Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1988 Nov;7(11):3465–3470. doi: 10.1002/j.1460-2075.1988.tb03221.x

The human Lyt-3 molecule requires CD8 for cell surface expression.

J P DiSanto 1, R W Knowles 1, N Flomenberg 1
PMCID: PMC454846  PMID: 3145196

Abstract

We have previously identified a monoclonal antibody, T8/2T8-5H7, which clustered serologically with CD8 monoclonal antibodies, but lacked reactivity with L cell transfectants expressing the human CD8 molecule (Lyt-2 homologue). Based on these observations, we postulated that T8/2T8-5H7 might recognize the human Lyt-3 gene product. To test this hypothesis, we have isolated a full-length cDNA encoding the human Lyt-3 molecule and have characterized its product in additional transfection experiments. The results of these studies indicate that the human Lyt-3 cDNA encodes a product recognized by the antibody T8/2T8-5H7. Interestingly, the human Lyt-3 molecule cannot be expressed alone, but requires the human Lyt-2 homologue for efficient cell surface expression. A heterodimer composed of the human Lyt-2 and Lyt-3 molecules may have importance in T cell-target cell interactions.

Full text

PDF
3468

Images in this article

3468Image
on p.3468
3469Image
on p.3469

Selected References

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

  1. Acres R. B., Conlon P. J., Mochizuki D. Y., Gallis B. Phosphorylation of the CD8 antigen on cytotoxic human T cells in response to phorbol myristate acetate or antigen-presenting B cells. J Immunol. 1987 Oct 1;139(7):2268–2274. [PubMed] [Google Scholar]
  2. Acres R. B., Conlon P. J., Mochizuki D. Y., Gallis B. Rapid phosphorylation and modulation of the T4 antigen on cloned helper T cells induced by phorbol myristate acetate or antigen. J Biol Chem. 1986 Dec 5;261(34):16210–16214. [PubMed] [Google Scholar]
  3. Blanc D., Bron C., Gabert J., Letourneur F., MacDonald H. R., Malissen B. Gene transfer of the Ly-3 chain gene of the mouse CD8 molecular complex: co-transfer with the Ly-2 polypeptide gene results in detectable cell surface expression of the Ly-3 antigenic determinants. Eur J Immunol. 1988 Apr;18(4):613–619. doi: 10.1002/eji.1830180419. [DOI] [PubMed] [Google Scholar]
  4. Blue M. L., Hafler D. A., Craig K. A., Levine H., Schlossman S. F. Phosphorylation of CD4 and CD8 molecules following T cell triggering. J Immunol. 1987 Dec 15;139(12):3949–3954. [PubMed] [Google Scholar]
  5. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  6. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Johnson P. A human homolog of the mouse CD8 molecule, Lyt-3: genomic sequence and expression. Immunogenetics. 1987;26(3):174–177. doi: 10.1007/BF00365908. [DOI] [PubMed] [Google Scholar]
  9. Johnson P., Gagnon J., Barclay A. N., Williams A. F. Purification, chain separation and sequence of the MRC OX-8 antigen, a marker of rat cytotoxic T lymphocytes. EMBO J. 1985 Oct;4(10):2539–2545. doi: 10.1002/j.1460-2075.1985.tb03968.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson P., Williams A. F. Striking similarities between antigen receptor J pieces and sequence in the second chain of the murine CD8 antigen. Nature. 1986 Sep 4;323(6083):74–76. doi: 10.1038/323074a0. [DOI] [PubMed] [Google Scholar]
  11. Kavathas P., Sukhatme V. P., Herzenberg L. A., Parnes J. R. Isolation of the gene encoding the human T-lymphocyte differentiation antigen Leu-2 (T8) by gene transfer and cDNA subtraction. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7688–7692. doi: 10.1073/pnas.81.24.7688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ledbetter J. A., Seaman W. E., Tsu T. T., Herzenberg L. A. Lyt-2 and lyt-3 antigens are on two different polypeptide subunits linked by disulfide bonds. Relationship of subunits to T cell cytolytic activity. J Exp Med. 1981 Jun 1;153(6):1503–1516. doi: 10.1084/jem.153.6.1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ledbetter J. A., Tsu T. T., Clark E. A. Covalent association between human thymus leukemia-like antigens and CD8(Tp32) molecules. J Immunol. 1985 Jun;134(6):4250–4254. [PubMed] [Google Scholar]
  14. Littman D. R. The structure of the CD4 and CD8 genes. Annu Rev Immunol. 1987;5:561–584. doi: 10.1146/annurev.iy.05.040187.003021. [DOI] [PubMed] [Google Scholar]
  15. Littman D. R., Thomas Y., Maddon P. J., Chess L., Axel R. The isolation and sequence of the gene encoding T8: a molecule defining functional classes of T lymphocytes. Cell. 1985 Feb;40(2):237–246. doi: 10.1016/0092-8674(85)90138-2. [DOI] [PubMed] [Google Scholar]
  16. Luescher B., Naim H. Y., MacDonald H. R., Bron C. The mouse Lyt-2/3 antigen complex--I. Mode of association of the subunits with the membrane. Mol Immunol. 1984 Apr;21(4):329–336. doi: 10.1016/0161-5890(84)90104-4. [DOI] [PubMed] [Google Scholar]
  17. Lüscher B., Rousseaux-Schmid M., Naim H. Y., MacDonald H. R., Bron C. Biosynthesis and maturation of the Lyt-2/3 molecular complex in mouse thymocytes. J Immunol. 1985 Sep;135(3):1937–1944. [PubMed] [Google Scholar]
  18. MacDonald H. R., Glasebrook A. L., Bron C., Kelso A., Cerottini J. C. Clonal heterogeneity in the functional requirement for Lyt-2/3 molecules on cytolytic T lymphocytes (CTL): possible implications for the affinity of CTL antigen receptors. Immunol Rev. 1982;68:89–115. doi: 10.1111/j.1600-065x.1982.tb01061.x. [DOI] [PubMed] [Google Scholar]
  19. Miller J., Germain R. N. Efficient cell surface expression of class II MHC molecules in the absence of associated invariant chain. J Exp Med. 1986 Nov 1;164(5):1478–1489. doi: 10.1084/jem.164.5.1478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nakauchi H., Nolan G. P., Hsu C., Huang H. S., Kavathas P., Herzenberg L. A. Molecular cloning of Lyt-2, a membrane glycoprotein marking a subset of mouse T lymphocytes: molecular homology to its human counterpart, Leu-2/T8, and to immunoglobulin variable regions. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5126–5130. doi: 10.1073/pnas.82.15.5126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Panaccio M., Gillespie M. T., Walker I. D., Kirszbaum L., Sharpe J. A., Tobias G. H., McKenzie I. F., Deacon N. J. Molecular characterization of the murine cytotoxic T-cell membrane glycoprotein Ly-3 (CD8). Proc Natl Acad Sci U S A. 1987 Oct;84(19):6874–6878. doi: 10.1073/pnas.84.19.6874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ratnofsky S. E., Peterson A., Greenstein J. L., Burakoff S. J. Expression and function of CD8 in a murine T cell hybridoma. J Exp Med. 1987 Dec 1;166(6):1747–1757. doi: 10.1084/jem.166.6.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Snow P. M., Terhorst C. The T8 antigen is a multimeric complex of two distinct subunits on human thymocytes but consists of homomultimeric forms on peripheral blood T lymphocytes. J Biol Chem. 1983 Dec 10;258(23):14675–14681. [PubMed] [Google Scholar]
  25. Snow P. M., Van de Rijn M., Terhorst C. Association between the human thymic differentiation antigens T6 and T8. Eur J Immunol. 1985 May;15(5):529–532. doi: 10.1002/eji.1830150520. [DOI] [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. Spurr N. K., Goodfellow P. N., Johnson P. CD8B, the human equivalent of the mouse Ly-3 gene is localized on chromosome 2. Immunogenetics. 1988;27(1):70–72. doi: 10.1007/BF00404448. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Walker I. D., Murray B. J., Hogarth P. M., Kelso A., McKenzie I. F. Comparison of thymic and peripheral T cell Ly-2/3 antigens. Eur J Immunol. 1984 Oct;14(10):906–910. doi: 10.1002/eji.1830141009. [DOI] [PubMed] [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES