Figure 6. Maternal WSD attenuates fetal monocyte and macrophage cytokine responses to ex vivo stimulation.

(A) NHP model of maternal diet-induced obesity. Fetal (GD130) PBMC, ileal leukocytes, and spleenocytes were obtained from rhesus macaque born to dams on a control chow diet (n=3) and western-style diet (n=4). Fetal macrophages from ileal lamina propria lymphocytes and splenocytes were FACS-sorted and cultured overnight with E. coli. Supernatants were collected and cytokines/chemokines were measured using Luminex. (B) Bar graph representing the frequency (mean and ± SEM) of responding fetal monocytes to LPS in PBMC isolated from GD130 fetal circulation measured by intracellular cytokine staining. (C) Frequencies of CD14 + HLA-DR+macrophages within live total ileal leukocytes (top) or live CD3-CD20- leukocytes (bottom) (D–E) Median fluorescence intensity (MFI) of (D) M1-associated HLA-DR and (E) M2-associated CD209 and CD206 within live macrophage population in ileum (top) and spleen (bottom). (F–G) Bubble plots comparing significantly different cytokine/chemokine levels in response to E. coli stimulation in (F) gut (ileum) and (G) splenic macrophages. Statistical differences between chow and WSD groups are highlighted. *-p<0.05.