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Stretch-induced human myometrial cytokines enhance immune cell recruitment via endothelial activation

Cellular & Molecular Immunology volume 12, pages 231–242 (2015)Cite this article

Abstract

Spontaneous term labour is associated with amplified inflammatory events in the myometrium including cytokine production and leukocyte infiltration; however, potential mechanisms regulating such events are not fully understood. We hypothesized that mechanical stretch of the uterine wall by the growing fetus facilitates peripheral leukocyte extravasation into the term myometrium through the release of various cytokines by uterine myocytes. Human myometrial cells (hTERT-HM) were subjected to static mechanical stretch; stretch-conditioned media was collected and analysed using 48-plex Luminex assay and ELISA. Effect of stretch-conditioned media on cell adhesion molecule expression of human uterine microvascular endothelial cells (UtMVEC-Myo) was detected by quantitative polymerase chain reaction (qPCR) and flow cytometry; functional assays testing leukocyte–endothelial interactions: adhesion of leukocytes to endothelial cells and transendothelial migration of calcein-labelled primary human neutrophils as well as migration of THP-1 monocytic cells were assessed by fluorometry. The current in vitro study demonstrated that mechanical stretch (i) directly induces secretion of multiple cytokines and chemokines by hTERT-HM cells (IL-6, CXCL8, CXCL1, migration inhibitory factor (MIF), VEGF, G-CSF, IL-12p70, bFGF and platelet-derived growth factor subunit B (PDGF-bb), P<0.05); stretch-induced cytokines (ii) enhance leukocyte adhesion to the endothelium of the surrounding uterine microvasculature by (iii) inducing the expression of endothelial cell adhesion molecules and (iv) directing the transendothelial migration of peripheral leukocytes. (vi) Chemokine-neutralizing antibodies and broad-spectrum chemokine inhibitor block leukocyte migration. Our data provide a proof of mechanical regulation for leukocyte recruitment from the uterine blood vessels to the myometrium, suggesting a putative mechanism for the leukocyte infiltrate into the uterus during labour and postpartum involution.

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Acknowledgements

The authors confirm that there are no conflicts of interest. This study was supported by a grant from the CIHR (#37775). We gratefully thank Mrs Anna Dorogin for her assistance with THP-1 chemotaxis assay.

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Authors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada

    Yu-Hui Lee, Oksana Shynlova & Stephen J Lye

  2. Department of Physiology, University of Toronto, Toronto, Canada

    Yu-Hui Lee & Stephen J Lye

  3. Department of Obstetrics & Gynecology, University of Toronto, Toronto, Canada

    Oksana Shynlova & Stephen J Lye

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  1. Yu-Hui Lee
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Correspondence to Oksana Shynlova.

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Lee, YH., Shynlova, O. & Lye, S. Stretch-induced human myometrial cytokines enhance immune cell recruitment via endothelial activation. Cell Mol Immunol 12, 231–242 (2015). https://doi.org/10.1038/cmi.2014.39

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