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. 2019 Nov 26;10:2749. doi: 10.3389/fimmu.2019.02749

Figure 2.

Figure 2

CPA160−189-specific immune responses in BALB/c mice post vaccination. (A) BALB/c mice were vaccinated thrice with 2 week intervals with CPA160−189-loaded p8-PLGA nanoparticles or p8-PLGA nanoparticles. (B) Two weeks after each vaccination, spleens were collected and splenocytes were stimulated in vitro with CPA160−189 for 72 h. CPA160−189-specific splenocyte proliferation was determined by [3H]-thymidine incorporation after another 18 h of culture. The culture supernatants from spleen cell cultures obtained 2 weeks after the last booster vaccination were assessed for production of (C) IL-2, (D) IFN-γ, (E) IL-4, and (F) TNFα by Luminex. Also, spleen cells were cultured in the presence of CPA160−189 for 48 h for the assessment of CPA160−189-specific (G) IFN-γ-producing CD4+ T cells and (H) IFN-γ-producing CD8+ T cells by flow cytometry. Each point depicts the data from a single sample (n = 5 per group). Results are shown as mean ± SD and levels of significance, indicated by p-values, were assessed by one-way ANOVA and Tukey's multiple comparison tests (*p < 0.05; **p < 0.01; ***p < 0.001). Experiments were conducted twice with similar results and representative results from one experiment are given.