Aging modifies endometrial dendritic cell function and unconventional double negative T cells in the human genital mucosa.

Immune function in the genital mucosa balances reproduction with protection against pathogens. As women age, genital infections, and gynecological cancer risk increase, however, the mechanisms that regulate cell-mediated immune protection in the female genital tract and how they change with aging remain poorly understood. Unconventional double negative (DN) T cells (TCRaß?+?CD4-CD8-) are thought to play important roles in reproduction in mice but have yet to be characterized in the human female genital tract. Using genital tissues from women (27–77 years old), here we investigated the impact of aging on the induction, distribution, and function of DN T cells throughout the female genital tract. Overall design: 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 immune cells, the single cell suspensions were filtered to remove cells larger than 30mm, followed by depletion of 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).