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. 2018 Mar 27;9:643. doi: 10.3389/fimmu.2018.00643

Figure 4.

Figure 4

In vivo immune responses induced by p24 recombinant Mycobacterium bovis BCG (rBCG) strains. (A) Schematic of the immunization performed for the in vivo immunological assays. With 4 weeks interval, each group (five mice/group) was immunized twice with wild-type BCG, two rBCG strains, and rSmeg strain. Four weeks after final immunization, mice were sacrificed and their spleens and blood samples were collected for immunological analyses. (B) IFN-γ secretion levels following in vitro stimulation of splenocytes from mice vaccinated with the p24 rBCG strains were detected using an ELISPOT analysis. Representative images of ELISPOT membrane in each group are shown below the graph. (−), negative control; (+), positive control. (C) Levels of the IL-2, IFN-γ, TNF-α, IL-6, and IL-10 cytokines following in vitro stimulation with p24 of splenocytes from mice vaccinated with the p24 rBCG strains were detected using enzyme-linked immunosorbent assay analyses. A total of five mice per group was analyzed. Data are representative of two independent experiments. Mean ± SD are shown. *P < 0.05; **P < 0.01; ***P < 0.001 (Student’s t-test).