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. 1985 Dec 16;4(13A):3447–3453. doi: 10.1002/j.1460-2075.1985.tb04103.x

Hormonal regulation of cell surface expression of the major histocompatibility antigen H-2Ld in transfected cells.

H Ponta, R Ball, M Steinmetz, B Groner
PMCID: PMC554683  PMID: 3004961

Abstract

The murine major histocompatibility antigens are cell surface glycoproteins which play an important role in the recognition of foreign antigens by cytotoxic T lymphocytes. Modulation of the level of expression of histocompatibility antigens could therefore be useful for the study of the interaction between the antigen presenting cells and T lymphocytes. The glucocorticoid hormone-inducible promoter, located in the long terminal repeat of mouse mammary tumor virus, was used to replace the promoter region of a cloned H-2Ld class I gene. The chimeric gene was introduced into cultured cells. Glucocorticoid induction of MMTV LTR H-2Ld mRNA could be shown by blot analysis. An S1 nuclease protection assay indicated that the transfected cells accurately initiate the chimeric mRNA. Immunoprecipitation of H-2Ld protein with a specific monoclonal antibody showed inducibility also at the cellular protein level. Fluorescence-activated cell sorter analysis monitored a 3-fold increase of H-2Ld on the cell surface when the transfected cells were grown in the presence of dexamethasone. This increase of H-2Ld expression was accompanied by a corresponding decrease on the cell surface of the endogenous H-2Kk.

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Selected References

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

  1. Bernards R., Schrier P. I., Houweling A., Bos J. L., van der Eb A. J., Zijlstra M., Melief C. J. Tumorigenicity of cells transformed by adenovirus type 12 by evasion of T-cell immunity. 1983 Oct 27-Nov 2Nature. 305(5937):776–779. doi: 10.1038/305776a0. [DOI] [PubMed] [Google Scholar]
  2. Chapman A. B., Costello M. A., Lee F., Ringold G. M. Amplification and hormone-regulated expression of a mouse mammary tumor virus-Eco gpt fusion plasmid in mouse 3T6 cells. Mol Cell Biol. 1983 Aug;3(8):1421–1429. doi: 10.1128/mcb.3.8.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Fasel N., Buetti E., Firzlaff J., Pearson K., Diggelmann H. Nucleotide sequence of the 5' noncoding region and part of the gag gene of mouse mammary tumor virus; identification of the 5' splicing site for subgenomic mRNAs. Nucleic Acids Res. 1983 Oct 25;11(20):6943–6955. doi: 10.1093/nar/11.20.6943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fasel N., Pearson K., Buetti E., Diggelmann H. The region of mouse mammary tumor virus DNA containing the long terminal repeat includes a long coding sequence and signals for hormonally regulated transcription. EMBO J. 1982;1(1):3–7. doi: 10.1002/j.1460-2075.1982.tb01115.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Groner B., Hynes N. E. Number and location of mouse mammary tumor virus proviral DNA in mouse DNA of normal tissue and of mammary tumors. J Virol. 1980 Mar;33(3):1013–1025. doi: 10.1128/jvi.33.3.1013-1025.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Groner B., Salmons B., Günzburg W. H., Hynes N. E., Ponta H. Expression of proviral DNA of mouse mammary tumour virus and its transcriptional control sequences. Oxf Surv Eukaryot Genes. 1984;1:87–110. [PubMed] [Google Scholar]
  9. Grove J. R., Ringold G. M. Selection of rat hepatoma cells defective in hormone-regulated production of mouse mammary tumor virus RNA. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4349–4353. doi: 10.1073/pnas.78.7.4349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huang A. L., Ostrowski M. C., Berard D., Hager G. L. Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus. Cell. 1981 Dec;27(2 Pt 1):245–255. doi: 10.1016/0092-8674(81)90408-6. [DOI] [PubMed] [Google Scholar]
  11. Hui K., Grosveld F., Festenstein H. Rejection of transplantable AKR leukaemia cells following MHC DNA-mediated cell transformation. Nature. 1984 Oct 25;311(5988):750–752. doi: 10.1038/311750a0. [DOI] [PubMed] [Google Scholar]
  12. Hynes N. E., Kennedy N., Rahmsdorf U., Groner B. Hormone-responsive expression of an endogenous proviral gene of mouse mammary tumor virus after molecular cloning and gene transfer into cultured cells. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2038–2042. doi: 10.1073/pnas.78.4.2038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hynes N., van Ooyen A. J., Kennedy N., Herrlich P., Ponta H., Groner B. Subfragments of the large terminal repeat cause glucocorticoid-responsive expression of mouse mammary tumor virus and of an adjacent gene. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3637–3641. doi: 10.1073/pnas.80.12.3637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jakobovits E. B., Majors J. E., Varmus H. E. Hormonal regulation of the Rous sarcoma virus src gene via a heterologous promoter defines a threshold dose for cellular transformation. Cell. 1984 Oct;38(3):757–765. doi: 10.1016/0092-8674(84)90271-x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Kress M., Glaros D., Khoury G., Jay G. Alternative RNA splicing in expression of the H-2K gene. Nature. 1983 Dec 8;306(5943):602–604. doi: 10.1038/306602a0. [DOI] [PubMed] [Google Scholar]
  17. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  18. Margulies D. H., Evans G. A., Ozato K., Camerini-Otero R. D., Tanaka K., Appella E., Seidman J. G. Expression of H-2Dd and H-2Ld mouse major histocompatibility antigen genes in L cells after DNA-mediated gene transfer. J Immunol. 1983 Jan;130(1):463–470. [PubMed] [Google Scholar]
  19. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moore K. W., Sher B. T., Sun Y. H., Eakle K. A., Hood L. DNA sequence of a gene encoding a BALB/c mouse Ld transplantation antigen. Science. 1982 Feb 5;215(4533):679–682. doi: 10.1126/science.7058332. [DOI] [PubMed] [Google Scholar]
  21. Murre C., Reiss C. S., Bernabeu C., Chen L. B., Burakoff S. J., Seidman J. G. Construction, expression and recognition of an H-2 molecule lacking its carboxyl terminus. Nature. 1984 Feb 2;307(5950):432–436. doi: 10.1038/307432a0. [DOI] [PubMed] [Google Scholar]
  22. Ozato K., Evans G. A., Shykind B., Margulies D. H., Seidman J. G. Hybrid H-2 histocompatibility gene products assign domains recognized by alloreactive T cells. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2040–2043. doi: 10.1073/pnas.80.7.2040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ozato K., Hansen T. H., Sachs D. H. Monoclonal antibodies to mouse MHC antigens. II. Antibodies to the H-2Ld antigen, the products of a third polymorphic locus of the mouse major histocompatibility complex. J Immunol. 1980 Dec;125(6):2473–2477. [PubMed] [Google Scholar]
  24. Ozato K., Mayer N., Sachs D. H. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J Immunol. 1980 Feb;124(2):533–540. [PubMed] [Google Scholar]
  25. Papkoff J., Ringold G. M. Use of the mouse mammary tumor virus long terminal repeat to promote steroid-inducible expression of v-mos. J Virol. 1984 Nov;52(2):420–430. doi: 10.1128/jvi.52.2.420-430.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ponta H., Günzburg W. H., Salmons B., Groner B., Herrlich P. Mouse mammary tumour virus: a proviral gene contributes to the understanding of eukaryotic gene expression and mammary tumorigenesis. J Gen Virol. 1985 May;66(Pt 5):931–943. doi: 10.1099/0022-1317-66-5-931. [DOI] [PubMed] [Google Scholar]
  27. Ponta H., Kennedy N., Skroch P., Hynes N. E., Groner B. Hormonal response region in the mouse mammary tumor virus long terminal repeat can be dissociated from the proviral promoter and has enhancer properties. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1020–1024. doi: 10.1073/pnas.82.4.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reiss C. S., Evans G. A., Margulies D. H., Seidman J. G., Burakoff S. J. Allospecific and virus-specific cytolytic T lymphocytes are restricted to the N or C1 domain of H-2 antigens expressed on L cells after DNA-mediated gene transfer. Proc Natl Acad Sci U S A. 1983 May;80(9):2709–2712. doi: 10.1073/pnas.80.9.2709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Satz M. L., Singer D. S. Effect of mouse interferon on the expression of a porcine major histocompatibility gene introduced into mouse L cells. J Immunol. 1984 Jan;132(1):496–501. [PubMed] [Google Scholar]
  30. Schrier P. I., Bernards R., Vaessen R. T., Houweling A., van der Eb A. J. Expression of class I major histocompatibility antigens switched off by highly oncogenic adenovirus 12 in transformed rat cells. 1983 Oct 27-Nov 2Nature. 305(5937):771–775. doi: 10.1038/305771a0. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Stroynowski I., Orn A., Goodenow R. S., McMillan M., Forman J., Brayton P. R., Frelinger J., Hood L. Cytotoxic T lymphocytes recognize determinants on the BALB/c-H-2Ld molecule controlled by alpha 1 and alpha 2 but not alpha 3 external domains. Immunogenetics. 1984;20(2):141–154. doi: 10.1007/BF00364486. [DOI] [PubMed] [Google Scholar]
  33. Tanaka K., Isselbacher K. J., Khoury G., Jay G. Reversal of oncogenesis by the expression of a major histocompatibility complex class I gene. Science. 1985 Apr 5;228(4695):26–30. doi: 10.1126/science.3975631. [DOI] [PubMed] [Google Scholar]
  34. Transy C., Lalanne J. L., Kourilsky P. Alternative splicing in the 5' moiety of the H-2Kd gene transcript. EMBO J. 1984 Oct;3(10):2383–2386. doi: 10.1002/j.1460-2075.1984.tb02143.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wallich R., Bulbuc N., Hämmerling G. J., Katzav S., Segal S., Feldman M. Abrogation of metastatic properties of tumour cells by de novo expression of H-2K antigens following H-2 gene transfection. Nature. 1985 May 23;315(6017):301–305. doi: 10.1038/315301a0. [DOI] [PubMed] [Google Scholar]
  36. Weis J. H., Seidman J. G. The expression of major histocompatibility antigens under metallothionein gene promoter control. J Immunol. 1985 Mar;134(3):1999–2003. [PubMed] [Google Scholar]
  37. 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]
  38. Wong G. H., Clark-Lewis I., Harris A. W., Schrader J. W. Effect of cloned interferon-gamma on expression of H-2 and Ia antigens on cell lines of hemopoietic, lymphoid, epithelial, fibroblastic and neuronal origin. Eur J Immunol. 1984 Jan;14(1):52–56. doi: 10.1002/eji.1830140110. [DOI] [PubMed] [Google Scholar]
  39. Zinkernagel R. M., Doherty P. C. MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Adv Immunol. 1979;27:51–177. doi: 10.1016/s0065-2776(08)60262-x. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Zuniga M. C., Malissen B., McMillan M., Brayton P. R., Clark S. S., Forman J., Hood L. Expression and function of transplantation antigens with altered or deleted cytoplasmic domains. Cell. 1983 Sep;34(2):535–544. doi: 10.1016/0092-8674(83)90386-0. [DOI] [PubMed] [Google Scholar]

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