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小C型凝集素受体在小鼠真菌性角膜炎先天性免疫反应中的作用

Mincle in the innate immune response of mice fungal keratitis.

作者信息

Yu Guo-Rong, Lin Jing, Zhang Jie, Che Cheng-Ye, Peng Xu-Dong, Li Cui, Hu Li-Ting, Zhu Guo-Qiang, He Kun, Zhao Gui-Qiu

机构信息

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

出版信息

Int J Ophthalmol. 2018 Apr 18;11(4):539-547. doi: 10.18240/ijo.2018.04.01. eCollection 2018.

Abstract

AIM

To investigate how macrophage inducible C-type lectin (Mincle) influences inflammation in mice fungal keratitis induced by ().

METHODS

C57BL/6 mice were infected with after pretreated with Mincle agonist TDB or Mincle neutralizing antibody (MincleAb), taking DMSO or IgG as control group respectively. The cornea lesions were monitored with slit-lamp microscope and evaluated by clinical score. Mincle expression was assessed using reverse transcription-ploymerase chain reaction (RT-PCR) and immunostaining. The expression of cytokines (IL-1β, TNF-α and IL-6) chemokines (CXCL-1 and MIP-2) was determined by RT-PCR and ELISA. Neutrophil infiltration was observed by immunostaining. The levels of nitric oxide (NO) generated by corneas were tested by Griess reaction.

RESULTS

Mincle mRNA and protein levels were higher in infected corneas than normal corneas of C57BL/6 mice, saving clinical scores revealed differences. When pretreated with Mincle agonist TDB, the mRNA and protein levels of IL-1β, TNF-α and IL-6 in infected corneas were significantly increased compared with the control group (<0.01). Results of the counterpart in corneas pretreated with Mincle neutralizing antibody was decreased consistently (<0.01). Expression of CXCL1 and MIP-2 mRNA levels were up-regulated in TDB group and down-regulated in MincleAb group (<0.01), coincide with neutrophil aggregation degree in corneas showed by immunostaining. As for the concentration of NO, it was promoted in TDB group compared with DMSO control group, and decreased in MincleAb group compared with IgG control group.

CONCLUSION

Mincle plays a dual role in mice fungal keratitis. It participates in the innate immune system by enhancing inflammation. What's more, Mincle can mediate cytotoxic effects by regulating the formation of NO.

摘要

目的

研究巨噬细胞诱导性C型凝集素(Mincle)如何影响小鼠由()诱导的真菌性角膜炎中的炎症反应。

方法

将C57BL/6小鼠分别用Mincle激动剂TDB或Mincle中和抗体(MincleAb)预处理后感染(),分别以二甲基亚砜(DMSO)或IgG作为对照组。用裂隙灯显微镜监测角膜病变情况,并通过临床评分进行评估。采用逆转录-聚合酶链反应(RT-PCR)和免疫染色法评估Mincle的表达。通过RT-PCR和酶联免疫吸附测定(ELISA)法测定细胞因子(IL-1β、TNF-α和IL-6)及趋化因子(CXCL-1和MIP-2)的表达。通过免疫染色观察中性粒细胞浸润情况。采用格里斯反应检测角膜产生的一氧化氮(NO)水平。

结果

感染角膜中Mincle的mRNA和蛋白水平高于C57BL/6小鼠的正常角膜,临床评分显示存在差异。用Mincle激动剂TDB预处理后,感染角膜中IL-1β、TNF-α和IL-6的mRNA和蛋白水平与对照组相比显著升高(<0.01)。用Mincle中和抗体预处理角膜的相应结果则持续降低(<0.01)。TDB组中CXCL1和MIP-2 mRNA水平上调,MincleAb组中下调(<0.01),这与免疫染色显示的角膜中中性粒细胞聚集程度一致。至于NO浓度,与DMSO对照组相比,TDB组中升高,与IgG对照组相比,MincleAb组中降低。

结论

Mincle在小鼠真菌性角膜炎中起双重作用。它通过增强炎症反应参与固有免疫系统。此外,Mincle可通过调节NO的形成介导细胞毒性作用。

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