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Immunogenetics

, Volume 18, Issue 6, pp 625–637 | Cite as

Structural polymorphism of the DC1 light chains from DR1-, DR2-, and DRw6-positive cell lines

  • Deborah A. Shackelford
  • Beatrice Eibl
  • Jack L. Strominger
Article

Abstract

The heavy and light chain subunits of the DC1 antigen from several cell lines expressing different DR specificities were compared by two-dimensional (2-D) gel electrophoresis. The DC1 light chains from cell lines typed as DR1, DR2, or DRw6 differed in their isoelectric points. No difference in charge was observed for the DC 1 heavy chain from the three cell lines. The DC1 light chains from different DRw6-positive cell lines were also found to be structurally polymorphic. The DRw6 cell lines examined did not all express the same DR-like light chains, indicating that the DRw6 specificity is biochemically complex, which is in agreement with the serologic studies of others.

Keywords

Light Chain Heavy Chain Isoelectric Point Serologic Study Structural Polymorphism 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations used in this paper

2-D

two-dimensional

Tm

tunicamycin

mAb

monoclonal antibody

SaCI

Staphylococcus aureus Cowan I strain bacteria

RaMIgG

rabbit anti-mouse IgG serum

SDS-PAGE

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

IEF

isoelectric focusing

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References

  1. Andrews, D. W., Bono, M. R., and Strominger, J. L.: A homozygous human cell line contains three subsets of HLA-DR-like antigens distinguishable by amino acid sequencing. Biochemistry 21: 6625–6628, 1982Google Scholar
  2. Bonner, W. M. and Laskey, R. A.: A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46: 83–88, 1974Google Scholar
  3. Bono, M.R. and Strominger, J.L.: Direct evidence of homology between human DC-1 antigen and murine I-A molecules. Nature 299: 836–838, 1982Google Scholar
  4. Bono, R., Hyafil, F., Kalil, J., Koblar, V., Wiels, J., Wollman, E., Mawas, C., and Fellous, M.: Monoclonal antibodies against HLA-DRw antigens. Transplant. Clin. Immunol. 11: 109–120, 1979Google Scholar
  5. Brodsky, F. M., Parham, P., and Bodmer, W. F.: Monoclonal antibodies to HLA-DRw determinants. Tissue Antigens 16: 30–48, 1980Google Scholar
  6. Charron, D. J. and McDevitt, H. O.: Characterization of HLA-DR region antigens by two-dimensional gel electrophoresis. Molecular genotyping. J. Exp. Med. 152, 18s-36s, 1980Google Scholar
  7. Corte, G., Calabi, F., Damiani, G., Bargellesi, A., Tosi, R., and Sorrentino, R.: Human Ia molecules carrying DO determinants differ in both α and β-subunits from la molecules carrying DR determinants. Nature 292: 357–360, 1981Google Scholar
  8. De Kretser, T. A., Crumpton, M. J., Bodmer, J. G., and Bodmer, W. F.: Two-dimensional gel analysis of the polypeptides precipitated by a polymorphic HLA-DR 1, 2, w6 monoclonal antibody: Evidence for a third locus. Eur. J. Immunol. 12: 600–606, 1982aGoogle Scholar
  9. De Kretser, T. A., Crumpton, M. J., Bodmer, J. G., and Bodmer, W. F.: Demonstration of two distinct light chains in HLA-DR-associated antigens by two-dimensional gel electrophoresis. Eur. J. Immunol. 12: 214–221, 1982bGoogle Scholar
  10. Doran, T. J., Honeyman, M., Zwinge, W., Edwards, C. M., and Bashir, H. V.: Heterogeneity of the DRw6 antigen. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 808–809, UCLA Press, Los Angeles, 1980Google Scholar
  11. Duquesnoy, R. J.: Definitionof three subdivisions of HLA-DRw6. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 817–818, UCLA Press, Los Angeles, 1980Google Scholar
  12. Duquesnoy, R. J., Marrari, M., and Armen, K.: Identification of an HLA-DR-associated system of B-cell alloantigens. Transplant. Proc. 11: 1757–1760, 1979Google Scholar
  13. Fauchet, R., Bouhallier, O., Jalais, E., Dejour, G., Genetet, B., and Simon, M.: Serological pattern of DRw6. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 814–816, UCLA Press, Los Angeles, 1980Google Scholar
  14. Finn, O. J. and Levy, R.: Multiple HLA-DR antigens: Detection with monoclonal antibodies and translation in vitro. Proc. Natl. Acad. Sci. U.S.A. 79: 2658–2662, 1982Google Scholar
  15. Freed, J. H.: Biochemistry of the murine la antigens. In S. Ferrone and C. David (eds.): Ia Antigens. Volume I: Mice, pp. 1–36, CRC Press, Boca Raton, Florida, 1982Google Scholar
  16. 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: 550–560, 1982Google Scholar
  17. Hurley, C. K., Shaw, S., Nadler, L., Schlossman, S., and Capra, J. D.: Alpha and beta chains of SB and DR antigens are structurally distinct. J. Exp. Med. 156: 1557–1562, 1982Google Scholar
  18. Karr, R. W., Kannapell, C. C., Stein, J. A., Fuller, T. C., Duquesnoy, R. J., Rodey, G. E., Mann, D. L., Gebel, H. M., and Schwartz, B. D.: Demonstration of a third structurally distinct human Ia beta chain by two-dimensional gel electrophoresis. J. Exp. Med. 156: 652–657, 1982Google Scholar
  19. Kaufman, J. F. and Strominger, J. L.: Both chains of HLA-DR bind to the membrane with a penultimate hydrophobic region and the heavy chain is phosphorylated at its hydrophilic carboxy terminus. Proc. Natl. Acad. Sci. U.S.A. 76: 6304–6308, 1979Google Scholar
  20. Kaufman, J. F. and Strominger, J. L.: HLA-DR light chain has a polymorphic N-terminal region and a conserved Ig-like C-terminal region. Nature 297: 694–696, 1982Google Scholar
  21. Kaufman, J. F., Andersen, 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, 1980Google Scholar
  22. Kaur, D. and Mittal, K. K.: Fine serological subdivisions of DRw6. In P. I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 810–811, UCLA Press, Los Angeles, 1980Google Scholar
  23. Kessler, S. W.: Rapid isolation of antigens from cells with a Staphylcoccal protein A-antibody adsorbent: Parameter of the interaction of antibody-antigen complexes with protein A. J. Immunol. 115: 1617–1624, 1975Google Scholar
  24. Korman, A. J., Auffray, C., Schamboeck, A., and Strominger, J. L.: The amino acid sequence and gene organization of the heavy chain of the HLA-DR antigen: Homology to immunoglobulins. Proc. Natl. Acad. Sci. U.S.A. 79: 6013–6017, 1982Google Scholar
  25. Kratzin, H., Yang, C. Y., Gotz, H., Pauly, E., Kolbel, S., Egert, G., Thinnes, F. P., Wernet, P., Altevogt, P., and Hilschmann, N.: Primary structure of class II human histocompatibility antigens. Hoppe Seylers Z. Physiol. Biochem. 362, 1665–1669, 1981Google Scholar
  26. Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970Google Scholar
  27. Lampson, L. A. and Levy, R.: Two populations of la-like molecules on a human B cell line. J. Immunol. 125: 293–299, 1980Google Scholar
  28. Larhammar, D., Wiman, K., Schenning, L., Claesson, L., Gustafsson, K., Peterson, P. A., and Rask, L.: Evolutionary relationship between HLA-DR antigen β chains, HLA-A, B, C antigen subunits and immunoglobulin chains. Scand. J. Immunol. 14: 617–622, 1981Google Scholar
  29. Larhammar, D., Schenning, L., Gustafsson, K., Wiman, K., Claesson, L., Rask, L., and Peterson, P. A.: Complete amino acid sequence of an HLA-DR antigen-like β chain as predicated from the nucleotide sequence: Similarities with immunoglobulins and HLA-A, -B and -C antigens. Proc. Natl. Acad. Sci. U.S.A. 79: 3687–3691, 1982Google Scholar
  30. Long, E. O., Wake, C. T., Strubin, M.; Gross, N., Accolla, R., Carrel, S., and Mach, B.: Isolation of distinct cDNA clones encoding HLA-DRβ chain by use of an expression assay. Proc. Natl. Acad. Sci. U.S.A. 79: 7465–7469, 1982Google Scholar
  31. Markert, M. L. and Cresswell, P.: Polymorphism of human B-cell alloantigens: Evidence for three loci within the HLA system. Proc. Natl. Acad. Sci. U.S.A. 77: 6101–6104, 1980Google Scholar
  32. 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 Press, Los Angeles, 1980Google Scholar
  33. 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, 1980Google Scholar
  34. Shackelford, D. A. and Strominger, J. L.: Analysis of the oligosaccharides on the HLA-DR and DC1 B cell antigens. J. Immunol. 130: 274–282, 1983Google Scholar
  35. 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, 1981aGoogle Scholar
  36. Shackelford, D. A., Lampson, L. A., and Strominger, J. L.: Analysis of HLA-DR antigens by using monoclonal antibodies: Recognition of conformational differences in biosynthetic intermediates. J. Immunol. 127: 1403–1410, 1981bGoogle Scholar
  37. Shackelford, D. A., Kaufman, J. F., Korman, A. J., and Strominger, J. L.: HLA-DR antigens: Structure, separation of subpopulations, gene cloning and function. Immunol. Rev. 66: 133–187, 1982Google Scholar
  38. Shackelford, D. A., Lampson, L. A., and Strominger, J. L.: Separation of three class II antigens from a homozygous human B cell line. J. Immunol. 130: 289–296, 1983Google Scholar
  39. Springer, T. A., Kaufman, J. F., Siddoway, L. A., Mann, D. L., and Strominger, J. L.: Purification of HLAlinked B lymphocyte alloantigens in immunologically active form by preparative sodium dodecyl sulfate-gel electrophoresis and studies on their subunit association. J. Biol. Chem 252: 6201–6207, 1977Google Scholar
  40. Tanigaki, N., Tosi, R., Duquesnoy, R. J., and Ferrara, G. B.: Three Ia species with different structures and alloantigenic determinants in an HLA-homozygous cell lines. J. Exp. Med. 157: 231–247, 1983Google Scholar
  41. 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: 140–141, 1980Google Scholar
  42. Tosi, R., Tanigaki, N., Centis, N., Ferrara, G. B., and Pressman, D.: Immunological dissection of human Ia molecules. J. Exp. Med. 148, 1592–1611, 1978Google Scholar
  43. Vives, J., Ercilla, G., and Nunez, A.: The HLA-DRw6 complex. In P.I. Terasaki (ed.): Histocompatibility Testing 1980, pp. 812–813, UCLA Press, Los Angeles, 1980Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • Deborah A. Shackelford
    • 1
  • Beatrice Eibl
    • 1
  • Jack L. Strominger
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyHarvard UniversityCambridge

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