Abstract
Recently several investigators have shown that the immune response of an animal immunized with hapten-carrier conjugates is significantly influenced by the nature of the carrier molecule. Siskind et at. (1) demonstrated in guinea pigs that the carrier to which DNP is coupled influences the amount, type, and functional activity (i.e., affinity) of anti-DNP antibodies, and Fronstin, Sage, and Vazquez (2) have shown that the degree of anti-DNP responsiveness elicited in mice is directly related to the antigenicity of the carrier.
The present study examines the antihapten antibody response of guinea pigs immunized with arsanilic acid conjugates of homologous and heterologous serum protein carriers, and compares the effect of natural and acquired tolerance to the carrier on these responses.
Materials and Methods. Random bred, adult male albino guinea pigs (300-360 g) were used in these experiments.
Reagents. Azobenzenearsonate (ABA) conjugates were made as before (3) by diazoti-zation and coupling in the ratio of 10-5 moles arsanilic acid to 10 mg protein. Human, bovine, rabbit, and guinea pig serum albumins (HSA, BSA, RSA, GSA) and gamma globulins (HGG, BGG, RGG, GGG), obtained from Pentex Inc., Kankakee, Ind., were utilized as carrier proteins. In addition, arsanilic acid conujgated to egg lysozyme (Worthington Lab.) was used as the test antigen in studies of the antihapten antibody response. All conjugates were dialyzed against 0.01 M borate buffer pH 8.5 before use.
Immunization procedure. Guinea pigs were immunized with 0.1 ml of complete Freund's adjuvant containing 100 fig of ABA-protein distributed equally among the four footpads. The complete Freund's adjuvant was prepared with 8.5 parts light mineral oil: 1.5 parts Arlacel A: 5 mg/ml killed, dried M. tuberculosis.
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