Marilen Benner

B CELLS CONTRIBUTE TO DECIDUAL IMMUNITY 103 4 (A) Gating strategy to select lymphocytes as starting population of all samples used. (B) Representative flow cytometry plots to assess general lymphocyte composition of single cell suspensions based on CD3/CD56 expression resulting in frequencies of total NK cells, T cells, and NKT cells. (C) NK cells separated in NKbright and NKdim subsets based on CD16 expression. (D) CD4/CD8 expression within all CD45 + CD3 + lymphocytes. (E +F) Naive and effector memory (EM) cells of all CD4 + lymphocytes. (G) T cells with CCR4 - CCR6 - CXCR3 + , CCR4 + CCR6 - CXCR3 + and CCR4 + CCR6 + CXCR3 - profile. representing respectively T helper 1, T helper 2 and T helper 17 cells (H) Gating strategy to assess regulatory T cells. (I) Frequencies of NKT cells. Except for the depicted gates of CD3/CD56 expression, gate settings across tissues were identical and representative plots show cell distribution within these gates as frequency of parent population. PB, peripheral blood; MMC, menstrual blood mononuclear cells; 1 st , first trimester decidual lymphocytes; ^ p < 0.05, ^^ p < 0.01, ^^^ p < 0.001 Kruskal-Wallis and post-hoc Dunn’s multiple comparison test (non-parametric) to assess difference between isolated mucosal tissues and peripheral blood; * p < 0.05, *** p<0.001 Kruskal-Wallis and post-hoc Dunn’s multiple comparison test (non-parametric) to compare menstrual blood mononuclear cells, 1 st trimester, 2 nd trimester and term decidua. Data are represented as mean±SEM.