Immune Response and Protective Efficacy of a Heterologous DNA-Protein Immunization with Leishmania Superoxide Dismutase B1

Figure 3

Prechallenge cytokine response of mice immunized with LdFeSODB1 antigen. The level of cytokine production was assessed in stimulated spleen cells. (a) IFN-γ, (b) IL-10, and (c) mean IFN-γ/IL-10 ratio. Mice were immunized twice with pcDNA-LdFeSODB1 or pcDNA-mGMCSF-LdFeSODB1 and boosted with rLdFeSODB1 protein. The control mice received three doses of pcDNA, pcDNA-mGMCSF, or CpG ODN alone. The level of IFN-γ and IL-10 was measured from stimulated spleen cells using cytokine ELISA kit (BD Biosciences). The concentration of the cytokines (ng/ml) was calculated by correlating the optical density to concentration of the protein standard included in the kit. The mean concentration and standard error of the mean (SEM) of five mice per group are shown. This is one of two experiments with similar result. The IFN-γ/IL-10 ratio was calculated from cells that were stimulated with rLdFeSODB1. Statistical comparison between groups was performed using Mann–Whitney test. Asterisks indicate statistically significant difference in the cytokine production between mice immunized with antigen and the respective controls ().

(a)

(b)

(c)