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. 2020 Jun 1;9(1):1771142. doi: 10.1080/2162402X.2020.1771142

Figure 8.

Figure 8.

ARG2 vaccination results on LL/2 tumor growth delay. (a) Murine IFNy ELISPOT screening of PBMCs from the spleens of C57BL/6 mice vaccinated with one of six different predicted ARG2 epitopes. 8*10^5 cells were plated pr. well and peptide and control stimulation were performed in triplicates. Specific spot counts (peptide-specific IFNy secreting cells) are given as the difference in number of IFNy spots between averages of the wells stimulated with peptide and control wells. (b) Murine IFNy ELISPOT of PBMCs from the spleen of C57BL/6 mice vaccinated with ARG2 peptide P4 (M1-M5) or a control vaccination (Ctrl1-4). 8*10^5 cells were plated pr. well and peptide and control stimulation were performed in triplicates. Specific spot counts (peptide-specific IFNy secreting cells) are given as the difference in number of IFNy spots between averages of the wells stimulated with peptide and control wells.(c) ARG2 expression in engrafted tumors of different origin in C57BL/6 background. 3 engrafted tumors were evaluated pr. tumor type. Data are represented as relative expression to the housekeeping gene Hprt1, mean+SD, n = 3. (d) Treatment schedule for ARG2 (P4) vaccination in Lewis lung carcinoma (LL/2) challenged mice. On day 0, 0.5*10^6 LL/2 tumor cells were injected subcutaneously in the right flank of female C57BL/6 mice. Hereafter, mice were left untreated or received either control vaccination or ARG2 vaccination on day 0 and 7. (e) Average growth of LL/2 tumors in untreated or ARG2 (P4) vaccinated mice (n = 15 for each group). Female C57BL/6 mice were inoculated with LL/2 cells and were left untreated or received ARG2 (P4) vaccination on day 0 and 7, as indicated in (d). Error bars indicate the standard error of the mean (SEM). * = p = .013 (Two-way ANOVA with Bonferroni correction). (f) Individual tumor sizes from the treatment groups described in (e). Error bars indicate the standard deviation (SD). *** = p ≤ 0.001 (Two-tailed Student t-test). (g) Average tumor growth of LL/2 tumors in control vaccinated or ARG2 (P4) vaccinated mice (n = 20 for each group). Female C57BL/6 mice were inoculated with LL/2 cells and received either control vaccination or ARG2 (P4) vaccination on day 0 and 7, as indicated in (d). Error bars indicate the standard error of the mean (SEM). **** = p ≤ 0.0001 (Two-way ANOVA with Bonferroni correction). (h) Individual tumor sizes from the treatment groups described in (g). Error bars indicate the standard deviation (SD). **** = p ≤ 0.0001 (Two-tailed Student t-test).