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Antimicrobial peptides expression in activated human corneal keratocytes
Author(s) -
MUSHTAQ F,
MOHAMMED I,
TINT NL,
HOPKINSON A,
DUA HS
Publication year - 2008
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2008.6233.x
Subject(s) - cathelicidin , antimicrobial peptides , fibroblast , biology , ethidium bromide , gene expression , microbiology and biotechnology , beta defensin , wound healing , receptor , innate immune system , cell culture , gene , immune system , immunology , antimicrobial , genetics , dna
Purpose The eye despite being a delicate structure,has evolved many protective mechanisms.Antimicrobial peptides(AMPs) and Toll‐like receptors(TLRs)have an important role in the ocular defense system.However during an infection or trauma, the innate defense system is breached.In this instance,keratocytes and fibroblasts aids in wound healing as well as provide a second line of defense.Little is known about AMP expression by TLR signalling pathways in corneal fibroblasts.This study therefore demonstrates the mRNA expression of both novel and known AMPs in various fibroblast phenotypes(myofibroblasts and keratocytes).In addition, AMPs were also shown in microbial ligand and/or pro‐inflammatory cytokine stimulated human corneal fibroblast cultures (HCFC). Methods Fresh Keratocytes and HCFC were cultured and collected at different stages of growth and/or after stimulation with microbial ligands. Total RNA was extracted and cDNA was synthesized for semi‐quantitative gene expression studies. Polymerase chain reaction (PCR) amplified product were analyzed on ethidium bromide stained 1% agarose gels. Results Constitutive expression of Thy‐1(CD‐90), vimentin, keratocan and alpha SMA was shown in all cultures.Keratocyte phenotype marker CD‐34 showed decreased expression from 1st to 4th passage.Positive expression of Human beta defensin 109, Liver‐expressed AMP(LEAP‐2) and cathelicidin/LL‐37 was demonstrated in cytokine and microbial ligand stimulated HCFC. Conclusion Reminiscent with previous studies,we have demonstrated the mRNA expression of novel and known AMPs in corneal fibroblast mediated by TLR dependent signalling mechanisms. This result warrants the further study to fully understand the role of AMPs in ocular wound healing.