Vaginal Product Formulation Alters the Innate Anti-viral Activity and Glycome of Cervicovaginal Fluids with Implications for Viral Susceptibility.

Glycosylated proteins (i.e. mucins, IgG) are important mediators of innate antiviral immunity in the vagina, however our current knowledge of the role that glycan themselves play in genital immunity is relatively low. Herein we evaluate the relationship between innate antiviral immunity and glycomic composition in cervicovaginal lavage fluid (CVL) collected as part of a phase-I clinical trial testing the impact of two distinct formulations of the antiretroviral drug dapivirine. Using lectin microarray technology, we discovered that formulation (hydrogel- versus film-based delivery) impacted the CVL glycome, with hydrogel formulations inducing more changes, including a loss of high mannose. The loss of this epitope correlated to a loss of anti-HIV-1 activity. Glycoproteomic identification of high-mannose proteins revealed a cohort of antiproteases shown to be important in HIV-1 resistance, whose expression co-varied with the high-mannose signature. Our data strongly suggests high-mannose as a marker for secreted proteins mediating innate antiviral immunity in vaginal fluids and that drug formulation may impact this activity as reflected in the glycome.