Anderson1, D., Nadolski1, A., Xu1, C., Pudney1, J., Politch1, J., Viglianti1, G., Bowman1, F., Ayehunie2, S., and Klausner2, M. 1Boston University School of Medicine, Boston, MA and 2MatTek Corporation, Ashland, MA.

This study by researchers at Boston University School of Medicine and MatTek Corp. demonstrated that the EpiVaginal full thickness human cervico-vaginal tissue equivalent can provide valuable information on molecular mechanisms of vaginal innate immunity, and serve as a test system to determine effects of microbicides on vaginal immune defense. Background: The MatTek EpiVaginal™ (EV) full thickness tissue model, grown from primary human ectocervical epithelial and fibroblast cells on tissue culture inserts, stratifies to resemble the normal human vaginal epithelium. We have studied the expression of mediators of innate immunity in EV cultures and in normal human vaginal and ectocervical tissue. Methodology: RT-PCR, immunohistology and Bio-Plex/ELISA techniques were used to quantify the expression of Toll-like receptors (TLRs), proinflammatory cytokines (IL-6, IL-1b), chemokines (IL-8, RANTES) and antimicrobial proteins (SLPI, lysozyme and lactoferrin) in cells and culture supernatants. TLR ligands were used to confirm TLR activity in the EV model. Results: Both EV and normal tissues expressed TLR-1, 2, 3, 5 and 6, as detected by RT-PCR; stimulation of EV cultures with ligands that activate these TLRs induced secretion of IL-8 and RANTES, as well as IL-6 and G-CSF, into the lower culture chamber. SLPI was abundantly expressed by epithelial cells in normal vaginal tissues and in the EVä model. Our earlier studies showed that proinflammatory cytokines are increased and SLPI is decreased in vaginal secretions from women following the use of the spermicide, Nonoxynol-9. Similarly, proinflammatory cytokine expression was upregulated and SLPI expression was downregulated following treatment of EV cultures with subtoxic levels of N-9. Lysozyme was produced by fibroblast cells in the lamina propria of normal and EV tissues. Lactoferrin was produced by macrophages residing in vaginal tissue, but not by epithelial cells or fibroblasts in normal or EV tissues; however, lactoferrin was passively absorbed by the superficial layer of the EV stratified squamous epithelium following exposure to semen, a fluid that contains high concentrations of lactoferrin. Conclusion: These findings indicate that the EpiVaginal model can provide valuable information on molecular mechanisms of vaginal innate immunity, and serve as a test system to determine effects of microbicides on vaginal immune defense.


Absorption antimicrobial proteins, Anti-microbial, Anti-microbial peptides, Anti-microbial proteins, Antimicrobial, Antimicrobial peptides, Antimicrobial proteins, CLO, EpiVaginal, FLAG, FSL, Full thickness tissue model, G-CSF, GSK, IL-1b, IL-6, IL-8, Innate immunity, LPS, Lactoferrin, Lactoferrin seminal plasma, Lamina propria, Ligand-induced, Lysozyme, Microbicide, Microbicides, Nonoxynol-9, Normal human vaginal epithelium, PAM, Poly IC, Primary human ectocervical epithelial cells, Primary human fibroblast cells, Proinflammatory chemokines , Proinflammatory cytokine secretion, Proinflammatory cytokines, RANTES, SLPI, Spermicide, TLR's, TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, Toll-like receptors, VEC-100-FT, Vaginal immune defense, Vaginal innate immunity, Vaginal secretions

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