Murine cytolytic CD8+ T cell clones generated in a high cloning efficiency, accessory cell-free culture system express a restricted lymphokine profile.
To determine whether cytolytic T lymphocyte activity is associated with a particular lymphokine profile, lymphokine synthesis was analyzed in a large panel of primary clones derived from single murine CD8+ lymph node T cells. The clones were generated at high efficiency (60-70%), under conditions that had been optimized for the induction of specific cytolytic activity, by culture with phorbol 12-myristate 13-acetate, ionomycin, IL-2, and IFN-gamma for 6 days and then with IL-2 and IL-6 for a further 2 days. When the clones were restimulated for 24 hr with IL-2 and immobilized antibodies to CD3, CD8, and LFA-1, most secreted IL-3 and IFN-gamma and about a third secreted TNF. Although levels of production of IL-3 and IFN-gamma were positively correlated with each other (r = 0.85) and weakly correlated with cytolytic activity (r = 0.68 and 0.55, respectively). TNF titers were unrelated to any other function measured. None of the clones tested secreted detectable IL-4 or IL-6. Similarly, analysis of several clones for lymphokine mRNA expression following cDNA amplification by the polymerase chain reaction revealed the presence of IL-2, IL-3, IFN-gamma, TNF-alpha, and GM-CSF but not IL-4 or IL-6 transcripts. Cytolytic CD8+ T cell clones generated in this high efficiency, accessory cell-independent cloning system therefore expressed a restricted lymphokine profile characterized by the synthesis of IL-3 and IFN-gamma, with variable production of TNF.
Maraskovsky
E
E
Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Victoria, Australia.
Rogers
L A
LA
Troutt
A B
AB
Kelso
A
A
D000818:Animals; D001483:Base Sequence; D016827:CD8 Antigens / Q000276:immunology*; D002999:Clone Cells / Q000187:drug effects; D003603:Cytotoxins / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism*; D007377:Interleukin-3 / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D010703:Phorbol Esters / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*
D016827:CD8 Antigens; D003603:Cytotoxins; D007377:Interleukin-3; D010703:Phorbol Esters; D012333:RNA, Messenger; D015759:Ionomycin; D007371:Interferon-gamma