420. BIOCOMPATIBILITY OF SOLID DOSAGE ANTI-HIV-1 MICROBICIDES AND VAGINAL PRODUCTS WITH THE MUCOSAL CYTOKINE NETWORK.

---

Trifonova, R., Pasicznyk, J-M., Fichorova, R. Laboratory of Genital Tract Biology, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women’s Hospital, Boston, MA. Antimicrobial Agents and Chemotherapy, 50, (12), 4005-4010, (2006).

Summary:
This study by researchers in the Obstetrics, Gynecology and Reproductive Biology Dept. at Brigham and Women’s Hospital (Boston) demonstrated how MatTek’s EpiVaginal human cervico-vaginal tissue equivalent can be used to evaluate the impact of full-strength microbicide formulations on multiple immune mediators and their balance in a modeled human cervico-vaginal environment.

Background: At the mucosal port of HIV-1 entry, cytokines and chemokines can be beneficial by stimulating appropriate immune responses and tissue repair or harmful by mediating inflammatory tissue destruction and viral replication.

To avoid pathogenic responses, anti-HIV-1 microbicides in final formulated forms that are compatible with the homeostatic cytokine balance in the human vagina should be selected in physiologically relevant human model systems prior to introduction in clinical trials.

Researchers in the Laboratory of Genital Tract Biology, Department of Obstetrics, Gynecology and Reproductive Biology at the Brigham and Women’s Hospital used human vaginal explants and a bioengineered human cervicovaginal epithelium (MatTek EpiVaginal VEC-100) to establish multiple immunoinflammatory endpoints for evaluation of undiluted vaginal formulations.

Methodology: Researchers tested vaginal gels and films available over the counter and newly developed film and gel formulations of the anti-HIV-1 candidate microbicide cellulose acetate 1,2-benzenedicarboxylate (CAP). MTT and LDH assays were used to assess tissue viability. Multiple cytokine/chemokine profiling was performed using Affymetrix gene chip analysis, traditional ELISA and protein multiplex platforms, MesoScale Discovery and Beadlyte Luminex.

Results: The sensitivity of cytokine responses in the EpiVaginal VEC model was validated by comparison to vaginal tissue explants. TNFá induced the expression of an array of proinflammatory genes in the VEC tissue model. In contrast to CAP formulations and KY® moisturizing gel, vaginal cleansing and contraceptive films (VCF) containing surface active agents induced significant cytotoxicity and release of interleukin (IL)-1á, IL-1â and chemokines that are capable of enhancing HIV-1 replication.

These responses were paralleled by the release of protective mediators e.g. IL-1RA and SLPI. The intracellular stores of IL-1RA were depleted following VCF treatment of both native and ex vivo reconstructed epithelial tissues. In the EpiVaginal VEC model, the VCF contraceptive film, which contains Nonoxynol-9, was associated with a significant imbalance in the IL-1RA:IL-1 ratio, elevated IL-8 and a previously unreported increase of interferon-ã-inducible protein (IP)-10 (p<0.01).

Conclusions: This study presents a methodological approach for evaluating the impact of full-strength microbicide formulations on a multitude of immune mediators and their balance in a modeled human cervicovaginal environment (EpiVaginal tissues). These findings confirm the lack of pro-inflammatory effects of gel and film formulations of the anti-HIV-1 candidate microbicide CAP.

====================

Request an Electronic Copy (PDF format) of this Technical Paper

====================

EpiVaginal Data Sheet

EpiVaginal Specifications

EpiVaginal Technical References

---

Applications: Cytokine expression/analysis, HIV/AIDS, Inflammation - vaginal, Microbicides

Keywords: Anti-HIV-1 microbicides, Cervicovaginal epithelium, Chemokines, Cytokines, EpiVaginal, HIV, HIV-1, IL-1ra, Inflammation, Inflammatory, Microbicide, Microbicides, Microbicides, Mucosal cytokine network, Pro-inflammatory, Pro-inflammatory response, TNF-a, Tissue viability, VEC model, VEC-100

Endpoints: Affymetriz gene chip analysis, ELISA, IL-1a, IL-1b, IL-8, IL-IRA:IL-1 ratio, Interferon-gamma-inducible protein (IP-10), LDH assay, MTT assay, Protein multiplex platforms

Materials Tested: 1,2-benzenedicarboxylate (CAP), KY® moisturizing gel, Microbicide cellulose acetate, Microbicides, Nonoxynol-9, TNF-a, VCF, Vaginal cleansing and contraceptive films (VCF), Vaginal gels and films

---