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. 2022 Dec 23;12(9):2202691. doi: 10.1002/adhm.202202691

Figure 4.

Figure 4

Immunogenicity detection of epitopes derived from ESCs. a) ELISPOT assay for immune cell activation of splenocytes in the ESC (C57) or ESC (129) + CpG + GM‐CSF vaccinated group (n = 5) compared to PBS (n = 5) group upon exposure to selected epitopes. Quantitative analysis of the ELISPOT assay for IFN‐γ secretion. b) C57BL/6 mice were inoculated with bladder cancer cells for 1 week, and injected with PBS, ESC (C57) or ESC (129) + CpG + GM‐CSF once a week for 2 weeks. ELISPOT assay was performed. Significant increase in number of IFN‐γ spots in the ESC (C57) and ESC (129) + CpG + GM‐CSF‐vaccinated C57BL/6 mice (n = 3) compared to PBS group (n = 3) upon exposure to prediction epitopes. Quantitative analysis of the ELISPOT assay for IFN‐γ secretion. c) Antitumor effects of each epitope were detected in bladder cancer‐inoculated mice. C57BL/6 mice (n = 5) were implanted with 3 x 105 MB49 bladder cancer cells s.c. in the right flank, after 1 week, mice were immunized with PBS or different epitope peptides on day 3, day 6, and day 9 and boosted on day 15. Tumor growth was measured every 2 d. d) Five days after the last vaccination, splenocytes (5 × 105) were cocultured with different peptides (10 µg) for the duration of 20 h, cellular immune responses were measured using ELISPOT assay (n = 4). Experiments were repeated three times. Quantitative analysis of the ELISPOT assay for IFN‐γ secretion. Data represent mean ± SEM, *p < 0.05, **p < 0.001, ***p < 0.001, ****p < 0.0001 (Student's t‐test).