title: Multi-parameter flow cytometric analysis of uterine immune cell fluctuations over the murine estrous cycle.
abstract: Investigating immune cell populations within various reproductive tissues commonly utilises flow cytometric methods. With advances in fluorophore technology and equipment capabilities, multiple cell types from a single tissue sample can be identified by using different combinations of cell surface markers to distinguish specific cell populations. Here a protocol optimized for mouse uterine tissue was used to show the proportional changes in dendritic cells, monocyte/macrophages, T and B cells, NK and NK T cells, and the granulocytes, neutrophils and eosinophils at each of the four stages of the estrous cycle. Importantly, we demonstrate that use of anti-SiglecF or assessment of FSC/SSC plots could be used to differentiate monocyte/macrophage and eosinophil populations that otherwise cannot be distinguished by use of the common combination of antibodies against F4/80 and CD11b. Our results clearly indicate that within the uterus a dynamic population of immune cells resides, with many cell types reaching peak abundance at estrus and metestrus phases of the cycle, consistent with their importance in the response to paternal antigens and/or pathogens encountered after insemination.
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute, Royal Adelaide Hospital, and Sansom Institute, School of Pharmacy and Medical Science, University of South Australia, Adelaide, S.A., 5000, Australia; Robinson Research Institute, Discipline of Obstetrics and Gynaecology, School of Medicine, University of Adelaide, Adelaide, S.A., 5005, Australia. Electronic address: kerrilyn.diener@adelaide.edu.au.
authors.forename: Kerrilyn R
authors.initials: KR
authors.lastname: Diener
authors.affiliation: Robinson Research Institute, Discipline of Obstetrics and Gynaecology, School of Medicine, University of Adelaide, Adelaide, S.A., 5005, Australia.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Robertson
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute, Royal Adelaide Hospital, and Sansom Institute, School of Pharmacy and Medical Science, University of South Australia, Adelaide, S.A., 5000, Australia; Robinson Research Institute, Discipline of Obstetrics and Gynaecology, School of Medicine, University of Adelaide, Adelaide, S.A., 5005, Australia.
authors.forename: John D
authors.initials: JD
authors.lastname: Hayball
authors.affiliation: Experimental Therapeutics Laboratory, Hanson Institute, Royal Adelaide Hospital, and Sansom Institute, School of Pharmacy and Medical Science, University of South Australia, Adelaide, S.A., 5000, Australia; Robinson Research Institute, Discipline of Obstetrics and Gynaecology, School of Medicine, University of Adelaide, Adelaide, S.A., 5005, Australia.
authors.forename: Erin L
authors.initials: EL
authors.lastname: Lousberg
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D030762:Estrous Cycle / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D007962:Leukocytes / Q000166:cytology / Q000276:immunology*; D051379:Mice; D014599:Uterus / Q000166:cytology / Q000276:immunology*
keywords: Eosinophils; Estrous cycle; Flow cytometry; Immune cells; SiglecF; Uterine tissue