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WO1990013644A2 - Domaine cytoplasmique soluble du recepteur de glycoproteine cd2 humaine - Google Patents

Domaine cytoplasmique soluble du recepteur de glycoproteine cd2 humaine Download PDF

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Publication number
WO1990013644A2
WO1990013644A2 PCT/US1990/002584 US9002584W WO9013644A2 WO 1990013644 A2 WO1990013644 A2 WO 1990013644A2 US 9002584 W US9002584 W US 9002584W WO 9013644 A2 WO9013644 A2 WO 9013644A2
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pro
cell
arg
amino acid
gln
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WO1990013644A3 (fr
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Ellis L. Reinherz
Hsiu-Ching Chang
Philippe Moingeon
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Dana Farber Cancer Institute Inc
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Dana Farber Cancer Institute Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70507—CD2

Definitions

  • T lymphocyte T cell
  • T11 1 and T11 2 epitopes are expressed on resting as well as activated T cells.
  • Anti-T11 3 antibodies recognize a spatially distinct epitope that is preferentially expressed on mitogen- or
  • Perturbation of the extracellular domain of CD2 by its ligand, lymphocyte function-associated antigen 3 (LFA-3), or certain anti-CD2 monoclonal antibodies provides signals that synergize to augment T cell antigen receptor-mediated stimulation. Following such membrane perturbation, a rapid turnover in
  • nuclear activation follows including IL-2 gene expression resulting in lymphokine secretion and proliferation.
  • the protein predicted by cDNA sequence is comprised of the following three domains: (i) a hydrophilic 185 amino acid extracellular domain accounting for more than half of the molecule and bearing only limited homology to members of the immunoglobulin supergene family including T4, the T cell surface glycoprotein; (ii) a single characteristic hydrophobic 25 amino acid transmembrane domain; and (iii) a 117 amino acid cytoplasmic domain, rich in proline and basic amino acid residues.
  • cDNAs encoding the murine and rat equivalent of CD2 have been cloned and sequenced to show >50% amino acid sequence identity (Clayton et al., Eur. J. Immunol. 17:1367-1370 (1987); Sewell et al., Eur. J. Immunol. 17 : 1015 - 1020 ( 1987 ) ; Will iams et al. , J. Exp. Med . 165:368-380 (1987); Clayton et al, J.
  • This invention pertains to soluble peptides which correspond to the cytoplasmic domain of human CD2 glycoprotein, or a portion of the cytoplasmic domain, which is necessary for transduction of a CD2-mediated T lymphocyte activation signal into a cell.
  • the soluble peptides can comprise the entire 117-residue amino acid sequence of the mature human CD2 cytoplasmic domain or 77 amino acid residues corresponding to the amino-terminus of the CD2 cytoplasmic domain.
  • Preferred peptides include a 35 amino acid sequence corresponding to a portion of the cytoplasmic domain which is necessary for transduction of a CD2- mediated T lymphocyte activation signal into a cell.
  • the invention also pertains to an isolated DNA sequence encoding the cytoplasmic domain of human CD2 glycoprotein or a portion thereof.
  • This invention further pertains to cells which normally do not bear CD2 receptors, which have been transformed by an expression vector containing a
  • CD2-encoding DNA to express CD2 protein on their surface.
  • the transformed cells can be used to screen for substances which are agonistic or antagonistic to CD2-mediated cellular responses.
  • the invention also pertains to chimeric cell surface receptors containing a CD2-derived cytoplasmic domain and a non-CD2 extracellular domain.
  • chimeric receptors are encoded by chimeric gene contructs comprising DNA encoding the extracellular domain of a non-CD2 cell surface receptor linked to DNA encoding the cytoplasmic domain of human CD2
  • chimeric gene can express the chimeric surface receptor and can be used to screen for substances which are agonistic or antagonistic to the non-CD2 receptor- mediated response.
  • Figure 1 is the DNA sequence of the human CD2 cytoplasmic domain.
  • Figure 2 is a comparison of predicted transmembrane and cytoplasmic domain sequences of human and mouse CD2. Amino acid residues are designated in single letter code with the transmembrane regions underlined. Identical residues in human and mouse CD2 sequences are starred.
  • Figure 3 is a schematic representation of the transmembrane and cytoplasmic regions of human CD2 and variant molecules. Constructs of full length, deletion and substitution mutants of CD2 are diagrammed. The region most conserved between human and mouse CD2 is stippled, and the two repeating PPPGHR segments are marked in black. The X denotes the histidine residues thought to form a putative binding site.
  • restriction sites that generate the truncated CD2 molecules are marked by arrows with numbers in
  • Figure 4 is a schematic representation of the structure of the DOL retroviral expression vector.
  • Figures 5a-f are graphic representations of flow cytometric analysis of CD2 expression on murine T cells. Indirect immunofluorescence assays were carried out using the anti-T11 1 monoclonal antibody (3T4-8B5) (thick line) and compared to an irrelevant antibody (1HT4-4E5) (thin line) as background.
  • Figures 6a and 6b are audioradiograms for
  • Figures 7a-f are histograms of various T cell lines for the measurement of cytosolic Ca +2 by indo-1 fluorescence.
  • Figures 8a-d represent indirect immunofluorescence detection of murine CD3 and human CD2 on the surface of modulated and unmodulated CD2 FL.
  • Figure 8e represents the measurement of [Ca +2 ] i vs. time in modulated CD2 FL cells after stimulation with anti-T11 2 and anti-T11 3 antibodies (black arrow) or calcium ionophore (open arrow).
  • Figure 9 is a graphic representation of ( 3 H) thymidine incorporation of stimulated or unactivated murine cells, CD2 FL or 3D054.8, which were grown on IL2-dependent murine CTLL-20 cells. Cells were
  • Figures 10a and 10b are histogram representations of IL-2 production by CD2 FL, CD2 ⁇ 98, CD2 ⁇ 77, CD2 ⁇ 43, CD2 ⁇ 18, 3D054.8 and murine T cell line CD2 M271-2 upon antigen (ovalbumin) or anti-T11 2 and anti-T11 3
  • the present invention pertains to soluble peptides having an amino acid sequence that corresponds to the cytoplasmic domain of human CD2 glycoprotein or a portion thereof, which is necessary for transduction of a CD2 -mediated T lymphocyte activation signal into a cell.
  • the peptides of this invention are soluble in aqueous medium.
  • the soluble peptides include an amino acid sequence of approximately 35 amino acid residues corresponding to a portion of the cytoplasmic domain of CD2 necessary for transduction of a CD2-mediated T cell activation signal into a cell.
  • This region contains four histines at amino acid position 264, 271, 278 and 282 and includes two tandomly repeated segments (PPPGHR, amino acids 260-265 and 274-279). Mutations at positions 278-279 alter the structure of the second repeat without affecting cell activation, suggesting that the more carboxy-terminal repeat is not required for cell activation.
  • the peptide has the following 35 amino acid residues.
  • the soluble peptides have an amino acid sequence that correspond to the entire amino acid sequence of the mature human CD2 cytoplasmic domain and have the following 117 amino acid residue sequence: 211 Thr Lys Arg Lys Lys Gln Arg Ser Arg Arg
  • the soluble peptides of this invention can be a functional portion of the cytoplasmic domain which is necessary for transduction of a CD2-mediated T cell activation signal into a cell.
  • the soluble peptide has about 77 amino acid residues corresponding to the amino-terminal portion of the CD2 cytoplasmic domain. Additional amino acid residues can be attached to the carboxy - terminus of the 77 amino acid sequence. Specifically, 21 amino acid residues corresponding to the carboxy-terminus of the CD2 cytoplasmic domain (i.e., amino acid position 288-308) can be attached to the soluble peptide of 77 amino acid residues.
  • the resulting 98 residue peptide is
  • the soluble CD2 peptides of this invention include analogous or homologous sequences which encode proteins capable of transducing a CD2-mediated activation signal into a cell.
  • These peptides can include sequences in which functionally equivalent amino acid residues are substituted for residues within the sequence resulting in a silent change.
  • one or more amino acid residues within the sequence can be substituted by another amino acid of a similar polarity which acts as a functional equivalent, resulting in a silent alteration.
  • Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs.
  • nonpolar (hydrophobic) amino acids include glycine, alanine, leucine, isoleucine, valine, proline, phenyl- alanine, tryptophan and methionine.
  • the polar neutral amino acids include serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The charged
  • (basic) amino acids include arginine, lysine and histidine.
  • the negatively charged (acidic) amino acids include aspartic and glutamic acid.
  • the peptide structure can be modified by deletions, additions, Inversion, insertions or substitutions of one or more amino. acid residues in the sequence which do not substantially detract from the functional properties of the peptide.
  • Naturally occurring allelic variations and modifications are included within the scope of the invention so long as the variation does not substantially reduce the ability of the peptide to transduce a CD2-mediated T cell activation signal into a cell.
  • Soluble peptides of this invention can be made by enzymatic fragmentation of human CD2 glycoprotein or a portion thereof, by peptide synthesis or recombinant DNA technology.
  • the peptides or modified equivalents thereof can be synthesized directly by standard solid or liquid phase chemistries for peptide synthesis.
  • the above amino acid sequence or modified equivalent thereof encoding the cytoplasmic domain of CD2 can be synthesized by the solid phase procedure of Merrifield.
  • the soluble CD2 peptides will be produced by inserting DNA encoding the peptide (i.e., CD2 DNA which encodes the desired amino acid sequence of the cytoplasmic domain of CD2) into an appropriate vector/host system where it is expressed.
  • DNA sequence which encodes the cytoplasmic domain of native human CD2 is shown in Figure 1. The entire DNA
  • CD2 has been described in U.S. Application Serial No. 06/932,871, filed November 18, 1986, by Reinherz et al.
  • This native sequence or a variant thereof can be used.
  • any of the possible DNA sequences which encode the amino acid sequences given above can be used.
  • a substantial coding equivalent is any nucleic acid sequence which encodes an amino acid sequence having essentially the same functional characteristics as the native human CD2 cytoplasmic domain.
  • the native DNA sequence of CD2 can be modified by deletion, insertion or substitution of nucleotides to yield peptides which exhibit substantially the same properties of the above-described soluble peptides.
  • the DNA sequences can be chemically synthesized or they can be obtained from natural sources using recombinant DNA technology.
  • Host-vector systems can be used to express the peptides of this invention. Primarily, the vector system must be compatible with the host cell used.
  • Host-vector systems include, but are not limited to, the following: bacteria transformed with bacteriophage DNA, plasmid DNA or cosmid DNA; micro-organisms, such as yeast containing yeast vectors; mammalian cell systems infection with virus (e.g., vaccinia virus, adenovirus, etc.); insect cell systems infected with virus (e.g., baculovirus).
  • the recombinant DNA vector can be introduced into appropriate host cells (bacteria, virus, yeast, mammalian cells or the like) by transformation, transduction or transfection (depending upon the vector/host cell system) and cultured to express the peptides of this invention.
  • This invention also pertains to cells which have been transformed to express CD2 protein on its surface.
  • the cells are derived from cells which do not normally express CD2 protein or do not express the CD2 protein at significant levels on its surface.
  • a continuous non-CD2 bearing cell line is transformed with an expression vector containing CD2-encoding DNA to thereby cause the cell line to express CD2 protein on its surface.
  • the expression vector will contain the DNA sequence encoding the extracellular, transmembrane and cytoplasmic domains of human CD2 glycoprotein. DNA encoding the native CD2 glycoprotein and portions of the protein have been previously described in U.S. Patent Application Serial No.
  • cytoplasmic domain of CD2 need only be a portion of cytoplasmic domain which is necessary for transduction of a CD2-mediated T lymphocyte activation signal into a cell, as described above.
  • Suitable expression vectors include the DOL retroviral vector, baculovirus transfer vectors, pAc373 and pCDVI. The DOL retroviral expression vector is preferred.
  • the transformed cells which express the CD2 surface receptor can be used to screen for substances which are agonistic or antagonistic to CD2-mediated cellular response, such as drugs, chemicals and
  • the transformed cell is contacted with a substance to be tested under
  • Stimulation of the CD2 receptor is determined by a change in intracellular calcium concentration.
  • calcium concentration is measured within the cell before and after the cell is contacted with the substance.
  • This system can be used to screen for substances which can block the CD2 surface receptor and inhibit CD2-mediated cellular response.
  • the transformed cell is contacted with a substance to be tested for blocking activity and with a CD2 activating agent (i.e., a compound known to mediate CD2 activation of the cell) under conditions which would permit CD2 stimulation by the activating agent.
  • a CD2 activating agent i.e., a compound known to mediate CD2 activation of the cell
  • Intracellular calcium concentration is then measured as indicative of CD2-mediated cellular response.
  • the cytoplasmic domain of the CD2 receptor can be exploited to provide screening assays for substances that interact with cell surface receptors other than CD2.
  • the CD2 cytoplasmic domain can be linked to a non-CD2 extracellular domain to make a chimeric receptor.
  • the chimeric receptor has the extracellular region of a receptor of choice, but the Intracellular domain of the CD2 receptor which, thus, produces a CD2-type intracellular response (calcium mediated).
  • the chimeric receptor is made by producing an expressible chimeric gene construct which can be used to transform an appropriate host cell.
  • the chimeric gene construct comprises DNA encoding the extracellular domain of a non-CD2 cell surface receptor; DNA encoding the transmembrane domain of a cell surface receptor (either CD2 or non-CD2); and DNA encoding the cytoplasmic domain of human CD2 glycoprotein or any portion of the domain which functions for transduction of a CD2-mediated T lympocyte activation signal into a cell.
  • DNA encoding the transmembrane domain can correspond to DNA encoding the transmembrane domain of any cell surface receptor.
  • a cell (preferably one which does not ordinarily express the receptor of choice, such as mammalian cells of various tissue origins) is transformed with the chimeric gene construct and caused to express the chimeric surface receptor.
  • These transformed cells can be used to screen for substances which are agonistic or antagonistic to the receptor-mediated cellular response in screening assays, as described above.
  • the CD2 cytoplasmic domain permits interaction with the
  • the human CD2 cDNA PB2 (Sayre, P.H. et al. , Proc. Natl. Acad. Sci. USA 84:2941-2945 (1987)), was digested with restriction enzyme Hphl, Banll, Fokl, StuI and Aval, respectively, and blunted with T4 DNA polymerase.
  • Hphl, Banll, Fokl, StuI and Aval was digested with restriction enzyme Hphl, Banll, Fokl, StuI and Aval, respectively, and blunted with T4 DNA polymerase.
  • Hphl and Banll digests were ligated to the linker 5'-CTAAGGATCCTTAG-3' while Fokl, StuI and Aval digests were ligated to the linker
  • substitution mutants were generated by oligonucleotide-directed in vitro mutagenesis as described by Taylor, J.W. et al., Nucl . Acids Res. 13: 8764-8785 (1985).
  • the synthetic oligonucleotides utilized for mutagenesis were 5'-CAGGCACCTAGTGATGAGCCCGCGCCTCCT-3' for CD2 M271-2 which changes the wild-type sequence CATCGT (His-Arg) into GATGAG (Asp-Glu) and 5'- CCGCCTCCTGGAGATGAGGTTCAGCACCAG-3' for CD2 M278-9 which changes the wild- type sequence CACCGT (His-Arg) into GATGAG (Asp-Glu).
  • the full length cDNAs as well as modified cDNAs were inserted into the retrovirus expression vector DOL (kindly provided by Dr. Thomas Roberts, Dana Farber Cancer Institute) ( Figure 4). Selected restriction sites on the vector shown are: B, BamHI; RI, EcoRI; RV, EcoRV; H3, Hindlll.
  • the vector contains two promoters: MLV-LTR to drive the expression of the CD2 cDNA and the SV40 promoter to express the neomycin resistance gene (Korman, A. et al., Proc. Natl. Acad.
  • the plasmids containing full length or modified CD2 cDNAs were isolated and sequenced around the modified region by double-stranded sequencing before
  • transfection by Ca precipitation into ⁇ -2, a helper- free retrovirus packaging cell line The murine T cell hybridoma 3D054.8 cell line specific for ovalbumin in the context of the H-2 (I-A d ) molecule (Haskins, K. et al., J. Exp. Med. 157:1149-1169 (1983)) and lacking the human CD2 glycoprotein was infected with the defective retroviruses. Both transiently expressed and permanent viral stocks were used to infect the hybridoma in the presence of 8 ⁇ g/ml polybrene (Aldrich Chemicals, Milwaukee, WI).
  • the G418 resistant clones were analyzed for surface CD2 expression using an indirect immunofluorescence assay with anti-T11 1 and anti-T11 2 antibodies and positive clones were further sorted on the Epics V cell sorter.
  • CD2 FL full length
  • CD2 ⁇ C98, CD2 ⁇ C77, CD2 ⁇ C43, CD2 ⁇ C18 and CD2 ⁇ C-3 resulted from infection with retroviruses containing the entire extracellular and transmembrane segment of CD2 but only 98 or fewer of the 117 cytoplasmic residues.
  • Cells were maintained in RPMI 1640 (Gibco) medium supplemented with 10% heat-inactivated fetal calf serum (Flow Labs, McLean, VA), 50 ⁇ M 2-mercaptoethanol (2-ME), 1mM sodium pyruvate, 2 mM L-glutamic acid, 1% penicillin-streptomycin (Whittaker M.A.
  • Flow cytometric analysis of CD2 expression on murine T cells was carried out by indirect immunofluorescence assays using anti-T11 1 monoclonal antibody (3T4-8B5) and compared to an irrelevant antibody (1HT4-4E5) as background. Ascites were used at a 1:200 dilution and 10 6 cells incubation for 30 minutes at 4oC. After washing in RPMI 1640 with 2% fetal calf serum (FCS), bound antibodies were detected with a 1:40 dilution of fluorescein coupled goat anti-mouse IgG as a second antibody (Meloy, Springfield, VA). 10,000 cells were analyzed per sample on an Epics V cell sorter.
  • FCS fetal calf serum
  • FIG. 5a-f A representative pattern of reactivity with the anti-CD2 monoclonal antibody is shown in Figures 5a-f. Histograms represent the number of cells (ordinate) vs. log 10 fluorescence intensity (abscissa). All of these transfe-ctants express comparable levels of CD2
  • CD2 ⁇ C18 and CD2 ⁇ C-3 which express on the order of 50% the copy number.
  • the surface expression of CD2 on ⁇ C-3 was unstable and did not allow functional analysis.
  • similar reactivities to those obtained with anti-T11 1 were found using the anti-T11 2 antibody while none of the unactivated clones were stained by the antl-T11 3 antibody.
  • Immunoprecipi tates were extensively washed with RIPA buffer and separated on a 10% SDS-PAGE after treatment with 5% 2-ME. The gel was dried and the autoradiograph exposed for two weeks at -70oC with intensifying screens. A contaminant band of 70KD was regularly detected in the autoradiograms despite extensive preclearing, as previously reported by
  • a 53KD band was identified of clone CD2 FL.
  • FIGS. 7a-f show analyses of alteration in
  • CD2 ⁇ C43 and CD2 ⁇ C18 clones were not triggered by anti-T11 2 and anti-T11 3 antibodies.
  • the non-transfected line 3D054.8 was also not stimulated. Given that an immediate [Ca 2+ ] rise was i
  • [Ca ] i can be induced through human CD2 structures expressed on the membrane of murine T cells even in the absence of the carboxy-terminal 40 amino acid residues of the CD2 cytoplasmic segment.
  • the present results with human CD2 are consistent with an independent report by Williams et al., (He, Q. et al ., Cell
  • Figures 8a-d represent indirect immunofluor- escence detection of murine CD3 and human CD2
  • Figure 8e represents measurement of [Ca ] i vs. time In modulated CD2 FL cells after stimulation with anti-T11 2 and anti-T11 3 antibodies (black arrow) or calcium ionophore (open arrow).
  • anti-CD3 modulated CD2 FL cells were no longer stimulated by anti-T11 2 and anti-T11 3 antibodies to increase [Ca 2+ ] i .
  • the incubation of CD2 FL cells under similar conditions with anti-human CD3 antibody (Leu4) did not have any effect. This result suggests that regulation of the
  • CD2 pathway by CD3-Ti is intact in the cellular model herein. Stimulation of IL-2 production through the human CD2 molecule and its vatiant forms on murine T cells.
  • IL-2 production upon antigen (ovalbumin) or anti-T11 2 and anti-T11 3 stimulation was determined using the above methods.
  • IL-2 production was obtained by running culture supernatants in parallel to a titration curve of recombinant IL-2 (Biogen Labs, Cambridge, MA). The limit of detection for IL-2 was ⁇ 4 U/ml.
  • M represents the murine T cell line CD2 M271-2. Analysis of IL-2 production by two independent clones of the CD2 M278-9 provided evidence that such cells could produce
  • CD2 ⁇ C77 clones express human
  • CD2 molecules lacking residues 289 to 316 corresponds to the segment most conserved among human and murine molecules with 24 out of 27 residues being identical ( Figures 2 and 3). Presumably, these conserved residues function in another facet of CD2 biology unrelated to IL-2 induction and/or secretion. In contrast, the ⁇ C43 truncated molecules, as well as shorter truncations are non-functional with respect to
  • CD2 cytoplasmic domain is involved in signal transduction and that one essential sequence of the cytoplasmic domain necessary for CD2-mediated activation is located between amino acids 253 and 287.
  • This region contains four histidines at amino acid positions 264, 271, 278 and 282 and includes two tandemly repeated segments (PPPGHR, amino acids 260-265 and 274-279) (see Figures 2 and 3). These histidine residues could represent a binding site for an ion, cyclic nucleotide or other small regulatory molecule.
  • substitution mutants of CD2 were produced by site- directed mutagenesis.
  • Two different categories of mutants with a stable CD2 surface expression were obtained: CD2 M172-2 and CD2 M278-9 in which the positively charged histidine and arginine residues at position 271 and 272 or 278 and 279 were replaced by negatively charged aspartic acid and glutamic acid, respectively (Figure 3).
  • Functional characterization of these two mutants shows that IL-2 production can be induced by anti-T11 2 and anti-T11 3 antibodies to a level comparable to that of the CD2 clones ( Figure 10).
  • a putative "cage" requiring four histidine residues, is not necessary for this CD2 activation.
  • DOL vectors containing CD2FL, CD2 ⁇ C77 or CD2 ⁇ C98 (CD2 truncated cDNA) have been deposited in the

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Abstract

L'invention se rapporte à des peptides solubles et à un ADN codant pour les peptides qui correspondent au domaine cytoplasmique de la glycoprotéine CD2 humaine ou d'une partie de cette glycoprotéine, nécessaire à la transduction d'un signal d'activation de lymphocytes T par CD2 dans une cellule. L'invention se rapporte également à des cellules transformées en vue d'exprimer un récepteur de surface de CD2, ainsi qu'à des procédés de criblage de substances qui stimulent ou bloquent la fonction réceptrice de CD2.
PCT/US1990/002584 1989-05-09 1990-05-09 Domaine cytoplasmique soluble du recepteur de glycoproteine cd2 humaine Ceased WO1990013644A2 (fr)

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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996013584A1 (fr) * 1994-11-01 1996-05-09 Targeted Genetics Corporation Recepteurs chimeres servant a produire des lymphocites t cytotoxique th, activables selectivement independants des cellules auxiliaires
US5610148A (en) * 1991-01-18 1997-03-11 University College London Macroscopically oriented cell adhesion protein for wound treatment
US5629287A (en) * 1991-01-18 1997-05-13 University College London Depot formulations
US5641748A (en) * 1995-06-07 1997-06-24 Biogen, Inc. Caip-like gene family
US5656438A (en) * 1995-06-07 1997-08-12 Biogen, Inc. CAIP-like gene family
US5837844A (en) * 1995-06-07 1998-11-17 Biogen, Inc. CAIP-like gene family
WO1999036534A1 (fr) * 1998-01-13 1999-07-22 Dana Farber Cancer Institute, Inc. CLONAGE ET CARACTERISATION D'UNE PROTEINE D'ADAPTATION SEMBLABLE A cdc-15 (CD2BP1)
WO2000034309A1 (fr) * 1998-12-04 2000-06-15 Dana-Farber Cancer Institute, Inc. Clonage et caracterisation d'une proteine de liaison de cd2 (cd2bp2)
US6171800B1 (en) 1995-06-07 2001-01-09 Biogen, Inc. Method of making and binding CAIP polypeptides
US6423824B1 (en) 1995-06-07 2002-07-23 Biogen, Inc. CAIP-like gene family
WO2020198413A1 (fr) * 2019-03-27 2020-10-01 The Trustees Of The University Of Pennsylvania Thérapie par lymphocytes t à récepteurs antigéniques chimériques (car) de tn-muc1

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Title
Cell, Volume 54, 23 September 1988, Cell Press, OI HE et al.: " A Role in Transmembrane Signaling for the Cytoplasmic Domain of CD2 T Lymphocyte Surface Antigen", pages 979-984 see the whole article *
Eur. J. Immunol. Volume 18, 1988, VCH Verlagsgesellschaft mbH, (Weinheim, DE) A. ALCOVER et al.: " The T11 (CD2) cDNA Encodes a Transmembrane Protein which Expresses T111, T112 and T113 Epitopes but which does not Independently Mediate Calcium Influx: Analysis by Gene Transfer in a Baculovirus System", pages 363-367 see the whole article *
Immunology Today, Volume 10, No. 1, 1989, Elsevier Science Publishers Ltd, (GB), A. TRAUNECKER et al.: "Solubilizing the T-Cell Receptor Problems in Solution", pages 29-32 see page 29, paragraph 2; page 31, paragraph 3 *
Letters to Nature, Volume 339, 25 May 1989 P. MOINGEON et al.: "CD2-Mediated Adhesion Facilitates T Lymphocyte Antigen Recognition Function", pages 312-314 see the whole article *
Proc. Natl. Acad Sci. USA, Volume 83, November 1986 W.A. SEWELL et al.: "Molecular Cloning of the Human T-Lymphocyte Surface CD2 (T11) Antigen", pages 8718-8722 see the whole article (cited in the application) *
Proc. Natl. Acad. Sci. USA, Volume 84, May 1987, P.H. SAYRE et al.: "Molecular Cloning and Expression of T11 cDNAs Reveal a Receptor-Like Structure on Human T Lymphocytes", pages 2941-2945 see the whole article (cited in the application) *
Proc. Natl. Acad. Sci. USA, Volume 85, March 1988, D.J. DIAMOND et al.: "Exon-Intron Organization and Sequence Comparison of Human and Murine T11 (CD2) Genes", pages 1615-1619 see the whole article (cited in the application) *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5610148A (en) * 1991-01-18 1997-03-11 University College London Macroscopically oriented cell adhesion protein for wound treatment
US5629287A (en) * 1991-01-18 1997-05-13 University College London Depot formulations
WO1996013584A1 (fr) * 1994-11-01 1996-05-09 Targeted Genetics Corporation Recepteurs chimeres servant a produire des lymphocites t cytotoxique th, activables selectivement independants des cellules auxiliaires
US6083751A (en) * 1994-11-01 2000-07-04 Targeted Genetics Corporation Chimeric receptors for the generation of selectively-activatable TH-independent cytotoxic T cells
US5656438A (en) * 1995-06-07 1997-08-12 Biogen, Inc. CAIP-like gene family
US5837844A (en) * 1995-06-07 1998-11-17 Biogen, Inc. CAIP-like gene family
US5641748A (en) * 1995-06-07 1997-06-24 Biogen, Inc. Caip-like gene family
US6171800B1 (en) 1995-06-07 2001-01-09 Biogen, Inc. Method of making and binding CAIP polypeptides
US6423824B1 (en) 1995-06-07 2002-07-23 Biogen, Inc. CAIP-like gene family
US7087390B2 (en) 1995-06-07 2006-08-08 Biogen Idec Ma Inc. CAIP-like gene family
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WO2020198413A1 (fr) * 2019-03-27 2020-10-01 The Trustees Of The University Of Pennsylvania Thérapie par lymphocytes t à récepteurs antigéniques chimériques (car) de tn-muc1

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