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02114_2 SPECTRUM OF HIV-1 INHIBITORY ACTIVITY OF SULFONATED POLYMERS, CANDIDATE TOPICAL MICROBICIDES Klotman, Mary* A number of candidate topical microbicides under development are within the same class of sulphated or sulfonated polysaccharides and include cellulose sulfate (CS), polysterene sulfonate (PSS), polymethelenehydroquinone sulfonate (PMHS) and Pro2000. In order to better understand the potential of this class of compounds, we compared these four compounds for their anti-HIV activity, cytotoxicity, activity in cervical lavage fluids and their antiviral mechanism(s) of action. METHODS: Primary macrophages and T-cells as well as U87 cells engineered to express single co-receptors were treated with CS, PSS, PMHS and Pro2000 and challenged with laboratory-adapted or primary HIV-1 isolates as well as replication defective viruses carrying the luciferase indicator gene. The latter challenges were done with the addition of the compounds or AZT or small molecule entry inhibitors either at the same time as viral infection or varying amounts of time following infection. Finally, compounds were tested for their ability to block gp120 binding to the cell and for their ability to bind directly to gp120 using surface plasmon resonance (SPR) analyses with a BIAcore 3000 system. RESULTS: All four compounds blocked HIV-1 replication in primary macrophages and CD4+ T-cells in the range of 10-100 ug/ml. The majority of the anti-HIV affect was during virus entry although there were some post-entry affects, particularly with PSS and CS. Each compound blocked binding of gp120 to CD4+/coreceptor positive U87 cells. PSS, CS and PMHS showed similar stable, high affinity binding to gp120 while CS could not be tested in this assay. Finally, all four compounds showed significant antiviral activity in the presence of cervical lavage fluid. CONCLUSIONS: The compounds CS, PSS, PMHS and Pro 2000 all inhibit HIV-1 in primary macrophages and CD4+ cells in the presence of cervical lavage fluid. Consistent with their blocking of HIV-1 entry is their ability to block gp120 binding to the cell and their high affinity stable binding to gp120. Dr. Mary E. Klotman |
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