Alloantiserum-mediated suppression of histocompatibility-linked Ir-gene-controlled immune responses. Suppressive effects of IgG fragments derived from alloantisera

Abstract

Fab, Fc, and F(ab)'(2) fragments were prepared by enzymatic hydrolysis of the IgG fraction of strain 13 antistrain 2 alloantisera. These fragments were not cytotoxic to lymphocytes bearing strain 2 histocompatibility antigens, but the Fab and F(ab)'(2) fragments retained functional combining sites as indicated by their ability to suppress the cytotoxicity mediated by the intact antistrain 2 antibodies. The F(ab)'(2) fragments were much more efficient as inhibitors in this system than the Fab fragments. F(ab)'(2) at 0.06 mg/ml and 0.45 mg/ml Fab produced comparable degrees of suppression. The F(ab)'(2) at 0.06 mg/ml completely suppressed DNP copolymer of L-glutamic acid and L-lysine (GL)-stimulated tritiated thymidine incorporation. The monovalent Fab at 0.45 mg/ml, however, had no significant effect on the in vitro responses to DNP-GL. Addition of the intact alloantisera can be delayed 3 h after initiation of the antigen-stimulated cultures with no loss of suppression. After a delay of 6 h 45% suppression was observed. The requirement for the divalent molecule and the observation that effective suppression of the in vitro responses is still obtained when the alloantiserum is added several hours after initiation of the cultures both suggest that the immunosuppression results from an active process affecting the lymphocyte membrane that renders the cell refractory to the antigenic stimulus.