Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1996 Aug;88(4):577–585. doi: 10.1046/j.1365-2567.1996.d01-682.x

Preparation of monoclonal anti-porcine CD3 antibodies and preliminary characterization of porcine T lymphocytes.

H Yang 1, C A Oura 1, P A Kirkham 1, R M Parkhouse 1
PMCID: PMC1456639  PMID: 8881760

Abstract

The CD3-T-cell receptor complex is the clonotypic surface structure by which T lymphocytes recognize foreign antigens and are subsequently activated. Because of the low immunogenicity of the CD3 molecules, anti-CD3 monoclonal antibodies (mAb) are difficult to prepare and have not been available in several species. Following isolation of porcine CD3, 14 anti-porcine CD3 mAb were prepared, which define six groups of CD3-epsilon epitopes, coprecipitate two types of TCR and reveal considerable heterogeneity of CD3 expression amongst lymphocyte subpopulations. Thus, both CD3 positive and negative subpopulations of CD2 or CD8 positive cells were found in the blood. The density of CD3 on CD2+ or CD8+ cells was relatively low and heterogeneous, whereas the CD2-, CD8- or MAC320+ T cells expressed CD3 at a higher and more homogeneous level. Finally, in the thymus, staining with anti-CD3 resolved large thymocytes into two subsets: one expressing a high level of CD3 and the other being negative. In contrast, small thymocytes expressed CD3 at a low and more homogeneous level. Immunohistological studies confirmed the presence of clearly detectable CD3 in thymus medulla and the T-cell regions of peripheral lymphoid tissues. Most of the mAb were mitogenic, when presented to peripheral blood mononuclear cells in immobilized form. The anti-CD3 mAb also induced redirected cytotoxicity which was shown to be Fc receptor dependent.

Full text

PDF
577

Images in this article

580Figure 2
on p.580
580Figure 3
on p.580
582Figure 5
on p.582

Selected References

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

  1. Allison J. P., Lanier L. L. Structure, function, and serology of the T-cell antigen receptor complex. Annu Rev Immunol. 1987;5:503–540. doi: 10.1146/annurev.iy.05.040187.002443. [DOI] [PubMed] [Google Scholar]
  2. Ashwell J. D., Klusner R. D. Genetic and mutational analysis of the T-cell antigen receptor. Annu Rev Immunol. 1990;8:139–167. doi: 10.1146/annurev.iy.08.040190.001035. [DOI] [PubMed] [Google Scholar]
  3. Banchereau J., de Paoli P., Vallé A., Garcia E., Rousset F. Long-term human B cell lines dependent on interleukin-4 and antibody to CD40. Science. 1991 Jan 4;251(4989):70–72. doi: 10.1126/science.1702555. [DOI] [PubMed] [Google Scholar]
  4. Binns R. M., Duncan I. A., Powis S. J., Hutchings A., Butcher G. W. Subsets of null and gamma delta T-cell receptor+ T lymphocytes in the blood of young pigs identified by specific monoclonal antibodies. Immunology. 1992 Oct;77(2):219–227. [PMC free article] [PubMed] [Google Scholar]
  5. Binns R. M. The Null/gamma delta TCR+ T cell family in the pig. Vet Immunol Immunopathol. 1994 Oct;43(1-3):69–77. doi: 10.1016/0165-2427(94)90122-8. [DOI] [PubMed] [Google Scholar]
  6. Ceuppens J. L., Bloemmen F. J., Van Wauwe J. P. T cell unresponsiveness to the mitogenic activity of OKT3 antibody results from a deficiency of monocyte Fc gamma receptors for murine IgG2a and inability to cross-link the T3-Ti complex. J Immunol. 1985 Dec;135(6):3882–3886. [PubMed] [Google Scholar]
  7. Chan A. C., Desai D. M., Weiss A. The role of protein tyrosine kinases and protein tyrosine phosphatases in T cell antigen receptor signal transduction. Annu Rev Immunol. 1994;12:555–592. doi: 10.1146/annurev.iy.12.040194.003011. [DOI] [PubMed] [Google Scholar]
  8. Clevers H., Alarcon B., Wileman T., Terhorst C. The T cell receptor/CD3 complex: a dynamic protein ensemble. Annu Rev Immunol. 1988;6:629–662. doi: 10.1146/annurev.iy.06.040188.003213. [DOI] [PubMed] [Google Scholar]
  9. Davis W. C., MacHugh N. D., Park Y. H., Hamilton M. J., Wyatt C. R. Identification of a monoclonal antibody reactive with the bovine orthologue of CD3 (BoCD3). Vet Immunol Immunopathol. 1993 Nov;39(1-3):85–91. doi: 10.1016/0165-2427(93)90167-3. [DOI] [PubMed] [Google Scholar]
  10. Hammerberg C., Schurig G. G. Characterization of monoclonal antibodies directed against swine leukocytes. Vet Immunol Immunopathol. 1986 Feb;11(2):107–121. doi: 10.1016/0165-2427(86)90092-9. [DOI] [PubMed] [Google Scholar]
  11. Hirt W., Saalmüller A., Reddehase M. J. Distinct gamma/delta T cell receptors define two subsets of circulating porcine CD2-CD4-CD8- T lymphocytes. Eur J Immunol. 1990 Feb;20(2):265–269. doi: 10.1002/eji.1830200206. [DOI] [PubMed] [Google Scholar]
  12. Jones M., Cordell J. L., Beyers A. D., Tse A. G., Mason D. Y. Detection of T and B cells in many animal species using cross-reactive anti-peptide antibodies. J Immunol. 1993 Jun 15;150(12):5429–5435. [PubMed] [Google Scholar]
  13. Jonjić S., Koszinowski U. H. Monoclonal antibodies reactive with swine lymphocytes. I. Antibodies to membrane structures that define the cytolytic T lymphocyte subset in the swine. J Immunol. 1984 Aug;133(2):647–652. [PubMed] [Google Scholar]
  14. Jung G., Honsik C. J., Reisfeld R. A., Müller-Eberhard H. J. Activation of human peripheral blood mononuclear cells by anti-T3: killing of tumor target cells coated with anti-target-anti-T3 conjugates. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4479–4483. doi: 10.1073/pnas.83.12.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kirkham P. A., Takamatsu H., Yang H., Parkhouse R. M. Porcine CD3 epsilon: its characterization, expression and involvement in activation of porcine T lymphocytes. Immunology. 1996 Apr;87(4):616–623. doi: 10.1046/j.1365-2567.1996.498566.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leiden J. M. Transcriptional regulation of T cell receptor genes. Annu Rev Immunol. 1993;11:539–570. doi: 10.1146/annurev.iy.11.040193.002543. [DOI] [PubMed] [Google Scholar]
  17. Leo O., Foo M., Sachs D. H., Samelson L. E., Bluestone J. A. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1374–1378. doi: 10.1073/pnas.84.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Oettgen H. C., Pettey C. L., Maloy W. L., Terhorst C. A T3-like protein complex associated with the antigen receptor on murine T cells. Nature. 1986 Mar 20;320(6059):272–275. doi: 10.1038/320272a0. [DOI] [PubMed] [Google Scholar]
  19. Parsons K. R., Crocker G., Sopp P., Howard C. J., Davis W. C. Identification of monoclonal antibodies specific for the gamma/delta TCR. Vet Immunol Immunopathol. 1993 Nov;39(1-3):161–167. doi: 10.1016/0165-2427(93)90177-6. [DOI] [PubMed] [Google Scholar]
  20. Pescovitz M. D., Lunney J. K., Sachs D. H. Murine anti-swine T4 and T8 monoclonal antibodies: distribution and effects on proliferative and cytotoxic T cells. J Immunol. 1985 Jan;134(1):37–44. [PubMed] [Google Scholar]
  21. Pescovitz M. D., Lunney J. K., Sachs D. H. Preparation and characterization of monoclonal antibodies reactive with porcine PBL. J Immunol. 1984 Jul;133(1):368–375. [PubMed] [Google Scholar]
  22. Pescovitz M. D., Sakopoulos A. G., Gaddy J. A., Husmann R. J., Zuckermann F. A. Porcine peripheral blood CD4+/CD8+ dual expressing T-cells. Vet Immunol Immunopathol. 1994 Oct;43(1-3):53–62. doi: 10.1016/0165-2427(94)90120-1. [DOI] [PubMed] [Google Scholar]
  23. Reinherz E. L., Kung P. C., Goldstein G., Schlossman S. F. A monoclonal antibody with selective reactivity with functionally mature human thymocytes and all peripheral human T cells. J Immunol. 1979 Sep;123(3):1312–1317. [PubMed] [Google Scholar]
  24. Saalmüller A., Hirt W., Reddehase M. J. Porcine gamma/delta T lymphocyte subsets differing in their propensity to home to lymphoid tissue. Eur J Immunol. 1990 Oct;20(10):2343–2346. doi: 10.1002/eji.1830201026. [DOI] [PubMed] [Google Scholar]
  25. Saalmüller A., Reddehase M. J., Bühring H. J., Jonjić S., Koszinowski U. H. Simultaneous expression of CD4 and CD8 antigens by a substantial proportion of resting porcine T lymphocytes. Eur J Immunol. 1987 Sep;17(9):1297–1301. doi: 10.1002/eji.1830170912. [DOI] [PubMed] [Google Scholar]
  26. Sachs D. H. The pig as a potential xenograft donor. Vet Immunol Immunopathol. 1994 Oct;43(1-3):185–191. doi: 10.1016/0165-2427(94)90135-x. [DOI] [PubMed] [Google Scholar]
  27. Samelson L. E., Harford J. B., Klausner R. D. Identification of the components of the murine T cell antigen receptor complex. Cell. 1985 Nov;43(1):223–231. doi: 10.1016/0092-8674(85)90027-3. [DOI] [PubMed] [Google Scholar]
  28. Santos-Argumedo L., Teixeira C., Preece G., Kirkham P. A., Parkhouse R. M. A B lymphocyte surface molecule mediating activation and protection from apoptosis via calcium channels. J Immunol. 1993 Sep 15;151(6):3119–3130. [PubMed] [Google Scholar]
  29. Sato K., Ohtsuka K., Watanabe H., Asakura H., Abo T. Detailed characterization of gamma delta T cells within the organs in mice: classification into three groups. Immunology. 1993 Nov;80(3):380–387. [PMC free article] [PubMed] [Google Scholar]
  30. Schlitt H. J., Kurrle R., Wonigeit K. T cell activation by monoclonal antibodies directed to different epitopes on the human T cell receptor/CD3 complex: evidence for two different modes of activation. Eur J Immunol. 1989 Sep;19(9):1649–1655. doi: 10.1002/eji.1830190920. [DOI] [PubMed] [Google Scholar]
  31. Schwinzer R., Franklin R. A., Domenico J., Renz H., Gelfand E. W. Monoclonal antibodies directed to different epitopes in the CD3-TCR complex induce different states of competence in resting human T cells. J Immunol. 1992 Mar 1;148(5):1322–1328. [PubMed] [Google Scholar]
  32. Snary D., Woods F. R., Crumpton M. J. Disruption of solid tissue for plasma membrane preparation. Anal Biochem. 1976 Aug;74(2):457–465. doi: 10.1016/0003-2697(76)90226-8. [DOI] [PubMed] [Google Scholar]
  33. Transy C., Moingeon P. E., Marshall B., Stebbins C., Reinherz E. L. Most anti-human CD3 monoclonal antibodies are directed to the CD3 epsilon subunit. Eur J Immunol. 1989 May;19(5):947–950. doi: 10.1002/eji.1830190525. [DOI] [PubMed] [Google Scholar]
  34. Tunnacliffe A., Olsson C., de la Hera A. The majority of human CD3 epitopes are conferred by the epsilon chain. Int Immunol. 1989;1(5):546–550. doi: 10.1093/intimm/1.5.546. [DOI] [PubMed] [Google Scholar]
  35. Verwilghen J., Baroja M. L., Van Vaeck F., Van Damme J., Ceuppens J. L. Differences in the stimulating capacity of immobilized anti-CD3 monoclonal antibodies: variable dependence on interleukin-1 as a helper signal for T-cell activation. Immunology. 1991 Feb;72(2):269–276. [PMC free article] [PubMed] [Google Scholar]
  36. Yang H., Binns R. M. Expression and regulation of the porcine CD44 molecule. Cell Immunol. 1993 Jun;149(1):117–129. doi: 10.1006/cimm.1993.1141. [DOI] [PubMed] [Google Scholar]
  37. Yang H., Hutchings A., Binns R. M. Preparation and reactivities of anti-porcine CD44 monoclonal antibodies. Scand J Immunol. 1993 Apr;37(4):490–498. doi: 10.1111/j.1365-3083.1993.tb03324.x. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES