Fig. 7: Sex-dimorphic impact of Maternal Zika Infection on Offspring Neutrophil Response to LPS.

a Experimental design. Created in BioRender. Alvero, A. (2025) http://BioRender.com/750w48I. Bone marrow neutrophils were isolated at PND35 from male and female offspring of control (n = 3–4 isolations) and ZIKV-exposed groups (n = 3–4 isolations) and treated with 100 ng/mL LPS to assess activation. Each isolation pooled neutrophils from 2–3 mice and was plated in duplicate or triplicate. b Representative scatter plots show gating of CD11b⁺Ly6G⁺ neutrophil subsets. c, d After 2 h LPS, the percentage of activated neutrophils (CD64⁺CD62L⁻) within CXCR2^int and CXCR2^high populations was analyzed by sex: control untreated and LPS groups had n = 3 isolations, ZIKV untreated and LPS groups had n = 4 isolations. *p = 0.0142, ****p < 0.0001. e, f After 24 h LPS, activated neutrophils were analyzed: for males, control untreated and LPS (n = 4 isolations), ZIKV untreated and LPS (n = 3 isolations); for females, control untreated and LPS (n = 1 isolation), ZIKV untreated and LPS (n = 2 isolations). **p = 0.0021, ***p = 0.0001, ****p < 0.0001. g, h Cytokine levels in conditioned media after 24 h LPS (Luminex) were measured with male groups n = 3 isolations each, female controls n = 1 isolation, and ZIKV n = 2 isolations. For males, IL-6 (*p = 0.0122, ***p = 0.0007), TNF-α (****p < 0.0001), G-CSF (**p = 0.0087, ***p = 0.0001), and MIP-1α (**p = 0.006, *p = 0.0144) showed significant changes. For females, IL-6 (*p = 0.0175, **p = 0.0033), IL-1β (***p = 0.0006, ****p < 0.0001), MIP-1β (**p = 0.0024, *p = 0.036 for ZIKV control vs. LPS, *p = 0.0447 for LPS control vs. ZIKV), and RANTES (**p = 0.0029 for ZIKV control vs. LPS, **p = 0.0019 for LPS control vs. ZIKV, ****p < 0.0001) were significant. Data are mean ± SD. Two-way ANOVA with Šídák’s multiple comparisons was used. Female and male mice symbols (panel c–h) were created in BioRender. Alvero, A. (2025) http://BioRender.com/u0w40ys. Source data are provided as a Source Data file.