Expression of CRAMP via PGN-TLR-2 and of alpha-defensin-3 via CpG-ODN-TLR-9 in corneal fibroblasts.
OBJECTIVE
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 alpha-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. alpha-Defensin 3 was not expressed in NIH murine corneal fibroblasts (which do not express the TLR-9 molecule) treated with CpG-ODN.


CONCLUSIONS
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.
Departamento de Microbiología, Laboratorio de Microbiología General, Escuela Nacional de Ciencias Biológicas, Carpio y Plan de Ayala s/n México, DF, 11340, México.
S
S
Rodriguez-Martinez
M E
ME
Cancino-Diaz
J C
JC
Cancino-Diaz
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism*; D054804:Cathelicidins; D002478:Cells, Cultured; D003315:Cornea / Q000276:immunology*; D005347:Fibroblasts / Q000276:immunology*; D007111:Immunity, Cellular; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008815:Mice, Inbred Strains; D009838:Oligodeoxyribonucleotides / Q000276:immunology; D010457:Peptidoglycan / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051217:Toll-Like Receptor 9 / Q000276:immunology; D023084:alpha-Defensins / Q000378:metabolism*
D023181:Antimicrobial Cationic Peptides; C408982:CPG-oligonucleotide; D054804:Cathelicidins; D008070:Lipopolysaccharides; D009838:Oligodeoxyribonucleotides; D010457:Peptidoglycan; D051195:Toll-Like Receptor 2; D051217:Toll-Like Receptor 9; D023084:alpha-Defensins