DOI: 10.5061/dryad.nzs7h44nq
title: Data from: Vaginal host immune-microbiome interactions in a cohort of primarily African-American women who ultimately underwent spontaneous preterm birth or delivered at term

description: Background: Recent studies suggest that alterations in the vaginal
 microbiome allow for the assessment of the risk for spontaneous preterm
 birth (PTB), the leading cause of neonatal morbidity and mortality
 worldwide. However, the associations between the local immune response and
 the vaginal microbiome are still poorly understood. Herein, we
 characterize the vaginal host immune-microbiome interactions in women who
 ultimately underwent PTB and in those who delivered at term. Methods:
 Vaginal fluid samples from 52 pregnant women (of whom 18 underwent PTB and
 34 delivered at term) were collected from 10-32 weeks in a case-control
 study. Concentrations of 33 immune mediators were determined using
 sensitive and specific immunoassays. The previously published 16S rRNA
 gene sequence and bacterial phylotype data of these subjects were utilized
 in this study. Linear mixed effects models were utilized to test
 associations between vaginal immune mediator concentrations and bacterial
 phylotype relative abundances. Results: 1) Specific immune mediators
 (β-defensins 2 and 3, IL-1β, CXCL10, CCL2, CCL3, SLPI, and VEGF)
 correlated with 18 different vaginal bacterial phylotypes in the overall
 study population; 2) vaginal concentrations of CXCL10, CCL2, CCL3, SLP1
 and VEGF negatively correlated with non-Lactobacillus members of the
 vaginal microbiome; 3) vaginal concentrations of CXCL10 were negatively
 correlated with 15 bacterial phylotypes, most of which are typical members
 of Community State Type IV of the vaginal microbiome, such as Gardnerella
 vaginalis, Megasphaera sp. type 1, and Atopobium vaginae; 4) Gemella spp.
 were negatively correlated with vaginal concentrations of VEGF, CCL2,
 CCL3, SLPI, and CXCL10; 5) when comparing PTB cases to term controls, five
 soluble immune mediators (CCL26, CCL22 and CCL2, CXCL10 and IL-16), and
 especially CCL26, were negatively correlated with five typical members of
 Community State Type IV: Sneathia sanguinegens, Parvimonas micra,
 Veillonellaceae, BVAB2, and Gemella spp; and 6) Sneathia sanguinegens had
 stronger negative associations with all five soluble immune mediators
 (CCL26, CCL22 and CCL2, CXCL10 and IL-16) in PTB cases than in term
 controls. Conclusions: The assessment of vaginal host immune-microbiome
 interactions revealed that specific soluble immune mediators, mainly
 CXCL10, negatively correlated with typical members of Community State Type
 IV of the vaginal microbiome. In addition, this assessment particularly
 showed that Sneathia sanguinegens had stronger negative associations with
 different immune mediators, including CXCL10 and CCL26, in women who
 ultimately had a PTB compared to those who delivered at term. These
 findings provide insight into the vaginal host immune-microbiome
 interactions in normal and complicated pregnancies.
description: A sample of vaginal fluid was collected under direct
 visualization from the posterior vaginal fornix using an E-Swab for immune
 mediator quantification. Vaginal swabs were stored at −80°C
 until analysis. Vaginal fluid samples were assessed using sensitive and
 specific V-PLEX immunoassays (Meso Scale Discovery, Gaithersburg, MD, USA)
 to measure vaginal fluid concentrations of several soluble immune
 mediators: the pro-inflammatory V-PLEX 10-spot assay K15049D-2 [IFN-γ,
 IL-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6], cytokine VPLEX 10-spot
 assay K15050D-2 [GM-CSF, IL-12/IL-23p40, IL-15, IL-16, IL-17A, IL-1α, 99
 IL-5, IL-7, TNFα, TNFβ, VEGF], and chemokine V-PLEX 10-spot assay
 K15047D-2 [CCL11(Eotaxin), CCL26(Eotaxin-3), CXCL8(IL-8), CXCL10(IP-10),
 CCL2(MCP-1), CCL13(MCP-4), CCL22(MDC), CCL3(MIP-1α), CCL4(MIP-1β),
 CCL17(TARC)], according to the manufacturer’s instructions. The plate
 signals were read by QuickPlex SQ 120 (Meso Scale Discovery). Standard
 curves were generated and the assay values  of the samples were
 interpolated from the curves. Secretory leukocyte protease inhibitor
 (SLPI) (R&D Systems, Minneapolis, MN, USA), β-defensin-1 and 3
 (Aviscera Bioscience, Santa Clara, CA, USA), and β-defensin 2 (ALPCO,
 Salem, NH, USA) concentrations were measured in vaginal fluid samples 
 using sensitive and specific single-analyte immunoassays, according to the
 manufacturers' instructions. The intensity of developed color was
 measured by a SpectraMax M5 Microplate Reader (San Jose, CA, USA). The
 concentrations of SLPI, and β-defensins 1, 2 and 3 were determined by
 interpolation from the standard curves. Total protein concentrations were
 measured by a BCA Protein Assay kit from 117 (ThermoFisher, catalogue
 number 23225).