relation: http://d-scholarship-dev.library.pitt.edu/31150/
title: Medroxyprogesterone acetate has minimal impact on HIV-1 replication in ectocervical tissue ex vivo
creator: Mariko, Hawa
description: A link between use of the contraceptive depomedroxyprogesterone acetate (DMPA) and risk of HIV-1 acquisition has been demonstrated in epidemiologic studies. Peak medroxyprogesterone-17-acetate (MPA) levels occur 4 days post-injection with serum progestin levels of 2 nM. However, other studies have evaluated its in vitro effects at supraphysiologic concentrations resulting in immune modulation of epithelial cell lines and peripheral blood mononuclear cells. To formally evaluate MPA effects on HIV-1 infection in a more physiologic model, we utilized our polarized ectocervical tissue explant model. Our hypothesis is physiologic concentrations of MPA will not affect HIV-1 replication in ectocervical tissue ex vivo.    Human ectocervical tissue was processed into polarized explants. Explants were cultured alone (control) or with MPA in the basolateral compartment at physiological (0.05nM, 0.5nM, 1nM, 5nM) (n=17) or higher concentrations (500nM, 5μM, 500μM) (n=10) for 48 h, infected  with either 5x10^2 or 5x10^3 TCID50 HIV-1BaL apically for 24 h, and then washed. Basolateral supernatants were collected and replenished with fresh medium containing MPA every 3-4 days up to 21 days. Supernatant collections at the 48 h time point (pre-infection) were tested for cytokine expression while all following supernatant collections post-infection were analyzed for viral p24gag concentrations via ELISA. PBMCs were cultured for 72 hours in media containing MPA at 5μM, 50μM, and500μM (n=5), including a negative and activated control, then analyzed for antibodies via flow cytometry.    Viral replication over 21 days in the presence of supraphysiologic concentrations of MPA indicated suppression, while that in the presence of physiologic concentrations was not significantly different from the control. Evaluation of cytokine markers indicated a hormone dose-dependent relationship between decreasing MPA concentrations (500μM-5nM) and increasing expression of pro-inflammatory cytokines approaching levels of the control, which were significant for IL-12 (p40), IL-6, and IL-8.    MPA does not significantly affect HIV-1 replication at physiologic concentrations but has an overall suppressive effect on replication at supraphysiologic concentrations in ectocervical tissue in cervical tissue ex vivo.     The public health significance of DMPA continuation requires balancing unintended pregnancy while diminishing HIV-1 acquisition risk, especially when the risks of infection do not outweigh the contraceptive benefits.
date: 2017-06-29
type: University of Pittsburgh ETD
type: PeerReviewed
format: application/pdf
language: en
identifier: http://d-scholarship-dev.library.pitt.edu/31150/1/mariko_hawa_ETDApril2017.pdf
identifier:   Mariko, Hawa  (2017) Medroxyprogesterone acetate has minimal impact on HIV-1 replication in ectocervical tissue ex vivo.  Master's Thesis, University of Pittsburgh.    (Unpublished)