Lyt-2 Negative and T Cell Growth Factor Independent Cytotoxic T Lymphocyte Hybridomas
Homogeneous populations of cytotoxic T lymphocytes (CTL) are essential for studying the CTL receptor and killing mechanism. Such populations are obtained mainly through continuous CTL lines. However, the limiting amounts of cells thus produced, the months required to establish these lines and the need for repeated stimuli to maintain CTL growth and activity represent major drawbacks. To circumvent these problems we developed functional CTL-hybridomas which proliferate autonomously both in culture and in vivo (1–3). Previous attempts to generate T cell growth factor (TCGF)-independent CTL-hybridomas were unsuccessful (4,5), possibly due to CTL-induced nonspecific lysis of the fusion partner during hybridization. Our approach was based on the assumption that nonspecific lysis during hybridization may be prevented by a transient inactivation of CTL with trypsin prior to fusion and cytotoxicity regenerated in the hybrid cells after fusion, as is the case with trypsin-inactivated parental CTL (1). Using this approach, we have generated functional CTL-hybridomas which constitutively express their killing potential.
KeywordsKilling Mechanism Specific Killing Killing Potential Cytofluorometric Analysis Nonspecific Lysis
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