Journal Article

Performance of Swabs, Lavage, and Diluents to Quantify Biomarkers of Female Genital Tract Soluble Mucosal Mediators

Louissaint, Nicolette, Mellors, John, Dezzutti, Charlene S., Hendrix, Craig W., Pan, Zhenyu, Kalyoussef, Sabah, Torres, N. Merna, Wang, Lei, Hillier, Sharon L., Marrazzo, Jeanne M., Hladik, Florian, Herold, Betsy C., Parikh, Urvi

2011

Overview

Measurement of immune mediators and antimicrobial activity in female genital tract secretions may provide biomarkers predictive of risk for HIV-1 acquisition and surrogate markers of microbicide safety. However, optimal methods for sample collection do not exist. This study compared collection methods. Secretions were collected from 48 women (24 with bacterial vaginosis [BV]) using vaginal and endocervical Dacron and flocked swabs. Cervicovaginal lavage (CVL) was collected with 10 mL of Normosol-R (n = 20), saline (n = 14), or water (n = 14). The concentration of gluconate in Normosol-R CVL was determined to estimate the dilution factor. Cytokine and antimicrobial mediators were measured by Luminex or ELISA and corrected for protein content. Endogenous anti-HIV-1 and anti-E. coli activity were measured by TZM-bl assay or E. coli growth. Higher concentrations of protein were recovered by CVL, despite a 10-fold dilution of secretions, as compared to swab eluents. After protein correction, endocervical swabs recovered the highest mediator levels regardless of BV status. Endocervical and vaginal flocked swabs recovered significantly higher levels of anti-HIV-1 and anti-E. coli activity than Dacron swabs (P<0.001). BV had a significant effect on CVL mediator recovery. Normosol-R tended to recover higher levels of most mediators among women with BV, whereas saline or water tended to recover higher levels among women without BV. Saline recovered the highest levels of anti-HIV-1 activity regardless of BV status. Endocervical swabs and CVL collected with saline provide the best recovery of most mediators and would be the optimal sampling method(s) for clinical trials.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adult / alpha-Defensins - analysis / alpha-Defensins - immunology / Analysis of Variance / Antiinfectives and antibacterials / Antimicrobial activity / Antimicrobial agents / Antiretroviral drugs / Antiviral activity / Bioavailability / Biological markers / Biology / Biomarkers / Biomarkers - analysis / Cell adhesion & migration / Cervix Uteri - metabolism / Cervix Uteri - microbiology / Chemokines / Clinical trials / Collection / Cytokines / Cytokines - analysis / Cytokines - immunology / Dacron / Diluents / Dilution / E coli / Eluents / Enzyme-linked immunosorbent assay / Escherichia coli / Excipients / Family medical history / Female / Females / Genital tract / Genitalia, Female - metabolism / Gluconates / Gynecology / Health risks / HIV / HIV Infections - diagnosis / HIV Infections - immunology / HIV Infections - prevention & control / HIV-1 - immunology / Human immunodeficiency virus / Humans / Immunoglobulins / Infections / Interleukin-8 - analysis / Interleukin-8 - immunology / Medical research / Medicine / Methods / Mucosa / Mucous Membrane - metabolism / Obstetrics / Pediatrics / Polyethylene terephthalate / Polypeptides / Protein folding / Recovery / Risk Factors / Saline water / Sampling methods / Secretions / Secretory Leukocyte Peptidase Inhibitor - analysis / Secretory Leukocyte Peptidase Inhibitor - immunology / Solubility / Specimen Handling - methods / Therapeutic Irrigation - methods / Vagina / Vagina - metabolism / Vagina - microbiology / Vaginal Smears - methods / Vaginosis / Vaginosis, Bacterial - immunology / Vaginosis, Bacterial - microbiology / Young Adult