Abstract
We describe here two new monoclonal antibodies that react with surface antigens of human lymphocytes. Antibody 7.2 identified a determinant on the framework region of the human Ia antigen. It was cytotoxic for all cultured B-cell lines, normal B cells, and monocytes. The antibody was not cytotoxic for normal T cells or for established T leukemic cell lines. In immune precipitation assays, the 7.2 antibody reacted with a bimolecular complex of two chains that resolved in polyacrylamide gels as polypeptides with molecular weights of 29000 and 34000 daltons. These precipitation results were analogous to those achieved with a rabbit antiserum prepared against human Ia antigens. Antibody 9.3 identified a determinant on the framework region of a T-cell antigen. It was cytotoxic for 50–80% of peripheral T cells and for 20–50% of thymocytes. It was not cytotoxic for cultured B-cell lines, normal B cells, or monocytes. In immune precipitation assays, the 9.3 antibody reacted with a single polypeptide with a molecular weight of 44000 daltons. Due to the expression of this antigen on a limited subpopulation of human T cells, we have designated the antigen HuLyt-1.
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Amos, D. G.: Cytotoxicity testing.NIAID Manual For Tissue Typing Techniques, pp. 25–28, USDHEW Publication No. (NIH) 76-545, 1976
Barnstable, C. J., Bodmer, W. F., Brown, G., Galfre, G., Milstein, C., Williams, A. F., and Ziegler, A.: Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens — new tools for genetic analysis.Cell 14: 9–20, 1978
Bonner, N. W. and Laskey, R.: A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels.Eur. J. Biochem. 46: 83–88, 1974
Evans, R. L., Faldetta, T. J., Humphreys, R. E., Pratt, D. M., Yunis, E. J., and Schlossman, S. F.: Peripheral human T cells sensitized in mixed leucocyte culture synthesize and express Ia-like antigen.J. Exp. Med. 148: 1440–1445, 1978
Fu, S. M., Chiorazzi, N., Wang, C. Y., Montazeri, G., Kunkel, H. G., Ko, H. S., and Gottlieb, A. B.: Ia-bearing T lymphocytes in man. Their identification and role in the generation of allogeneic helper activity.J. Exp. Med. 148: 1423–1428, 1978
Halper, J., Fu, S. M., Wang, C. Y., Winchester, R., and Kunkel, H. G.: Patterns of expression of human “Ia-like” antigens during the terminal stages of B cell development.J. Immunol. 120: 1480–1484, 1977
Hansen, J. A., Fu, S. M., Antonelli, P., Kamoun, M., Hurley, J. N., Winchester, R. J., Dupont, B., and Kunkel, H. G.: B-lymphoid cell lines derived fromHLA-D homozygous donors.Immunogenetics 8: 51–64, 1979
Ko, H. S., Fu, S. M., Yu, D. T. Y., and Kunkel, H. G.: Ia determinants on stimulated T lymphocytes: Occurrence on mitogen- and antigen-activated T cells.J. Exp. Med. 150: 246, 1979
Kohler, G. and Milstein, C.: Continuous cultures of fused cells secreting antibody of predefined specificity.Nature 256: 495–497, 1975
Kohler, G., Hengartner, H., and Schulman, M.: Immunoglobulin production by lymphocyte hybridomas.Eur. J. Immunol. 8: 82–88, 1978
Laemmli, A. K.: Cleavage of structural protein during the assembly of the head of bacteriophage.Nature 277: 680–685, 1970
Loken, M. R. and Herzenberg, L. A.: Analysis of cell populations with a fluorescence — activated cell sorter.Ann. NY. Acad. Sci. 254: 163–171, 1975
McMichael, A. J., Pilch, J. R., Galfre, G., Mason, D. Y., Fabre, J. W., and Milstein, C.: A human thymocyte antigen defined by a hybrid myeloma monoclonal antibody.Eur. J. Immunol. 9: 205–210, 1979
NIAID Manual Of Tissue Typing Techniques: NIH lymphocyte microcytotoxicity technique. USDHEW No. (NIH) 76-545: 22–24, 1976
Nowinski, R. C., Lostrom, M. E., Tarn, M. R., Stone, M. R., and Burnette, W. N.: The isolation of hybrid cell lines producing monoclonal antibodies against the p15(E) protein of ecotropic murine leukemic viruses.Virology 93: 111–126, 1979
O'Farrell, P. H.: High resolution two-dimensional electrophoresis of proteins.J. Biol. Chem. 250: 4007–4021, 1975
Reinherz, E. L., Kung, P. C., Goldstein, G., and Schlossman, S. F.: A monoclonal antibody with selective reactivity with functionally mature human thymocytes and all peripheral human T cells.J. Immunol. 123: 1312–1317, 1979a
Reinherz, E. L., Kung, P. C., Goldstein, G., and Schlossman, S. F.: Separation of functional subsets of human T cells by a monoclonal antibody.Proc. Natl. Acad. Sci. U.S.A. 76: 4061–4065, 1979b
Stocker, J. W., Garotta, G., Hausmann, B., Trucco, M., and Ceppellini, R.: Separation of human B cells bearing HLA-DR antigens using a monoclonal antibody rosetting method.Tissue Antigens 13: 212–222, 1979
Swanstrom, R. and Shank, P. R.: X-ray intensifying screens greatly enhance the detection by autoradiography of the radioactive isotopes 32p and125I.Anal. Biochem. 86: 184–192, 1978
Trucco, M. M., Stocker, J. W., and Ceppellini, R.: Monoclonal antibodies against human lymphocyte antigens.Nature 273: 666–668, 1978
Vitetta, E. S., Baur, S., and Uhr, J.: Cell surface immunoglobulin. Isolation and characterization of immunoglobulin from mouse splenic lymphocytes.J. Exp. Med. 134: 242–264, 1971
Winchester, R. J., Wang, C. Y., Halper, J., and Hoffman, T.: Studies with B cell allo- and heteroantisera: Parallel reactivity and special properties.Scand. J. Immunol. 5: 745, 1976
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Hansen, J.A., Martin, P.J. & Nowinski, R.C. Monoclonal antibodies identifying a novel T-Cell antigen and Ia antigens of human lymphocytes. Immunogenetics 10, 247–260 (1980). https://doi.org/10.1007/BF01561573
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DOI: https://doi.org/10.1007/BF01561573