title: New candidate biomarkers in the female genital tract to evaluate microbicide toxicity.
abstract: Vaginal microbicides hold great promise for the prevention of viral diseases like HIV, but the failure of several microbicide candidates in clinical trials has raised important questions regarding the parameters to be evaluated to determine in vivo efficacy in humans. Clinical trials of the candidate microbicides nonoxynol-9 (N9) and cellulose sulfate revealed an increase in HIV infection, vaginal inflammation, and recruitment of HIV susceptible lymphocytes, highlighting the need to identify biomarkers that can accurately predict microbicide toxicity early in preclinical development and in human trials. We used quantitative proteomics and RT-PCR approaches in mice and rabbits to identify protein changes in vaginal fluid and tissue in response to treatment with N9 or benzalkonium chloride (BZK). We compared changes generated with N9 and BZK treatment to the changes generated in response to tenofovir gel, a candidate microbicide that holds promise as a safe and effective microbicide. Both compounds down regulated mucin 5 subtype B, and peptidoglycan recognition protein 1 in vaginal tissue; however, mucosal brush samples also showed upregulation of plasma proteins fibrinogen, plasminogen, apolipoprotein A-1, and apolipoprotein C-1, which may be a response to the erosive nature of N9 and BZK. Additional proteins down-regulated in vaginal tissue by N9 or BZK treatment include CD166 antigen, olfactomedin-4, and anterior gradient protein 2 homolog. We also observed increases in the expression of C-C chemokines CCL3, CCL5, and CCL7 in response to treatment. There was concordance in expression level changes for several of these proteins using both the mouse and rabbit models. Using a human vaginal epithelial cell line, the expression of mucin 5 subtype B and olfactomedin-4 were down-regulated in response to N9, suggesting these markers could apply to humans. These data identifies new proteins that after further validation could become part of a panel of biomarkers to effectively evaluate microbicide toxicity.
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Scott
authors.initials: S
authors.lastname: Fields
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Benben
authors.initials: B
authors.lastname: Song
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Bareza
authors.initials: B
authors.lastname: Rasoul
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Julie
authors.initials: J
authors.lastname: Fong
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Melissa G
authors.initials: MG
authors.lastname: Works
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Shew
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yiu
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Jon
authors.initials: J
authors.lastname: Mirsalis
authors.affiliation: Biosciences Division, SRI International, Menlo Park, California, United States of America.
authors.forename: Annalisa
authors.initials: A
authors.lastname: D'Andrea
mesh_terms: D000282:Administration, Intravaginal; D000818:Animals; D000890:Anti-Infective Agents / Q000009:adverse effects; D001548:Benzalkonium Compounds / Q000009:adverse effects; D015415:Biomarkers / Q000378:metabolism; D002482:Cellulose / Q000009:adverse effects / Q000031:analogs & derivatives; D016207:Cytokines / Q000096:biosynthesis*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005836:Genitalia, Female / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis*; D015658:HIV Infections / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D051379:Mice; D055257:Mucin-5B / Q000096:biosynthesis*; D017137:Nonoxynol / Q000009:adverse effects; D011817:Rabbits; D014621:Vagina / Q000187:drug effects / Q000378:metabolism
chemical_list: D000890:Anti-Infective Agents; D001548:Benzalkonium Compounds; D015415:Biomarkers; D016207:Cytokines; C524579:Muc5b protein, mouse; D055257:Mucin-5B; C454127:OLFM4 protein, human; C113582:Pglyrp1 protein, mouse; D016179:Granulocyte Colony-Stimulating Factor; D017137:Nonoxynol; D002482:Cellulose; C020254:cellulose sulfate