTTCAGCCT GGTTAAG 27 (2) INFORMATION FOR SEQ ID NO: 79:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 39 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 79

ACCTCGAGGA GGTGGCTTAT GAGTATTTCT TCCAGGGTA (2) INFORMATION FOR SEQ ID NO: 80:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 33 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 80:

GGTGCGGGAA AGGTGACTCT CAGGATCCGG AAT 33 (2) INFORMATION FOR SEQ ID NO: 81:

(i) SEOUENCE CHARACTERISTICS:

(A) LENGTH: 33 base pairs (B) ΤΥΡΕ: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE ΤΥΡΕ: cDNA (xi) SEOUENCE DESCRIPTION: SEQ ID NO: 81:

ATTCCGGATC CTGAGAGTCA CCTTTCCCGC ACC 33

Claims (59)

PATENT APPLICATIONS A composition for treating multiple sclerosis in a mammal comprising at least one peptide, said peptide being selected from the following group of peptides: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12) ), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2, or its modifications or analogs or peptidomimetics . Composition according to claim 1, characterized in that said at least one peptide is selected from the group consisting of MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 ( SEO ID NO: 14) and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2. Composition according to claim 1, characterized in that said at least one peptide is MBP-4 (SEQ ID NO: 14). A composition for treating multiple sclerosis in a mammal comprising at least one peptide derived from MBP selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2, with reference to the composition comprises at least 40% of the total T cell reactivity to MBP in a population of individuals having T cells that respond to MBP. 5. A composition for treating multiple sclerosis in a mammal comprising at least two MBP peptides selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3) , MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEO ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEO ID NO: 13), MBP-4 (SEO ID NO: 14) and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2. Composition according to claim 5, characterized in that said at least two peptides are selected from the group consisting of MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 ( SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2. Composition according to claim 5, characterized in that one of the at least two peptides comprises MBP-4. Composition according to claim 5, characterized in that it further comprises at least one peptide selected from the group consisting of amino acid sequences: 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32), 82-96 (SEQ ID NO: 33), 8297 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100 P> Y] (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83-101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 88-100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111-135 ( SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141-160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50) , 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153-170 (SEQ ID NO: 53), all as shown in Figure 14. Composition according to claim 5, characterized in that it further comprises at least one peptide selected from the group consisting of amino acid sequences: 13-25 (SEQ ID NO: 31), 87-99 (SEQ ID NO: 43), 87 -99 [91K> A] (SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 82-100 [100P> Υ] (SEQ ID NO: 46), all as shown in Figure 14. A composition for treating multiple sclerosis in a mammal comprising at least two peptides derived from MBP, with at least one peptide being MBP-4 (SEQ ID NO: 14) and at least one peptide is selected from the group consisting of: MBP-1 (SEQ ID NO: 2) MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP2.4 (SEQ ID NO: 9) ), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-5 ( SEQID: 15), 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32), 82 -96 (SEQ ID NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100 P> Y] (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40) ), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 88-100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111-135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141 -160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153-170 (SEQ ID NO. : 53), all as shown in Figures 2 and 14. Composition according to claim 10, characterized in that at least one peptide is MBP-4 (SEQ ID NO: 14) and at least one peptide is selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP -3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-5 (SEQ ID NO: 15), 13-25 (SEQ ID NO: 31), 87-99 (SEQ ID NO) : 43), 87-99 [91K> A] (SEQ ID NO: 36), 82-100 (SEQ ID NO: 30), 82-100 [100P> Y] (SEQ ID NO: 46). A composition for treating multiple sclerosis in a mammal comprising at least two MBP peptides selected from the following peptide group: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP -1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO. : 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP- 3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2, said composition comprising at least 40% of the total T cell reactivity to MBP in a population of individuals who have T cells that respond to MBP. Composition according to claim 5, characterized in that it comprises a sufficient percentage of the total T cell reactivity to MBP such that administration of the composition to an individual with MS results in downregulation of the MS autoimmune response. A composition for treating multiple sclerosis in a mammal, characterized in that it comprises at least two MBP peptides comprising a T cell epitope, said composition being selected from the group of compositions consisting of: MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-3 (SEQ ID NO: 12), and MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEO ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-3 (SEQ ID NO: 12); MBP-1 (SEQ ID NO: 2) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11); MBP-1.1 (SEQ ID NO: 3) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11); MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11); MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-1 (SEQ ID NO: 2) or MBP-1.1 (SEQ ID NO: 3); MBP-1.1 (SEQ ID NO: 3), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO: 30), 82-100 [100P> Yj (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3) and MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO) : 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10) or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEO ID NO : 30), 82-100 [100P> Y] (SEO ID NO: 46), 87-99 (SEQ ID NO: 43), and 87-99 [91K> A] (SEQ ID NO: 36), all as are shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all of which are shown in FIG. 2; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO. : 30), 82-100 [100P> Yj (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all as shown in FIG. 2. The composition of claim 5, further comprising at least one peptide having a T cell activity derived from human myelin oligodendrocyte protein (MOG). 16. The composition of claim 15, wherein said MOG derived peptide is selected from the group consisting of: Human MOG 1-13 (SEQ ID NO: 54) Human MOG 103-115 (SEQ ID NO: 55) Human MOG 1-121 (SEQ ID NO: 56) Human MOG 1-20 (SEQ ID NO: 57) Human MOG 11-30 (SEQ ID NO: 58) Human MOG 21 -40 (SEQ ID NO: 59) Human MOG 31-50 (SEQ ID NO: 60) Human. MOG 141-160 (SEQ ID NO: 61) Human MOG 151-170 (SEQID: 62) Human MOG 161-180 (SEQ ID NO: 63) Human MOG 199-218 (SEQ ID NO: 64) G Q F R V I G P R H P I R HSYQEEAAMELKV GQFRVIGPRHPIRALVGDEV ELPCRTSPGKNATGMEVGWY RPPFSRVVHLYRNGKDQDGD QAPEYRGRTELLKDAIGEGK VTLRIRNVRFSDEGGFTCFF RDHSYQEEAAMELKVEDPFYW GQFRVIGPRHPIRALVGDEV; PIRALVGDEVELPCRISPGK; ELPCR1SPGKNATGMEVGWY; NATGMEVGWYRPPFSRVVHL; TVGLVFLCLQYRLRGKLRAE; YRLRGKLRAEIENLHRTFDP; I EN LH RT F DPH FL R VPC W K I; and YNWLHRRLAGQFLEELRNPF. The composition of claim 14, further comprising at least one peptide having T cell activity derived from MOG. 18. Isolated peptide derived from MBP, characterized in that said peptide is selected from the group consisting of: MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2.1 (SEQ ID NO: 6); MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEO ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15), all as shown in Figure 2, or a modification thereof or an analog. An isolated peptide according to claim 18, characterized in that said peptide is MBP-4 (SEQ ID NO: 14) or is a modification or analogue thereof. 20. An peptide analogue of claim 18, wherein the at least one amino acid residue of said peptide is substituted with alanine, glutamic acid or methylamino acid. 21. Peptide selected from the group consisting of MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO) : 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), all as shown in Figure 2, characterized in that lysine (K) is substituted with alanine (A). 22. MBP-2.1 peptide analogue (SEQ ID NO: 6), characterized in that lysine (K) at position 10 is substituted with alanine (A), said peptide analogue having the amino acid sequence DENPVVHFFANIVTPRTPPPSQGK (SEQ ID NO: 71 ). 23. A peptidomimetic peptide selected from the group consisting of: MBP-1 (SEO ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP- 2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), characterized in that at least one normal peptide bond is substituted by a non-peptide bond, a peptide bond analogue or a reduced bond analog. 24. A peptide selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEO ID NO: 9), MBP -2.5 (SEO ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEO ID NO: 13), MBP-4 (SEQ ID NO) : 14) and MBP-5 (SEQ ID NO: 15), characterized in that it has been modified by the addition of at least one charged amino acid residue to the amino terminal end, the carboxy terminal end, or to both terminal ends of said peptide to increase solubility of said peptide in aqueous solution. 25. A peptide selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEO ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14) and MBP-5 (SEQ ID NO: 15), characterized in that it has been modified by the addition of at least one amino acid residue to the amino terminal end, the carboxy terminal end, or to both terminal ends of said peptide to increase T cell activity of said peptide, wherein at least one amino acid residue is derived from the protein sequence of native MBP. Use of a composition according to claim 1 for the manufacture of a medicament for treating or preventing multiple sclerosis. Use according to claim 26, characterized in that said medicament is adapted for at least one of the following routes of administration: i.v. injection in non-immunogenic form, subcutaneous injection in non-immunogenic form, oral administration, inhalation administration, sublingual administration, transdermal administration, rectal administration or any combination thereof. Use according to claim 26, characterized in that said medicament is adapted for subcutaneous administration in a non-immunogenic form. The use of the composition, however, of claims 5, 10, 14 or 15 for the manufacture of a medicament for the treatment of multiple sclerosis. Use of at least two different compositions according to claim 1 for the manufacture of a medicament for the treatment of multiple sclerosis. 31. The use of a combination of peptides derived from MBP selected from the group of peptide combinations in which: MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-3 (SEQ ID NO: 12), and MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1 (SEQ ID NO: 2), MBP-2 (SEQ ID NO: 5), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1 (SEQ ID NO: 2), MBP-2.1 (SEQ ID NO: 6) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2 (SEQ ID NO: 5) and MBP-4 (SEQ ID NO: 14); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-5 (SEQ ID NO: 15); MBP-1.1 (SEQ ID NO: 3), MBP-2.1 (SEQ ID NO: 6) and MBP-3 (SEO ID NO: 12); MBP-1 (SEQ ID NO: 2) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2-6 (SEQ ID NO: 11); MBP-1.1 (SEQ ID NO: 3) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11); MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11); MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-1 (SEO ID NO: 2) or MBP-1.1 (SEO ID NO: 3); MBP-1.1 (SEQ ID NO: 3), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEO ID NO: 30), 82-100 [100P> Y ] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3) and MBP-4 (SEO ID NO: 14) and a peptide selected from the group, 100 consisting of: MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP -2.6 (SEQ ID NO: 11), all as shown in FIG. 2; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO. : 30), 82-100 [100P> Yj (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all as are shown in FIG. 2; MBP-1.1 (SEQ ID NO: 3), MBP-5 (SEQ ID NO: 15), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: 82-100 (SEQ ID NO. : 30), 82-100 [100P> Y] (SEQ ID NO: 46), 87-99 (SEQ ID NO: 43) and 87-99 [91K> A] (SEQ ID NO: 36), all as shown in FIG. 14; MBP-1.1 (SEQ ID NO: 3), MBP-3 (SEQ ID NO: 12), MBP-4 (SEQ ID NO: 14) and a peptide selected from the group consisting of: MBP-2.2 (SEQ ID NO) : 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), or MBP-2.6 (SEQ ID NO: 11), all as are shown in FIG. 2, for the manufacture of a medicament for the treatment of multiple sclerosis in a mammal. 32. A multipeptide formulation for pharmaceutical administration to individuals with MS, characterized in that it comprises at least two MBP peptides, each peptide being soluble and stable at a physiologically acceptable pre-determined pH, said peptides being selected from the group consisting of : MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ) ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEO ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), and MBP-5 (SEQ ID NO: 15). 33. Use of a therapeutic composition comprising at least one peptide having a T cell activity derived from a myelin antigen or an analogue of said peptide in an amount effective to reduce the symptoms of multiple sclerosis for the manufacture of a medicament for the treatment of an advanced multiple sclerosis condition in a mammal. 101 Use according to claim 33, characterized in that the myelin antigen is selected from the group consisting of myelin basic protein (MBP), myelin oligodendrocyte protein (MOG), proteolipid protein (PLP), and myelin-related glycoprotein (MAG) . Use according to claim 33, characterized in that said peptide comprises a peptide analogue having an MHC binding affinity greater than the MHC binding affinity of the peptide from which the analogue is derived. 36. Use of a therapeutic composition comprising at least one MBP peptide, said peptide being selected from the group consisting of: MBP-1 (SEQ ID NO: 2) MBP-1.1 (SEQ ID NO: 3), MBP-1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP -3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14). MBP-5 (SEQ ID NO: 15), 13-25 (SEQ ID NO: 31), 31-50 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 8292 (SEQ ID NO: 32), 82-96 (SEQ ID NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100 P> Y] (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83-101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 85-100 (SEQ ID NO: 41), 86-105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 8799 [91K> A] (SEQ ID NO: 36), 88-100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141-160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) ), and 153-170 (SEQ ID NO: 53), all as shown in Figures 2 and 14, or an analogue of said peptide, for the manufacture of a medicament for treating an advanced condition of multiple sclerosis in a mammal. Use according to claim 36, characterized in that said at least one peptide is selected from the group consisting of: MBP-1 (SEQ ID NO: 2), MBP-1.1 (SEQ ID NO: 3), MBP -1.2 (SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5), MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEQ ID NO: 7), MBP-2.3 (SEQ ID NO: 8) ), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 ( SEO ID NO: 13), MBP-5 (SEQ ID NO: 15), 13-25 (SEQ ID NO: 31), 87-99 (SEO ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 82-100 (SEO ID NO: 30), 82100 [100P> Y] (SEQ ID NO: 46) or its analogue. 102 Use according to claim 33, characterized in that said at least one peptide is derived from human MOG and is selected from the group consisting of: HumaniMOG 1-13 (SEQ ID NO: 54) Human.MOG 103-115 (SEQ ID NO: 55) HumaniMOG 1-121 (SEQIDNO: 56) HumaniMOG 1-20 (SEQ ID NO: 57) HumaniMOG 11-30 (SEQ ID NO: 58) HumaniMOG 21-40 (SEQ ID NO: 59) Humar »MOG 31-50 (SEQ ID NO: 60) HumaniMOG 141-160 (SEQ ID NO: 61) Human MOG 151-170 (SEQ ID NO: 62) Human MOG 161-180 (SEQ ID NO: 63) Human MOG 199-218 (SEQ ID NO: 64) G Q F R V I G P R H P I R H S YQEEAAMELKV GQFRV IG PRH P I R A LVGDE V ELPCRTSPGKNATGMEVGWY RPPFSRVVHLYRNGKDQDGD QAPEYRGRTELLKDAIGEGK VTLRIRNVRFSDEGGFTCFF RDHSYQEEAAMEIKVEDPFYW GQFRVIGPRHPIRALVGDEV; PIRALVGDEVELPCRISPGK; ELPCRISPGKNATGMEVGWY. NATGMEVGWYRPPFSRVVHL; TVGLVFLCLQYRLRGKLRAE; YRLRGKLRAEIENLHRTFDP; I E N L H R T F D P H F L R V P C W K I; and YNWLHRRLAGQFLEELRNPF. Use according to claim 36, characterized in that said medicament further comprises at least one human MOG peptide selected from the group consisting of: Human MOG 1-13 (SEQ ID NO: 54) HumanMOG 103-115 (SEQ ID NO: 55) Human MOG 1-121 (SEQ ID NO: 56) Human MOG 1-20 (SEQ ID NO: 57) Human MOG 11-30 (SEQ ID NO: 58) Human MOG 21-40 (SEQ ID NO: 59) Human MOG 31-50 (SEQ ID NO: 60) HumaniMOG 141 -160 (SEQ ID NO: 61) Human MOG 151-170 (SEQ ID NO: 62) HumaniMOG 161-180 (SEQ ID NO: 63) HumaniMOG 199-218 (SEQ ID NO: 64) G Q F R V I G P R H P I R HSYQEEAAM ELKV GQFRVIGPRHPIRALVGDEV ELPCRTSPGKNATGMEVGWY RPPFSRVVHLYRNGKDQDGD QAPEYRGRTELLKDAIGEGK VTLRIRNVRFSDEGGFTCFF RDHSYQEEAAMELKVEDPFYW GQFRVIGPRHP1RALVGDEV; PIRALVGDEVELPCRISPGK; ELPCRISPGKNATGMEVGWY; NATCMEVGWYRPPFSRVVHL; TVGLVFLCLQYRLRGKLRAE; YRLRGKLRAEIENLHRTFDP; I EN L H RT F D P H F L R V PC W K I; and YNWLHRRLAGQFLEELRNPF. Use according to claim 33, characterized in that said medicament is adapted for such a route of administration comprising subcutaneous or intravenous injection of said peptide. 41. The use of at least one therapeutic composition comprising at least one isolated and purified peptide, said peptide having a defined length, a defined sequence of amino acid residues and T cell activity, said composition capable of down-regulating multiple sclerosis symptoms in a mammalian population with advanced stages of multiple sclerosis when administered in a non-immunogenic form, 103 and wherein said composition is soluble in aqueous solution and is stable at physiologically acceptable pH, for the manufacture of a medicament for the treatment of a mammal in the advanced state of multiple sclerosis. Use according to claim 41, characterized in that the administration comprises the simultaneous or sequential administration of at least two of said therapeutic compositions. Use of a therapeutic composition according to claim 5, 10, 14 or 15 in an amount effective to down-regulate the symptoms of multiple sclerosis for the manufacture of a medicament for the treatment of an advanced condition of multiple sclerosis in a mammal. Use according to claim 33, characterized in that the advanced stage of multiple sclerosis is a returning-contracting MS, chronic progressive MS, primary progressive MS or benign MS. Use according to claim 33, characterized in that said medicament is adapted for administration in combination with / 3-interferon Use of at least one peptide of myelin autoantigen having T cell activity for the manufacture of a medicament for the treatment of multiple sclerosis in an individual, characterized in that said medicament is adapted for administration in combination with a therapeutically effective amount of IFN- / 3. Use according to claim 46, characterized in that said medicament is adapted for the simultaneous administration of said peptide and said IFN- / 3. 48. A composition comprising a therapeutically effective amount of at least one peptide having T cell activity of human MBP and a therapeutically effective amount of IFN- / 3 in a pharmaceutically acceptable carrier or diluent. The composition of claim 48, wherein said at least one peptide is selected from the group consisting of: MBP-1 (SEQ ID NO: 2) MBP-1.1 (SEQ ID NO: 3), MBP-1.2 ( SEQ ID NO: 4), MBP-2 (SEQ ID NO: 5). MBP-2.1 (SEQ ID NO: 6), MBP-2.2 (SEO ID NO: 7), MBP-2.3 (SEQ ID NO: 8), MBP-2.4 (SEQ ID NO: 9), MBP-2.5 (SEQ ID NO: 10), MBP-2.6 (SEQ ID NO: 11), MBP-3 (SEQ ID NO: 12), MBP-3.1 (SEQ ID NO: 13), MBP-4 (SEQ ID NO: 14), MBP-5 (SEQ ID NO: 15). 13-25 (SEQ ID NO: 31), 3150 (SEQ ID NO: 18), 61-80 (SEQ ID NO: 21), 82-92 (SEQ ID NO: 32), 82-96 (SEQ ID 104 NO: 33), 82-97 (SEQ ID NO: 34), 82-98 (SEQ ID NO: 35), 82-100 (SEQ ID NO: 30), 82-100 [100 P> Y] (SEQ ID NO: 46), 83-100 (SEQ ID NO: 37), 83-101 (SEQ ID NO: 38), 84-97 (SEQ ID NO: 39), 84-100 (SEQ ID NO: 40), 85 -100 (SEQ ID NO: 41), 86105 (SEQ ID NO: 42), 87-99 (SEQ ID NO: 43), 87-99 [91K> A] (SEQ ID NO: 36), 88100 (SEQ ID NO: 44), 88-99 (SEQ ID NO: 45), 111-135 (SEQ ID NO: 47), 122-140 (SEQ ID NO: 48), 139-170 (SEQ ID NO: 49), 141 -160 (SEQ ID NO: 28), 142-166 (SEQ ID NO: 50), 142-168 (SEQ ID NO: 51), 146-160 (SEQ ID NO: 52) and 153-170 (SEQ ID NO. : 53), all as shown in Figures 2 and 14. 50. The use of at least one peptide having T cell activity for the manufacture of a medicament for preventing the onset of multiple sclerosis in a susceptible individual, characterized in that said medicament is adapted for administration in combination with IFN-β. Use of at least one composition according to claim 1 for the manufacture of a medicament for preventing the onset of multiple sclerosis in an individual who is susceptible to it, characterized in that said medicament is adapted for administration in combination with IFN-β. Use of at least one composition according to claim 5, 10, 14 or 15 for the manufacture of a medicament for the treatment of multiple sclerosis in a mammal, characterized in that said medicament is adapted for administration in combination with IFN-β. 53. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a MHC class II peptide complex capable of binding T cell receptor and inducing anergy in a T-bearing receptor cell, wherein the complex consists essentially of: a MHC class II component comprising extracellular regions of a MHC class II molecule sufficient to form an antigen-binding pocket, said component being encoded by an autoimmune disease related allele, the component being soluble under physiological conditions in the absence of detergent or lipid; and the autoantigen peptide of claim 1, wherein the autoantigen peptide is bound to an antigen-binding pocket. 54. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a MHC class II peptide complex capable of binding the T cell receptor and inducing anergy in the T cell bearing the receptor, wherein the complex consists essentially of: 105 of the MHC class II component comprising extracellular regions of the MHC class II molecule sufficient to form an antigen-binding pocket, said component being encoded by an autoimmune disease related allele, the component being soluble under physiological conditions in the absence of detergent or lipid; and the autoantigen peptide according to claim 8, wherein the autoantigen peptide is bound to an antigen-binding pocket. 55. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a MHC class II peptide complex capable of binding the T cell receptor and inducing anergy in the T cell bearing the receptor, wherein the complex consists essentially of: a MHC class II component comprising extracellular regions of a MHC class II molecule sufficient to form an antigen-binding pocket, said component being encoded by an autoimmune disease related allele, the component being soluble under physiological conditions in the absence of detergent or lipid; and the autoantigen peptide of claim 16, wherein the autoantigen peptide is bound to an antigen-binding pocket. The composition of claim 53, wherein the autoimmune disease is multiple sclerosis. The composition of claim 1, comprising tandem copies of said peptide. Composition according to claim 8, characterized in that it comprises tandem copies of said peptide. 59. The composition of claim 16, comprising the tandem copies of said peptide. For Immulogic Pharmaceutical Corporation: 25833-VJI-97 / LZ 106 ABSTRACT Ingredients and treatment of multiple sclerosis The present invention provides isolated peptides and peptide combinations derived from myelin autoantigens such as MBP, MOG, PLP and MAG suitable for the treatment of multiple sclerosis, including prophylactic and therapeutic compositions and methods for preventing or treating multiple sclerosis. Preferred compositions of the invention comprise at least one isolated purified peptide which is free of all other polypeptides or contaminants, the peptide comprising an amino acid sequence, a myelin autoantigen having T cell activity. The therapeutic composition of the invention is capable of down-regulating an autoantigen-specific immune response to a myelin autoantigen in a population of people with or susceptible to multiple sclerosis by reducing, eliminating or preventing disease symptoms and / or preventing or delaying the onset or progression of disease . Additionally, the compositions and methods of the present invention when administered in the advanced stage of the disease, eradicate ongoing paralysis or other signs of the disease when administered during the acute phase of the disease, or prevent disease recurrence when administered during remission.