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T Cell Depletion by Cytotoxic Elimination

Karen S. Hathcock1

1National Cancer Institute, Bethesda, Maryland

Publication Name: 
Current Protocols in Immunology
Unit Number: 
UNIT 3.4
DOI: 
10.1002/0471142735.im0304s00
Online Posting Date: 
May, 2001
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Abstract

This unit describes the complete removal of T cells from lymphocyte preparations based on the presence of the glycoprotein Thy-1 on the cell surface of T lymphocytes. As in unit 3.3, cytotoxic elimination is employed; however, Thy-1-specific antibodies are used rather than MHC class II-specific antibodies so that T cells are eliminated rather than B cells and accessory cells. Murine T cells can be further fractionated into subpopulations based on their expression of cell differentiation markers CD4 (Leu3, L3T4, T4) and CD8 (Leu2, Lyt2,3). A protocol in this unit describes removal of T cell subsets by cytotoxic elimination using CD-specific antibodies. An alternate but technically more demanding method of enriching T cell subsets is the panning procedure described in unit 3.5.

     
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Table of Contents

  • Unit Introduction
  • Basic Protocol: Complete T Cell Removal Using Thy-1-Specific Monoclonal Antibodies
  • Basic Protocol: Removal of T Cell Subsets Using CD4/CD8-Specific Monoclonal Antibodies
  • Commentary
  • Bibliography
  • Tables
     
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Literature Cited

 Literature Cited
    Ceredig, R., Lowenthal, J.N., Nabholz, M., and MacDonald, H.R. 1985. Expression of interleukin-2 receptors as a differentiation marker on intrathymic stem cells. Nature (Lond.) 314:98-100.
    Dialynas, D.P., Wilde, D.B., Marrack, P., Pierres, A., Wall, K.A., Havran, W., Otten, G., Loken, M.R., Pierres, M., Kappler, J., and Fitch, F.W. 1983. Characterization of the murine antigenic determinant designated L3T4a, recognized by monoclonal antibody GK1.5: Expression of L3T4a by functional T cell clones appears to correlate primarily with class II MHC antigen-reactivity. Immunol. Rev. 74:29-58.
    Lake, P., Clark, E.A., Khorshidi, M., and Sunshine, G.H. 1979. Production and characterization of cytotoxic Thy-1 antibody-secreting hybrid cell lines: Detection of T cell subsets. Eur. J. Immunol. 9:875-886.
    Ledbetter, J.A. and Herzenberg, L.A. 1979. Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol. Rev. 47:63-90.
    Logberg, L., Wassmer, P., and Shevach, E.M. 1985. Role of the L3T4 antigen in T cell activation. I. Description of a monoclonal IgM antibody to a distinct epitope (L3T4b) of the L3T4 antigen and its effect on interleukin-2 induced thymocyte proliferation. Cell. Immunol. 94:299-311.
    Marshak-Rothstein, A., Fink, P., Gridley, T., Raulet, D.H., Beven, M.J., and Gefter, M.L. 1979. Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. J. Immunol. 122:2491-2499.
    Sarmiento, M., Glasebrook, A.L., and Fitch, F. 1980. IgG or IgM monoclonal antibodies reactive to different determinant on the molecular complex bearing Lyt2 antigen block T cell mediated cytolysis in the absence of complement. J. Immunol. 125:2665-2672.
    Shen, F.W. 1981. Monoclonal antibodies and T cell hybridomas. Res. Monogr. Immunol. 3:25-37.
 Key References
    Dialynas et al., 1983. See above.
    Pierres, A., Naquet, P., Agthoven, A.V., Bekkahoucha, F., Denzot, D., Mishal, Z., Schmitt-Verhulst, A.M., and Pierres, M. 1984. A rat anti-mouse monoclonal antibody (H1 29-19) inhibits the proliferation of Ia-reactive T cell clones and delineates two phenotypically distinct (T4+Lyt2,3– and T4– Lyt2,3+) subsets among anti-Ia cytolytic T cell clones. J. Immunol. 131:2775-2783.
    Swain, S.L. 1981. Significance of Lyt phenotypes: Lyt2 antibodies block activation of T cells that recognize class I MHC antigens regardless of their function. Proc. Natl. Acad. Sci. U.S.A. 78:7101-7105.

These articles describe the concepts and methods of fractionating T cell subsets on the basis of CD4 (L3T4) and CD8 (Lyt2) expression.

     
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