Objectives
To quantify and compare cervical immune cell populations in women with normal and abnormal vaginal microbiota. Our hypothesis was that women with non-normal vaginal flora by Nugent score and women with key BV microbes by quantitative PCR (qPCR) would have increased numbers of cervical CD4 and CD8 T lymphocytes expressing co-receptors for HIV (CCR5) and activation (CD69).
Methods
Women aged 18-34 who screened negative for sexually transmitted infections (GC, CT, TV, and HIV) and were confirmed by mass spectrometry to be free of exogenous hormone use and in the follicular phase of menses were enrolled. Cervical cytobrushes and biopsies were collected for immune cell quantification and vaginal swabs were collected for microbiota assessment. Cytobrushes and biopsies were processed and stained to quantify CD4, CD4CCR5, CD4CD69, CD8, CD8CCR5 and CD8CD69 cells using flow cytometric analysis. Cell counts from cervical biopsies were adjusted per gram of tissue. Women were stratified by Nugent score (score of 0-3 vs 4-10) and by vaginal microbiota as determined by qPCR for key microbes. Differences in cell quantities were evaluated using the Mann Whitney U test.
Methods
Women aged 18-34 who screened negative for sexually transmitted infections (GC, CT, TV, and HIV) and were confirmed by mass spectrometry to be free of exogenous hormone use and in the follicular phase of menses were enrolled. Cervical cytobrushes and biopsies were collected for immune cell quantification and vaginal swabs were collected for microbiota assessment. Cytobrushes and biopsies were processed and stained to quantify CD4, CD4CCR5, CD4CD69, CD8, CD8CCR5 and CD8CD69 cells using flow cytometric analysis. Cell counts from cervical biopsies were adjusted per gram of tissue. Women were stratified by Nugent score (score of 0-3 vs 4-10) and by vaginal microbiota as determined by qPCR for key microbes. Differences in cell quantities were evaluated using the Mann Whitney U test.
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