Lytic Effect of Bacterial Products on Lymphocytes of Tuberculous Animals

Abstract

Suspensions of white blood cells predominantly lymphocytes were prepared from blood buffy coats and teased splenic pulp. Advantage was taken of the clumping tendency of granulocytes to allow them to settle out of suspensions at a more rapid rate than other cell types, leaving lymphocytes in the supernatant fluid for further study. Red cells were removed from these supernatants by floating the white cells free on the interface of isotonic concentrated albumin solutions. Within an hour, in the presence of fresh normal serum, 20-50% of the lymphocytes from tuberculous mice were lysed by 50[gamma] PPD/ml. Mouse granulocytes and monocytes were not affected. PPD did not cause lympholy-sis of normal mouse lymphocytes or lymphocytes from mice infected Salmonella enteriditis. Broth filtrate of S. enteri-ditis caused specific lympholysis for mice infected S. enteriditis, and did not lyse cells from normal or tuberculous mice. Lymphocytes from tuberculous guinea pigs were also specifically lysed by PPD and not by 5. enteriditis filtrate. Guinea pig lympholysis was promptly followed by destruction of neutrophilic granulocytes. Mouse and guinea pig-specific cell lysis is blocked or greatly slowed by phosphate buffer at pH 7.2, or by suspending the cells in saline or albumin sol-lutions. It should be recalled that the guinea pig develops a typical delayed type skin reaction to PPD; the mouse does not.