Inflammatory Cytokine-Mediated Regulation of Thrombospondin-1 and CD36 in Conjunctival Cells

Laura Soriano-Romaní

Laura Contreras-Ruiz

Laura García-Posadas

Antonio López-García

Sharmila Masli

Yolanda Diebold

^{Ocular Surface Group-IOBA, University of Valladolid, Valladolid, Spain.}

^{Biomedical Research Networking Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain.}

^{Department of Ophthalmology, Boston University School of Medicine, Boston, Massachusetts.}

^{Corresponding author.}

Address correspondence to:, Dr. Yolanda Diebold, Ocular Surface Group-IOBA,, University of Valladolid, Edificio IOBA, Campus Miguel Delibes,, Paseo de Belén 17, 47011 Valladolid, Spain,
E-mail:Email: se.avu.dem.aboi@loy

Received 2015 Mar 6; Accepted 2015 Jun 1.

Abstract

Purpose: Increased expression of transforming growth factor-β2 (TGF-β2) is reported in the conjunctiva of dry eye patients with no increase of anti-inflammatory activity of TGF-β2. Our aim was to compare the expression of molecules involved in TGF-β2 activation, thrombospondin-1 (TSP-1) and CD36, during murine and human conjunctival inflammation.

Methods: Human conjunctival tissue from cadaveric donors, human conjunctival epithelial primary cells and fibroblasts, and murine conjunctivas were immunostained for TSP-1, CD36, or TGF-β2. Inflamed conjunctival tissues were obtained from C57BL/6 wild-type (WT) mice induced to develop experimental dry eye (EDE) with 10 days of desiccating conditions and scopolamine injections and TSP-1-deficient (TSP1^{) mice, which spontaneously develop Sjögren's syndrome-associated conjunctival inflammation with age. Immunostaining intensities were compared using ImageJ software. Cultures of human conjunctival fibroblasts were stimulated with IL-1β and both secreted protein and message levels of TSP-1, CD36, and TGF-β2 were analyzed.}

Results: TSP-1 and CD36 were detectable in human and murine conjunctival tissues as well as primary conjunctival epithelial cells and fibroblasts. Increased conjunctival immunostaining of TGF-β2 and reduced CD36 were detected in EDE mice compared with WT mice. Interestingly, increased TGF-β2 and CD36 conjunctival immunostaining was detected in TSP1^{mice. The expression of TSP-1 and CD36 was downregulated in IL-1β-stimulated conjunctival fibroblasts at both the protein and message level, while active TGF-β2 was undetected.}

Conclusions: The absence or reduced expression of either of the molecules involved in TGF-β2 activation supports proinflammatory conditions in the conjunctiva. Changes in TSP-1 and CD36 may serve as potential biomarkers of conjunctival inflammation.

Abstract

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