Coordinate expression of cytolytic activity and cytotoxic T cell-specific carbohydrate antigens in a T cell hybridoma.

The expression of cytotoxic T lymphocyte (CTL)-specific carbohydrate antigens (termed CT antigens) was studied by using a cytolytically inducible T cell hybridoma, KSH4.13.6. Expression of the CT determinants occurred concomitantly with the expression of cytolytic activity after induction of the hybrid with supernatants from Con A-activated rat spleen cells. Purified IL 2 was also proven to be effective in inducing cytolytic activity and CT antigen expression, but the time course of activation by IL 2 was prolonged as compared to activation by crude supernatants. Furthermore, the activation process was reversible because removal of the hybrid from inducing medium resulted in the loss of cytolytic capability and CT antigen expression. By separating the low and high expressors of CT antigen from an induced hybrid population, it was shown that the level of CT antigen expression correlated with the cytolytic ability of the hybrid. High expressors of CT antigen exhibited four- to 50-fold greater lytic activity than populations with low CT antigen levels. Binding experiments using lectins indicated that an increase in GalNAc-containing oligosaccharides also occurred on activation of the hybrid. This finding agrees with our results which indicated that the CT carbohydrate antigens are probably associated with O-linked glycans. Because our previous results with CTL clones indicated that the CT antigens were associated with the T200 glycoproteins, we performed immunoprecipitation experiments with surface-labeled induced and uninduced KSH4.13.6. The T200 glycoproteins were precipitated by the CT1 monoclonal antibody from the induced population, but not from the uninduced population. Furthermore, precipitation by the GalNAc-recognizing lectin from Vicia villosa revealed marked differences in the GalNAc-containing proteins between the induced and uninduced populations. Thus, the results indicate that the T cell-derived polypeptide hormone IL 2 is able to influence the glycosylation of specific proteins in CTL, which results in the appearance of carbohydrate antigens whose expression is linked to the activation state of the CTL.