Abstract
The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-colour immunofluorescent studies showed that CVS5 recognized all T lymphocytes and that CVS4 and CVS8 recognized two mutually exclusive subsets of CVS5-positive cells. In the thymus there was a large population of CVS4/CVS8 double-positive cells. Immunohistochemical staining with these mAb was restricted to T-lymphocyte areas. CVS4 and CVS5 precipitated molecules of 58,000 and 69,000 MW, respectively, in both reducing and non-reducing conditions. CVS8 precipitated two molecules of 32,000 and 39,000 MW in reducing conditions, and one molecule of 69,000 MW in non-reducing conditions. This evidence suggests that CVS5, CVS4 and CVS8 recognize the equine homologues of CD5, CD4 and CD8, and that the characteristics of these antigens are similar to those of other species.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baldwin C. L., Teale A. J., Naessens J. G., Goddeeris B. M., MacHugh N. D., Morrison W. I. Characterization of a subset of bovine T lymphocytes that express BoT4 by monoclonal antibodies and function: similarity to lymphocytes defined by human T4 and murine L3T4. J Immunol. 1986 Jun 15;136(12):4385–4391. [PubMed] [Google Scholar]
- Chan M. M., Chen C. L., Ager L. L., Cooper M. D. Identification of the avian homologues of mammalian CD4 and CD8 antigens. J Immunol. 1988 Apr 1;140(7):2133–2138. [PubMed] [Google Scholar]
- Clevers H., MacHugh N. D., Bensaid A., Dunlap S., Baldwin C. L., Kaushal A., Iams K., Howard C. J., Morrison W. I. Identification of a bovine surface antigen uniquely expressed on CD4-CD8- T cell receptor gamma/delta+ T lymphocytes. Eur J Immunol. 1990 Apr;20(4):809–817. doi: 10.1002/eji.1830200415. [DOI] [PubMed] [Google Scholar]
- Crump A. L., Davis W., Antczak D. F. A monoclonal antibody identifying a T-cell marker in the horse. Anim Genet. 1988;19(4):349–357. doi: 10.1111/j.1365-2052.1988.tb00826.x. [DOI] [PubMed] [Google Scholar]
- Donaldson W. L., Crump A. L., Zhang C. H., Kornbluth J., Kamoun M., Davis W., Antczak D. F. At least two loci encode polymorphic class I MHC antigens in the horse. Anim Genet. 1988;19(4):379–390. doi: 10.1111/j.1365-2052.1988.tb00829.x. [DOI] [PubMed] [Google Scholar]
- Ellis J. A., Baldwin C. L., MacHugh N. D., Bensaid A., Teale A. J., Goddeeris B. M., Morrison W. I. Characterization by a monoclonal antibody and functional analysis of a subset of bovine T lymphocytes that express BoT8, a molecule analogous to human CD8. Immunology. 1986 Jul;58(3):351–358. [PMC free article] [PubMed] [Google Scholar]
- Fowlkes B. J., Pardoll D. M. Molecular and cellular events of T cell development. Adv Immunol. 1989;44:207–264. doi: 10.1016/s0065-2776(08)60643-4. [DOI] [PubMed] [Google Scholar]
- Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
- Howard C. J., Parsons K. R., Jones B. V., Sopp P., Pocock D. H. Two monoclonal antibodies (CC17, CC29) recognizing an antigen (Bo5) on bovine T lymphocytes, analogous to human CD5. Vet Immunol Immunopathol. 1988 Sep;19(2):127–139. doi: 10.1016/0165-2427(88)90004-9. [DOI] [PubMed] [Google Scholar]
- Huang H. J., Jones N. H., Strominger J. L., Herzenberg L. A. Molecular cloning of Ly-1, a membrane glycoprotein of mouse T lymphocytes and a subset of B cells: molecular homology to its human counterpart Leu-1/T1 (CD5). Proc Natl Acad Sci U S A. 1987 Jan;84(1):204–208. doi: 10.1073/pnas.84.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knapp W., Rieber P., Dörken B., Schmidt R. E., Stein H., vd Borne A. E. Towards a better definition of human leucocyte surface molecules. Immunol Today. 1989 Aug;10(8):253–258. doi: 10.1016/0167-5699(89)90135-7. [DOI] [PubMed] [Google Scholar]
- Lanier L. L., Federspiel N. A., Ruitenberg J. J., Phillips J. H., Allison J. P., Littman D., Weiss A. The T cell antigen receptor complex expressed on normal peripheral blood CD4-, CD8- T lymphocytes. A CD3-associated disulfide-linked gamma chain heterodimer. J Exp Med. 1987 Apr 1;165(4):1076–1094. doi: 10.1084/jem.165.4.1076. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lunney J. K., Pescovitz M. D. Phenotypic and functional characterization of pig lymphocyte populations. Vet Immunol Immunopathol. 1987 Dec;17(1-4):135–144. doi: 10.1016/0165-2427(87)90134-6. [DOI] [PubMed] [Google Scholar]
- Mackay C. R., Maddox J. F., Brandon M. R. Lymphocyte antigens of sheep: identification and characterization using a panel of monoclonal antibodies. Vet Immunol Immunopathol. 1987 Dec;17(1-4):91–102. doi: 10.1016/0165-2427(87)90130-9. [DOI] [PubMed] [Google Scholar]
- Maddox J. F., Mackay C. R., Brandon M. R. Surface antigens, SBU-T4 and SBU-T8, of sheep T lymphocyte subsets defined by monoclonal antibodies. Immunology. 1985 Aug;55(4):739–748. [PMC free article] [PubMed] [Google Scholar]
- McClure S. J., Hein W. R., Yamaguchi K., Dudler L., Beya M. F., Miyasaka M. Ontogeny, morphology and tissue distribution of a unique subset of CD4-CD8- sheep T lymphocytes. Immunol Cell Biol. 1989 Aug;67(Pt 4):215–221. doi: 10.1038/icb.1989.33. [DOI] [PubMed] [Google Scholar]
- McGuire T. C., Poppie M. J. Hypogammaglobulinemia and thymic hypoplasia in horses: a primary combined immunodeficiency disorder. Infect Immun. 1973 Aug;8(2):272–277. doi: 10.1128/iai.8.2.272-277.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parnes J. R. Molecular biology and function of CD4 and CD8. Adv Immunol. 1989;44:265–311. doi: 10.1016/s0065-2776(08)60644-6. [DOI] [PubMed] [Google Scholar]
- Perryman L. E., Bue C. M., Magnuson N. S., Mottironi V. D., Ochs H. S., Wyatt C. R. Immunologic reconstitution of foals with combined immunodeficiency. Vet Immunol Immunopathol. 1987 Dec;17(1-4):495–508. doi: 10.1016/0165-2427(87)90165-6. [DOI] [PubMed] [Google Scholar]
- Perryman L. E., Wyatt C. R., Magnuson N. S., Mason P. H. T lymphocyte development and maturation in horses. Anim Genet. 1988;19(4):343–348. doi: 10.1111/j.1365-2052.1988.tb00825.x. [DOI] [PubMed] [Google Scholar]
- Tompkins M. B., Gebhard D. H., Bingham H. R., Hamilton M. J., Davis W. C., Tompkins W. A. Characterization of monoclonal antibodies to feline T lymphocytes and their use in the analysis of lymphocyte tissue distribution in the cat. Vet Immunol Immunopathol. 1990 Dec;26(4):305–317. doi: 10.1016/0165-2427(90)90115-9. [DOI] [PubMed] [Google Scholar]
- Wyatt C. R., Davis W. C., McGuire T. C., Perryman L. E. T lymphocyte development in horses. I. Characterization of monoclonal antibodies identifying three stages of T lymphocyte differentiation. Vet Immunol Immunopathol. 1988 Feb;18(1):3–18. doi: 10.1016/0165-2427(88)90032-3. [DOI] [PubMed] [Google Scholar]
- Wyatt C. R., Magnuson N. S., Perryman L. E. Defective thymocyte maturation in horses with severe combined immunodeficiency. J Immunol. 1987 Dec 15;139(12):4072–4076. [PubMed] [Google Scholar]
- van Agthoven A., Terhorst C., Reinherz E., Schlossman S. Characterization of T cell surface glycoproteins T 1 and T 3 present on all human peripheral T lymphocytes and functionally mature thymocytes. Eur J Immunol. 1981 Jan;11(1):18–21. doi: 10.1002/eji.1830110105. [DOI] [PubMed] [Google Scholar]