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Spatiotemporal regulation of human IFN-ε and innate immunity in the female reproductive tract
Nollaig M. Bourke, Sharon L. Achilles, Stephanie U-Shane Huang, Helen E. Cumming, San S. Lim, Irene Papageorgiou, Linden J. Gearing, Ross Chapman, Suruchi Thakore, Niamh E. Mangan, Sam Mesiano, Paul J. Hertzog
Nollaig M. Bourke, Sharon L. Achilles, Stephanie U-Shane Huang, Helen E. Cumming, San S. Lim, Irene Papageorgiou, Linden J. Gearing, Ross Chapman, Suruchi Thakore, Niamh E. Mangan, Sam Mesiano, Paul J. Hertzog
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Research Article Immunology Reproductive biology

Spatiotemporal regulation of human IFN-ε and innate immunity in the female reproductive tract

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Abstract

Although published studies have demonstrated that IFN-ε has a crucial role in regulating protective immunity in the mouse female reproductive tract, expression and regulation of IFN-ε in the human female reproductive tract (hFRT) have not been characterized to our knowledge. We obtained hFRT samples from a well-characterized cohort of women to enable us to comprehensively assess ex vivo IFN-ε expression in the hFRT at various stages of the menstrual cycle. We found that among the various types of IFNs, IFN-ε was uniquely, selectively, and constitutively expressed in the hFRT epithelium. It had distinct expression patterns in the surface and glandular epithelia of the upper hFRT compared with basal layers of the stratified squamous epithelia of the lower hFRT. There was cyclical variation of IFN-ε expression in the endometrial epithelium of the upper hFRT and not in the distal FRT, consistent with selective endometrial expression of the progesterone receptor and regulation of the IFNE promoter by progesterone. Because we showed IFN-ε stimulated important protective IFN-regulated genes in FRT epithelium, this characterization is a key element in understanding the mechanisms of hormonal control of mucosal immunity.

Authors

Nollaig M. Bourke, Sharon L. Achilles, Stephanie U-Shane Huang, Helen E. Cumming, San S. Lim, Irene Papageorgiou, Linden J. Gearing, Ross Chapman, Suruchi Thakore, Niamh E. Mangan, Sam Mesiano, Paul J. Hertzog

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Figure 5

Exclusive expression of IFN-ε in hFRT regulates immune-protective IRGs.

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Exclusive expression of IFN-ε in hFRT regulates immune-protective IRGs.
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(A) Spearman’s correlation analysis of the expression of IFNE with the IRGs MX1, CXCL10, and OAS2 across hFRT biopsy samples. (B) Primary uterine epithelial cells were isolated from endometrial biopsy specimens and cultured for 3 days prior to stimulation with IFN-β (n = 9) or IFN-ε (n = 20). (C) Expression of type I IFN (IFNA1, IFNA2, IFNA4, IFNB, IFNE), type II IFN (IFNG), and type III IFN (IL28A, IL28B, IL29) was quantified by qPCR in matched vaginal, ectocervical, and endometrial biopsy samples from 32 women regardless of phase of menses. In vaginal samples, IFNA1 was not detectable (N/D) in 10, IFNA2 was N/D in 24, IFNA4 was N/D in 31, IFNB was N/D in 13, IFNG was N/D in 2, IFNL1and IFNL2 were N/D in 31, and IFNL3 was N/D in 30 specimens. In ectocervical samples, IFNA1 was N/D in 6, IFNA2 was N/D in 24, IFNA4 was N/D in 28, IFNB was N/D in 7, IFNG was N/D in 1, IFNL1 was N/D in 30, IFNL2 was N/D in 31, and was IFNL3 was N/D in 27 specimens. In endometrial samples, IFNA1 was N/D in 10, IFNA2 was N/D in 24, IFNA4 was N/D in 31, IFNB was N/D in 13, IFNG was N/D in 2, IFNL1 and IFNL2 was N/D in 31, and IFNL3 was N/D in 30 specimens. Gene expression was quantified using qPCR, normalized to 18S expression, and expressed relative to untreated control cells. The box plots depict the minimum and maximum values (whiskers), the upper and lower quartiles, and the median. The length of the box represents the interquartile range. Data were analyzed using Kruskal-Wallis testing with Dunn’s multiple-comparison analysis: ****P < 0.0001; or Mann-Whitney U test: *P < 0.05, **P < 0.01, ***P < 0.001. US, unstimulated.

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