A microsomal fraction of Cryptococcus neoformans induces lymphocyte blastogenesis in infected guinea pigs.
Differential centrifugation of a homogenate from a mechanically disrupted, acapsular isolate of Cryptococcus neoformans resulted in a 105,000 x g supernatant (105 K) and a microsomal fraction (MS), both of which were capable of eliciting specific delayed cutaneous hypersensitivity and in vitro blastogenesis in infected guinea pigs. Polyacrylamide gel electrophoresis revealed two major proteins in the MS and seven proteins in the 105 K fractions. Electron microscopy of the MS showed both membranes and ribosomes. In vitro lymphocyte blastogenesis elicited by 1 to 10 micrograms/ml of antigens was maximal after 4 days of incubation; the reacting populations were peripheral blood leukocytes (PBL) and peritoneal exudate cells (PEC). Spleen cells of infected animals were unresponsive to in vitro antigenic stimulation. A simplified schedule of priming animals was infection with a single dose of virulent cryptococci. Under these conditions 3 of 6 animals' PBL responded with stimulation ratios of 6.55, 21.1, 35.42 to the MS and 1.41, 9.33, 17.39 to the 105 K antigens at 1 microgram/ml. Four of six animals' PEC response were positive with stimulation ratios of 2.62, 2.72, 4.02 and 7.20 towards MS, and 2.62, 5.13, 5.71, 10.01 to the 105 K antigens at 1 microgram/ml. When small capsule and large capsule isolates were used for infection, the small capsule form was not isolated from the brain, in contrast to its isolation from 2 of 3 animals receiving large capsule forms. Two of three animals in each group responded with blastogenic indices more vigorous in the PBL, and the most potent antigen was MS. There was no obvious difference in lymphocyte reactivity between the two groups.