Multi-omic analysis of dendritic cell populations in the female genital tract.

The female genital tract (FGT) represents a complex and dynamic environment with specialized immune mechanisms uniquely designed to maintain a delicate balance between protection against invading pathogens and accommodating the unique physiological changes associated with reproductive function. Dendritic cells (DCs) are critical in shaping mucosal immunity against pathogens and maintaining tissue homeostasis. The unique ability of DCs to recognize invading pathogens through pattern recognition receptors (PRRs), and prime naive T cell function, make DCs ideal targets for vaccination and therapeutic strategies against cancers and infections. DC subsets and mononuclear phagocyte populations at human mucosal surfaces remain poorly defined. Local characterization of these populations is important, since DCs and mononuclear phagocyte populations are highly specialized depending on the tissue of residence. Hysterectomy samples of endometrium and endocervix of female donor, determined to be healthy according to study criteria, (no cancer or active infections) were enzymatically digested to generate single-cell suspensions. To enrich for myeloid immune cells, the single cell suspensions were filtered to remove cells larger than 30mm, followed by depletion of CD3+ T cells, CD19+ B cells, CD235a+ red blood cells and fibroblasts through magnetic bead selection. The cells were subsequently stained with CITE-seq antibodies (AbSeq) and sample multiplexing antibodies (hs_SampleTag). Single-cell suspensions were processed on the BD Rhapsody system according to manufacturer’s protocols. cDNA libraires were constructed using BD Rhapsody Whole Transcriptome (WTA) Amplification KIT (BD Biosciences, Catalog# 633801).