Cytotoxic T cell clone‐specific monoclonal antibodies used to select clonotypic antigen‐specific cytotoxic T cells

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Abstract
Two rat monoclonal antibodies (mAb) have been produced which recognize a clonespecific determinant on the alloreactive cytotoxic T lymphocyte (CTL) clone 3F9. CTL clone 3 F9 of BALB/c origin is specific for H‐2Db and can be grown by weekly restimulation with irradiated stimulator spleen cells expressing H‐2Db in the presence of interleukin 2. Two mAb against T cell clone 3F9, 44‐22‐l(IgG2a) and 46‐6 B5(IgM), have been proven to be clone specific: they inhibit cytotoxic activity of 3F9 only and bind specifically to 3 F9 when compared in a panel of different CTL clones, or cells from different mixed lymphocyte cultures (MLC), BALB/c thymus and spleen cells. The mAb 44‐22‐1 has been used to sort cells from a primary MLC BALB/c anti‐ H‐2Db by fluorescence‐activated cell sorter (FACS) to select CTL expressing 3 F9 clonotype‐specific determinants. The lymphocytes reactive with 44‐22‐1 represent a minor subpopulation of the CTL of the primary MLC. The specific alloreactive cytotoxicity of unsorted lymphocytes of the bulk primary MLC could not be inhibited by the mAb 44‐22‐1 and 46‐6 B5 whereas the sorted 3 F9 clonotype‐positive cultures could be inhibited very effectively. All the CTL clones derived from the FACS‐sorted clonotype‐positive culture show all the same properties and are identical with clone 3F9 with respect to antigen‐specific cytotoxicity, inhibition of cytotoxicity by the mAb and surface markers.

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