Factors affecting 7S and 17S antibody concentrations and affinities in chickens

Yamaga, Karen
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The formation of different immunoglobin classes and binding constants were determined during the immune response in chickens injected with highly substituted dinitrophenyl-bovine gamma globulin (DNP-BGG). A comparison between intravenous immunization and intramuscular injections with adjuvants was made. Antisera were assessed for hemagglutination and precipitating anti-DNP antibody, and the presence of 78 and 178 antibodies was detected by radioinmunoe1ectrophoresis (RIE). The binding constants were measured from 78 and 17S globulin preparations using extensive equilibrium dialysis determinations. Following an intravenous injection of DNP-BGG, the primary response was characterized by a transient synthesis of low amounts of 7S and 178 anti-INP antibodies. A single intravenous injection did not induce sufficient amounts of antibody to perform equilibrium dialysis studies. However, a second intravenous injection resulted in a more vigorous production of both classes of antibodies. Hem agglutinating titers were high and precipitins were present within 4 days after the second injection. Furthermore, the 178 antibody was detected for at least 4 months by RIE. In contrast to the response to intravenous injections, the response of chickens given a single intranus0l1ar injection of antigen in either Freund's complete (FCA) or incomplete (FIA) adjuvants was characterized by an initial synthesis of 78 and 178 antibodies followed by the exc1usive and persistent production of 7S antibodies. Hemagglutinating titers were low even after 2 intramuscular injections of antigen in adjuvant. After a single injection of antigen in FCA with doses ranging from 0.02 to 20 mg, binding of the 7S anti-DNP antibody was detected by RIE for at least 490 days. As a result of either intravenous or intramuscular injections of INP-BGG, the anti-BGG responses were more transient in nature than the anti-hapten responses. After two intravenous injections little or no anti -BGG antibodies were detected by RIE one month later. Persistent synthesis of 7S anti-BGG antibodies generally required 2 intramuscular injections of DNP-BGG in FCA. Injections of low doses of antigen in either FCA or FIA in chickens induced 7S antibodies of high affinity (-ΔF° > 10 kcal/mole) by 3 months. The non-linearity of the Sips plots generated from the binding data indicated that a shift in the distribution of antibody affinities occurred due to the production of high affinity antibodies. In birds immunized with FCA the high affinity antibodies had restricted heterogeneity as indicated by heterogeneity indices of 1. Chickens, given multiple intravenous injections, however, failed to produce high affinity antibodies. Despite altering the priming dose, the interval between injections and the number of injections, chickens injected intravenously produced 7S antibodies with -ΔF° values that were consistently less than 10 kcal/mole. Sips plots of the binding data indicated that the antibody affinities were distributed in a normal array. It appears that at least two important conditions are required for eliciting high affinity antibody, namely, limited concentrations of antigen and the adjuvant effects. Nearly identical affinities and heterogeneity indices were obtained for both the 7S and 17S antibodies isolated from 12 individual chickens given 2 intravenous injections of antigen. Thus, intravenous injections of antigen failed to elicit high affinity 7S and 17S antibodies. The conditions required to generate circulating 17S antibody of high affinity are not known. In chickens given 2 intravenous injections of DNP-BGG, more 7S antibody was produced when the interval between intravenous injections was short than when the interval was longer. Intravenous injections apparently failed to induce large numbers of long-lived memory cells. Less antibody was synthesized in birds given either low (0.02 mg) or high (20 mg) doses of priming antigen than in birds injected with intermediate doses. Specifically purified low affinity 7S antibodies had valences of less than 2, whereas a valence close to 2 was found for antibodies of higher affinities. Similarly l7S antibody of low affinity gave valences less than 10. Obtaining valences less than the expected value is best explained on the basis of contamination of the preparation with nonbinding proteins or the presence of antibody with very low affinity.
Thesis (Ph. D.)--University of Hawaii at Manoa, 1974.
Bibliography: leaves 131-141.
xiii, 141 leaves ill
Poultry -- Physiology, Immunoglobulins, Gamma globulins -- Metabolism
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Theses for the degree of Doctor of Philosophy (University of Hawaii at Manoa). Microbiology; no. 685
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