IL-2 and IFN-gamma are two necessary lymphokines in the development of cytolytic T cells.
A filler cell-free limiting-dilution microculture system has been developed for the expansion and differentiation of a high proportion of single CD4-CD8+ T cells into cytolytic T cell (CTL) clones. The stimulus used was PMA together with the calcium ionophore ionomycin. The growth and differentiation factors were rIL-2, together with either a Con A-stimulated spleen cell supernatant (CAS) or rIFN-gamma. CTL activity was monitored by an autoradiographic 111In-release assay. With CAS and rIL-2 present, 50% of all potential precursors (CTL-p) produced cytolytic clones. Substitution of rIFN-gamma for CAS gave a similar efficiency with up to 42% of CTL-p producing cytolytic clones. rIL-2 alone allowed only a small proportion (6%) of CD4-CD8+ T cells to become cytolytic clones. Addition of rIL-2 and rIFN-gamma at various stages of the culture demonstrated that IL-2 was required throughout, but exogenous IFN-gamma was required only during the early stages. It is concluded that for at least 40% of all CTL-p, the lymphokines IL-2 and IFN-gamma are essential and act in synergy to induce proliferation and differentiation into CTL.
Maraskovsky
E
E
Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
Chen
W F
WF
Shortman
K
K
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D002454:Cell Differentiation; D002474:Cell-Free System; D002478:Cells, Cultured; D002999:Clone Cells / Q000276:immunology / Q000502:physiology; D003602:Cytotoxicity, Immunologic; D007371:Interferon-gamma / Q000502:physiology*; D007376:Interleukin-2 / Q000502:physiology*; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D010641:Phenotype; D013602:T-Lymphocytes, Cytotoxic / Q000145:classification / Q000276:immunology* / Q000502:physiology
D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D007376:Interleukin-2; D007371:Interferon-gamma