UBC Theses and Dissertations
Characterization of T cell clones derived from a mixed lymphocyte reaction Kwong, Pearl C.
Monoclonal T cells with distinct function and specificity were isolated and characterized. The method used to obtain monoclonal T cells involved four steps: First, C57BL/6 spleen cells were cultured with irradiated D-BA/2 spleen cells in vitro. Second, the activated C57BL/6 cells were maintained for several months in a medium containing T Cell Growth Factor(TCGF) , irradia'ted DBA/2 and C57BL/6 spleen cells. Third, the activated cells were cloned in microtiter wells by limiting dilution. Fourth, wells detected to contain growth were expanded and tested for their functional activities. Using this method, three classes of clones were derived. The first class of clones were cytotoxic clones, presumably of H-2[sup b] phenotype, which have different specificities against foreign histocompatibility antigens. The second class of clones was an H-2[sup b], Thy 1⁺, Lyt 2⁺ and Lyt 1 low cytotoxic clone whose specificity was against cells carrying H-2D[sup b], antigens. The supernatant derived from this clone nonspecifically supressed CTL generation and mitogen activation, but was weakly toxic in the ⁵¹Cr release assay. The third class of clones was an H-2[sup d], Thy 1⁺ Lyt 1⁻ and Lyt 2⁻ clone which helped augment the CTL responses of cells carrying the H-2D[sup b] antigens. This clone required accessory cells for its helper function and was effective only when added during the early phase of the CTL response. The supernatant derived from this clone exhibited the same properties as the clone and acted in synergy with TCGF in the augmentation of CTL responses.
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