title: Effects of TGF-beta2 on immune response-related gene expression profiles in the human corneal endothelium.
abstract: OBJECTIVE
  To determine the effects of transforming growth factor (TGF)-beta2 on immune-response-related gene expression profiles in the stimulated human corneal endothelium (HCE).
  
  
  METHODS
  A human complementary DNA (cDNA) expression array analysis was used to investigate the effects of TGF-beta2 on cultured HCE incubated with interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha. Gene-specific semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to examine the gene expression patterns revealed by the cDNA expression array analysis. Moreover, the expression of newly identified genes in HCE was confirmed by RT-PCR in human donor corneas.
  
  
  RESULTS
  cDNA expression array analysis and semiquantitative RT-PCR revealed that TGF-beta2 downregulated the expression of IL-6, growth-related (Gro)-alpha (CXCL1), monocyte chemotactic protein (MCP)-1 (CCL2), granulocyte-colony stimulating factor (G-CSF), and insulin-like growth factor binding protein (IGFBP)-5 and upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-1. ELISA confirmed TGF-beta2-mediated changes in the expression of IL-6, CXCL1, CCL2, G-CSF, IGFBP-5, and TIMP-1 at the protein level. CXCL1, G-CSF, and IGFBP-5 mRNAs were detected for the first time in the HCE of donor corneas.
  
  
  CONCLUSIONS
  TGF-beta2 downregulates IL-6, CXCL1, CCL2, G-CSF, and IGFBP-5, and upregulates TIMP-1 in cultured HCE stimulated with proinflammatory cytokines, suggesting the immunomodulatory role of TGF-beta2 in the aqueous humor and the pathophysiological significance of TGF-beta2 in the anterior chamber of the eye.
authors.affiliation: Department of Corneal Tissue Regeneration, Tokyo University Graduate School of Medicine, Tokyo, Japan. syamagami-tky@umin.ac.jp
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yokoo
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Mimura
authors.forename: Shiro
authors.initials: S
authors.lastname: Amano
mesh_terms: D018929:Cell Culture Techniques; D018932:Chemokine CCL2 / Q000378:metabolism; D054360:Chemokine CXCL1; D019743:Chemokines, CXC / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis; D015536:Down-Regulation; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects*; D005802:Genes, MHC Class II / Q000187:drug effects*; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D018975:Insulin-Like Growth Factor Binding Protein 5 / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D053781:Transforming Growth Factor beta2; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C475829:CXCL1 protein, human; D018932:Chemokine CCL2; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D018076:DNA, Complementary; D018975:Insulin-Like Growth Factor Binding Protein 5; D036341:Intercellular Signaling Peptides and Proteins; D007375:Interleukin-1; D015850:Interleukin-6; C509041:TGFB2 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D014409:Tumor Necrosis Factor-alpha; D016179:Granulocyte Colony-Stimulating Factor