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H-2 Antigens pp 191-200 | Cite as

The Previously Defined Rd Class I Molecule is a Non-ß2M Associated Conformational Variant of Ld

  • Ted H. Hansen
  • Nancy B. Myers
  • Jill B. Keeney
  • David R. Lee
Part of the NATO ASI Series book series (NSSA, volume 144)

Abstract

Post-translational differences between murine class I molecules were compared. As determined in a pulse-chase experiment, the rate of N-linked oligosaccharide processing of Ld molecules was found to be 4–5 times slower than that of Dd, Kd or Ddml molecules. Furthermore, the relative molar ratio of ß2m to class I was found to be significantly lower with Ld than with Dd, Kd or Ddml molecules. Thus, both the slower processing and weaker ß2m association correlate with the previously reported lower cell surface expression of Ld compared to Dd, Kd or Ddml molecules. Since Ddml is a chimeric Dd/Ld molecule these results demonstrate that the amino terminus of the class I molecule influences processing, ß2m-association and cell surface expression.

The Rd molecule was initially defined by sequential immunoprecipitation using anti-Ld reactive mAbs 30–5–7 (α2 specific) and 28–14–8 (α3 specific). In contrast to Ld, the Rd molecule is only positive with mAb 28–14–8. In this communication we demonstrate that both Ld and Rd are products of the H-2L d gene using an H-2L d gene transfectant. Furthermore, the 30–5–7- Ld molecule (Rd) was found not to be ß2m associated, in contrast to the 30–5–7+ Ld molecule where ß2m was readily apparent. As much as 40% of the biosynthetically-labelled Ld molecules were found in this non-ß2m associated conformation. Thus, these studies suggest that Ld molecules have several unique properties that could all be related to an aberrant ß2m association. These properties include slower oligosaccharide processing, lower cell surface expression and detection of a non-ß2m associated conformational variant.

Keywords

Cell Surface Expression Sequential Precipitation Antigen Preparation Daudi Cell Complex Type Oligosaccharide 
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.

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References

  1. Allen, H., Fraser, J., Flyer, D., Calvin, S., and Flavell, R.: ß2-Microglobulin is not required for cell surface expression of the murine class I histocompatibility antigen H-2Db or of a truncated H-2Db. Proc. Natl. Acad. Sci. USA 83: 7447–7451, 1986.Google Scholar
  2. Anderson, M., Paabo, S., Nilsson, T., and Peterson, P.A.: Impaired intracellular transport of class I MHC antigens as a possible meansGoogle Scholar
  3. for adenoviruses to evade immune surveillance. Cell 43: 215–222, 1985.Google Scholar
  4. Beck, J.C., Hansen, T.H., Cullen, S.E., and Lee, D.R.: Slower processingGoogle Scholar
  5. weaker 82m association and lower surface expression of H-2Ld are influenced by its amino terminus. J. Immunol. 137:.916–922, 1986.Google Scholar
  6. Goodfellow, P.N., Jones, E.A., van Heyningen, V., Solomon, E., Bobrow, M., Miggiano, V., and Bodmer, W.F.: The 82-microglobulin genes is on chromosome 15 and not in the HL-A region. Nature 254: 267–269, 1975.Google Scholar
  7. Hansen, T.H., Ozato, K., Melino, M.R., Coligan, J.E., Kindt, T.J., Jandinski, J.J., and Sachs, D.H.: Immunochemical evidence in two haplotypes for at least three D region encoded molecules, D, L, and R. J. Immunol. 126: 1713–1716.Google Scholar
  8. Ivanyi, D., and Demant, P.: Capping experiments fail to reveal H-2Rd molecules in Dd region. Transplant. Proc. 15: 2039–2041, 1983.Google Scholar
  9. Koch, S., Robinson, P.J., Koch, N., and Hammerling, G.J.: Separation of H-2Dd, H-2Ld and H-2Rd by two-dimensional gel electrophoresis. Immunogenetics 17: 215–218, 1983.PubMedCrossRefGoogle Scholar
  10. Krangel, M.S., Orr, H.T., and Strominger, J.L.: Assembly and maturation of HLA-A and HLA-B antigens in vivo. Cell 18: 979–991, 1979.Google Scholar
  11. Lillehoj, E.P., Hansen, T.H., Sachs, D.H., and Coligan, J.E.: Primary structural evidence that the H-2Dq encodes at least three distinct gene products: Dq, Lq and Rq. Proc. Natl. Acad. Sci. USA 81: 2499 2503, 1984.Google Scholar
  12. Martinko, J.M., Anderson, S.J., and Potter, T.: A murine cell line defective in expression of several class I molecules. Immunogenetics 23: 64–66, 1986.Google Scholar
  13. Nichols, E.A., Krakauer, T., and Hansen, T.H.: Two-dimensional gel comparisons of murine H-2D region-associated antigens of different H-2 haplotypes. J. Immunol. 131: 2440–2444, 1983.PubMedGoogle Scholar
  14. Parnes, J.R., and Seidman, J.G.: Structure of wild-type and mutant mouse ß2-microglobulin. Cell 29: 661–669, 1982.Google Scholar
  15. Ploegh, H.L., Cannon, L.E., and Strominger, J.L.: Cell-free translation of the mRNAs for the heavy and light chains of HLA-A and HLA-B antigens. Proc. Natl. Acad. Sci. USA 76: 2273–2277, 1979.Google Scholar
  16. Potter, T.A., Hansen, T.H., Habbersett, R., Ozato, K., and Ahmed, A.:Google Scholar
  17. Flow microfluorometric analysis of H-2L expression. J. Immunol. 127:580–584, 1981.Google Scholar
  18. Potter, T.A., Boyer, C., Schmitt-Verhulst, A.M., Golstein, P., andGoogle Scholar
  19. Rajan, T.V.: Expression of H-2Db on the cell surface in the absence of detectable 132-microglobulin. J. Exp. Med. 160: 317–322, 1984.PubMedCrossRefGoogle Scholar
  20. Rosa, F., Berissi, H., Weissenbach, J., Maroteaux, L., Fellous, M., and Revel, M.: The ß2-microglobulin mRNA in human Daudi cells has a mutated initiation codon but is still inducible by interferon. EMBO J. 2: 239–243, 1983.PubMedGoogle Scholar
  21. Weiss, J.H., and Murre, C.: Differential expression of H-2Dd and H-2Ld histocompatibility antigens. J. Exp. Med. 161: 356–365, 1985.CrossRefGoogle Scholar
  22. Yokoyama, K., and Nathenson, S.G.: Intramolecular organization of thealloantigenic determinants and the 82-microglobulin binding site to different regions of the H-2Kb glycoprotein. J. Immunol. 130: 1419–1425, 1983.Google Scholar

Copyright information

© Springer Science+Business Media New York 1987

Authors and Affiliations

  • Ted H. Hansen
    • 1
  • Nancy B. Myers
    • 1
  • Jill B. Keeney
    • 1
  • David R. Lee
    • 1
  1. 1.Department of GeneticsUniversity School of MedicineWashingtonUSA

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