Preformed purified peptide/major histocompatibility class I complexes are potent stimulators of class I‐restricted T cell hybridomas

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Abstract
A panel of antigen‐specific, major histocompatibility complex class I‐restricted T cell hybridomas has been generated to examine the capacity of peptide/class I complexes to stimulate T cells at the molecular level. Peptide/class I complexes were generated in detergent solution, purified and quantitated. Latex particles were subsequently coated with known amounts of preformed complexes and used to stimulate the T cell hybridomas. Stimulation was specific, i.e. only the appropriate peptide/class I combination were stimulatory, and quite sensitive, i.e. as little as 300 complexes per bead could be detected by the T cells. Preformed complexes were about 500000 times more potent than free peptide in terms of T cell stimulation, demonstrating the physiological relevancy of the biochemically generated complexes. Surprinsingly, the majority (including the most sensitive of the hybridomas) had lost CD8 expression, suggesting that antigen‐specific stimulation of class I‐restricted T cell hybridomas, as assessed by IL‐2 release, does not depend on CD8.