title: Searching for lower female genital tract soluble and cellular biomarkers: defining levels and predictors in a cohort of healthy Caucasian women.
abstract: BACKGROUND
  High concentrations of pro-inflammatory cytokines have been previously observed in the genital fluids of women enrolled in microbicide trials and may explain observed increased HIV transmission in some of these trials. Although the longitudinal nature of these studies allows within-subject comparisons of post-product levels to baseline levels, the fact that the physiologic variations of these cytokines and other markers of immune activation are not fully defined in different populations, makes it difficult to assess changes that can be directly attributed to microbicide use as opposed to other biological and behavioural factors.
  
  
  METHODS
  Cervicovaginal lavage samples were collected from 30 healthy Caucasian and assayed for concentrations of ten cytokines/chemokines, total protein content and two antimicrobial proteins using a multiplex immunoassay and ELISA. Cellular markers were characterized by flow cytometry on mononuclear cells collected from the endocervix using flocked swabs. Bacterial quantification was performed using quantitative PCR.
  
  
  RESULTS
  Ectopy, menstrual cycle phase, prostate-specific antigen and presence of leucocytes in endocervical cells' supernatant were associated with the concentrations of cyto-/chemokines in cervicovaginal secretions. Approximately 3% of endocervical cells collected were monocytes of which a median of 52% (SD  = 17) expressed both CD4 and CCR5 markers. Approximately 1% of the total cells were T-cells with a median of 61% (SD  = 10) CD4 and CCR5 expression. Around 5% of the monocytes and 16% of the T-cells expressed the immune activation marker HLA-DR. Higher percentages of T-cells were associated with greater quantities of IL-1RA, GM-CSF and elafin.
  
  
  CONCLUSIONS
  We demonstrate the presence of selected soluble and cellular immune activation markers and identify their predictors in the female genital tract of healthy women. Future clinical trials should consider ectopy, sexual activity, menstrual cycle phase and presence of bacterial species as possible confounders when evaluating the possible inflammatory effects of microbicide compounds.
authors.affiliation: Virology Unit, Division of Microbiology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors.forename: Jordan K
authors.initials: JK
authors.lastname: Kyongo
authors.forename: Vicky
authors.initials: V
authors.lastname: Jespers
authors.forename: Odin
authors.initials: O
authors.lastname: Goovaerts
authors.forename: Johan
authors.initials: J
authors.lastname: Michiels
authors.forename: Joris
authors.initials: J
authors.lastname: Menten
authors.forename: Raina N
authors.initials: RN
authors.lastname: Fichorova
authors.forename: Tania
authors.initials: T
authors.lastname: Crucitti
authors.forename: Guido
authors.initials: G
authors.lastname: Vanham
authors.forename: Kevin K
authors.initials: KK
authors.lastname: Ariën
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D002584:Cervix Uteri / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology; D018925:Chemokines / Q000378:metabolism; D015331:Cohort Studies; D005260:Female; D005786:Gene Expression Regulation; D006684:HLA-DR Antigens / Q000378:metabolism; D006262:Health; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D054892:Metagenome; D009000:Monocytes / Q000166:cytology; D011480:Protease Inhibitors / Q000378:metabolism; D019713:Receptors, CCR5 / Q000378:metabolism; D012995:Solubility; D013601:T-Lymphocytes / Q000166:cytology; D014621:Vagina / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D044465:White People; D023083:beta-Defensins / Q000378:metabolism
chemical_list: D015415:Biomarkers; D018925:Chemokines; D006684:HLA-DR Antigens; D036341:Intercellular Signaling Peptides and Proteins; D011480:Protease Inhibitors; D019713:Receptors, CCR5; D023083:beta-Defensins