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. 2019 Dec 19;4(24):e130540. doi: 10.1172/jci.insight.130540

Figure 4. Longitudinal analysis reveals that BCG revaccination significantly enhances Ag85A- and BCG-specific IFN-γ and/or IL-2 CD8+ T cell in whole blood.

Figure 4

Left panel shows ICS data for Ag85A, and right panel shows data for BCG following in vitro stimulation of whole blood from BCG-revaccinated and control subjects. Representative flow cytometry plots show IFN-γ and/or IL-2 cytokine–positive CD8+ T cells after in vitro stimulation with either Ag85A (A) or BCG (E) at T0 and T2 (A) or T0 and T5 (E) Total frequencies of Ag85A- (B) and BCG-specific (F) IFN-γ and/or IL-2 cytokine–positive CD8+ T cells in all participants are shown after background subtraction. Line graphs show changes in frequencies of specific CD8+ T cells over time in BCG-revaccinated (Group 1 [IGRA+, red, n = 19] and Group 2 [IGRA–, blue, n = 18]) subjects and unvaccinated controls (Group 3 [IGRA+, black, n = 18] and Group 4 [IGRA–, brown, n = 18]). P values for longitudinal samples were calculated by comparing each time point with baseline using the Freidman test and corrected for multiple comparisons using Dunn’s test. Further, significant differences were determined using the Wilcoxon paired, 2-tailed t test before and after vaccination. Bonferroni adjusted P value threshold of 0.02 was considered statistically significant. Median frequencies of Ag85A (C) and BCG (G) specific IFN-γ and/or IL-2 cytokine–positive CD8+ T cells over time were compared between unvaccinated and revaccinated IGRA+ and IGRA– subjects (Group 1 versus Group 3 and Group 2 versus Group 4). Mann-Whitney U test was used to determine significant differences between groups. P < 0.05 was considered significant. Analysis of CD45RA and CD27 expression was used to determine percentage distribution of naive (CD45RA+CD27+), central memory (CM, CD45RA–CD27+), effector memory (EM, CD45RA–CD27–), and terminally differentiated T effector memory cells (TD, CD45RA+CD27–) Ag85A- (D) and BCG-specific (H) IFN-γ and/or IL-2 CD8+ T cells in BCG-revaccinated (Group 1 [IGRA+] and Group 2 [IGRA–]) subjects. Wilcoxon paired, 2-tailed t test was used to determine significant differences before and after vaccination. P < 0.05 was considered significant.