Expression of IFN-epsilon in the Female Reproductive Tract
Condition: Interferon Expression in the Female Reproductive Tract
Study type: Observational

Summary: 
      The proposed study aims to determine the expression of Interferon-ε (IFNε) in the female
      genital tract, which has never been mapped before in humans. High expression in other
      mammalian female reproductive tract (FRT) has been suggested to be important in anti-sexually
      transmitted infection (STI)/HIV immunity and its expression decreases with progestogen
      treatment. Future studies are planned to understand if hormonal contraceptives affect the
      presence of interferon in the healthy female genital tract, potentially changing the risk of
      for STI and HIV.
    

Detailed description: 
      Objectives:

        1. To optimize measurement protocols for IFNε and related biomarkers in samples from the
           female reproductive tract.

        2. To quantify and characterize IFNε expression in the upper and lower genital tract and
           blood in normally cycling women during the follicular and luteal phases of the menstrual
           cycle.

      Study design: This is a single site, parallel cross-sectional study

      Specific Aims:

      The aim of this study is to determine which "IFNε -related assays" can be performed on
      different types of FRT samples. The possible "IFNε-related assays" are:

        -  IFNε messenger ribonucleic acid (mRNA) levels by quantitative reverse transcriptase
           polymerase chain reaction (qRT PCR) (requiring frozen tissue processed for RNA)

        -  IFN Regulated Gene expression by qRT PCR, microarray or targeted gene set using Fluidigm
           platform (frozen tissue)

        -  PR and ER gene expression (as above)

        -  IFNε tissue protein levels* (by immunoassay) and progesterone and estrogen receptors
           (PR/ER) by Western blotting (frozen tissue)

        -  IFNε and Neopterin (a well characterized compound whose production can be stimulated by
           IFNs and whose specificity in this setting needs validation) or other IFNε-biomarkers**
           by immunoassay in cervicovaginal lavage (CVL) fluids

        -  Immunohistochemistry of IFNε, PR/ER, + other (e.g. cells, cytokines) (fixed tissue)

             -  Studies of FRT samples from wild type and IFNε-/- mice would be used to first
                validate Neopterin as a "biomarker" of IFNε activity in the FRT; and microarray
                analyses may reveal other secreted proteins that may be used as "biomarkers" of
                IFNε activity.
    

Intervention name(s): Reproductive tract and blood sampling
Intervention type(s): Procedure
Intervention description(s): Reproductive tract and blood sampling will be performed at enrollment only in this cross-sectional parallel cohort study.
Observational model: Cohort
Time perspective: Cross-Sectional
Sampling method: Non-Probability Sample
Study population: 
        Healthy females aged18 through 35 years (inclusive) at screening
      
Biospecimen description: 
      whole blood plasma vaginal swabs cervical swabs Vaginal fluid collected by sponge
      cervicovaginal lavage fluid cervical cells collected by cytobrush vaginal biopsies cervical
      biopsies endometrial biopsies
    
Keywords: interferon-epsilon; female reproductive tract; neopterin; menstrual cycle