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Ellen B. Cook, James L. Stahl, Elizabeth D. Schwantes, Sameer K. Mathur, Neal P. Barney; Human Conjunctival Epithelial Cells Express the Receptor for TSLP. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1120.
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The conjunctival epithelium forms a barrier that isolates the eye from the external environment and represents one of the first lines of immune defense against allergens, bacterial components and macromolecules. Thymic stromal lymphopoetin (TSLP) is an epithelial-derived cytokine that plays a critical role in allergic diseases and has recently been shown to affect epithelial barrier function. The purpose of this study was to examine whether human conjunctival epithelial cells express the receptor for TSLP (TSLPR) and whether its expression is affected by pro-inflammatory cytokines.
Primary human conjunctival epithelial cells were purified from cadaveric conjunctival tissues. Conjunctival epithelial cells (< 5 passages) were challenged for 24 hrs with a combination of TNFα (5 ng/ml), IL-1β (5 ng/ml), and IFNγ (0.5 ng/ml). Expression of mRNA for TSLPR and TSLP was examined by real-time quantitative PCR using beta-glucuronidase (GUS) as a reference gene. Immunostaining for surface expression of TSLPR was evaluated using flow cytometry.
Baseline expression of both TSLPR and TSLP was low but expression of both genes was inducible by stimulation with TNFα, IL-1β and IFNγ. This induction was greater for TSLPR (16-fold) than for TSLP (1.6-fold). Baseline surface expression of TSLPR was detectable by flow cytometry (compared to isotype control) but was not upregulated by incubation with cytokines at the time point measured.
Expression of both TSLPR and TSLP by human conjunctival epithelial cells suggests that TSLP may play an autocrine role in maintenance of conjunctival epithelial cell homeostasis as well as an important role in ocular allergic inflammation.
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