Alloantiserum-Mediated Suppression of Ir Gene Controlled Antigen-Induced Lymphocyte Activation: Effect on Uridine and Leucine Incorporation

Abstract

Abstract Antisera to guinea pig histocompatibility antigens specifically suppress Ir gene-controlled antigen-stimulated, DNA-synthetic responses in vitro. To define further the mechanisms of alloantiserum-mediated suppression and to utilize this suppression as a probe of the cellular events occurring during lymphocyte activation, we have examined the effects of alloantisera on 14C-leucine and 3H-uridine incorporation, earlier events after antigen stimulation. The RNA and protein synthetic responses of peritoneal exudate lymphocytes from immunized guinea pigs to DNP-GL (controlled by a strain 2-linked Ir gene) are 40 to 50% of maximum by 24 hr in culture and at or near maximum by 48 hr. DNA synthesis was only 2% of maximum at 24 hr and maximum at 72 hr. Comparisons of the degree of stimulation of the incorporation of all three precursors reveals an excellent correlation between leucine and uridine but poor correlation between leucine and thymidine. Despite the observed differences in time course of incorporation and magnitude of response, the susceptibility to suppression by alloantisera of all three responses was virtually identical. Anti-2 sera, added at the initiation of culture, completely suppress all three responses. If addition was delayed 8 hr, 50 to 60% suppression was still demonstrated, whereas only minimal suppression was evidenced if addition was delayed 18 hr. These results suggest that by 8 to 10 hr of incubation with antigen, the responding cells have undergone the necessary biochemical and structural changes to eventuate in an immune response; and, further - more, that alloantisera do not suppress by blocking antigen access to, or release from, its lymphocyte membrane receptor, but rather by a dynamic alteration of the lymphocyte membrane surface which interferes with the stimulus being generated by the antigen-receptor complex.