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02624 MINIMALLY INVASIVE ASSESSMENT OF MUCOSAL INFLAMMATION USING RECTAL SWABS Elliott, Julie* Rationale: Previous studies have linked mucosal inflammation and disruption to increased risk of HIV-1 infection. Nonoxynol-9 has been linked to increased HIV transmission, possibly via IL-1b mediated activation in vaginal mucosa. For assessment of safety of topical microbicides, minimally invasive, sensitive techniques are needed to assess biologic correlates of mucosal inflammation. We report pilot data on the measurement of IL-1b and calprotectin expression by RT-PCR in cells eluted from rectal swabs. Both IL-1b and calprotectin are known to be increased in mucosal inflammation Methods: Investigations using cell lines were performed to optimize cell number and RNA recovery from swabs. Rectal swabs (n=29) were collected from five antiretroviral naïve Peruvian HIV+ subjects (median CD4 492 cells/?L; range (313-581), enrolled in an ongoing randomized, double blind, placebo controlled trial evaluating the effect of valacyclovir on herpes and HIV shedding. Swabs were inserted 3 cm from the anal margin and rotated once 360o and immediately stored in RNAlater. IL-1b and calprotectin expression were determined by real time PCR and expressed relative to normal, uninflamed colon RNA. Results: b-actin signals were seen for all the samples processed. IL-1b expression was determined in 20/29 subjects with a range of 0.2-2000 fold increase over normal; calprotectin in 27/29 subjects with a range of 3-5000 fold normal. Conclusions: These preliminary data demonstrate the feasibility of using rectal swabs to determine the presence of biologic markers of rectal inflammation in clinical trial subjects. Future studies are required to prospectively evaluate the correlation between the expression levels of IL-1b and calprotectin in rectal swabs with endoscopic, histological, and fecal calprotectin derived evidence of inflammation. Julie Elliott |
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