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CBA spleen cells have been stimulated in vitro with A/Jap influenza virus-infected CBA spleen cells to generate a ‘primary’ cytotoxic lymphocyte (CL) response. The culture conditions were devised to allow the segregation of individual clones of CL and cytotoxicity measured by the lysis of infected or non-infected L-929 cells. The specificity was assessed by splitting clones and measuring the ability of the clones to discriminate between pairs of targets. Influenza A/FMI and A/Jap strains were used. Subsets of clones were detected which could lyze either A/Jap-infected or A/FMI-infected target cells. In addition CL clones were found which lyzed uninfected L-929 cells and a fourth category were clones which could not discriminate between A/FMI- and A/Jap-infected targets.