Immortalization of human T cell clones by Herpesvirus saimiri. Signal transduction analysis reveals functional CD3, CD4, and IL-2 receptors.
Investigation of human activated T cells has been complicated by the need for periodic restimulation with Ag/mitogen and accessory cells and by the limited life span of most human T cell clones. To overcome these problems, we have transformed established human T cell clones to permanent growth with Herpesvirus saimiri, a lymphoma-inducing virus of nonhuman primates. Three human CD4+ T cell clones were investigated in detail. They have been growing in the presence of exogenous IL-2 but without restimulation with mitogen or feeder cells for more than 11 mo with doubling times between 2 and 4 days. In contrast, their nontransformed parent clones needed to be restimulated with PHA and feeder cells every 14 to 21 days. To compare responses of H. saimiri-transformed clones with those of their parent clones, we stimulated the cells with IL-2 or with anti-CD3 and/or anti-CD4 mAb with and without cross-linking on the cell surface. Transformed and nontransformed T cell clones were strikingly similar in parameters of early signal transduction, namely, tyrosine phosphorylation and mobilization of calcium. Ligation of their TcR/CD3 complexes by mAb or by Ag in the presence of autologous accessory cells increased the proliferation and the secretion of IFN-gamma. Taken together, we have shown that human T cell clones immortalized with H. saimiri express functional CD3, CD4, and IL-2R. They constitute a simple, stable, reproducible and accessory cell-free model system for the investigation of signal transduction events in activated human T cells.
Bröker
B M
BM
Max-Planck-Gesellschaft, Arbeitsgruppen für Rheumatologie am Institut für Klinische Immunologie, Universität Erlangen-Nürnberg, Germany.
Tsygankov
A Y
AY
Müller-Fleckenstein
I
I
Guse
A H
AH
Chitaev
N A
NA
Biesinger
B
B
Fleckenstein
B
B
Emmrich
F
F
D000942:Antigens, Bacterial / Q000276:immunology; D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002472:Cell Transformation, Viral; D002999:Clone Cells; D006567:Herpesvirus 2, Saimiriine / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D010750:Phosphoproteins / Q000378:metabolism; D019000:Phosphotyrosine; D013601:T-Lymphocytes / Q000166:cytology*; D013745:Tetanus Toxoid / Q000276:immunology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D000942:Antigens, Bacterial; D010750:Phosphoproteins; D013745:Tetanus Toxoid; D019000:Phosphotyrosine; D014443:Tyrosine; D002118:Calcium