Immune response in rabbits to surface components of extracellular and intracellular forms of vaccinia virus

The development of cellular as well as humoral immune response to extracellular and intracellular forms of vaccinia virus (ECV and ICV, respectively) and their surface antigens were studied in rabbits. Direct lymphocyte cytotoxicity and peripheral blood leukocyte migration inhibition tests were used to measure cell-mediated immune response, while neutralizing and hemagglutination-inhibiting antibodies were assayed for measuring humoral immune response. Direct cytotoxicity of lymphocytes from rabbits immunized with ECV or its surface proteins was demonstrable by day 7 after immunization, and by the end of week 3 it almost declined to pre-immunization levels. Inoculation with ICV or its surface proteins failed to induce lymphocyte cytotoxicity. In contrast, migration inhibition of peripheral blood leukocytes from rabbits immunized with ECV, ICV, or their surface proteins was observed with homologous antigens. However, leukocytes from rabbits immunized with ECV or its surface proteins also showed migration inhibition in the presence of ICV. Similarly, in the humoral immune response, neutralizing antibodies were produced against homologous as well as heterologous forms of virus despite immunization with purified preparations of ECV, ICV, or their surface proteins. Adsorption with purified ICV preparations abolished the neutralizing activity of these antisera against heterologous forms of virus. Hemagglutination-inhibiting antibodies, on the other hand, were produced only after immunization with ECV or its surface proteins. In addition, antibody-dependent cell-mediated cytotoxicity was employed to detect specific antibody response after immunization of rabbits with live virus, ECV, and ICV. Antisera raised against ECV or live virus supported antibody-dependent cell-mediated cytotoxicity, whereas ICV-antiserum failed to do so. The antibody activity present in the former antisera was abolished by absorption with cell membranes from vaccinia-infected cells but not with purified ICV. The data suggest that immunization with inactivated ECV seems to bring about interaction between host immune response (cellular and humoral) and virus-infected cells, which may, perhaps, be necessary for protection against pox virus infection. A similar interaction is unlikely to occur after immunization with inactivated ICV.