Antibody 12‐15 cross‐reacts with mouse Feγ receptors and CD2: study of thymus expression, genetic polymorphism and biosynthesis of the CD2 protein

Previously we described a monoclonal antibody (mAb 12–15) that reacted with murine Fc receptor proteins (betal, beta2 and alpha) and an undefined molecule of 37 kDa (beta3) on certain types of cells. Here we present serological and biochemical evidence that the beta3 chain is expressed on mouse thymocytes and that it is identical to the CD2 antigen. By immunofluorescence staining and cytofluorographic analysis >95% of thymocytes were positive. Brightly staining cells coincided with cortison‐resistant thymocytes suggesting that mature thymocytes expressed higher levels of the antigen. Biosynthetic labeling of DBA/2 thymocytes with [35S]methionine showed that the size of the CD2 precursor molecule was 43 kDa which was processed to approximately 55–65 kDa in the mature molecule. mAb 12–15 was also reactive with the tunicamycin‐treated form of the CD2 antigen suggesting that the cross‐reactive epitope was of protein nature. Comparison of different mouse strains indicated that two molecular forms of CD2 exist. On BALB/c thymocytes, the relative mass of the native molecule was approximately 60–70 kDa (CD2.1) and slightly larger than in DBA/2 (CD2.2). Following endoglycosidase F treatment both proteins still showed a slight difference in electrophoretic mobility. Several inbred mouse strains were analyzed for expression of CD2 forms. When mAb 12–15 was used in cyotoxic T lymphocyte inhibition experiments using specific CTL and tumor target cells it was found that the antibody could specifically inhibit CTL‐mediated lysis presumably by interfering with CD2 function.

[1]  M. Crumpton,et al.  Species conservation of the T cell lymphocyte CD2 cell surface antigen , 1988, European journal of immunology.

[2]  E. Reinherz,et al.  Adhesion domain of human T11 (CD2) is encoded by a single exon. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Paterson,et al.  Activation of rat T lymphocytes by anti-CD2 monoclonal antibodies , 1988, The Journal of experimental medicine.

[4]  P. Altevogt,et al.  Murine Fcγ receptor proteins: identification of a previously unrecognized molecule with a monoclonal antibody (12‐15) , 1988 .

[5]  E. Reinherz,et al.  Exon-intron organization and sequence comparison of human and murine T11 (CD2) genes. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[6]  M. Crumpton,et al.  Murine CD2 gene expression during fetal thymus ontogeny , 1988, European journal of immunology.

[7]  P. Altevogt,et al.  Structural basis for altered soybean agglutinin lectin binding between a murine metastatic lymphoma and an adhesive low malignant variant. , 1987, Experimental cell research.

[8]  P. Altevogt,et al.  New antigens presented on tumor cells can cause immune rejection without influencing the frequency of tumor-specific cytolytic T cells. , 1987, Cellular immunology.

[9]  M. Reed,et al.  Direct evidence for a receptor-ligand interaction between the T-cell surface antigen CD2 and lymphocyte-function-associated antigen 3. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Seed,et al.  Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselection procedure. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[11]  E. Reinherz,et al.  Molecular cloning and expression of T11 cDNAs reveal a receptor-like structure on human T lymphocytes. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[12]  S. Meuer,et al.  Alternative pathway activation of T cells by binding of CD2 to its cell-surface ligand , 1987, Nature.

[13]  Michael Loran Dustin,et al.  Rosetting of activated human T lymphocytes with autologous erythrocytes. Definition of the receptor and ligand molecules as CD2 and lymphocyte function-associated antigen 3 (LFA-3) , 1987, The Journal of experimental medicine.

[14]  Michael Loran Dustin,et al.  Purified lymphocyte function-associated antigen 3 binds to CD2 and mediates T lymphocyte adhesion , 1987, The Journal of experimental medicine.

[15]  D. Paterson,et al.  Similarities in sequences and cellular expression between rat CD2 and CD4 antigens , 1987, The Journal of experimental medicine.

[16]  B. Haynes,et al.  Thymocyte binding to human thymic epithelial cells is inhibited by monoclonal antibodies to CD-2 and LFA-3 antigens. , 1987, Journal of immunology.

[17]  M. Crumpton,et al.  Molecular cloning of the human T-lymphocyte surface CD2 (T11) antigen. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[18]  T. Springer,et al.  Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones , 1986, Nature.

[19]  H. Morse,et al.  Alleles of the Ly-17 alloantigen define polymorphisms of the murine IgG Fc receptor. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[20]  P. Brottier,et al.  T cell activation via CD2 [T, gp50] molecules: accessory cells are required to trigger T cell activation via CD2-D66 plus CD2-9.6/T11(1) epitopes. , 1985, Journal of immunology.

[21]  E. Reinherz.  A molecular basis for thymic selection: regulation of T11 induced thymocyte expansion by the T3-Ti antigen/MHC receptor pathway. , 1985, Immunology today.

[22]  P. Altevogt,et al.  Resistance of metastatic tumor variants to tumor-specific cytotoxic T-lymphocytes not due to defects in expression of restricting major histocompatibility complex molecules in murine cells. , 1984, Cancer research.

[23]  E. Reinherz,et al.  An alternative pathway of T-cell activation: A functional role for the 50 kd T11 sheep erythrocyte receptor protein , 1984, Cell.

[24]  F. Sánchez‐Madrid,et al.  The functional significance, distribution, and structure of LFA-1, LFA-2, and LFA-3: cell surface antigens associated with CTL-target interactions. , 1983, Journal of immunology.

[25]  G. Longton,et al.  Identification and functional characterization of two distinct epitopes on the human T cell surface protein Tp50. , 1983, Journal of immunology.

[26]  P. Altevogt,et al.  Metastatic potential severely altered by changes in tumor cell adhesiveness and cell-surface sialylation , 1983, The Journal of experimental medicine.

[27]  J. Strominger,et al.  Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[28]  W. Decock,et al.  Suppression of human T-cell mitogenesis and E-rosette formation by the monoclonal antibody OKT11A. , 1981, Immunology.

[29]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[30]  P. Anderson,et al.  Regulatory interactions between members of the immunoglobulin superfamily. , 1988, Immunology today.

[31]  E. Reinherz,et al.  Murine and human T11 (CD2) cDNA sequences suggest a common signal transduction mechanism , 1987, European journal of immunology.

[32]  M. Crumpton,et al.  The murine homologue of the T lymphocyte CD2 antigen: molecular cloning, chromosome assignment and cell surface expression , 1987, European journal of immunology.

[33]  A. F. Williams,et al.  A year in the life of the immunoglobulin superfamily. , 1987, Immunology today.

[34]  Michael Loran Dustin,et al.  The lymphocyte function-associated LFA-1, CD2, and LFA-3 molecules: cell adhesion receptors of the immune system. , 1987, Annual review of immunology.

[35]  P. Altevogt,et al.  Different expression of Lyt differentiation antigens and cell surface glycoproteins by a murine T lymphoma line and its highly metastatic variant , 1982, European journal of immunology.