Cellular basis for a hapten-specific state of tolerance induced in vitro

A hapten-specific unresponsive state was induced in vitro by the incubation of normal murine spleen cells with highly conjugated dinitrophenylated bovine gamma-globulin (DNP-BGG) or a dinitrophenylated copolymer of D-glutamic acid and D-lysine (DNP-D-GL) for 24 hr. After this incubation period spleen cells were washed and cultured for 4 days with the thymic-independent antigen dinitrophenylated polyacrylamide beads (DNP-PAA) or the thymic-dependent antigen trinitrophenylated burro the erythrocytes (TNP-BRBC). Preincubation with either DNP-BGG or DNP-D-GL led to a specific depression of the in vitro anti-hapten plaque-forming cell response. The degree of depression was dependent upon the concentration of the tolerogen and the duration of preincubation. The response to DNP-PAA or TNP-PAA beads was depressed to a greater degree than was the response to TNP-BRBC. The cellular basis of the immunologic unresponsiveness induced by DNP-BGG was attributable to an inhibition of B cell function whereas the unresponsive state induced with DNP-D-GL was due to both a specific inhibition of B cell function and the activation of antigen-specific suppressor T cells.