Constitutive and mitogen-induced production of T cell growth factor by stable T cell hybridoma lines.
Stable T cell growth factor- (TCGF; IL 2) producing cloned T cell hybridoma lines were constructed by fusing murine alloantigen-activated T cells with the 8-azaguanine-resistant lymphoma line, BW5147. Many, but not all, clones of one of these hybridomas, i.e., hybridoma 24, secreted TCGF constitutively, but production was markedly enhanced by stimulation with T cell mitogens. Large numbers of TCGF-secreting hybridoma cells in a stable functional state could be obtained from histocompatible mice inoculated with cloned T cell hybridomas. Moreover, such in vivo-derived hybridoma cells could be stimulated sequentially with mitogen at least twice to secrete their biologically-active product, resulting in larger TCGF yields from the same cells. The secreted product of these T cell hybridoma lines resembled TCGF isolated from other cellular sources in that it: a) supported the growth of a TCGF-dependent T cell line; b) provided help for the induction of alloantigen-reactive cytotoxic T lymphocytes from thymocyte precursors; c) facilitated concanavalin A-induced mitogenic responses of low thymocyte numbers; d) had an apparent m.w. of 30,000 to 40,000 by gel filtration chromatography; and e) was eluted from DEAE-Sephacel ion-exchange chromatography columns by salt concentrations of 30 to 150 mM NaCl. The ability of these T cell hybridomas to grow in vivo and retain their functional characteristics in a stable form should prove useful in terms of providing large numbers of TCGF-secreting cells and studying in vivo aspects of the production of TCGF as well as other immunoregulatory mediators.
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D002459:Cell Fusion; D002999:Clone Cells / Q000378:metabolism; D003208:Concanavalin A / Q000494:pharmacology*; D006183:H-2 Antigens / Q000276:immunology; D006825:Hybridomas / Q000032:analysis / Q000378:metabolism*; D007376:Interleukin-2 / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D008222:Lymphokines / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008809:Mice, Inbred C3H; D008811:Mice, Inbred DBA; D013601:T-Lymphocytes / Q000378:metabolism*; D018800:Thy-1 Antigens; D013997:Time Factors
D000954:Antigens, Surface; D006183:H-2 Antigens; D007376:Interleukin-2; D008222:Lymphokines; D008565:Membrane Proteins; D018800:Thy-1 Antigens; D003208:Concanavalin A