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

Abstract

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.

  • AD-3, α-defensin 3
  • BD-1, β-defensin-1
  • CpG-ODN, cytosine-phosphorous-guanine oligonucleotide
  • CRAMP, cathelin related antimicrobial peptide
  • HNP, human neutrophil peptides
  • LPS, lipopolysaccharide
  • NF-kB, nuclear factor kappa B
  • PAMPs, pathogen associated molecular patterns
  • PCCF, primary cultures of corneal fibroblast
  • PCR, polymerase chain reaction
  • PGNs, peptidoglycans
  • RT, reverse transcriptase
  • TIR, TLR/IL-1 receptor
  • TLRs, toll-like receptors
  • TNF, tumour necrosis factor
  • CRAMP
  • α-defensin 3
  • corneal fibroblast
  • toll-like receptors
  • antimicrobial peptides
  • AD-3, α-defensin 3
  • BD-1, β-defensin-1
  • CpG-ODN, cytosine-phosphorous-guanine oligonucleotide
  • CRAMP, cathelin related antimicrobial peptide
  • HNP, human neutrophil peptides
  • LPS, lipopolysaccharide
  • NF-kB, nuclear factor kappa B
  • PAMPs, pathogen associated molecular patterns
  • PCCF, primary cultures of corneal fibroblast
  • PCR, polymerase chain reaction
  • PGNs, peptidoglycans
  • RT, reverse transcriptase
  • TIR, TLR/IL-1 receptor
  • TLRs, toll-like receptors
  • TNF, tumour necrosis factor
  • CRAMP
  • α-defensin 3
  • corneal fibroblast
  • toll-like receptors
  • antimicrobial peptides
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