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CD4+ CTL are not Ca2+ dependent and may use the Fas-mediated lytic pathway. Addition of selected concentrations of Mg2+ and EGTA (Ca2+ chelator) to the CD4+ CTL assay mixtures did not inhibit BHV-specific cytolytic activity, suggesting that antigen-specific CD4+ CTL are not calcium dependent. Medium was added to macrophage target cells as a negative control for miscellaneous lytic activity in CTL assays. Similar results were obtained in five separate experiments.
