BACKGROUND: The authors previously reported that human ocular surface epithelium expressed TLR3 and that its ligand polyI:C stimulated the secretion of IL-6, IL-8 and IFN-β. In this study, to examine comprehensive effects of polyI:C stimulation of primary human conjunctival epithelial cells (PHCjECs), the authors performed a gene-expression analysis of the polyI:C-stimulated PHCjECs using oligonucleotide microarrays, GeneChip. METHODS: The transcripts upregulated upon polyI:C stimulation in PHCjECs from two individuals were examined using GeneChip. Eleven new upregulated transcripts of interest were confirmed by quantitative real-time PCR (RT-PCR), and seven proteins produced by those transcripts were examined by ELISA or immunoblot analysis in PHCjECs from three other individuals, respectively. RESULTS: GeneChip analysis showed that 150 transcripts were upregulated more than threefold and that 47 transcripts were upregulated more than 10-fold upon polyI:C stimulation in the PHCjECs. Eleven of the 47 upregulated transcripts (CXCL11, RIG-I, IL28A, CXCL10, CCL5, CCL4, MDA5, IL7R, TSLP, CCL20 and ICAM-1) were significantly upregulated upon polyI:C stimulation by quantitative RT-PCR, and the levels of seven proteins of the transcripts CXCL11, CXCL10, CCL5, CCL20, TSLP, RIG-I and MDA5 were confirmed by ELISA or immunoblot analysis to increase significantly in polyI:C-stimulated PHCjECs. CONCLUSIONS: Our results might show that TLR3 of conjunctival epithelium could not only induce antiviral innate immune responses but also regulate the allergic reactions.
BACKGROUND: The authors previously reported that human ocular surface epithelium expressed TLR3 and that its ligand polyI:C stimulated the secretion of IL-6, IL-8 and IFN-β. In this study, to examine comprehensive effects of polyI:C stimulation of primary human conjunctival epithelial cells (PHCjECs), the authors performed a gene-expression analysis of the polyI:C-stimulated PHCjECs using oligonucleotide microarrays, GeneChip. METHODS: The transcripts upregulated upon polyI:C stimulation in PHCjECs from two individuals were examined using GeneChip. Eleven new upregulated transcripts of interest were confirmed by quantitative real-time PCR (RT-PCR), and seven proteins produced by those transcripts were examined by ELISA or immunoblot analysis in PHCjECs from three other individuals, respectively. RESULTS: GeneChip analysis showed that 150 transcripts were upregulated more than threefold and that 47 transcripts were upregulated more than 10-fold upon polyI:C stimulation in the PHCjECs. Eleven of the 47 upregulated transcripts (CXCL11, RIG-I, IL28A, CXCL10, CCL5, CCL4, MDA5, IL7R, TSLP, CCL20 and ICAM-1) were significantly upregulated upon polyI:C stimulation by quantitative RT-PCR, and the levels of seven proteins of the transcripts CXCL11, CXCL10, CCL5, CCL20, TSLP, RIG-I and MDA5 were confirmed by ELISA or immunoblot analysis to increase significantly in polyI:C-stimulated PHCjECs. CONCLUSIONS: Our results might show that TLR3 of conjunctival epithelium could not only induce antiviral innate immune responses but also regulate the allergic reactions.
Authors: Daniel R Saban; Virginia Calder; Chuan-Hui Kuo; Nancy J Reyes; Darlene A Dartt; Santa J Ono; Jerry Y Niederkorn Journal: Curr Eye Res Date: 2013-01-02 Impact factor: 2.424
Authors: Korneliusz Golebski; Joost van Tongeren; Danielle van Egmond; Esther J de Groot; Wytske J Fokkens; Cornelis M van Drunen Journal: PLoS One Date: 2016-04-06 Impact factor: 3.240
Authors: Radhika Susarla; Lei Liu; Elizabeth A Walker; Iwona J Bujalska; Jawaher Alsalem; Geraint P Williams; Sreekanth Sreekantam; Angela E Taylor; Mohammad Tallouzi; H Susan Southworth; Philip I Murray; Graham R Wallace; Saaeha Rauz Journal: PLoS One Date: 2014-04-15 Impact factor: 3.240