Subpopulations of Mouse Spleen Lymphocytes. II. Immunological Reactivity of Spleen Cells Fractionated on BSA Density Gradients

We found in previous experiments that fractionation of non-immune mouse spleen cells on bovine serum albumin density gradients yields two subpopulations of T cells, one of high, the other of low density. Both subopopulations could be stimulated in corporate thymidine by the T cell-specific mitogen concanavalin A (con A). In the present investigation, spleen cells of mice immunized to sheep red cells (SRC) were similarly fractionated and the fractions recovered were assayed for: (a) reactivity to con A; (B) REACTIVITY TO SRC and (c) capacity to function as helper cells when stimulated with the homologous (SRC) or with a heterologous (donkey red cells) (DRC) antigen. Two subpopulations of cells reacting to con A were found in the spleens of the primed mice, corresponding to the subpopulations found in the non-immune mice. Both subpopulations contained cells responding to SRC (as measured by thymidine incorporation) and cells endowed with helper activity. The two subpopulations appeared to differ, however, in their specificity: while the denser cells could only exert their helper effect when stimulated by the specific antigen, the light cells could be effectively stimulated by both the specific (SRC) and the nonspecific (DRC) antigen.