title: Inhibition of RelA expression via RNA interference induces immune tolerance in a rat keratoplasty model.
abstract: RelA, the most important regulator of NF-kB activity, and its mechanisms in keratoplasty immune rejection have not been fully investigated. In the present study, lentivirus-mediated silencing of RelA expression in a bone marrow-derived dendritic cell (BMDC) model was tested. The BMDCs were transfected with RelA-shRNA to induce an immature, maturation-resistant and tolerogenic phenotype, while not significantly changing IFN-γ, IL-10 and IL-17 expression. A fully allogeneic rat cornea transplant model was established for in vivo studies. The allograft mean survival time (MST) of lv-shRelA-DC injection groups were significantly longer than the untreated BMDC group and control group. The corneal opacity and neovascularization scale of the lv-shRelA-DC injection groups were slight compared to pair control others. Postoperative flow cytometric analysis revealed that the percentage of Treg positive cells was dramatically increased in animals that received an lv-shRelA-DC injection. ELISA and qRT-PCR analyses of serum showed that IFN-γ and IL-17 expression were suppressed by lv-shRelA-DC treatement. In vivo experiments demonstrated that IL-10 induced immunosuppression was partly attributed to injection of lv-shRelA-DC throughout the experiment, differing from the general anti-inflammatory factors. Luciferase and Chromatin IP evaluation showed that RelA knockdown in BMDCs significantly reduces DNA binding to IFN-γ, IL-10 and the IL-17 promoter and inhibited of transcriptional activity. Taken together, this study illustrates a significant role of RelA in mediating the corneal neovascularization by affecting IL-17 expression. Our comprehensive analysis shows that the significant role of RelA provides a novel and feasible therapeutic approach for the prevention of corneal allograft rejection.
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Jize
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Songfu
authors.initials: S
authors.lastname: Feng
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Guoguo
authors.initials: G
authors.lastname: Yi
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Ruiwen
authors.initials: R
authors.lastname: Yi
authors.affiliation: Department of Ophtalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. Electronic address: Luxh63@163.com.
authors.forename: Xiaohe
authors.initials: X
authors.lastname: Lu
authors.affiliation: Guangzhou Women and Children's Medical Center, Guangzhou 510623, China.
authors.forename: Wanfu
authors.initials: W
authors.lastname: Xu
authors.affiliation: Department of Ophtalmology, Guangzhou first people's Hospital, Guangzhou 510180, China.
authors.forename: Haijiang
authors.initials: H
authors.lastname: Qiu
mesh_terms: D064591:Allografts; D000818:Animals; D047369:Chromatin Immunoprecipitation; D016510:Corneal Neovascularization / Q000276:immunology*; D016039:Corneal Transplantation; D003713:Dendritic Cells / Q000276:immunology; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D006084:Graft Rejection / Q000276:immunology; D006085:Graft Survival / Q000276:immunology; D007108:Immune Tolerance / Q000276:immunology*; D007150:Immunohistochemistry; D016753:Interleukin-10 / Q000276:immunology; D020381:Interleukin-17 / Q000276:immunology; D008297:Male; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D051996:Transcription Factor RelA / Q000276:immunology* / Q000378:metabolism; D014181:Transplantation Immunology / Q000276:immunology*
chemical_list: D020381:Interleukin-17; C498145:Rela protein, rat; D051996:Transcription Factor RelA; D016753:Interleukin-10
keywords: Bone marrow-derived dendritic cell (BMDC); Cornea; Immune tolerance; NF-kB; RelA; RelA-shRNA