title: Analysis of Ag-presenting cells in the conjunctiva during the development of experimental immune-mediated blepharoconjunctivitis.
abstract: OBJECTIVE
  The aim of this study was to investigate the phenotypes of antigen (Ag) presenting cells (APCs) in the conjunctiva during the development of experimental immune-mediated blepharoconjunctivitis (EC), which serves as a model for investigation of severe types of human allergic conjunctivitis.
  
  
  METHODS
  Brown Norway rats treated by ovalbumin (OVA) were used in this study. To confirm the restriction of MHC class II by OVA-specific T cells, monoclonal Abs against MHC class II were added to the conventional proliferation assay. To evaluate the MHC class II expression in the conjunctiva during the development of EC, an immunohistochemical analysis, either as the single or double staining, was performed. Conjunctival fibroblast cell lines were established from naive rats and the MHC class II expression was evaluated by flow cytometric analysis. To examine the roles of costimulatory molecules, OVA-specific T cells were stimulated with anti-TcR Ab and anti-CD28 Ab and then subjected for Western blotting to evaluate the ERK phosphorylation. Finally, in vivo expression of B7 molecules was examined immunohistochemically.
  
  
  RESULTS
  OVA-specific T cells recognized OVA in the context of MHC class II. MHC class II was expressed in conjunctival macrophages but not in fibroblasts. EC induction was accompanied by abundant infiltration of macrophages positive for MHC class II. MHC class II was also expressed in conjunctival epithelial cells by EC induction. Stimulation from CD28 was necessary for ERK phosphorylation. B7-2, but not B7-1, was expressed in the conjunctiva.
  
  
  CONCLUSIONS
  Conjunctival macrophages may represent a major source of APCs for the induction of EC in the conjunctiva.
authors.affiliation: Department of Ophthalmology, Kochi Medical School, Kohasu, Oko-cho, Nankoku City 783-8505, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Ozaki
authors.forename: Atsuki
authors.initials: A
authors.lastname: Fukushima
authors.forename: Waka
authors.initials: W
authors.lastname: Ishida
authors.forename: Olga
authors.initials: O
authors.lastname: Zinchuk
authors.forename: Kazuyo
authors.initials: K
authors.lastname: Fukata
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Hideaki
authors.initials: H
authors.lastname: Enzan
authors.forename: Masaru
authors.initials: M
authors.lastname: Takata
authors.forename: Hideo
authors.initials: H
authors.lastname: Yagita
authors.forename: Hisayuki
authors.initials: H
authors.lastname: Ueno
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D015703:Antigens, CD / Q000378:metabolism; D051940:B7-2 Antigen; D001762:Blepharitis / Q000276:immunology*; D015153:Blotting, Western; D002452:Cell Count; D003228:Conjunctiva / Q000276:immunology*; D003233:Conjunctivitis, Allergic / Q000276:immunology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005434:Flow Cytometry; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D007124:Immunoenzyme Techniques; D016130:Immunophenotyping; D008213:Lymphocyte Activation; D008264:Macrophages / Q000276:immunology*; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008962:Models, Theoretical; D010047:Ovalbumin / Q000276:immunology; D010766:Phosphorylation; D051381:Rats; D011914:Rats, Inbred BN; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D015703:Antigens, CD; D051940:B7-2 Antigen; C497966:Cd86 protein, rat; D000949:Histocompatibility Antigens Class II; D008562:Membrane Glycoproteins; D010047:Ovalbumin; D048049:Extracellular Signal-Regulated MAP Kinases