Understanding the effects of sex and sex hormones on markers of human macrophage polarization through streamlined analysis of large flow cytometric datasets using R 9023

Abstract Description Lung macrophages (MΦ) expressing M2 markers contribute to asthma pathogenesis and are more abundant in asthmatic females (F) than males (M). Understanding how sex and sex hormones affect MΦ polarization and function is key for developing new asthma therapies for both sexes. In this study, we investigated the effect of sex hormones on MΦ polarization and implemented an R-based flow cytometry and statistical analysis pipeline to reduce time and errors associated with manual analysis of large datasets. CD14+ blood monocyte-derived MΦ from healthy control (HC) and allergic asthmatic (AA) M and F were treated with or without the respective sex hormones Testosterone (T) or Estrogen (E2), polarized with IL-4 or IFN-γ + LPS, and analyzed by flow cytometry. We found that IL-4 upregulated M2-like markers CD206 and CD209 in all MΦ, while the M1-like marker, CD80, was not induced by IFN-γ + LPS but was greater on AA vs HC M MΦ. Surprisingly, the M1 marker CD64 was greater on IL-4-treated AA F vs M MΦ, and on IL-4-treated AA M vs HC M MΦ treated with or without T. CD25 was increased by IFN-γ + LPS on HC M MΦ, and on AA F MΦ by IFN-γ + LPS with E2. No differences were observed in CD163 and HLA-DR. Overall, we found asthma-related elevations in CD64 and CD80, and F AA sex differences in CD64. Sex hormones did not significantly change MΦ surface markers although we only tested one concentration. Adopting R-based analysis allows faster more accurate analysis of large flow cytometric datasets. Funding Sources Supported by NIH HL124477 Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)