Novel Vaginal Microflora Colonization Model Providing New Insight into Microbicide Mechanism of Action
Several broad-spectrum microbicides, including cellulose sulfate (CS), have passed conventional preclinical and phase I clinical safety evaluation and yet have failed to protect women from acquiring IDV -1 in phase II/Ill trials. Concerns have been raised that current preclinical algorithms are deficient in addressing the complexity of the microflora-regulated vaginal mucosal barrier. We applied a novel microflora-colonized model to evaluate CS and hydroxyethylcellulose (HEC), which is used as a “universal placebo” in microbicide trials. Cervicovaginal epithelial cultures were colonized with normal vaginal microflora isolates representing common Lactobacillus species used as probiotics (L acidophilus and L crispatus) or Prevotella bivia and Atopobium vaginae, most prevalent in the disturbed microflora of bacterial vaginosis (BV). At baseline, all strains maintained constant epithelium-associated CFUs without inducing cytotoxicity and apoptosis. CS selectively reduced epithelium-associated CFUs and (to a lesser extent) planktonic CFUs, most significantly affecting L crispatus. Inducing only minor changes in sterile epithelial cultures, CS-induced expression of innate immunity mediators (RANTES, interleukin-8 [lL-8], and secretory leukocyte protease inhibitor [SLPI]) in microflora-colonized epithelia. most significantly potentiating effects of bacteria causing BV. In the absence of CS, all bacterial strains except L acidophilus activated NF-KB, although IL-8 and RANTES levels were increased by the presence of BV causing bacteria only. CS enhanced NF-KB activation in a dose-dependent manner under all conditions, including L. acidophilus colonization. HEC remained inert. These results offer insights into possible mechanisms of CS clinical failure. The bacterially colonized cervicovaginal model reveals unique aspects of microflora-epithelium-drug interactions and innate immunity in the female genital tract and should become an integral part of preclinical safety evaluation of anti-HIV microbicides and other vaginal formulations.
EpiVaginal (VEC-100), bacterial adherence, microflora colonization, Lactobacillus. acidophilus, L crispatus, Prevotella bivia, Atopobium vaginae, cellulose sulfate, microbicides, HIV-1, probiotics
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