TOLL-LIKE RECEPTOR EXPRESSION AND FUNCTION
type: CreativeWork
description: Purpose: Ocular surface inflammation presents from various etiologies (e.g. dry eye and Pseudomonas aeruginosa (PA) infection), yet little is known about the role of toll-like receptors (TLR) which, upon activation, stimulate inflammation. The objective of this research was to determine: (1) baseline expression of TLRs in ocular surface cells, if TLR activation or products thereof (antimicrobial peptides, AMPs) can modulate TLR expression, and if TLR agonists can stimulate the expression of AMPs human β–defensin (hBD-2) or LL-37 which can kill PA; (2) if ocular surface cell TLR expression is modulated in response to dry eye associated conditions, desiccation and in dry eye patients; (3) TLR expression in mice with experimental dry eye (EDE), AMP expression in EDE mice and dry eye subjects and if topical TLR agonist application modulates the inflammatory response in mice with EDE in vivo. Methods: (1) TLR expression was examined by RT-PCR, flow cytometry or immunostaining. Cells were exposed to AMPs or TLR agonists to determine TLR mRNA expression or the antimicrobial activity of the culture media against PA. (2) TLR mRNA or protein expression was examined by real-time RT- PCR or western blotting in cells cultured with hyperosmolar stress (HOS), or dry eye associated cytokines (IL-1α/β, TNFα, TGFβ), in a human corneal organ culture model following desiccation and in conjunctival impression cytology (CIC) samples from human dry eye patients. (3) TLR and/or AMP expression was determined at the ocular surface and/or lacrimal gland in mice with EDE using quantitative RT-PCR and immunostaining. AMP mRNA expression was examined in human dry eye CIC samples. A TLR agonist cocktail was applied to the ocular surface of mice with EDE. Central corneal thickness (CCT), recruitment of inflammatory cells into to the corneal stroma and corneal fluorescein staining were determined by in vivo imaging and corneal epithelial AMP expression was determined by quantitative RT-PCR and immu…
abstract: Purpose: Ocular surface inflammation presents from various etiologies (e.g. dry eye and Pseudomonas aeruginosa (PA) infection), yet little is known about the role of toll-like receptors (TLR) which, upon activation, stimulate inflammation. The objective of this research was to determine: (1) baseline expression of TLRs in ocular surface cells, if TLR activation or products thereof (antimicrobial peptides, AMPs) can modulate TLR expression, and if TLR agonists can stimulate the expression of AMPs human β–defensin (hBD-2) or LL-37 which can kill PA; (2) if ocular surface cell TLR expression is modulated in response to dry eye associated conditions, desiccation and in dry eye patients; (3) TLR expression in mice with experimental dry eye (EDE), AMP expression in EDE mice and dry eye subjects and if topical TLR agonist application modulates the inflammatory response in mice with EDE in vivo. Methods: (1) TLR expression was examined by RT-PCR, flow cytometry or immunostaining. Cells were exposed to AMPs or TLR agonists to determine TLR mRNA expression or the antimicrobial activity of the culture media against PA. (2) TLR mRNA or protein expression was examined by real-time RT- PCR or western blotting in cells cultured with hyperosmolar stress (HOS), or dry eye associated cytokines (IL-1α/β, TNFα, TGFβ), in a human corneal organ culture model following desiccation and in conjunctival impression cytology (CIC) samples from human dry eye patients. (3) TLR and/or AMP expression was determined at the ocular surface and/or lacrimal gland in mice with EDE using quantitative RT-PCR and immunostaining. AMP mRNA expression was examined in human dry eye CIC samples. A TLR agonist cocktail was applied to the ocular surface of mice with EDE. Central corneal thickness (CCT), recruitment of inflammatory cells into to the corneal stroma and corneal fluorescein staining were determined by in vivo imaging and corneal epithelial AMP expression was determined by quantitative RT-PCR and immu…
author: Redfern, Rachel
language: eng
keywords: Toll-like receptors, Dry eye diseases, Inflammation
date published: 2010-12
date modified: 2026-07-24T02:31:47Z
provider: University of Houston
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Houston

PropertyValue
property ID: Handle
value: http://hdl.handle.net/10657/895