Update on the Mouse/Chamber Vaccine Model

by David Abraham

The diffusion chamber/mouse model was used to answer several questions regarding the induction of protective immunity to larval Onchocerca volvulus.

The first question related to the route of administration of immunizations. Mice were immunized subcutaneously, intraperitoneally, or intramuscularly with a killed preparation of O. volvulus third-stage larvae (L3). A reduction in the survival of challenge L3 of approximately 60% was seen regardless of the route of administration of the vaccine.

The second question addressed concurred the choice of adjuvant to be used with the recombinant antigens. To answer this question 5 recombinant antigens (OI3, OV7, C27, paramyosin, and RAL2) were selected based on their occurrence in L3. These antigens were combined at equal concentrations into a cocktail which was then used with 4 different adjuvants to test the effect that the adjuvants had on the development of protective immunity. Immunization with the cocktail of antigens with the adjuvants Ribi, BCG, and QS21 (a plant saponin adjuvant; Cambridge Biosciences) did not induce protective immunity. Immunization, however, with the antigens with the adjuvant block copolymer (Robert Hunter, Cytrx Corporation) using the water in oil formulation (BCWO) caused a 67% reduction in challenge parasite survival.

It was of interest to note that the percent eosinophils and the absolute number of eosinophils found in the diffusion chambers of animals immunized with antigen and BCWO was higher than in animals which had not been immunized or that had been inoculated with BCWO without antigen. Evidence is accumulating in the mouse-chamber model system that eosinophils are an integral component of the protective immune response. Therefore, the finding of elevated eosinophil numbers only in diffusion chambers recovered from animals immunized with antigen and BCWO suggests that the finding of a decrease in parasite survival in these animals was not an experimental artifact but was, indeed, a manifestation of a protective immune response.

A second trial was set up to test two formulations of the block copolymer, water in oil vs. oil in water, for their relative efficacies. This experiment failed to demonstrate statistical difference between control and immunized groups because of large variations in parasite recovery within all of the groups, although the appropriate trends were seen. We are currently retesting the two adjuvant systems and will then proceed to testing individual antigens with the adjuvant system of choice.