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A monoclonal antibody recognizing HLA-DR2 on malignant and activated cells

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Abstract

A monoclonal antibody, designated NDS15.38, which recognizes a polymorphic determinant of HLA-DR, was produced from a fusion in which mice were immunized with the human B lymphoblastoid cell line GIR2 (HLA type A1, B8, 27, Cw2, DR2, 7). NDS15.38 functions efficiently as an affinity column and purifies a two-chain complex of molecular weight 33 000 and 30 000 under reducing conditions. The monoclonal antibody reacts with HLA-DR2-positive B lymphoblastoid cell lines and B lymphocytes from patients with chronic lymphatic leukemia in an indirect radioactive binding assay. However, NDS15.38 does not appear to react with peripheral blood B lymphocytes from normal individuals. Using a peroxidase staining technique, NDS15.38 was shown to react with phytohemagglutin (PHA)-stimulated lymphocytes and with apparently activated B cells in the germinal centers of lymph nodes from individuals who were tissue typed as HLA-DR2. Thus it appears that NDS15.38 recognizes a polymorphic determinant of HLA-DR on malignant and stimulated cells, but not on resting cells.

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Abbreviations

BSA:

bovine serum albumin

CLL:

chronic lymphatic leukemia

CPM:

counts per minute

DAB:

diaminobenzidine tetrahydrochloride

DC:

sodium deoxycholate

HAT:

hypoxanthine, aminopterin, thymidine

IP:

intraperitoneal

PAGE:

polyacrylamide gel electrophoresis

PAP:

peroxidase-anti-peroxidase

PBL:

peripheral blood lymphocyte

PBS:

phosphate-buffered saline

PHA:

phytohemagglutinin

RAM:

immunoadsorbent-purified rabbit F(ab)2 anti-mouse F(ab)2

SDS:

sodium dodecyl sulphate

References

  • Accolla, R. S., Gross, N., Carrel, S., and Corte, G.: Distinct forms of both α and β subunits are present in the human la molecular pool. Proc. Natl. Acad. Sci. U.S.A. 78: 4549–4551, 1981

    Google Scholar 

  • Allison, J. P., Walker, L. E., Russell, W. A., Pellegrino, M. A., Ferrone, S., Reisfeld, R. A., Frelinger, J. A., and Silver, J.: Murine Ia and human DR antigens: homology of amino-terminal sequences. Proc. Natl. Acad. Sci. U.S.A. 75: 3953–3956, 1978

    Google Scholar 

  • Altevogt, P., Fohlman, J., Kurnick, J. T., Peterson, P., and Wigzell, H.: Biochemical comparison of HLA-DR molecules derived from autologous human T and B lymphocytes. Eur. J. Immunol. 10: 908–914, 1980

    Google Scholar 

  • Bhan, A. K., Nadler, L. M., Stashenko, P., McCluskey, R. T., and Schlossman, S. F.: Stages of B cell differentiation in human lymphoid tissues. J. Exp. Med. 154: 737–749, 1981

    Google Scholar 

  • Brodsky, F. M., Parham, P., and Bodmer, W. F.: Monoclonal antibodies to HLA-DRw determinants. Tissues Antigens 16: 30–48, 1980

    Google Scholar 

  • Corte, G., Calabi, F., Damiani, G., Bargellesi, A., Tosi, R., and Sorrentino, R.: Human Ia molecules carrying DC1 determinants differ in both α- and β-subunits from Ia molecules carrying DR determinants. Nature 292: 357–360, 1981

    Google Scholar 

  • de Kretser, T. A., Crumpton, M. J., Bodmer, J. G., and Bodmer, W. F.: Two-dimensional gel analysis of polypeptides precipitated by a polymorphic HLA-DR1, 2, w6 monoclonal antibody: evidence for a third locus. Eur. J. Immunol. 12: 600–606, 1982

    Google Scholar 

  • de Wolfe, W. C., Schlossman, S. F., and Yunis, E. J.: DRw antisera react with activated T cells. J. Immunol. 122: 1780–1784, 1979

    Google Scholar 

  • Dupont, B., Braun, D. W., Yunis, E. J., and Carpenter, C. B.: HLA-D by cellular typing. In P. I. Terasaki, (ed.): Histocompatibility Testing 1980, pp. 229–267, UCLA Tissue Typing Laboratory, Los Angeles, 1980

    Google Scholar 

  • Duquesnoy, R. J., Marrari, M., and Annen, K.: Identification of an HLA-DR-associated system of B-cell alloantigens. Transplant. Proc. 11: 1757–1760, 1979

    Google Scholar 

  • Fuggle, S. V., Errasti, P., Daar, A. S., Fabre, J. W., Ting, A., and Morris, P. J.: Localisation of MHC (HLA-ABC and DR) antigens in 46 kidneys. Differences in HLA-DR staining of tubules between kidneys. Transplantation, 35: 385–390, 1983

    Google Scholar 

  • Goodfellow, P. N., Jones, E. A., van Heynigen, V., Solomon, E., Bobrow, M., Miggiano, V., and Bodmer, W. F.: The B2- microglobulin gene is on chromosome 15 and not in the HL-A region. Nature 254: 267–269, 1975

    Google Scholar 

  • Hansen, J. A., Martin, P. J., Kamoun, M., Nisperos, B., and Thomas, E. D.: A supertypic HLA-DR specificity DR4+5 defined by murine monoclonal antibody. Hum. Immunol. 2: 103–111, 1981

    Google Scholar 

  • Johnson, J. P., Meo, T., Riethmüller, G., Schendel, D. J., and Wank, R.: Direct demonstration of an HLA-DR allotypic determinant on the low molecular (beta) subunit using a mouse monoclonal antibody specific for DR3. J. Exp. Med. 156: 104–111, 1982

    Google Scholar 

  • Kohler, G. and Milstein, C.: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495–497, 1975

    Google Scholar 

  • Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    Google Scholar 

  • Lakin, K. H. and Fabre, J. W.: Identification with a monoclonal antibody of a phylogenetically conserved brain-specific determinant on a 130 000 molecular weight glycoprotein of human brain. J. Neurochem. 37: 1170–1178, 1981

    Google Scholar 

  • Lampson, L. A. and Levy R.: Two populations of Ia-like molecules on a human B cell line. J. Immunol. 125: 293–299, 1980

    Google Scholar 

  • McKenzie, J. L. and Fabre, J. W.: Studies with a monoclonal antibody on the distribution of Thy-1 in the lymphoid and connective tissues of the dog. Transplantation 31: 275–282, 1981

    Google Scholar 

  • McMaster, W. R. and Williams, A. F.: Identification of la glycoproteins in rat thymus and purification from rat spleen. Eur. J. Immunol. 6: 426–433, 1979

    Google Scholar 

  • McMaster, W. R., Winearls, B., and Parham, P.: A monoclonal anti-rat Ia antibody which cross-reacts with a human HLA-DRw determinant. Tissue Antigens 14: 453–458, 1979

    Google Scholar 

  • 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 antibody. Eur. J. Immunol. 9: 200–204, 1979

    Google Scholar 

  • Makgoba, M. W., Fuggle, S. V., McMichael, A. J., and Morris, P. J.: HLA-DR products are a subset of human Ia antigens. Nature 301: 531–533, 1983

    Google Scholar 

  • Morris, R. J. and Williams, A. F.: Antigens on mouse and rat lymphocytes recognised by rabbit antiserum to rat brain: the quantitative analysis of zenogeneic antiserum. Eur. J. Immunol. 5: 274–281, 1975

    Google Scholar 

  • Park, M. S., Terasaki, P. I., Nakata, S., and Aoki, D.: Supertypic DR groups: MT1, MT2 and MT3. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 854–857, Munksgaard, Copenhagen, 1980

    Google Scholar 

  • Shackleford, D. A., Mann, D. L., van Rood, J. J., Ferrara, G. B., and Strominger, J. L.: Human B-cell alloantigens DCI, MT1 and LB12 are identical to each other but distinct from the HLA-DR antigen. Proc. Natl. Acad. Sci. U.S.A. 78: 4566–4570, 1981

    Google Scholar 

  • Silver, J., Walker, L. E., Reisfeld, R. A., Pellegrino, M. A., and Ferrone, S.: Structural studies of murine I-E and human DR antigens. Immunochemistry 16: 37–42, 1979

    Google Scholar 

  • Snary, D., Barnstable, C. J., Bodmer, W. F., Goodfellow, P., and Crumpton, M. J.: Human Ia antigens—purification and molecular structure. Cold Spring Harbor Symp. Quant. Biol. 41: 379–386, 1976

    Google Scholar 

  • Terasaki, P. I.: Nomenclature for factors of the HLA System1980. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 18–20, UCLA Tissue Typing Laboratory, Los Angeles, 1980

    Google Scholar 

  • Termijtelen, A., Boettcher, B., Bradley, B. A., D'Amaro, J., van Leeuwen, A., and van Rood, J. J.: DR typing in Australian aborigines. An indication for a second locus in the HLA-D region defined by serology. Tissue Antigens 16: 140–146, 1980

    Google Scholar 

  • Tosi, R., Tanigaki, N., Centis, D., Ferrara, G. B., and Pressman, D.: Immunological dissection of human Ia molecules. J. Exp. Med. 148: 1592–1611, 1978

    Google Scholar 

  • Trucco, M., de Petris, S., Garotta, G., and Cepellini, R.: Quantitative analysis of cell surface HLA structures by means of monoclonal antibodies. Hum. Immunol. 3: 233–243, 1980

    Google Scholar 

  • Werner, C. H., Klouda, P. J., Correa, M. C., Vassalli, P., and Jeannet M.: Isolation of B and T lymphocytes by nylon fiber columns. Tissue Antigens 9: 227–229, 1977

    Google Scholar 

  • Zola, H., Bradley, J., Macardle, P., McEvoy, R., and Thomas, M.: A hybridoma antibody which reacts with human cells carrying the DRw4 tissue type. Evidence for heterogeneity of DRw4. Transplantation 29: 72–73, 1980

    Google Scholar 

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Authors and Affiliations

  1. Nuffield Department of Surgery, John Radcliffe Hospital, University of Oxford, OX3 9DU, England

    Susan V. Fuggle, Jean Kirkley, Alan Ting & Peter J. Morris

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  1. Susan V. Fuggle
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  2. Jean Kirkley
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  3. Alan Ting
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  4. Peter J. Morris
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Fuggle, S.V., Kirkley, J., Ting, A. et al. A monoclonal antibody recognizing HLA-DR2 on malignant and activated cells. Immunogenetics 18, 415–428 (1983). https://doi.org/10.1007/BF00372473

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  • DOI: https://doi.org/10.1007/BF00372473

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