Fig. 5. Pregnancy affects T cell response to 3′UTR-Δ10-LAV vaccination in A129 mice.

a Experiment scheme. Top scheme: 12-week-old female A129 mice (n = 4 or 5)were mated with male mice; at E0.5, the pregnant mice were subcutaneously immunized with 10³ FFU 3′UTR-Δ10-LAV (Δ10) or PBS; at E18.5, they were sacrificed and splenocytes were harvested for T cell analysis. Bottom scheme: age-matched non-pregnant female mice (n = 3 or 6) were immunized and analyzed for T cell response to Δ10 vaccination, as described above for the pregnant group. b Percentages of CD4⁺IFN-γ⁺, CD8⁺IFN-γ⁺, CD4⁺IFN-γ⁺TNF-α⁺, and CD8⁺IFN-γ⁺TNF-α⁺ cells. Spleen cells were harvested and stained for IFN-γ⁺, TNF-α⁺, CD3⁺, and CD4⁺ or CD8⁺. Samples were acquired with BD Accuri C6 Flow Cytometer instrument. Total splenocytes were first gated to exclude, debris, cell fragments, and dead cells based on forward (FSC) and side scatter (SSC). CD3⁺CD4⁺ or CD3⁺CD8⁺ T cells were next gated for analysis of cytokine production. c Total numbers of CD4⁺ T cell subsets per spleen. Splenocytes were stimulated by ZIKV. d Total numbers of CD8⁺ T cell subsets. Splenocytes were stimulated by a ZIKV E peptide. Cytokines IL-2 (e) and IFN-γ (f) in cell culture media were measured after splenocytes had been stimulated by the viral E peptide for 3 days. An unpaired t test was performed to analyze statistical significance between indicated groups. **p < 0.01, ***p < 0.001, non-significant (n.s.) p > 0.5. Triple technical replicates were performed for (c), (d), (e) and (f). Placebo-P PBS placebo pregnant mice, placebo-NP PBS placebo non-pregnant mice, vaccine-P vaccinated pregnant mice, vaccine-NP vaccinated non-pregnant mice. Error bars represent standard deviations. Source data are provided as a Source Data file.