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Published on: August 20, 2016
Effects of TGF-beta2 on immune response-related gene expression profiles in the human corneal endothelium
Satoru Yamagami1, Seiichi Yokoo, Tatsuya Mimura
1Department of Corneal Tissue Regeneration, Tokyo University Graduate School of Medicine, Tokyo, Japan. syamagami-tky@umin.ac.jp
Purpose:
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.
