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Differences in gene regulation by TLR3 and IPS-1 signaling in murine corneal epithelial cells
Seitaro Komai1, Mayumi Ueta2, Hiromi Nishigaki1
1Department of Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kajii-Cho, Kamigyo-Ku, Kyoto, Japan.
Abstract:
Toll-like receptor 3 (TLR3) and interferon-beta promoter stimulator-1 (IPS-1) are associated with antiviral responses to double-stranded RNA viruses and contribute to innate immunity. We previously reported that conjunctival epithelial cell (CEC) TLR3 and IPS-1 pathways respond to the common ligand polyinosinic:polycytidylic acid (polyI:C) to regulate different gene expression patterns as well as CD11c + cell migration in murine-model corneas. However, the differences in the functions and the roles of TLR3 and IPS-1 remain unclear. In this study, we investigated the differences of TLR3 or IPS-1-induced gene expression in corneal epithelial cells (CECs) in response to polyI:C stimulation using cultured murine primary CECs (mPCECs) derived from TLR3 and IPS-1 knockout mice via comprehensive analysis. The genes associated with viral responses were upregulated in the wild-type mice mPCECs after polyI:C stimulation. Among these genes, Neurl3, Irg1, and LIPG were dominantly regulated by TLR3, while interleukin (IL)-6 and IL-15 were dominantly regulated by IPS-1. CCL5, CXCL10, OAS2, Slfn4, TRIM30α, and Gbp9 were complementarily regulated by both TLR3 and IPS-1. Our findings suggest that CECs may contribute to immune responses and that TLR3 and IPS-1 possibly have different functions in the corneal innate immune response.
Insights
Toll-like receptor 3 (TLR3) and interferon-beta promoter stimulator-1 (IPS-1) pathways in corneal epithelial cells differentially regulate antiviral gene expression. This study clarifies their distinct roles in the innate immune response to viral stimulation.
Area of Science:
- Immunology
- Ophthalmology
- Molecular Biology
Background:
- Toll-like receptor 3 (TLR3) and interferon-beta promoter stimulator-1 (IPS-1) are key mediators of innate immunity against double-stranded RNA viruses.
- Conjunctival epithelial cells (CECs) express TLR3 and IPS-1, responding to polyinosinic:polycytidylic acid (polyI:C) and influencing gene expression and cell migration.
Purpose of the Study:
- To investigate and delineate the distinct functional roles of TLR3 and IPS-1 in regulating gene expression within corneal epithelial cells (CECs) upon polyI:C stimulation.
- To identify specific genes predominantly or complementarily regulated by TLR3 and IPS-1 pathways in the context of viral response.
Main Methods:
- Utilized cultured murine primary CECs (mPCECs) derived from TLR3 and IPS-1 knockout mice.
- Stimulated mPCECs with polyI:C and performed comprehensive gene expression analysis.
- Compared gene expression patterns between wild-type and knockout mPCECs to determine pathway-specific regulation.
Main Results:
- PolyI:C stimulation upregulated antiviral response genes in wild-type mPCECs.
- TLR3 predominantly regulated Neurl3, Irg1, and LIPG.
- IPS-1 predominantly regulated interleukin (IL)-6 and IL-15.
- CCL5, CXCL10, OAS2, Slfn4, TRIM30α, and Gbp9 were regulated by both pathways.
Conclusions:
- CECs play a significant role in corneal innate immune responses.
- TLR3 and IPS-1 exhibit distinct, yet complementary, functions in regulating antiviral gene expression in CECs.
- Findings provide insight into the specific contributions of TLR3 and IPS-1 to corneal immunity.
