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02603 EFFECT OF 0.5% PRO 2000/5 GEL ON INFECTIOUS HIV-1 AND INFLAMMATORY MEDIATORS IN CERVICOVAGINAL SECRETIONS Keller, Marla1 PRO 2000/5 has been shown to inhibit HIV and other STI in cell culture and in animal models. However, there is as yet no data showing that any microbicide exhibits anti-viral activity in vivo. We are conducting a prospective double-blind placebo-controlled study among 20 HIV-infected women to assess whether a single 2-g intravaginal dose of 0.5% PRO 2000/5 Gel (P) reduces levels of infectious HIV present in cervicovaginal lavage (CVL) fluid collected one hour after application. A secondary objective is to assess the acute inflammatory response to a single application of 0.5% PRO 2000/5 Gel (P) by testing the CVL for inflammatory cells, IL-1b and IL-8. Inclusion criteria include age between 18 and 45, HIV-1 RNA &Mac179; 50,000 copies/ml, and a normal Pap smear within 6 months of screening. Exclusions include pregnancy, recent UTI or STI, and sexual intercourse or use of other vaginal products within 48 h of screening. Given the variability in the ability to reproducibly recover a subjects own infectious HIV from CVL, the anti-viral activity in CVL will be assessed by spiking with serial dilutions of HIV and quantifying the virus recovered. Serial two-fold dilutions of CVL supernatants collected at screening (2-3 days before) and 1h after application of PRO 2000/5 or placebo gel will be spiked with dilutions of HIV-1 and then inoculated onto susceptible cells. A dilution of CVL that inhibits or neutralizes virus will be determined. Additionally, the subjects own virus will be cultured directly from supernatant and cell pellets obtained before and after PRO 2000/5 or placebo gel application. A peroxidase test on the cell suspension will be used to quantify PMNs; IL-1b and IL-8 will be measured using commercial ELISA kits. In preliminary studies, infectious virus has been reproducibly and successfully recovered after spiking CVL samples with serial dilutions of virus. In addition, the anti-HIV activity of serial two-fold dilutions of PRO 2000/5 diluted in vitro in CVL has been established starting with an initial concentration of 500 mg/ml and spiked with 20 ng of p24/ml of replication defective JRFL-pseudotyped virus expressing a luciferase indicator gene. A 1:512 dilution or 0.977 mg/ml of PRO 2000 in CVL inhibited infection by the JRFL-pseudotyped virus. This strategy is a mechanism for testing in vivo the efficacy of a microbicide before embarking on large-scale clinical trials. Assistant Professor, Marla Keller, MD |
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