Endothelial cell apicobasal polarity coordinates distinct responses to luminally versus abluminally delivered TNF-α in a microvascular mimetic

Alec T. Salminen, Jeffrey Tithof, Yara Izhiman, Elysia A. Masters, Molly C. Mccloskey, Thomas R. Gaborski, Douglas H. Kelley, Anthony P. Pietropaoli, Richard E. Waugh, James L. Mcgrath

Research output: Contribution to journalArticlepeer-review

Abstract

Endothelial cells (ECs) are an active component of the immune system and interact directly with inflammatory cytokines. While ECs are known to be polarized cells, the potential role of apicobasal polarity in response to inflammatory mediators has been scarcely studied. Acute inflammation is vital in maintaining healthy tissue in response to infection; however, chronic inflammation can lead to the production of systemic inflammatory cytokines and deregulated leukocyte trafficking, even in the absence of a local infection. Elevated levels of cytokines in circulation underlie the pathogenesis of sepsis, the leading cause of intensive care death. Because ECs constitute a key barrier between circulation (luminal interface) and tissue (abluminal interface), we hypothesize that ECs respond differentially to inflammatory challenge originating in the tissue versus circulation as in local and systemic inflammation, respectively. To begin this investigation, we stimulated ECs abluminally and luminally with the inflammatory cytokine tumor necrosis factor alpha (TNF-α) to mimic a key feature of local and systemic inflammation, respectively, in a microvascular mimetic (μSiM-MVM). Polarized IL-8 secretion and polymorphonuclear neutrophil (PMN) transmigration were quantified to characterize the EC response to luminal versus abluminal TNF-α. We observed that ECs uniformly secrete IL-8 in response to abluminal TNF-α and is followed by PMN transmigration. The response to abluminal treatment was coupled with the formation of ICAM-1-rich membrane ruffles on the apical surface of ECs. In contrast, luminally stimulated ECs secreted five times more IL-8 into the luminal compartment than the abluminal compartment and sequestered PMNs on the apical EC surface. Our results identify clear differences in the response of ECs to TNF-α originating from the abluminal versus luminal side of a monolayer for the first time and may provide novel insight into future inflammatory disease intervention strategies.

Original languageEnglish (US)
Pages (from-to)275-289
Number of pages15
JournalIntegrative Biology
Volume12
Issue number11
DOIs
StatePublished - Nov 1 2020
Externally publishedYes

Bibliographical note

Funding Information:
Thisworkwas supported by theUS PublicHealth Service through the National Institutes of Health: [R01 HL125265 to R.W. and R43 GM137651 to J.L.M., R35GM119623 to T.G., T32 HL066988 to A.S., 1RF1AG057575-01 to D.H.K.].

Publisher Copyright:
© 2020 The Author(s) 2020. Published by Oxford University Press. All rights reserved.

Keywords

  • apicobasal polarity
  • endothelial cells
  • inflammation
  • neutrophils
  • sepsis
  • tissue-chip

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

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