title: Activation of toll-like receptor (TLR)2, TLR4, and TLR9 in the mammalian cornea induces MyD88-dependent corneal inflammation.
abstract: OBJECTIVE
  Toll-like receptors (TLRs), which recognize microbial products, have an important role in the host innate immune response. The purpose of the present study was to determine whether activation of these receptors leads to development of keratitis and to assess the role of the common adaptor molecule myeloid differentiation factor-88 (MyD88).
  
  
  METHODS
  Corneal epithelium of C57BL/6, TLR2(-/-), TLR9(-/-), and MyD88(-/-) mice was abraded and treated with Pam(3)Cys, LPS, or CpG DNA, which bind TLR2, -4, and -9, respectively, and neutrophil recruitment to the corneal stroma, development of corneal haze, and chemokine production were measured.
  
  
  RESULTS
  Activation of TLR2 and -9 stimulated neutrophil recruitment to the corneal stroma of C57BL/6 mice, but not TLR2(-/-) or -9(-/-) mice, respectively. In marked contrast, neutrophil migration to the corneal stroma of MyD88(-/-) mice challenged with Pam(3)Cys, LPS, or CpG DNA was completely ablated. Activation of TLR2, -4, and -9 also caused a significant increase in corneal thickness and haze, indicative of disruption of corneal clarity; however, this response was ablated in MyD88(-/-) mice, which were not significantly different from untreated corneas. Production of CXC chemokines MIP-2 and KC, which mediate neutrophil recruitment to the corneal stroma, was elevated in the corneal epithelium and stroma of control, but not MyD88(-/-) mice.
  
  
  CONCLUSIONS
  Together, these findings demonstrate that the corneal epithelium has functional TLR2 and -9, and that TLR2, -4, and -9 signal through MyD88. This pathway is therefore likely to have an important role in the early events leading to microbial keratitis.
authors.affiliation: Department of Ophthalmology, Case Western University, Cleveland, OH 44106-7286, USA.
authors.forename: Angela C
authors.initials: AC
authors.lastname: Johnson
authors.forename: Fred P
authors.initials: FP
authors.lastname: Heinzel
authors.forename: Eugenia
authors.initials: E
authors.lastname: Diaconu
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Amy G
authors.initials: AG
authors.lastname: Hise
authors.forename: Douglas
authors.initials: D
authors.lastname: Golenbock
authors.forename: Jonathan H
authors.initials: JH
authors.lastname: Lass
authors.forename: Eric
authors.initials: E
authors.lastname: Pearlman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000943:Antigens, Differentiation / Q000502:physiology*; D019743:Chemokines, CXC / Q000378:metabolism; D003319:Corneal Stroma / Q000276:immunology; D018899:CpG Islands; D003545:Cysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000187:drug effects / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D007634:Keratitis / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008074:Lipoproteins / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053594:Myeloid Differentiation Factor 88; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000276:immunology; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051217:Toll-Like Receptor 9
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000943:Antigens, Differentiation; D019743:Chemokines, CXC; D004268:DNA-Binding Proteins; D008070:Lipopolysaccharides; D008074:Lipoproteins; C507780:Myd88 protein, mouse; D053594:Myeloid Differentiation Factor 88; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; C493486:Tlr2 protein, mouse; C493487:Tlr4 protein, mouse; C495370:Tlr9 protein, mouse; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051217:Toll-Like Receptor 9; C069172:2,3-bis(palmitoyloxy)-2-propyl-1-palmitoylcysteine; D003545:Cysteine