Lymphokine requirements for the development of specific cytotoxic T cells from single precursors

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Abstract
A high cloning efficiency, filler cell-free culture system was developed for the growth of single murine cytotoxic T lymphocyte precursors (CTLp) and their differentiation into cytotoxic T lymphocytes (CTL). The system used nonspecific stimulation with phorbol ester and calcium ionophore in the presence of recombinant lymphokines. The optimal lymphokine combination was interleukin 2 throughout, together with interferon-γ during the first 6 days and interleukin 6 during the last 2 days of culture. Under these conditions half of all CD4−CD8+ T cells became CTL clones. The CTL were CD4−CD8+CD3+ TcR α/β+ and were derived from CD4−CD8+ Pgp-1− precursors.