Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Oct 1;168(4):1237–1245. doi: 10.1084/jem.168.4.1237

Functional consequences of anti-sense RNA-mediated inhibition of CD8 surface expression in a human T cell clone

PMCID: PMC2189074  PMID: 2459296

Abstract

An experimental approach for defining the function of CD8 has been developed by linking anti-sense RNA mutagenesis and T cell cloning technologies. We have transfected an anti-sense CD8 episomal expression vector into a CD8+ nontransformed human T cell clone that is specific for the human class I alloantigen HLA-B35. Expression of CD8 on this T cell clone, JH.ARL.1, was selectively and efficiently inhibited. Stimulation of this CD8- variant with specific alloantigen resulted in a marked loss of a number of functional responses, including cytotoxicity, proliferation, IL-2 secretion, and IL-2-R expression. However, these same functional responses could be elicited with stimuli that do not require antigen recognition to activate the T cell (anti- CD3 mAbs, PHA). The results of our study support the hypothesis that CD8 is required for recognition of class I MHC alloantigens that results in activation of T cell functional responses.

Full Text

The Full Text of this article is available as a PDF (605.5 KB).

Selected References

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

  1. Aparicio P., Jaraquemada D., López de Castro J. A. Alloreactive cytolytic T cell clones with dual recognition of HLA-B27 and HLA-DR2 antigens. Selective involvement of CD8 in their class I--directed cytotoxicity. J Exp Med. 1987 Feb 1;165(2):428–443. doi: 10.1084/jem.165.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blanchard D., van Els C., Borst J., Carrel S., Boylston A., de Vries J. E., Spits H. The role of the T cell receptor, CD8, and LFA-1 in different stages of the cytolytic reaction mediated by alloreactive T lymphocyte clones. J Immunol. 1987 Apr 15;138(8):2417–2421. [PubMed] [Google Scholar]
  3. Dembić Z., Haas W., Weiss S., McCubrey J., Kiefer H., von Boehmer H., Steinmetz M. Transfer of specificity by murine alpha and beta T-cell receptor genes. Nature. 1986 Mar 20;320(6059):232–238. doi: 10.1038/320232a0. [DOI] [PubMed] [Google Scholar]
  4. Dembić Z., Haas W., Zamoyska R., Parnes J., Steinmetz M., von Boehmer H. Transfection of the CD8 gene enhances T-cell recognition. Nature. 1987 Apr 2;326(6112):510–511. doi: 10.1038/326510a0. [DOI] [PubMed] [Google Scholar]
  5. Doyle C., Strominger J. L. Interaction between CD4 and class II MHC molecules mediates cell adhesion. Nature. 1987 Nov 19;330(6145):256–259. doi: 10.1038/330256a0. [DOI] [PubMed] [Google Scholar]
  6. Emmrich F., Strittmatter U., Eichmann K. Synergism in the activation of human CD8 T cells by cross-linking the T-cell receptor complex with the CD8 differentiation antigen. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8298–8302. doi: 10.1073/pnas.83.21.8298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fleischer B., Schrezenmeier H., Wagner H. Function of the CD4 and CD8 molecules on human cytotoxic T lymphocytes: regulation of T cell triggering. J Immunol. 1986 Mar 1;136(5):1625–1628. [PubMed] [Google Scholar]
  8. Gabert J., Langlet C., Zamoyska R., Parnes J. R., Schmitt-Verhulst A. M., Malissen B. Reconstitution of MHC class I specificity by transfer of the T cell receptor and Lyt-2 genes. Cell. 1987 Aug 14;50(4):545–554. doi: 10.1016/0092-8674(87)90027-4. [DOI] [PubMed] [Google Scholar]
  9. Geppert T. D., Lipsky P. E. Accessory cell-T cell interactions involved in anti-CD3-induced T4 and T8 cell proliferation: analysis with monoclonal antibodies. J Immunol. 1986 Nov 15;137(10):3065–3073. [PubMed] [Google Scholar]
  10. Goldstein S. A., Mescher M. F. Cytotoxic T cell activation by class I protein on cell-size artificial membranes: antigen density and Lyt-2/3 function. J Immunol. 1987 Apr 1;138(7):2034–2043. [PubMed] [Google Scholar]
  11. Hambor J. E., Hauer C. A., Shu H. K., Groger R. K., Kaplan D. R., Tykocinski M. L. Use of an Epstein-Barr virus episomal replicon for anti-sense RNA-mediated gene inhibition in a human cytotoxic T-cell clone. Proc Natl Acad Sci U S A. 1988 Jun;85(11):4010–4014. doi: 10.1073/pnas.85.11.4010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hünig T. Monoclonal anti-Lyt-2.2 antibody blocks lectin-dependent cellular cytotoxicity of H-2-negative target cells. J Exp Med. 1984 Feb 1;159(2):551–558. doi: 10.1084/jem.159.2.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaplan D. R., Bergmann C. A., Gould D., Landmeier B. Membrane-associated interleukin 2 epitopes on the surface of human T lymphocytes. J Immunol. 1988 Feb 1;140(3):819–826. [PubMed] [Google Scholar]
  14. Kaplan D. R., Braciale V. L., Braciale T. J. Antigen-dependent regulation of interleukin 2 receptor expression on cloned human cytotoxic T lymphocytes. J Immunol. 1984 Oct;133(4):1966–1969. [PubMed] [Google Scholar]
  15. Kaplan D. R., Griffith R., Braciale V. L., Braciale T. J. Influenza virus-specific human cytotoxic T cell clones: heterogeneity in antigenic specificity and restriction by class II MHC products. Cell Immunol. 1984 Oct 1;88(1):193–206. doi: 10.1016/0008-8749(84)90064-9. [DOI] [PubMed] [Google Scholar]
  16. Landegren U., Ramstedt U., Axberg I., Ullberg M., Jondal M., Wigzell H. Selective inhibition of human T cell cytotoxicity at levels of target recognition or initiation of lysis by monoclonal OKT3 and Leu-2a antibodies. J Exp Med. 1982 May 1;155(5):1579–1584. doi: 10.1084/jem.155.5.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
  18. Mentzer S. J., Smith B. R., Barbosa J. A., Crimmins M. A., Herrmann S. H., Burakoff S. J. CTL adhesion and antigen recognition are discrete steps in the human CTL-target cell interaction. J Immunol. 1987 Mar 1;138(5):1325–1330. [PubMed] [Google Scholar]
  19. Meuer S. C., Schlossman S. F., Reinherz E. L. Clonal analysis of human cytotoxic T lymphocytes: T4+ and T8+ effector T cells recognize products of different major histocompatibility complex regions. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4395–4399. doi: 10.1073/pnas.79.14.4395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moldwin R. L., Havran W. L., Nau G. J., Lancki D. W., Kim D. K., Fitch F. W. Antibodies to the L3T4 and Lyt-2 molecules interfere with antigen receptor-driven activation of cloned murine T cells. J Immunol. 1987 Aug 1;139(3):657–664. [PubMed] [Google Scholar]
  21. Platsoucas C. D. Human T cell antigens involved in cytotoxicity against allogeneic or autologous chemically modified targets. Association of the Leu 2a/T8 antigen with effector-target cell binding and of the T3/Leu 4 antigen with triggering. Eur J Immunol. 1984 Jun;14(6):566–577. doi: 10.1002/eji.1830140615. [DOI] [PubMed] [Google Scholar]
  22. Reinherz E. L., Schlossman S. F. The differentiation and function of human T lymphocytes. Cell. 1980 Apr;19(4):821–827. doi: 10.1016/0092-8674(80)90072-0. [DOI] [PubMed] [Google Scholar]
  23. Shaw S., Luce G. E., Quinones R., Gress R. E., Springer T. A., Sanders M. E. Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones. Nature. 1986 Sep 18;323(6085):262–264. doi: 10.1038/323262a0. [DOI] [PubMed] [Google Scholar]
  24. Shimonkevitz R., Luescher B., Cerottini J. C., MacDonald H. R. Clonal analysis of cytolytic T lymphocyte-mediated lysis of target cells with inducible antigen expression: correlation between antigen density and requirement for Lyt-2/3 function. J Immunol. 1985 Aug;135(2):892–899. [PubMed] [Google Scholar]
  25. Spits H., Yssel H., Voordouw A., de Vries J. E. The role of T8 in the cytotoxic activity of cloned cytotoxic T lymphocyte lines specific for class II and class I major histocompatibility complex antigens. J Immunol. 1985 Apr;134(4):2294–2298. [PubMed] [Google Scholar]
  26. Spits H., van Schooten W., Keizer H., van Seventer G., van de Rijn M., Terhorst C., de Vries J. E. Alloantigen recognition is preceded by nonspecific adhesion of cytotoxic T cells and target cells. Science. 1986 Apr 18;232(4748):403–405. doi: 10.1126/science.3485822. [DOI] [PubMed] [Google Scholar]
  27. Spits H., van Schooten W., Keizer H., van Seventer G., van de Rijn M., Terhorst C., de Vries J. E. Alloantigen recognition is preceded by nonspecific adhesion of cytotoxic T cells and target cells. Science. 1986 Apr 18;232(4748):403–405. doi: 10.1126/science.3485822. [DOI] [PubMed] [Google Scholar]
  28. Springer T. A., Dustin M. L., Kishimoto T. K., Marlin S. D. The lymphocyte function-associated LFA-1, CD2, and LFA-3 molecules: cell adhesion receptors of the immune system. Annu Rev Immunol. 1987;5:223–252. doi: 10.1146/annurev.iy.05.040187.001255. [DOI] [PubMed] [Google Scholar]
  29. Swain S. L., Dialynas D. P., Fitch F. W., English M. Monoclonal antibody to L3T4 blocks the function of T cells specific for class 2 major histocompatibility complex antigens. J Immunol. 1984 Mar;132(3):1118–1123. [PubMed] [Google Scholar]
  30. Swain S. L. Significance of Lyt phenotypes: Lyt2 antibodies block activities of T cells that recognize class 1 major histocompatibility complex antigens regardless of their function. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7101–7105. doi: 10.1073/pnas.78.11.7101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Swain S. L. T cell subsets and the recognition of MHC class. Immunol Rev. 1983;74:129–142. doi: 10.1111/j.1600-065x.1983.tb01087.x. [DOI] [PubMed] [Google Scholar]
  32. Takada S., Engleman E. G. Evidence for an association between CD8 molecules and the T cell receptor complex on cytotoxic T cells. J Immunol. 1987 Nov 15;139(10):3231–3235. [PubMed] [Google Scholar]
  33. Van Seventer G. A., Van Lier R. A., Spits H., Ivanyi P., Melief C. J. Evidence for a regulatory role of the T8 (CD8) antigen in antigen-specific and anti-T3-(CD3)-induced lytic activity of allospecific cytotoxic T lymphocyte clones. Eur J Immunol. 1986 Nov;16(11):1363–1371. doi: 10.1002/eji.1830161109. [DOI] [PubMed] [Google Scholar]
  34. Welte K., Platzer E., Wang C. Y., Rinnooy Kan E. A., Moore M. A., Mertelsmann R. OKT8 antibody inhibits OKT3-induced IL 2 production and proliferation in OKT8+ cells. J Immunol. 1983 Nov;131(5):2356–2361. [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES