Establishment of dose-response relationships in BALB/c mice, using Brucella cell surface protein and lipopolysaccharide

Abstract

SUMMARY A study was conducted to determine the immune (increased antibody) and protective (reduced colony-forming units) responses induced in mice by a: (i) single vaccinal inoculation, using various concentrations of Brucella cell surface protein (bcsp) or lipopolysaccharide (lps); (ii) primary inoculation, using various concentrations of bcsp, followed by a secondary inoculation, using a standard concentration of bcsp; and (iii) primary inoculation, using 1 concentration of bcsp or lps, followed by a secondary inoculation, using various concentrations of bcsp or lps. Four weeks after the primary inoculation, mice were challenge exposed with approximately 1 × 104 colony-forming units of Brucella abortus strain 2308 and all mice were euthanatized at 6 weeks. Reduced splenic weights and reduced colony-forming units in the spleens of vaccinated mice, compared with nonvaccinated mice, were the criteria of protection. Increase in serum IgM and IgG was defined as immunity. Both bcsp and lps induced protective and immune responses that were proportional to the dose given up to an optimal limit. However, concentrations higher than optimal decreased the protective and immune responses. This was true for mice given either 1 or 2 vaccinal inoculations. Enhanced secondary protective responses were seen only when suboptimal doses were used in the primary inoculation. Excessive or optimal doses in the secondary inoculations prevented or obscured the protectiveness and immunity by primary inoculations. The protective effects appeared to be additive when suboptimal doses were used in the primary and secondary inoculations. Inoculation of subimmunogenic doses induced a relative reduction in the antibody concentration after challenge exposure, compared with nonvaccinated mice. The overall results indicated that the protective responses induced by bcsp were probably attributable to lps. The results also indicated a linear increase in protection and immune response corresponding to increasing doses up to an optimal dose, and this stoichiometric optimum may be achieved by the use of 1 or more vaccinal inoculations. However, once this optimum was obtained, additional amounts of bcsp or lps cause perturbation of both the protective and serologic responses.