Abstract
Two-dimensional (2D) gel electrophoresis of immunoprecipitated HLA-DR antigens from eight homozygous typing cells (HTC) expressing the HLA-DRw8 specificity revealed a clustering of polymorphic β chain patterns into distinct electrophoretic variants. The variant patterns correlate with three discrete HLA-D clusters that are defined in the mixed leukocyte culture reaction (MLR) using DRw8-positive HTC. These HLA-D clusters have been provisionally designated Dw“8.1”, detected primarily in Caucasoids, Dw“8.2”, detected primarily in American Indians, and Dw“8.3”, detected predominantly in Orientals. All three HLA-Dw“8.1” cell lines express a single DR-locus product as defined by immunoprecipitation with a DR-specific monoclonal antibody, P4.1. This DR β chain is identical among the Dw“8.1” cell lines and different from the DR β chains of the Dw“8.2” and Dw“8.3” cell lines. Two separate Dw“8.2” HTC express a shared DR β chain that is slightly more basic than the 8.1 DR molecule; interestingly, one of these lines also expresses an additional DR-like β chain not found in the other cells. Thus, the two lines defining the Dw“8.2” cluster share one distinct class 11 molecule, but differ in another and therefore are not biochemically HLA-identical. Cells from the Dw“8.3” cluster are likewise distinct from all other Dw8 clusters. One additional DRw8-positive HTC has been analyzed and found to be distinct from the Dw“8.1”, “8.2” and “8.3” clusters by both MLR and 2D gels. lmmunoprecipitates using monoclonal antibody 1B5 [anti-DR and anti-DQ(DS)] identify additional polymorphic class II variants among the cell lines tested. These data indicate that HLA-DRw8 is a public serologic specificity present on class II molecules expressed on multiple distinct haplotypes. These haplotypes differ from each other in expression of polymorphic class II molecules encoded by at least two HLA loci. They also differ in HLA-D, even though they all type as HLA-DRw8 homozygous. In Dw“8.2”, variation in expressed β chains is not reflected in variation in HLA-D, indicating that MLR, as well as serologic typing, does not detect the full degree of allelic polymorphism within HLA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bodmer, W. F., Batchelor, J. R., Bodmer, J. G., Festenstein, H., and Morris, P. J. (eds.).: Histocompatibility Testing 1977, Munksgaard, Copenhagen, 1977
Crumpton, M., Bodmer, J., Bodmer, W., Heyes, J., Lindsay, J., and Rudd, C.: Biochemistry of class II antigens: Workshop report. In: Histocompatibility Testing, in press, 1984
Danilovs, J. A., Ayoub, G., and Terasaki, P. I.: Joint report: B lymphocyte isolation by thrombin-nylon wool. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 287–288, UCLA Press, Los Angeles, 1980
Dupont, B., Braun, D. W., Jr., Yunis, E. J., and Carpenter, C. B.: HLA-D by cellular typing. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 229–267, UCLA Press, Los Angeles, 1980
Gladstone, P. and Pious, D. A.: Stable variants affecting B cell alloantigens in human lymphoid cells. Nature 271: 459–461, 1978
Goyert, S. M., Shively, J. E., and Silver, J.: Biochemical characterization of a second family of human la molecules, HLA-DS, equivalent to murine I-A subregion molecules. J. Exp. Med. 156: 550–566, 1982
Groner, J. P., Watson, A. J., and Bach, F. H.: Dw/LD related molecular polymorphism of DR4 β-chains. J. Exp. Med. 157: 1687–1691, 1983
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 from HLA-D homozygous donors. Immunogenetics 8: 51–64, 1979
Jacobsen, S., Nepom, G. T., Richert, J. R., Biddison, W. E., and McFarland, H. F.: Identification of a specific HLA DR2 Ia molecule as a restriction element for measles virus specific HLA class II restricted cytotoxic T cell clones. J. Exp. Med., in press, 1984
Kaneko, T., Ishigamori, E., Otani, F., Watanabe, F., and Kashiwagi, N.: Characteristics of DYT and DKT2 specificities. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 891–892, UCLA Press, Los Angeles, 1980
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970
Lampson, L. A. and Levy, R.: Two populations of Ia-like molecules on a human B cell line. J. Immunol. 125: 293–299, 1980
Mickelson, E. M., Nisperos, B., Layrisse, Z., Kim, S. J., Thomas, E. D., and Hansen, J. A.: Analysis of the HLA-DRw8 haplotype: Recognition by HTC typing of three distinct antigen complexes in Caucasians, Native Americans, and Orientals. Immunogenetics 17: 399–410, 1983
Nepom, B. S., Nepom, G. T., Mickelson, E., Antonelli, 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
Nepom, B. S., Nepom, G. T., Mickelson, E., Schaller, J. G., Antonelli, P., and Hansen, J. A.: Specific HLA-DR4-associated histocompatibility molecules characterize patients with seropositive juvenile rheumatoid arthritis. J. Clin. Invest. 74: 287–291, 1984
Nepom, G. T., Nepom, B. S., Antonelli, P., Mickelson, E., Silver, J., Goyert, S. M., and Hansen, J. A.: The HLA-DR4 family of haplotypes consists of a series of distinct DR and DS molecules. J. Exp. Med. 159: 394–404, 1984a
Nepom, G. T., Nepom, B. S., Wilson, M. E., Mickelson, E., Amonelli, P., and Hansen, J. A.: Multiple Ia-like molecules characterize HLA-DR2-associated haplotypes which differ in HLA-D. Hum. Immunol. 10: 143–151, 1984b
Nose, Y., Sato, K, Nakagawa, N., Kondoh, K, Inouye, H., and Tsuji, K.: HLA-D clusters associated with DR4 in the Japanese population. Hum. Immunol. 5: 199–203, 1982
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: 1133–1140, 1977
Reinsmoen, N. L. and Bach, F. H.: Five HLA-D clusters with HLA-DR4 Hum. Immunol. 4: 249–258, 1982
Richiardi, R, Belvedere, M., Barelli, I., DeMaechi, M., and Curtoni, E. S.: Split of HLA-DRw2 into subtypic specificities closely correlated to two HLA-D products. Immunogenetics 7: 57–61, 1978
Sasazuki, T., Kaneoka, H., Tsuji, K., Nose, Y., Kashiwagi, N., and Kaneko, T.: HLA-D in Japan. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 274–276, UCLA Press, Los Angeles, 1980
Shackelford, D. A., Tompson, T. A., and Strominger, J. Y.: Analysis of HLA-DR antigens by using monoclonal antibodies: Recognition of conformational differences in biosynthetic intermediates. J. Immunol. 127: 1403–1410, 1981
Shackelford, D. A., Lampson, L. A., and Strominger, J. A.: Separation of three class II antigens from a homozygous human B cell line. J. Immunol. 130: 289–296, 1983
Van Rood, J. J., van Leeuwen, A., Keuning, J. J., and van Oud Ablas, A. B.: The serological recognition of the human MLC determinants using a modified cytotoxicity technique. Tissue Antigens 5: 73–79, 1975
Winchester, R. J. and Kunkel, H. G.: The human Ia system. Adv. Immunol. 28: 221–292, 1979
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Baldwin, G.C., Mickelson, E.M., Hansen, J.A. et al. Electrophoretic variation between class II molecules expressed on HLA-DRw8 homozygous typing cells reveals multiple distinct haplotypes. Immunogenetics 21, 49–60 (1985). https://doi.org/10.1007/BF00372241
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00372241