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Home / Equine Research Bank / Immunology / Characterization of monoclonal antibodies specific for equin...
Immunology1994; 82(4); 548-554;

Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5.

Abstract: Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was effective at inducing interleukin-2 receptor (IL-2R) expression on T lymphocytes, a feature consistent with antibodies specific for the epsilon chain of human and murine CD3. Three populations of cells in the thymus were distinguishable by UC F6G-3 target antigen density, suggesting increasing stages of T-cell maturation. UC F13C-5 precipitated a 67,000 MW protein, consistent with reported values for CD5 in multiple species. While this antibody exhibited characteristics of a pan-T-cell marker, low numbers of B lymphocytes also expressed the target antigen. Phorbol esters induced variable increases in target antigen density on B lymphocytes. These two antibodies, taken together with the few equine CD markers currently available, represent a substantial resource for further defining the equine immune system in health and disease.
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Publication Date: 1994-08-01 PubMed ID: 7530685PubMed Central: PMC1414917
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research involves the characterization of two monoclonal antibodies that specifically target equine versions of CD3 and CD5 – proteins present on immune cells. The study indicates that these antibodies could be used in studying the horse immune system in both healthy and diseased conditions.

Overview of antibodiy classes: UC F6G-3 and UC F13C-5

  • Two monoclonal antibodies, UC F6G-3 and UC F13C-5, were studied. These antibodies were found to target equine versions of CD3 and CD5 proteins respectively.
  • Both antibodies exhibited properties of pan-T-lymphocyte markers meaning they could bind to most T-lymphocytes, a type of immune cell.

Characterization of UC F6G-3

  • UC F6G-3 precipitated a protein complex with molecular weights ranging from 18,000 to 42,000, indicative of the human and murine CD3 complex.
  • Upon further breakdown of the complex, two proteins with molecular weights of 22,000 and 27,000 were identified.
  • Immobolized UC F6G-3 was effective at inducing interleukin-2 receptor (IL-2R) expression on T lymphocytes, which is consistent with antibodies that target the epsilon chain of human and murine CD3.
  • Three distinct populations of thymus cells could be identified based on the density of the target antigen, suggesting the potential of this antibody in recognizing different stages of T-cell maturation.

Characterization of UC F13C-5

  • UC F13C-5 precipitated a 67,000 MW protein, a value that is consistent with the CD5 protein in multiple species.
  • This antibody showed characteristics of a pan-T-cell marker, but in low numbers, it also targeted B lymphocytes, another type of immune cell.
  • The use of phorbol esters led to a variable increase in target antigen density on B lymphocytes.

Implications of the Findings

  • Together, the two antibodies offer a substantial resource for the further investigation of the equine immune system both in health and disease.
  • They add to the few equine CD markers currently available, offering more opportunities to broaden our understanding of horse immunology and potentially opening new pathways for equine disease treatment and prevention.

Cite This Article

APA
Blanchard-Channell M, Moore PF, Stott JL. (1994). Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5. Immunology, 82(4), 548-554.

Publication

Immunology
ISSN: 0019-2805
NlmUniqueID: 0374672
Country: England
Language: English
Volume: 82
Issue: 4
Pages: 548-554

Researcher Affiliations

Blanchard-Channell, M
  • Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616.
Moore, P F
    Stott, J L

      MeSH Terms

      • Animals
      • Antibodies, Monoclonal / biosynthesis
      • Antibodies, Monoclonal / immunology
      • Antigens, CD / chemistry
      • Antigens, CD / immunology
      • B-Lymphocytes / immunology
      • CD3 Complex / chemistry
      • CD3 Complex / immunology
      • CD5 Antigens
      • Horses / immunology
      • Immunoenzyme Techniques
      • Lymph Nodes / immunology
      • Lymphocyte Activation / immunology
      • Mice
      • Mice, Inbred BALB C
      • Molecular Weight
      • Precipitin Tests
      • Species Specificity
      • Thymus Gland / immunology

      References

      This article includes 28 references
      1. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 1970 Aug 15;227(5259):680-5.
        pubmed: 5432063doi: 10.1038/227680a0google scholar: lookup
      2. Taylor BC, Choi KY, Scibienski RJ, Moore PF, Stott JL. Differential expression of bovine MHC class II antigens identified by monoclonal antibodies.. J Leukoc Biol 1993 May;53(5):479-89.
        pubmed: 7684762doi: 10.1002/jlb.53.5.479google scholar: lookup
      3. Borst J, Prendiville MA, Terhorst C. Complexity of the human T lymphocyte-specific cell surface antigen T3.. J Immunol 1982 Apr;128(4):1560-5.
        pubmed: 6174606
      4. Borst J, Alexander S, Elder J, Terhorst C. The T3 complex on human T lymphocytes involves four structurally distinct glycoproteins.. J Biol Chem 1983 Apr 25;258(8):5135-41.
        pubmed: 6220014
      5. Pessano S, Oettgen H, Bhan AK, Terhorst C. The T3/T cell receptor complex: antigenic distinction between the two 20-kd T3 (T3-delta and T3-epsilon) subunits.. EMBO J 1985 Feb;4(2):337-44.
        pubmed: 2410254doi: 10.1002/j.1460-2075.1985.tb03634.xgoogle scholar: lookup
      6. Samelson LE, Harford JB, Klausner RD. Identification of the components of the murine T cell antigen receptor complex.. Cell 1985 Nov;43(1):223-31.
        pubmed: 3878228doi: 10.1016/0092-8674(85)90027-3google scholar: lookup
      7. Hurley WL, Finkelstein E, Holst BD. Identification of surface proteins on bovine leukocytes by a biotin-avidin protein blotting technique.. J Immunol Methods 1985 Dec 17;85(1):195-202.
        pubmed: 4078309doi: 10.1016/0022-1759(85)90287-xgoogle scholar: lookup
      8. Oettgen HC, Pettey CL, Maloy WL, Terhorst C. A T3-like protein complex associated with the antigen receptor on murine T cells.. Nature 1986 Mar 20-26;320(6059):272-5.
        pubmed: 2938011doi: 10.1038/320272a0google scholar: lookup
      9. Ceuppens JL, Baroja ML. Monoclonal antibodies to the CD5 antigen can provide the necessary second signal for activation of isolated resting T cells by solid-phase-bound OKT3.. J Immunol 1986 Sep 15;137(6):1816-21.
        pubmed: 3091691
      10. Weissman AM, Samelson LE, Klausner RD. A new subunit of the human T-cell antigen receptor complex.. Nature 1986 Dec 4-10;324(6096):480-2.
        pubmed: 3785426doi: 10.1038/324480a0google scholar: lookup
      11. Leo O, Foo M, Sachs DH, Samelson LE, Bluestone JA. Identification of a monoclonal antibody specific for a murine T3 polypeptide.. Proc Natl Acad Sci U S A 1987 Mar;84(5):1374-8.
        pubmed: 2950524doi: 10.1073/pnas.84.5.1374google scholar: lookup
      12. Scibienski RJ, Stott ML, Stott JL. Establishment and initial characterization of continuous in vitro cultures of bovine T lymphocytes.. Vet Immunol Immunopathol 1987 Mar;14(3):209-22.
        pubmed: 3109112doi: 10.1016/0165-2427(87)90090-0google scholar: lookup
      13. Freedman AS, Boyd AW, Bieber FR, Daley J, Rosen K, Horowitz JC, Levy DN, Nadler LM. Normal cellular counterparts of B cell chronic lymphocytic leukemia.. Blood 1987 Aug;70(2):418-27.
        pubmed: 3496927
      14. Wyatt CR, Davis WC, McGuire TC, Perryman LE. 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.
        pubmed: 2967580doi: 10.1016/0165-2427(88)90032-3google scholar: lookup
      15. Nishimura Y, Bierer BE, Jones WK, Jones NH, Strominger JL, Burakoff SJ. Expression and function of a CD5 cDNA in human and murine T cells.. Eur J Immunol 1988 May;18(5):747-53.
        pubmed: 2454193doi: 10.1002/eji.1830180514google scholar: lookup
      16. Baniyash M, Garcia-Morales P, Bonifacino JS, Samelson LE, Klausner RD. Disulfide linkage of the zeta and eta chains of the T cell receptor. Possible identification of two structural classes of receptors.. J Biol Chem 1988 Jul 15;263(20):9874-8.
        pubmed: 3290215
      17. Crump AL, Davis W, Antczak DF. A monoclonal antibody identifying a T-cell marker in the horse.. Anim Genet 1988;19(4):349-57.
        pubmed: 3069011doi: 10.1111/j.1365-2052.1988.tb00826.xgoogle scholar: lookup
      18. Freedman AS, Freeman G, Whitman J, Segil J, Daley J, Levine H, Nadler LM. Expression and regulation of CD5 on in vitro activated human B cells.. Eur J Immunol 1989 May;19(5):849-55.
        pubmed: 2472277doi: 10.1002/eji.1830190511google scholar: lookup
      19. Transy C, Moingeon PE, Marshall B, Stebbins C, Reinherz EL. Most anti-human CD3 monoclonal antibodies are directed to the CD3 epsilon subunit.. Eur J Immunol 1989 May;19(5):947-50.
        pubmed: 2472280doi: 10.1002/eji.1830190525google scholar: lookup
      20. Miescher GC, Schreyer M, MacDonald HR. Production and characterization of a rat monoclonal antibody against the murine CD3 molecular complex.. Immunol Lett 1989 Dec;23(2):113-8.
        pubmed: 2534389doi: 10.1016/0165-2478(89)90122-3google scholar: lookup
      21. Raveche ES. Possible immunoregulatory role for CD5 + B cells.. Clin Immunol Immunopathol 1990 Aug;56(2):135-50.
        pubmed: 1696187doi: 10.1016/0090-1229(90)90136-egoogle scholar: lookup
      22. Kasaian MT, Ikematsu H, Casali P. CD5+ B lymphocytes.. Proc Soc Exp Biol Med 1991 Jul;197(3):226-41.
        pubmed: 1712492doi: 10.3181/00379727-197-43250google scholar: lookup
      23. Blanchard-Channell M, Stott JL. Characterization of Borrelia coriaceae antigens with monoclonal antibodies.. Infect Immun 1991 Aug;59(8):2790-8.
        pubmed: 1855995doi: 10.1128/iai.59.8.2790-2798.1991google scholar: lookup
      24. Lunn DP, Holmes MA, Duffus WP. Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets.. Immunology 1991 Oct;74(2):251-7.
        pubmed: 1748472
      25. Nishimura T, Nakamura Y, Takeuchi Y, Tokuda Y, Iwasawa M, Kawasaki A, Okumura K, Habu S. Generation propagation, and targeting of human CD4+ helper/killer T cells induced by anti-CD3 monoclonal antibody plus recombinant IL-2. An efficient strategy for adoptive tumor immunotherapy.. J Immunol 1992 Jan 1;148(1):285-91.
        pubmed: 1345787
      26. Taylor BC, Stott JL, Thurmond MA, Picanso JP. Alteration in lymphocyte subpopulations in bovine leukosis virus-infected cattle.. Vet Immunol Immunopathol 1992 Feb 15;31(1-2):35-47.
        pubmed: 1570682doi: 10.1016/0165-2427(92)90085-5google scholar: lookup
      27. Taylor BC, Stott JL, Scibienski RJ, Redelman D. Development and characterization of a monoclonal antibody specific for the bovine low-affinity interleukin-2 receptor, BoCD25.. Immunology 1992 Sep;77(1):150-4.
        pubmed: 1398763
      28. Van Wauwe JP, De Mey JR, Goossens JG. OKT3: a monoclonal anti-human T lymphocyte antibody with potent mitogenic properties.. J Immunol 1980 Jun;124(6):2708-13.
        pubmed: 6966296

      Citations

      This article has been cited 20 times.
      1. Jaworska J, de Mestre AM, Wiśniewska J, Wagner B, Nowicki A, Kowalczyk-Zięba I, Wocławek-Potocka I. Populations of NK Cells and Regulatory T Cells in the Endometrium of Cycling Mares-A Preliminary Study.. Animals (Basel) 2022 Nov 30;12(23).
        doi: 10.3390/ani12233373pubmed: 36496894google scholar: lookup
      2. Patel RS, Tomlinson JE, Divers TJ, Van de Walle GR, Rosenberg BR. Single-cell resolution landscape of equine peripheral blood mononuclear cells reveals diverse cell types including T-bet(+) B cells.. BMC Biol 2021 Jan 22;19(1):13.
        doi: 10.1186/s12915-020-00947-5pubmed: 33482825google scholar: lookup
      3. Arthurs C, Suarez-Bonnet A, Willis C, Xie B, Machulla N, Mair TS, Cao K, Millar M, Thrasivoulou C, Priestnall SL, Ahmed A. Equine penile squamous cell carcinoma: expression of biomarker proteins and EcPV2.. Sci Rep 2020 May 12;10(1):7863.
        doi: 10.1038/s41598-020-64014-3pubmed: 32398763google scholar: lookup
      4. Saldinger LK, Nelson SG, Bellone RR, Lassaline M, Mack M, Walker NJ, Borjesson DL. Horses with equine recurrent uveitis have an activated CD4+ T-cell phenotype that can be modulated by mesenchymal stem cells in vitro.. Vet Ophthalmol 2020 Jan;23(1):160-170.
        doi: 10.1111/vop.12704pubmed: 31441218google scholar: lookup
      5. Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8(+) T and CD21(+) B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract.. J Virol 2017 Jul 1;91(13).
        doi: 10.1128/JVI.00418-17pubmed: 28424285google scholar: lookup
      6. Prieto JMB, Tallmadge RL, Felippe MJB. Developmental expression of B cell molecules in equine lymphoid tissues.. Vet Immunol Immunopathol 2017 Jan;183:60-71.
        doi: 10.1016/j.vetimm.2016.12.004pubmed: 28063478google scholar: lookup
      7. Kol A, Wood JA, Carrade Holt DD, Gillette JA, Bohannon-Worsley LK, Puchalski SM, Walker NJ, Clark KC, Watson JL, Borjesson DL. Multiple intravenous injections of allogeneic equine mesenchymal stem cells do not induce a systemic inflammatory response but do alter lymphocyte subsets in healthy horses.. Stem Cell Res Ther 2015 Apr 15;6(1):73.
        doi: 10.1186/s13287-015-0050-0pubmed: 25888916google scholar: lookup
      8. Ramsay JD, Ueti MW, Johnson WC, Scoles GA, Knowles DP, Mealey RH. Lymphocytes and macrophages are infected by Theileria equi, but T cells and B cells are not required to establish infection in vivo.. PLoS One 2013;8(10):e76996.
        doi: 10.1371/journal.pone.0076996pubmed: 24116194google scholar: lookup
      9. Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
        doi: 10.3727/215517912X647217pubmed: 23152950google scholar: lookup
      10. Go YY, Zhang J, Timoney PJ, Cook RF, Horohov DW, Balasuriya UB. Complex interactions between the major and minor envelope proteins of equine arteritis virus determine its tropism for equine CD3+ T lymphocytes and CD14+ monocytes.. J Virol 2010 May;84(10):4898-911.
        doi: 10.1128/JVI.02743-09pubmed: 20219931google scholar: lookup
      11. Mealey RH, Leib SR, Littke MH, Wagner B, Horohov DW, McGuire TC. Viral load and clinical disease enhancement associated with a lentivirus cytotoxic T lymphocyte vaccine regimen.. Vaccine 2009 Apr 21;27(18):2453-68.
        doi: 10.1016/j.vaccine.2009.02.048pubmed: 19368787google scholar: lookup
      12. Mealey RH, Stone DM, Hines MT, Alperin DC, Littke MH, Leib SR, Leach SE, Hines SA. Experimental Rhodococcus equi and equine infectious anemia virus DNA vaccination in adult and neonatal horses: effect of IL-12, dose, and route.. Vaccine 2007 Oct 23;25(43):7582-97.
        doi: 10.1016/j.vaccine.2007.07.055pubmed: 17889970google scholar: lookup
      13. Mealey RH, Littke MH, Leib SR, Davis WC, McGuire TC. Failure of low-dose recombinant human IL-2 to support the survival of virus-specific CTL clones infused into severe combined immunodeficient foals: lack of correlation between in vitro activity and in vivo efficacy.. Vet Immunol Immunopathol 2008 Jan 15;121(1-2):8-22.
        doi: 10.1016/j.vetimm.2007.07.011pubmed: 17727961google scholar: lookup
      14. Mealey RH, Littke MH, Leib SR, Davis WC, McGuire TC. Cloning and large-scale expansion of epitope-specific equine cytotoxic T lymphocytes using an anti-equine CD3 monoclonal antibody and human recombinant IL-2.. Vet Immunol Immunopathol 2007 Jul 15;118(1-2):121-8.
        doi: 10.1016/j.vetimm.2007.04.001pubmed: 17498813google scholar: lookup
      15. Mérant C, Messouak A, Cadoré JL, Monier JC. PNA-binding glycans are expressed at high levels on horse mature and immature T lymphocytes and a subpopulation of B lymphocytes.. Glycoconj J 2005 Feb;22(1-2):27-34.
        doi: 10.1007/s10719-005-0228-2pubmed: 15864432google scholar: lookup
      16. Patton KM, McGuire TC, Fraser DG, Hines SA. Rhodococcus equi-infected macrophages are recognized and killed by CD8+ T lymphocytes in a major histocompatibility complex class I-unrestricted fashion.. Infect Immun 2004 Dec;72(12):7073-83.
        doi: 10.1128/IAI.72.12.7073-7083.2004pubmed: 15557631google scholar: lookup
      17. De Guise S, Erickson K, Blanchard M, Dimolfetto L, Lepper H, Wang J, Stott JL, Ferrick DA. Characterization of a monoclonal antibody that recognizes a lymphocyte surface antigen for the cetacean homologue to CD45R.. Immunology 1998 Jun;94(2):207-12.
        doi: 10.1046/j.1365-2567.1998.00483.xpubmed: 9741342google scholar: lookup
      18. Drummer HE, Reubel GH, Studdert MJ. Equine gammaherpesvirus 2 (EHV2) is latent in B lymphocytes.. Arch Virol 1996;141(3-4):495-504.
        doi: 10.1007/BF01718313pubmed: 8645091google scholar: lookup
      19. Lunn DP, McClure JT, Schobert CS, Holmes MA. Abnormal patterns of equine leucocyte differentiation antigen expression in severe combined immunodeficiency foals suggests the phenotype of normal equine natural killer cells.. Immunology 1995 Mar;84(3):495-9.
        pubmed: 7751035
      20. Schrenzel MD, Ferrick DA. Horse (Equus caballus) T-cell receptor alpha, gamma, and delta chain genes: nucleotide sequences and tissue-specific gene expression.. Immunogenetics 1995;42(2):112-22.
        doi: 10.1007/BF00178585pubmed: 7607702google scholar: lookup
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