Optimising the collection of female genital tract fluid for cytokine analysis in pregnant women
File(s)
Published version
Author(s)
Short, CS
Quinlan, Rachael
Bennett, Philip
Shattock, robin
Taylor, graham
Type
Journal Article
Abstract
Introduction:
To 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.
Methods:
A 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.
Results
All 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 0.5g) 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.
Conclusion:
Both 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.
To 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.
Methods:
A 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.
Results
All 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 0.5g) 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.
Conclusion:
Both 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.
Date Issued
2018-07-01
Date Acceptance
2018-03-28
Citation
Journal of Immunological Methods, 2018, 458 (7), pp.15-20
ISSN
0022-1759
Publisher
Elsevier
Start Page
15
End Page
20
Journal / Book Title
Journal of Immunological Methods
Volume
458
Issue
7
Copyright Statement
© 2018 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/)
Sponsor
Wellcome trust
Wellcome Trust
March of Dimes Foundation
Grant Number
WT102757/Z/13/Z
102757/Z/13/Z
MOD European PRC Imperial Colllege London
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Immunology
Biochemistry & Molecular Biology
Female genital tract fluid
Cervicovaginal fluid
Pregnancy
Cytokines
Menstrual cup
Ophthalmic sponge
SPONTANEOUS PRETERM BIRTH
CERVICAL-VAGINAL FLUID
CERVICOVAGINAL SECRETIONS
INFLAMMATORY CYTOKINES
BACTERIAL VAGINOSIS
NONPREGNANT WOMEN
IMMUNE MARKERS
IMMUNOGLOBULINS
MUCUS
INTERLEUKIN-1-BETA
Cervicovaginal fluid
Cytokines
Female genital tract fluid
Menstrual cup
Ophthalmic sponge
Pregnancy
Adult
Body Fluids
Cervix Uteri
Cytokines
Female
Fibroblast Growth Factors
Humans
Pregnancy
Prospective Studies
Specimen Handling
Vagina
Cervix Uteri
Vagina
Body Fluids
Humans
Fibroblast Growth Factors
Cytokines
Specimen Handling
Prospective Studies
Pregnancy
Adult
Female
1107 Immunology
1108 Medical Microbiology
1113 Opthalmology and Optometry
Immunology
Publication Status
Published
Date Publish Online
2018-04-03