Abstract
IntroductionTo better understand the immunology of pregnancy, study of female genital tract fluid (FGF) is desirable. However the optimum method of collection of FGF in pregnant women for immunological methods, specifically cytokine measurement, is unknown. MethodsA prospective study of HIV-uninfected pregnant women comparing two methods of FGF collection: polyvinyl acetal sponge collection of cervical fluid (CF) and menstrual cup collection of cervicovaginal fluid (CVF). Samples were collected at 3 time points across the second and third trimesters: 14–21, 22–25 and 26–31 weeks. Multiplex chemi-luminescent assays were used to measure: IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13 and TNF-α. Optimal methodology for cytokine normalisation (sample weight, volume and total protein) was explored. ResultsAll cytokines were measurable in both fluid types. IL-1β, IL-8 and IL-6 were detected at the highest concentrations (ranking order CF > CVF > plasma). CVF collection was simpler, provided the largest volume of sample (median 0.5 g) with the potential for undiluted usage, and allowed for self-insertion. CF cytokine concentrations were intrinsically associated with sample weight and protein concentration however CVF cytokines were independent of these. ConclusionBoth methods of collection are robust for measurement of FGF cytokines during pregnancy. We recommend CVF collection using a menstrual cup as a viable option in pregnant women for high dimensional biological techniques.
Highlights
To better understand the immunology of pregnancy, study of female genital tract fluid (FGF) is desirable
In the context of pregnancy, the study of the FGF cytokine milieu has enabled the identification of IL-1β, IL-6, IL-8 and TNF-α as correlates of preterm delivery (Wei et al, 2010; Goldenberg et al, 2005; Tanaka et al, 1998; Discacciati et al, 2011; Goepfert et al, 2001; Jun et al, 2000)
In this study we found both polyvinyl acetal (PVA) sponges and the menstrual cup to be valid methods of collection for the measurement of FGF cytokines with all the multiplex cytokines being measureable in each fluid
Summary
To better understand the immunology of pregnancy, study of female genital tract fluid (FGF) is desirable. FGF comprises of cervical mucous/fluid (CF) and vaginal secretions or a combination of the two (cervicovaginal fluid (CVF)) An advantage of this fluid type is that, in addition to studying the local vaginal immune compartment, it can provide information about the upstream uterine and cervical environment (Dasari et al, 2007; Zegels et al, 2010). Another benefit is that it is obtained without the need for invasive procedures. In the context of pregnancy, the study of the FGF cytokine milieu has enabled the identification of IL-1β, IL-6, IL-8 and TNF-α as correlates of preterm delivery (Wei et al, 2010; Goldenberg et al, 2005; Tanaka et al, 1998; Discacciati et al, 2011; Goepfert et al, 2001; Jun et al, 2000)
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