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. 1982 Oct;79(20):6327–6331. doi: 10.1073/pnas.79.20.6327

Identification of human genomic clones coding the major histocompatibility antigens HLA-a2 and HLA-B7 by DNA-mediated gene transfer.

J A Barbosa, M E Kamarck, P A Biro, S M Weissman, F H Ruddle
PMCID: PMC347114  PMID: 6959120

Abstract

We have screened a large number of isolated human genomic clones that hybridize to a cloned HLA cDNA probe for their ability to direct the synthesis of HLA-A, -B, and -C surface antigens on mouse L cells following DNA-mediated gene transfer. The surface expression of human histocompatibility antigens, monitored by indirect immunofluorescence and the fluorescence-activated cell sorter, was examined at 60 hr after transfection and on hypoxanthine/aminopterin/thymidine-resistant (HATR) populations derived from cotransfer with the herpes simplex virus thymidine kinase gene. Two unique genomic clones designated JY B3.2 and JY 158, isolated from the human lymphoblastoid cell line JY (homozygous HLA-A2, -B7), were shown to contain gene sequences capable of directing expression of an HLA-A, -B, -C determinant. By using allo-specific antibodies, the gene products of these clones were identified as HLA-A2 and HLA-B7, respectively. HATR clonal populations isolated from cotransfections with these genomic clones displayed varying levels of surface HLA expression that correlated with the number of intact donor HLA sequences present in the cells. In general, these levels of expression were stable during 3 months in culture. This system provides a powerful tool for the study of human surface antigen gene structure, expression, and function on a mouse cell background.

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Selected References

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

  1. Arce-Gomez B., Jones E. A., Barnstable C. J., Solomon E., Bodmer W. F. The genetic control of HLA-A and B antigens in somatic cell hybrids: requirement for beta2 microglobulin. Tissue Antigens. 1978 Feb;11(2):96–112. doi: 10.1111/j.1399-0039.1978.tb01233.x. [DOI] [PubMed] [Google Scholar]
  2. Bach F. H., van Rood J. J. The major histocompatibility complex--genetics and biology. (First of three parts). N Engl J Med. 1976 Oct 7;295(15):806–813. doi: 10.1056/NEJM197610072951504. [DOI] [PubMed] [Google Scholar]
  3. Barnstable C. J., Bodmer W. F., Brown G., Galfre G., Milstein C., Williams A. F., Ziegler A. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Cell. 1978 May;14(1):9–20. doi: 10.1016/0092-8674(78)90296-9. [DOI] [PubMed] [Google Scholar]
  4. Brodsky F. M., Parham P., Barnstable C. J., Crumpton M. J., Bodmer W. F. Monoclonal antibodies for analysis of the HLA system. Immunol Rev. 1979;47:3–61. doi: 10.1111/j.1600-065x.1979.tb00288.x. [DOI] [PubMed] [Google Scholar]
  5. Cami B., Brégégère F., Abastado J. P., Kourilsky P. Multiple sequences related to classical histocompatibility antigens in the mouse genome. Nature. 1981 Jun 25;291(5817):673–675. doi: 10.1038/291673a0. [DOI] [PubMed] [Google Scholar]
  6. Chang L. J., Gamble C. L., Izaguirre C. A., Minden M. D., Mak T. W., McCulloch E. A. Detection of genes coding for human differentiation markers by their transient expression after DNA transfer. Proc Natl Acad Sci U S A. 1982 Jan;79(1):146–150. doi: 10.1073/pnas.79.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dausset J. The major histocompatibility complex in man. Science. 1981 Sep 25;213(4515):1469–1474. doi: 10.1126/science.6792704. [DOI] [PubMed] [Google Scholar]
  8. Evans G. A., Margulies D. H., Camerini-Otero R. D., Ozato K., Seidman J. G. Structure and expression of a mouse major histocompatibility antigen gene, H-2Ld. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1994–1998. doi: 10.1073/pnas.79.6.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goodenow R. S., McMillan M., Orn A., Nicolson M., Davidson N., Frelinger J. A., Hood L. Identification of a BALB/c H-2Ld gene by DNA-mediated gene transfer. Science. 1982 Feb 5;215(4533):677–679. doi: 10.1126/science.7058331. [DOI] [PubMed] [Google Scholar]
  10. Hood L., Campbell J. H., Elgin S. C. The organization, expression, and evolution of antibody genes and other multigene families. Annu Rev Genet. 1975;9:305–353. doi: 10.1146/annurev.ge.09.120175.001513. [DOI] [PubMed] [Google Scholar]
  11. Huttner K. M., Scangos G. A., Ruddle F. H. DNA-mediated gene transfer of a circular plasmid into murine cells. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5820–5824. doi: 10.1073/pnas.76.11.5820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jordan B. R., Bregegere F., Kourilsky P. Human HLA gene segment isolated by hybridization with mouse H-2 cDNA probes. Nature. 1981 Apr 9;290(5806):521–523. doi: 10.1038/290521a0. [DOI] [PubMed] [Google Scholar]
  14. Kamarck M. E., Barbosa J. A., Ruddle F. H. Somatic cell genetic analysis of HLA-A, B, C and human beta 2-microglobulin expression. Somatic Cell Genet. 1982 May;8(3):385–402. doi: 10.1007/BF01538895. [DOI] [PubMed] [Google Scholar]
  15. Klein G., Terasaki P., Billing R., Honig R., Jondal M., Rosén A., Zeuthen J., Clements G. Somatic cell hybrids between human lymphoma lines. III. Surface markers. Int J Cancer. 1977 Jan;19(1):66–76. doi: 10.1002/ijc.2910190110. [DOI] [PubMed] [Google Scholar]
  16. Kvist S., Bregegere F., Rask L., Cami B., Garoff H., Daniel F., Wiman K., Larhammar D., Abastado J. P., Gachelin G. cDNA clone coding for part of a mouse H-2d major histocompatibility antigen. Proc Natl Acad Sci U S A. 1981 May;78(5):2772–2776. doi: 10.1073/pnas.78.5.2772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. López de Castro J. A., Strominger J. L., Strong D. M., Orr H. T. Structure of crossreactive human histocompatibility antigens HLA-A28 and HLA-A2: possible implications for the generation of HLA polymorphism. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3813–3817. doi: 10.1073/pnas.79.12.3813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Margulies D. H., Evans G. A., Flaherty L., Seidman J. G. H-2-like genes in the Tla region of mouse chromosome 17. Nature. 1982 Jan 14;295(5845):168–170. doi: 10.1038/295168a0. [DOI] [PubMed] [Google Scholar]
  19. McMichael A. J., Ting A., Zweerink H. J., Askonas B. A. HLA restriction of cell-mediated lysis of influenza virus-infected human cells. Nature. 1977 Dec 8;270(5637):524–526. doi: 10.1038/270524a0. [DOI] [PubMed] [Google Scholar]
  20. Milman G., Herzberg M. Efficient DNA transfection and rapid assay for thymidine kinase activity and viral antigenic determinants. Somatic Cell Genet. 1981 Mar;7(2):161–170. doi: 10.1007/BF01567655. [DOI] [PubMed] [Google Scholar]
  21. Ohta T. Allelic and nonallelic homology of a supergene family. Proc Natl Acad Sci U S A. 1982 May;79(10):3251–3254. doi: 10.1073/pnas.79.10.3251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Orr H. T., Bach F. H., Ploegh H. L., Strominger J. L., Kavathas P., DeMars R. Use of HLA loss mutants to analyse the structure of the human major histocompatibility complex. Nature. 1982 Apr 1;296(5856):454–456. doi: 10.1038/296454a0. [DOI] [PubMed] [Google Scholar]
  23. Parham P., Bodmer W. F. Monoclonal antibody to a human histocompatibility alloantigen, HLA-A2. Nature. 1978 Nov 23;276(5686):397–399. doi: 10.1038/276397a0. [DOI] [PubMed] [Google Scholar]
  24. Ploegh H. L., Orr H. T., Strominger J. L. Major histocompatibility antigens: the human (HLA-A, -B, -C) and murine (H-2K, H-2D) class I molecules. Cell. 1981 May;24(2):287–299. doi: 10.1016/0092-8674(81)90318-4. [DOI] [PubMed] [Google Scholar]
  25. Ploegh H. L., Orr H. T., Strominger J. L. Molecular cloning of a human histocompatibility antigen cDNA fragment. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6081–6085. doi: 10.1073/pnas.77.10.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Singer D. S., Camerini-Otero R. D., Satz M. L., Osborne B., Sachs D., Rudikoff S. Characterization of a porcine genomic clone encoding a major histocompatibility antigen: expression in mouse L cells. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1403–1407. doi: 10.1073/pnas.79.5.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sood A. K., Pereira D., Weissman S. M. Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer. Proc Natl Acad Sci U S A. 1981 Jan;78(1):616–620. doi: 10.1073/pnas.78.1.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Steinmetz M., Frelinger J. G., Fisher D., Hunkapiller T., Pereira D., Weissman S. M., Uehara H., Nathenson S., Hood L. Three cDNA clones encoding mouse transplantation antigens: homology to immunoglobulin genes. Cell. 1981 Apr;24(1):125–134. doi: 10.1016/0092-8674(81)90508-0. [DOI] [PubMed] [Google Scholar]
  29. Steinmetz M., Moore K. W., Frelinger J. G., Sher B. T., Shen F. W., Boyse E. A., Hood L. A pseudogene homologous to mouse transplantation antigens: transplantation antigens are encoded by eight exons that correlate with protein domains. Cell. 1981 Sep;25(3):683–692. doi: 10.1016/0092-8674(81)90175-6. [DOI] [PubMed] [Google Scholar]
  30. Steinmetz M., Winoto A., Minard K., Hood L. Clusters of genes encoding mouse transplantation antigens. Cell. 1982 Mar;28(3):489–498. doi: 10.1016/0092-8674(82)90203-3. [DOI] [PubMed] [Google Scholar]
  31. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  32. Wilson J. T., Wilson L. B., deRiel J. K., Villa-komaroff L., Efstratiadis A., Forget B. G., Weissman S. M. Insertion of synthetic copies of human globin genes into bacterial plasmids. Nucleic Acids Res. 1978 Feb;5(2):563–581. doi: 10.1093/nar/5.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zinkernagel R. M., Doherty P. C. Restriction of in vitro T cell-mediated cytotoxicity in lymphocytic choriomeningitis within a syngeneic or semiallogeneic system. Nature. 1974 Apr 19;248(5450):701–702. doi: 10.1038/248701a0. [DOI] [PubMed] [Google Scholar]

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