Abstract
We have analyzed the HLA class II gene products from HLA-DRw6 homozygous cells. Epstein-Barr virus-transformed B-cell lines were internally labeled with [35S]-methionine. An NP-40 lysate of the cells was subjected to immunoprecipitation, first with a DRw52-like-specific monoclonal antibody and subsequently with a DR-specific framework antibody. The DR region-encoded gene products were analyzed by one-dimensional gel isoelectric focusing and two-dimensional gel electrophoresis. It is shown that DRw6 homozygous cell lines contain at least two nonallelic DR β chains, one carrying a DRw52 determinant and one DRw52-negative population. Both chains appear to be polymorphic between the cellularly defined subtypes of DRw6. The determinant responsible for the differential mixed lymphocyte culture reactivity of Dw18 and Dw19 cells resides on the DRw52-positive population, whereas the Dw6-Dw9 differences are attributed to determinants on both populations of DR light chains. The Dw16-derived DRw52+ chain much resembles the Dw18 DRw52+ light chain whereas there is a clear-cut difference between these two subtypes in the DRw52− population. We conclude that, for DRw6 homozygous cells, the cellularly recognized D determinants are probably located on DR-encoded molecules, both DRw52+ and DRw52−, and that charge shift of these chains is at least partly responsible for differential recognition of these cells in mixed lymphocyte cultures.
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Abbreviations
- MLC:
-
mixed lymphocyte culture
- 1D-IEF:
-
one-dimensional isoelectric focusing
- 2D:
-
two-dimensional
- moab:
-
monoclonal antibody
References
Bach, F. H.: The HLA class II genes and products: The HLA-D region. Immunol. Today 6: 89–94, 1985
Benacerraf, B.: Role of MHC gene products in immune regulation. Science 212: 1229–1238, 1981
Bodmer, J. and Bodmer, W.: Histocompatibility 1984. Immunol. Today 4: 219–225, 1984
Bodmer, J., Kennedy, L., Aizawa, M., Dawkins, R., Lepage, V., Mazilli, M. C., and Richiardi, P.: D region monoclonal antibodies. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 217–236, Springer Verlag, Berlin, Heidelberg, 1984
Bontrop, R. E., Koning, F., von Miltenburg, R., Gerrets, R., Thompson, A., and Giphart, M. J.: Heterogeneity of HLA class II gene products as studied in a family by sequential immunoprecipitation. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 526–530, Springer Verlag, Berlin, Heidelberg, 1984
Bosch, M. L., Schreuder, I., Spits, H., Termijtelen, A., Tilanus, M. G. J., and Giphart, M. J.: Polymorphisms within the HLA-DRw6 haplotype. I. Restriction fragment length variation and its correlation with serology. J. Immunol. 134: 3212–3217, 1985
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
Groner, J. P., Watson, A. J., and Bach, F. H.: DW/LD related molecular polymorphism of DR4 chains. J. Exp. Med. 157: 1687–1691, 1983
Hendriks, G. F. J., Persijn, G. G., Lansbergen, Q., D'Amaro, J., Schreuder, G. M. T., Cohen, B., and van Rood, J. J.: Excellent outcome after transplantation of renal allografts from HLA-DRw6-positive donors even in HLA-DR mismatches. Lancet 2: 187–189, 1983
Hurley, C. K., Giles, R. C., and Capra, J. D.: The human MHC: Evidence for multiple HLA D region genes. Immunol. Today 4: 219–225, 1983
Hurley, C. K., Giles, R. C., Nunez, G., DeMars, R., Nadler, L., Winchester, R., Stastny, P., and Capra, J. D.: Molecular localization of human class II MT2 and MT3 determinants. J. Exp. Med. 160: 472–493, 1984
Karr, R. W., Kannapell, C. C., Stein, J. A., Gebel, H. M., Mann, D. L., Duquesnoy, R. J., Fuller, T. C., Rodey, G. E., and Schwartz, B. D.: Molecular relationships of the human B cell alloantigens MT2, MB3, MT4, and DR5. J. Immunol. 128: 1809–1818, 1982
Kaufman, J. F., Anderson, R. L., and Strominger, J. L.: HLA DR antigens have polymorphic light chains and invariant heavy chains as assessed by lysine containing tryptic peptide analysis. J. Exp. Med. 152: 37s-53s, 1980
Kessler, S. W.: Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: Parameters of the interaction of antibody-antigen complexes with a protein A. J. Immunol. 115: 1617–1624, 1975
Koning, F., Schreuder, G. M. T., Giphart, M. J., and Bruning, J. W.: A monoclonal antibody with MT2-specificity: Serology and biochemistry. Hum. Immunol. 9: 221–230, 1984
Larhammar, D., Servenieus, B., Rask, L., and Peterson, P. A.: Characterization of an HLA-DR β pseudogene. Proc. Natl. Acad. Sci. U.S.A. 82: 1475–1479, 1985
Long, E. O., Wake, C. T., Gorski, J., and Mach,B.: Complete sequence of an HLA DR chain deduced from a cDNA clone and identification of multiple non-allelic DR β chain genes. EMBO J. 2: 389–394, 1983
Markert, M. L. and Cresswell, P.: Human B cell alloantigens: Expression of MB and MT determinants. J. Immunol. 128: 2004–2008, 1982
Mölders, H. H., Breuning, M. H., Ivanyi, P., and Ploegh, H. L.: Biochemical analysis of variant HLA-B27 antigens. Hum. Immunol. 6: 111–117, 1983
Nepom, B. S., Nepom, G. T., Michelson, E., Antarelli, P., and Hansen, J. A.: Electrophoretic analysis of human HLA-DR antigens from HLA-DR4 homozygous cell lines: Correlation between β-chain diversity and HLA-D. Proc. Natl. Acad. Sci. U.S.A. 80: 6962–6966, 1983
Nishimura, Y., Tsukamoto, K., Sone, T., Hirayama, K., Takenouchi, T., Ogasawara, K., Kasahara, M., Wakisaka, A., Aizawa, M., and Sasazuki, T.: Molecular and functional analysis of class II molecules characteristic to HLA-Dw2 and Dw12. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 504–508, Springer Verlag, Berlin, Heidelberg, 1984
O'Farrell, P. Z., Goodman, H. M., and O'Farrell, P. H.: High resolution two dimensional electrophoresis of basic as well as acidic proteins. Cell 12: 1133–1142, 1975
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, UCLA Tissue Typing Laboratory, Los Angeles, 1980
Pulleyblank, D. E. and Booth, G. M.: Improved methods for the fluorographic detection of weak β-emitting radioisotopes in agarose and acrylamide gel-electrophoresis media. J. Biomed. Biophys. Methods 4: 339–346, 1981
Rebai, N., Malissen, B., Dieres, M., Accolla, R. S., Corte, G., and Mawas, C.: Distinct HLA-DR epitopes and distinct families of HLA-DR molecules defined by 15 monoclonal antibodies either anti-DR or allo-anti-Ia cross reacting with human DR molecules. Eur. J. Immunol. 13: 106–111, 1983
Reinsmoen, N. L., Sazazuki, T., Kaneoka, H., Ohta, N., Noreen, H. J., Greenberg, L., and Kersey, J. H.: Two distinct HLA-D specificities (DHO and Dw2) in linkage with HLA-DRw2 as defined in white and Japanese populations]. Transplant. Proc. 10: 789–791, 1979
Schreuder, G. M. T., Parlevliet, J., Termijtelen, A., and van Rood, J. J.: A reanalysis of the HLA-DRw6 complex. Tissue Antigens 21: 62–74, 1983
Schreuder, G. M. T., Doxiadis, I., Parlevliet, J., and Grosse-Wilde, H.: HLA-DR, DQ, LB and TA10 spec cities of Ninth Workshop Homozygous typing cells. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 243–248, Springer Verlag, Berlin, Heidelberg, 1984
Schwartz, R. H.: The role of gene products of the major histocompatibility complex in T cell activation and cellular interactions. In W. E. Paul (ed.): Fundamental Immunology, pp. 379–438, Raven Press, New York, 1984
Shackelford, D. A. and Strominger, J. L.: Demonstration of structural polymorphism among HLA-DR light chains by two dimensional gel-electrophoresis. J. Exp. Med. 151: 144–165, 1980
Shackelford, D. A., Mann, D. L., van Rood, J. J., Ferrara, G. B., and Strominger, J. L.: Human B cell alloantigens DC1, 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
Strominger, J. L., Engelhard, V. H., Fuks, A., Guild, B. C., Hyafil, F., Kaufman, J. F., Korman, A. J., Kostyk, T. G., Krangel, M. S., Lancet, D., Lopez de Castro, J. A., Mann, D. L., Orr, H. T., Parham, P., Parker, K. C., Ploegh, H. L., Pober, J. S., Robb, R. J., and Shackelford, D. A.: The biochemical analysis of products of the major histocompatibility complex. In B. Benacerraf and M. Dorf (eds.): The Role of the Major Histocompatibility Complex in Immunobiology, pp. 115–172, Gorland Publishing Inc., New York, 1981
Tanigaki, N., Tosi, R., Sagawa, K., Minowada, J., and Ferrara, G. B.: The distribution of DR5, MT2, and MB3 specificities on human la subsets. Immunogenetics 17: 371–386, 1983
Termijtelen, A. and Mickelson, E.: First level testing of Dw6 HTCs. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 274–276, Springer Verlag, Berlin, Heidelberg, 1984
Termijtelen, A., Schreuder, G. M. T., Mickelson, E. M., and van Rood, J. J.: Ninth International Histocompatibility Workshop pretesting of Dw6 HTCs: Two subtypes of Dw6. Tissue Antigens 24: 10–17, 1984
Thorsby, E. and Piazza, A.: Typing for HLA D (CD-1 or MLC) determinants. In F. Kissmeyer Nielsen (ed.): Histocompatibility Testing 1975, pp. 414–434, Munksgaard, Copenhagen, 1975
Van Rood, J. J., van Leeuwen, A., Keuning, J. J., and Blussé van Oud Alblas, A.: The serological recognition of the human MLC determinants using a modified cytotoxicity technique. Tissue Antigens 5: 73–79, 1975
Van Rood, J. J., van Leeuwen, A., Jonker, M., Termijtelen, A., and Bradley, B. A.: Polymorphic B cell determinants in man. Cold Spring Harbor Symp. Quant. Biol. 4: 417–427, 1977
WHO Nomenclature Report 1984: Nomenclature for factors of the HLA system 1984. Tissue Antigens 24: 73–80, 1984
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Bosch, M.L., Termijtelen, A., Gerrets, R. et al. Polymorphisms within the HLA-DRw6 haplotype. Immunogenetics 22, 23–33 (1985). https://doi.org/10.1007/BF00430591
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DOI: https://doi.org/10.1007/BF00430591