title: Vaginal microbicides: detecting toxicities in vivo that paradoxically increase pathogen transmission.
abstract: BACKGROUND
  Microbicides must protect against STD pathogens without causing unacceptable toxic effects. Microbicides based on nonoxynol-9 (N9) and other detergents disrupt sperm, HSV and HIV membranes, and these agents are effective contraceptives. But paradoxically N9 fails to protect women against HIV and other STD pathogens, most likely because it causes toxic effects that increase susceptibility. The mouse HSV-2 vaginal transmission model reported here: (a) Directly tests for toxic effects that increase susceptibility to HSV-2, (b) Determines in vivo whether a microbicide can protect against HSV-2 transmission without causing toxicities that increase susceptibility, and (c) Identifies those toxic effects that best correlate with the increased HSV susceptibility.
  
  
  METHODS
  Susceptibility was evaluated in progestin-treated mice by delivering a low-dose viral inoculum (0.1 ID50) at various times after delivering the candidate microbicide to detect whether the candidate increased the fraction of mice infected. Ten agents were tested - five detergents: nonionic (N9), cationic (benzalkonium chloride, BZK), anionic (sodium dodecylsulfate, SDS), the pair of detergents in C31G (C14AO and C16B); one surface active agent (chlorhexidine); two non-detergents (BufferGel, and sulfonated polystyrene, SPS); and HEC placebo gel (hydroxyethylcellulose). Toxic effects were evaluated by histology, uptake of a 'dead cell' dye, colposcopy, enumeration of vaginal macrophages, and measurement of inflammatory cytokines.
  
  
  RESULTS
  A single dose of N9 protected against HSV-2 for a few minutes but then rapidly increased susceptibility, which reached maximum at 12 hours. When applied at the minimal concentration needed for brief partial protection, all five detergents caused a subsequent increase in susceptibility at 12 hours of approximately 20-30-fold. Surprisingly, colposcopy failed to detect visible signs of the N9 toxic effect that increased susceptibility at 12 hours. Toxic effects that occurred contemporaneously with increased susceptibility were rapid exfoliation and re-growth of epithelial cell layers, entry of macrophages into the vaginal lumen, and release of one or more inflammatory cytokines (Il-1beta, KC, MIP 1alpha, RANTES). The non-detergent microbicides and HEC placebo caused no significant increase in susceptibility or toxic effects.
  
  
  CONCLUSIONS
  This mouse HSV-2 model provides a sensitive method to detect microbicide-induced toxicities that increase susceptibility to infection. In this model, there was no concentration at which detergents provided protection without significantly increasing susceptibility.
authors.affiliation: Mucosal Protection Laboratory, Dept. of Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA. cone@jhu.edu
authors.forename: Richard A
authors.initials: RA
authors.lastname: Cone
authors.forename: Timothy
authors.initials: T
authors.lastname: Hoen
authors.forename: Xixi
authors.initials: X
authors.lastname: Wong
authors.forename: Raed
authors.initials: R
authors.lastname: Abusuwwa
authors.forename: Deborah J
authors.initials: DJ
authors.lastname: Anderson
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Moench
mesh_terms: D000818:Animals; D002482:Cellulose / Q000031:analogs & derivatives* / Q000627:therapeutic use / Q000633:toxicity; D003902:Detergents / Q000627:therapeutic use / Q000633:toxicity*; D004195:Disease Models, Animal; D064420:Drug-Related Side Effects and Adverse Reactions; D005260:Female; D006558:Herpes Genitalis / Q000517:prevention & control / Q000635:transmission* / Q000821:virology; D018258:Herpesvirus 2, Human / Q000187:drug effects* / Q000472:pathogenicity*; D006801:Humans; D051379:Mice; D017137:Nonoxynol / Q000627:therapeutic use / Q000633:toxicity; D011137:Polystyrenes / Q000627:therapeutic use / Q000633:toxicity; D013501:Surface-Active Agents / Q000627:therapeutic use / Q000633:toxicity*; D014621:Vagina / Q000821:virology*
chemical_list: D003902:Detergents; D011137:Polystyrenes; D013501:Surface-Active Agents; D017137:Nonoxynol; D002482:Cellulose; C002283:hydroxyethylcellulose