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. 1982 Apr;79(8):2658–2662. doi: 10.1073/pnas.79.8.2658

Multiple HLA-DR antigens: detection with monoclonal antibodies and translation in vitro.

O J Finn, R Levy
PMCID: PMC346260  PMID: 6979750

Abstract

L203 and L227 are mouse monoclonal antibodies specific for human HLA-DR antigens. Their reactivity with multiple, structurally different, HLA-DR antigens was examined by using radioiodinated human lymphoblastoid cells and cell lines, heterozygous or homozygous at the HLA-D locus. Antibody L203 precipitates DR antigens composed of 34,000 Mr heavy chains (alpha), and 28,000-29,000 Mr light chains (beta). Antibody L227 recognizes two other species of DR antigens. Their heavy chains have similar molecular weights of 34,000 but their light chains are different, one (beta) being 28,000-29,000 Mr and the other (beta') 23,000-25,000 Mr. Whereas L203 is directed against a combinatorial determinant generated by both DR chains, L227 is able to recognize each of the two light chains alone and reprecipitate them after their separate elution from sodium dodecyl sulfate gels. Poly(A)-containing mRNA was isolated from Raji cells and translated in vitro by microinjection in Xenopus oocytes. The oocytes translated and fully assembled all the DR molecules recognized by L203 and L227 antibodies.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allison J. P., Walker L. E., Russell W. A., Pellegrino M. A., Ferrone S., Reisfeld R. A., Frelinger J. A., Silver J. Murine Ia and human DR antigens: homology of amino-terminal sequences. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3953–3956. doi: 10.1073/pnas.75.8.3953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carrel S., Tosi R., Gross N., Tanigaki N., Carmagnola A. L., Accolla R. S. Subsets of human Ia-like molecules defined by monoclonal antibodies. Mol Immunol. 1981 May;18(5):403–411. doi: 10.1016/0161-5890(81)90102-4. [DOI] [PubMed] [Google Scholar]
  3. Charron D. J., McDevitt H. O. Analysis of HLA-D region-associated molecules with monoclonal antibody. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6567–6571. doi: 10.1073/pnas.76.12.6567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cook R. G., Siegelman M. H., Capra J. D., Uhr J. W., Vitetta E. S. Structural studies on the murine Ia alloantigens. IV. NH2-terminal sequence analysis of allelic products of the I-A and I-E/C subregions. J Immunol. 1979 Jun;122(6):2232–2237. [PubMed] [Google Scholar]
  5. Cook R. G., Uhr J. W., Capra J. D., Vitetta E. S. Structural studies on the murine IA alloantigens. II. Molecular weight characterization of the products of the I-A and I-E/C subregions. J Immunol. 1978 Dec;121(6):2205–2212. [PubMed] [Google Scholar]
  6. Dumont J. N. Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals. J Morphol. 1972 Feb;136(2):153–179. doi: 10.1002/jmor.1051360203. [DOI] [PubMed] [Google Scholar]
  7. Ferrone S., Allison J. P., Pellegrino M. A. Human DR (Ia-like) antigens: biological and molecular profile. Contemp Top Mol Immunol. 1978;7:239–281. doi: 10.1007/978-1-4757-0779-3_8. [DOI] [PubMed] [Google Scholar]
  8. Goyert S. M., Silver J. Isolation of I-A subregion-like molecules from subhuman primates and man. Nature. 1981 Nov 19;294(5838):266–268. doi: 10.1038/294266a0. [DOI] [PubMed] [Google Scholar]
  9. Gurdon J. B., Lane C. D., Woodland H. R., Marbaix G. Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells. Nature. 1971 Sep 17;233(5316):177–182. doi: 10.1038/233177a0. [DOI] [PubMed] [Google Scholar]
  10. Ikeman R. L., Sullivan A. K., Kositsky R., Bartok K., Jerry L. M. Molecular variants of human Ia-like antigens. Nature. 1978 Mar 16;272(5650):267–268. doi: 10.1038/272267a0. [DOI] [PubMed] [Google Scholar]
  11. Jones P. P. Analysis of H-2 and Ia molecules by two-dimensional gel electrophoresis. J Exp Med. 1977 Nov 1;146(5):1261–1279. doi: 10.1084/jem.146.5.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jones P. P., Murphy D. B., Hewgill D., McDevitt H. O. Detection of a common polypeptide chain in I--A and I--E sub-region immunoprecipitates. Mol Immunol. 1979 Jan;16(1):51–60. doi: 10.1016/0161-5890(79)90027-0. [DOI] [PubMed] [Google Scholar]
  13. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lampson L. A., Levy R. Two populations of Ia-like molecules on a human B cell line. J Immunol. 1980 Jul;125(1):293–299. [PubMed] [Google Scholar]
  16. Mann D. L., Abelson L., Harris S., Amos D. B. Second genetic locus in the HLA region for human B-cell alloantigens. Nature. 1976 Jan 15;259(5539):145–146. doi: 10.1038/259145a0. [DOI] [PubMed] [Google Scholar]
  17. Markert M. L., Cresswell P. Polymorphism of human B-cell alloantigens: evidence for three loci within the HLA system. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6101–6104. doi: 10.1073/pnas.77.10.6101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McMillan M., Cecka J. M., Murphy D. B., McDevitt H. O., Hood L. Structure of murine Ia antigens: partial NH2-terminal amino acid sequences of products of the I-E or I-C subregion. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5135–5139. doi: 10.1073/pnas.74.11.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  20. Park M. S., Terasaki P. I., Bernoco D., Iwaki Y. Evidence for a second B-cell locus separate from the DR locus. Transplant Proc. 1978 Dec;10(4):823–828. [PubMed] [Google Scholar]
  21. Pierres M., Rebouah J. P., Kourilsky F. M., Dosseto M., Mercier P., Mawas C., Malissen B. Cross-reactions between mouse Ia and human HLA-D/DR antigens analyzed with monoclonal alloantibodies. J Immunol. 1981 Jun;126(6):2424–2429. [PubMed] [Google Scholar]
  22. Quaranta V., Walker L. E., Pellegrino M. A., Ferrone S. Purification of immunologically functional subsets of human Ia-like antigens on a monoclonal antibody (Q5/13) immunoadsorbent. J Immunol. 1980 Oct;125(4):1421–1425. [PubMed] [Google Scholar]
  23. Shackelford D. A., Strominger J. L. Demonstration of structural polymorphism among HLA-DR light chains by two-dimensional gel electrophoresis. J Exp Med. 1980 Jan 1;151(1):144–165. doi: 10.1084/jem.151.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Silver J., Walker L. E., Reisfeld R. A., Pellegrino M. A., Ferrone S. Structural studies of murine I-E and human DR antigens. Mol Immunol. 1979 Jan;16(1):37–42. doi: 10.1016/0161-5890(79)90025-7. [DOI] [PubMed] [Google Scholar]
  25. Springer T. A., Kaufman J. F., Terhorst C., Strominger J. L. Purification and structural characterisation of human HLA-linked B-cell antigens. Nature. 1977 Jul 21;268(5617):213–218. doi: 10.1038/268213a0. [DOI] [PubMed] [Google Scholar]
  26. Suciu-Foca N., Godfrey M., Rohowsky C., McKiernan P., Susinno E., Broell J. Evidence that HLA-D is different from DR and that two different HLA loci code for B-cell antigens. Transplant Proc. 1979 Dec;11(4):1781–1787. [PubMed] [Google Scholar]
  27. Tanigaki N., Tosi R., Pressman D., Ferrara G. B. Molecular identification of human Ia antigens coded for by a gene locus closely linked to HLA-DR locus. Immunogenetics. 1980;10(2):151–167. doi: 10.1007/BF01561564. [DOI] [PubMed] [Google Scholar]
  28. Tsuji K., Nose Y., Komori K., Tajika R., Nakagawa M., Inouye H., Yamamura M. Probable existence of four subloci in the HLA-DR (or D) region. Transplant Proc. 1979 Dec;11(4):1792–1795. [PubMed] [Google Scholar]
  29. Winchester R. J., Kunkel H. G. The human Ia system. Adv Immunol. 1979;28:221–292. [PubMed] [Google Scholar]

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