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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 3

Expression of PR selective and cyclic changes only in upper hFRT.

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Expression of PR selective and cyclic changes only in upper hFRT.
(A)PR ...
(A)PR gene (PGR) mRNA expression, as determined by qPCR, in vaginal, ectocervical, and endometrial biopsy samples. (B) Representative images of PR expression in sections from matched biopsy samples from 33 women. Sections from the endometrium, ectocervix, and vagina were stained for expression of PR (brown) or IgG control. Scale bar, 200 μm. (C) PGR mRNA expression, as determined by qPCR, in vaginal, ectocervical, and endometrial biopsy samples stratified into follicular (n = 16) and luteal (n = 16) stages of the menstrual cycle. (D) Quantification and (E) representative IHC images of cytoplasmic PR staining intensity in endometrial epithelial cells from women in the follicular or luteal stage of the menstrual cycle. H-scores for staining were generated using the Aperio cytoplasm algorithm, which classifies cytoplasmic staining intensity scoring as 0, none; 1+, weak; 2+ moderate; or 3+, strong, and uses this to generate an H-score using the following formula: 1 × (%1+) + 2 × (%2+) + 3 × (%3+). PR staining is highlighted with red arrows. Significance determined using either Kruskal-Wallis testing with Dunn’s multiple-comparison analysis (A and C) or Mann-Whitney U test (D). **P < 0.01, ****P < 0.0001. EP, epithelium, GE, glandular epithelium; LE, luminal epithelium; ST, stroma.

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