Kinetics of lymphocyte transformation in mice immunized with viable avirulent forms of Cryptococcus neoformans

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Abstract
A murine model was developed to study the cell-mediated immune response of mice immunized with 1 of 2 live, avirulent forms of C. neoformans: a nonencapsulated mutant and a thinly encapsulated pseudohyphal variant. A lymphocyte transformation assay was used to evaluate the cellular response of control and sensitized spleen cells after in vitro incubation with 3 merthiolate-killed whole-cell antigens [Ag] of C. neoformans. An Ag-to-spleen cell ratio of 10:1 and 5 days of incubation of Ag-spleen cell mixtures were established as optimal conditions for maximum lymphocyte transformation. Maximum responses occurred from 2-3 wk after the last of 8 weekly i.p. inoculations of C. neoformans. This assay provided an accurate, reproducible method of studying cell-mediated immunity to C. neoformans and applications to the study of cryptococcal pathogenesis are proposed.