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  • Title: Signal requirements for the in vitro differentiation of cytotoxic T lymphocytes (CTL): distinct soluble mediators promote preactivation of CTL-precursors, clonal growth and differentiation into cytotoxic effector cells.
    Author: Hardt C, Diamantstein T, Wagner H.
    Journal: Eur J Immunol; 1985 May; 15(5):472-8. PubMed ID: 3158532.
    Abstract:
    We describe here that the requirement of accessory cells for the polyclonal activation of high-density (resting) murine T lymphocytes can be bypassed by soluble mediators present in culture supernatants of concanavalin A (Con A)-activated murine spleen cells. Induction of responsiveness is confined to Lyt-2+ T cells; GK 1.5+ T helper cells require signals provided by accessory cells. Using this system the lymphokine signal requirements for the polyclonal activation of Lyt-2+ cytotoxic T lymphocyte (CTL) precursors could be defined. We show that Con A fails to trigger in Lyt-2+ responder T cells the expression of interleukin 2 (IL 2) receptors and assume that this explains why recombinant (rec) DNA-derived IL 2 fails to induce proliferative responses. Complementation of rec IL 2 with an IL 2 receptor-inducing factor (RIF) induces proliferative responses. RIF alone triggers IL 2 receptor expression in 10-12% of Lyt-2+ T cells exposed to Con A. This lymphokine appears to be distinct from colony-stimulating factor 1, IFN-gamma and IL 1. Resting Lyt-2+ T cells cultured in limiting numbers in the presence of Con A, RIF plus rec IL 2 do proliferate, yet exhibit no cytolytic activity. Differentiation into CTL can be brought about by addition of cytotoxic T cell differentiation factor (CTDF). We conclude that the polyclonal activation pathway of CTL from resting CTL precursors can be subdivided into three stages: preactivation, clonal growth and CTL differentiation. Each of these stages appears to be controlled by a distinct lymphokine, RIF, IL 2 and CTDF, respectively.
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