Production of human suppressor T cell hybridomas

J Immunol. 1982 Nov;129(5):1986-92.

Abstract

To study human T cell suppression of immunoglobulin (Ig) synthesis with homogeneous populations of immunoregulatory cells, human suppressor T cell hybridomas were prepared by somatic cell fusion of concanavalin A-activated peripheral blood T cells with hypoxanthine-guanine phosphoribosyltransferase-(HGPRT, EC 2.4.2.8) deficient human leukemic CEM T cells. After selection in hypoxanthine-aminopterin-thymidine (HAT) medium and cloning by limiting cell dilution, two human T cell hybridomas were identified that produced 60 to 80% suppression of in vitro polyclonal immunoglobulin production when cocultured with pokeweed mitogen- (PWM) stimulated peripheral blood lymphocytes. Further, one of the suppressor T cell hybridomas constitutively secreted a soluble suppressor factor(s) (TsF) of m.w. 70,000 to 85,000 daltons, which produced reversible noncytotoxic inhibition of lectin-activated B cell Ig production. In contrast, this TsF did not inhibit lectin- or antigen-induced T cell proliferation, nor did it interfere with the generation or effector function of cytotoxic T cells. Additional studies indicated that this Tsf acts directly on B cells or monocytes rather than indirectly modulating the activity of immunoregulatory T cells. In summary, these studies suggest that techniques of somatic cell fusion may provide a valuable approach to further study human immunoregulatory cell-cell interactions as well as provide a source of sufficient quantities of important lymphokines for further purification and characterization.

MeSH terms

  • Cell Division
  • Cell Fusion*
  • Clone Cells / cytology
  • Clone Cells / immunology
  • Humans
  • Hybridomas / cytology
  • Hybridomas / immunology*
  • Immunoglobulins / biosynthesis
  • Immunologic Techniques*
  • Lymphokines / analysis
  • Lymphokines / biosynthesis
  • Phenotype
  • Pokeweed Mitogens / pharmacology
  • Suppressor Factors, Immunologic
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology
  • T-Lymphocytes, Regulatory / cytology*
  • T-Lymphocytes, Regulatory / immunology

Substances

  • Immunoglobulins
  • Lymphokines
  • Pokeweed Mitogens
  • Suppressor Factors, Immunologic