A B Supplemental Figure 1 PBMC Cervical Cytobrush

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A B Supplemental Figure 1 PBMC Cervical Cytobrush CD4+ CD8+ (CD3+/CD4-) PBMC Cervical Cytobrush B Participant 18 Participant 19 UV-mock UV-HSV-2 Supplemental Figure 1 A. Gating strategy to phenotype total T cells derived from a cervical cytobrush and PBMC sample from a representative HSV-2 infected participant. Gating focused on live CD3+ cells in the lymphocyte forward/side scatter regions that were CD4+ and CD4-negative, the latter of which was considered CD8+ for quantitation. The expression of CCR7/CD45RA and CD69/CD103 are displayed for the CD3+CD4+ and CD3+CD4- (CD8+) populations. B. Measuring cervical cytobrush-derived HSV-2 reactive CD4+ T cell responses from 2 HSVneg participants. Cells isolated from cervical cytobrushes obtained from 2 HSVneg participants were cultured with UV-mock (bottom graphs) or UV-HSV-2 (top graphs) and the resultant CD4+ T cells were analyzed by ICS and flow cytometry for the expression of IFN-g. Net % CD4+/IFNg+ (UV-HSV-2 minus UV-mock) was 0.0% for Participant 18 and 0.01% for Participant 19. In both cases, the proportion of CD4+/IFN-g+ cells stimulated with UV-HSV-2 was not statistically significantly different than those stimulated with UV-mock (Fisher’s Exact Test P>0.05).