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Updated: May 30, 2025

Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples
Published on: July 6, 2014
Bacterial Vaginosis Is Associated With Transcriptomic Changes but Not Higher Concentrations of Cervical Leukocytes in
Michelle C Sabo1, Salwa Mustafa1, Aparajita Saha1
1Department of Medicine, University of Washington, Seattle, Washington.
Background:
The association between bacterial vaginosis (BV) and increased human immunodeficiency virus (HIV) acquisition risk may be related to concentrations of HIV-susceptible immune cells in the cervix.
Methods:
Participants (31 with BV and 30 with normal microbiota) underwent cervical biopsy at a single visit. Immune cells were quantified and sorted using flow cytometry (n = 55), localization assessed by immunofluorescence (n = 16), and function determined by bulk RNA sequencing (RNA-seq) of live CD45+ cells (n = 21).
Results:
Linear regression analyses demonstrated no differences in mean log2 (cells/mg tissue) between women with BV versus normal microbiota for antigen-presenting cell (APC) subtypes linked to HIV risk (including CD1a+HLA-DR+ Langerhans cells, CD11c+CD14+ dendritic cells [DCs], and CD11c+HLA-DR+ DCs) and CD4+ T cells. Women with BV had a higher median proportion of CD11c+HLA-DR+ APCs (out of total cells) in cervical epithelium (0.1% vs 0.0%; P = .03 using Mann-Whitney test). RNA-seq identified 1032 differentially expressed genes (adjusted P < .05) in CD45+ cells between women with BV versus normal microbiota. Women with BV demonstrated downregulation of pathways linked to translation, metabolism, cell stress, and immune signaling.
Conclusions:
BV alters immune cell localization and function; future studies are needed to address how these changes may mediate HIV acquisition risk.
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