Expression of CRAMP via PGN-TLR-2 and of α-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts

• Published in: British journal of ophthalmology 2006-03, Vol.90 (3), p.378-382
• Main Authors: Rodriguez-Martinez, S, Cancino-Diaz, M E, Cancino-Diaz, J C
• Format: Article
• Language: English
• Subjects: AD-3; alpha-Defensins - metabolism; Animals; Antimicrobial Cationic Peptides - genetics; Antimicrobial Cationic Peptides - metabolism; antimicrobial peptides; BD-1; Biological and medical sciences; Cathelicidins; cathelin related antimicrobial peptide; Cells, Cultured; Cornea - immunology; corneal fibroblast; CpG-ODN; CRAMP; cytosine-phosphorous-guanine oligonucleotide; Escherichia coli; Fibroblasts - immunology; HNP; human neutrophil peptides; Immunity, Cellular; Laboratory Science - Extended Report; lipopolysaccharide; Lipopolysaccharides - immunology; LPS; Medical sciences; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Miscellaneous; NF-kB; nuclear factor kappa B; Oligodeoxyribonucleotides - immunology; Ophthalmology; PAMPs; pathogen associated molecular patterns; PCCF; PCR; Peptidoglycan - immunology; peptidoglycans; PGNs; polymerase chain reaction; primary cultures of corneal fibroblast; reverse transcriptase; Reverse Transcriptase Polymerase Chain Reaction - methods; Staphylococcus aureus; TIR; TLR/IL-1 receptor; TLRs; TNF; Toll-Like Receptor 2 - immunology; Toll-Like Receptor 9 - immunology; toll-like receptors; tumour necrosis factor; α-defensin 3; β-defensin-1
• ISSN: 0007-1161; 1468-2079
• DOI: 10.1136/bjo.2005.082289

Aims: To search for the induction of the expression of antimicrobial peptides in corneal fibroblasts treated with bacterial components. Methods: RT-PCR was performed to search for mRNAs expression of antimicrobial peptides and toll-like receptors (TLRs) in murine primary cultures of corneal fibroblast (PCCF) treated with lipopolysaccharide (LPS) from Escherichia coli, peptidoglycan from Staphylococcus aureus, and cytosine-phosphorous-guanine oligonucleotide (CpG-ODN). Cellular activation was blocked with anti-TRL antibodies. Results: LPS did not induce expression of antimicrobial peptide in corneal fibroblasts. Cathelin related antimicrobial peptide (CRAMP) and α-defensin 3 were overexpressed in a time and dose dependent manner in corneal fibroblasts treated with peptidoglycan and with CpG-ODN, respectively. CRAMP expression was blocked when PCCF were treated with anti-TLR-2 antibodies. α-Defensin 3 was not expressed in NIH murine corneal fibroblasts (which do not express the TLR-9 molecule) treated with CpG-ODN. Conclusion: Results suggest that corneal fibroblasts, which are the second cellular barrier of the cornea, can play an important part in the innate immunity of the eye via TLR stimulation.