Effects of dimethyl itaconate on secretion of pro-inflammatory factors in den-dritic cells and on T helper 17 cells in mice with experimental autoimmune uveitis
10.13389/j.cnki.rao.2024.0067
- VernacularTitle:衣康酸二甲酯对树突状细胞分泌促炎因子以及实验性自身免疫性葡萄膜炎小鼠辅助性T细胞17的影响
- Author:
Jiali WANG
1
;
Chao YANG
;
Sisi CHEN
;
Kailang ZHANG
;
Ruihua WEI
;
Hong NIAN
Author Information
1. 300384 天津市,天津医科大学眼科医院、眼视光学院、眼科研究所,国家眼耳鼻喉疾病临床医学研究中心天津市分中心,天津市视网膜功能与疾病重点实验室
- Keywords:
dimethyl itaconate;
dendritic cells;
T helper 17 cells;
experimental autoimmune uveitis
- From:
Recent Advances in Ophthalmology
2024;44(5):346-349
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of dimethyl itaconate(DMI)on the secretion of pro-inflammatory factors in dendritic cells(DCs)and on the interphotoreceptor retinoid-binding protein(IRBP)1-20-specific T helper 17(Th17)cells in mice with experimental autoimmune uveitis(EAU).Methods Bilateral femur and tibia of C57BL/6J mice were isolated to obtain bone marrow cells,and these bone marrow cells were directionally induced with granulocyte macrophage-colony-stimulating factor(GM-CSF)and interleukin(IL)-4 to differentiate DCs.After 6 days,DCs were ran-domly divided into the DMI group and the phosphate-buffered saline(PBS)group.Cells in the DMI group were pretreated with 250 μmol·L-1 DMI,and cells in the PBS group were pretreated with the same volume of PBS for 3 hours.After-wards,100 μg·L-1 lipopolysaccharide was added in the every group to stimulate cells for 24 hours.The relative mRNA ex-pression levels of IL-6,IL-1 β,and IL-23 in DCs were measured by quantitative real-time polymerase chain reaction(qRT-PCR).The EAU model was constructed by actively immunizing mice with IRBP1-20,Freund's incomplete adjuvant,and my-cobacterium tuberculosis H37RA.Thirteen days after immunization,T cells in the spleen and lymph node isolated from EAU mice were cocultured with DMI-treatedor PBS-treated DCs in the medium containing IRBP1-20.They polarized toward Th17 cells.The percentage of Th17 cells in the cocultured cells was detected by flow cytometry.The IL-17 level in the coculture supernatant was detected by enzyme-linked immunosorbent assay(ELISA).qRT-PCR was performed to detect the relative mRNA expression levels of retinoid-related orphan receptor gamma t(RORγt),IL-17,IL-23R,and GM-CSF in the cocultured cells.Results qRT-PCR analysis revealed that the relative mRNA expression levels of IL-6,IL-1β,and IL-23 in the DMI group were significantly lower thanthose in the PBS group(all P＜0.05).Flow cytometry analysis showed that the proportion of Th17 cells in the cocultured cells in the DMI group was significantly lower than that in the PBS group(P＜0.05).ELISA analysis exhibited that the IL-17 level in the coculture supernatant in the DMI group was significantly lower than that in the PBS group(P＜0.05).The relative mRNA expression levels of IL-17,ROR-γt,IL-23R and GM-CSF in cocultured cells in the DMI group significantly decreased compared with the PBS group(all P＜0.05).Conclusion DMI can reduce the expression of IL-6,IL-1β and IL-23 in DCs,thus negatively modulating the responses of IRBP1-20-specific Th17 cells.
