[1]苏亚茹,白浪,汤明芳,等.敲除TLR2基因对同种异体小鼠角膜移植排斥反应的影响[J].眼科新进展,2016,36(11):1006-1010.[doi:10.13389/j.cnki.rao.2016.0269]
 SU Ya-Ru,BAI Lang,TANG Ming-Fang,et al.Variation of immune response after allogeneic corneal transplantation in TLR2 knock-out mice[J].Recent Advances in Ophthalmology,2016,36(11):1006-1010.[doi:10.13389/j.cnki.rao.2016.0269]
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敲除TLR2基因对同种异体小鼠角膜移植排斥反应的影响
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年11期
页码:
1006-1010
栏目:
实验研究
出版日期:
2016-11-05

文章信息/Info

Title:
Variation of immune response after allogeneic corneal transplantation in TLR2 knock-out mice
作者:
苏亚茹白浪汤明芳于健刘晶孙宇飞孟婷
510000 广东省广州市,南方医科大学南方医院眼科
Author(s):
SU Ya-Ru BAI Lang TANG Ming-Fang YU Jian LIU Jing SUN Yu-Fei MENG Ting
Department of Ophthatmology , Nanfang Hospital of Southern Medical University , Guangzhou 510000 , Guangdong Province , China
关键词:
Toll样受体2基因敲除MyD88角膜移植免疫排斥CD4+T细胞干扰素-γ
Keywords:
toll-like receptor 2 gene knock-out Myd88 corneal transplantation immune rejection CD4+T cell IFN-7
分类号:
R772.2
DOI:
10.13389/j.cnki.rao.2016.0269
文献标志码:
A
摘要:
目的 探讨敲除Toll样受体2(toll-likereceptor2,TLR2)基因后是否抑制同种异体小鼠角膜排斥反应的发生。方法 以BALB/c为供体,各取30只C57BL/6和同背景TLR2基因敲除鼠为受体行右眼角膜移植术,设为野生组(WT)和基因敲除组(KO);取19只C57BL/6行右眼自体角膜移植术,为自体组(ISO);各取9只C57BL/6和TLR2基因敲除鼠分别设为野生对照组(WTcontrol)和基因敲除对照组(KOcontrol)。术后每周两次观察植片并记录免疫排斥的发生时间;术后14d,收集术眼同侧颈部淋巴结,行流式细胞术分析CD4+T细胞百分比;收集术眼角膜,行免疫组织化学染色和实时荧光定量PCR检测角膜干扰素(inter-feron,IFN)-γ、TLR2和MyD88的表达。结果 WT组和KO组小鼠角膜移植术后中位生存时间KO组(35.0±3.8)d长于WT组(21.0±1.5)d(P<0.05)。流式细胞术分析显示,WT组同侧颈部淋巴结CD4+T细胞百分比较WTcontrol组明显增高(P<0.05),而ISO和KO组相对于其对应的control组(WT/KOcontrol)差异均无统计学意义(均为P>0.05);免疫组织化学结果显示,ISO和KO组间角膜IFN-γ和MyD88分子表达无差异,但两者均低于WT组。KO组角膜几乎不表达TLR2分子,ISO组有微量表达,WT组表达明显增高;PCR检测显示,ISO和KO组角膜IFN-γ和TLR2mRNA相对表达量差异均无统计学意义(均为P>0.05),但两者均低于WT组(均为P<0.05);KO组角膜MyD88mRNA相对表达量比ISO组高(P<0.05),但两者均低于WT组(均为P<0.05)。结论 敲除TLR2基因可以一定程度抑制同种异体小鼠角膜排斥反应的发生。
Abstract:
Objective To explore whether toll-like receptor 2 ( TLR2 ) knock-out can attenuate immune rejection after allogeneic corneal transplantation. Methods BALB/c mice were used as donors.30 C57BL/6 mice and 30 TLR2 knock-out mouse were chosen as receptors to establish allograft corneal transplantation models, which were grouped as the wide type ( WT) mouse group and knock-out ( KO) mouse group. Another 19 C57BL/6 mice were performed isograft corneal transplantation and were termed as isograft ( ISO) group. Nine normal mice of wild-type C57BL/6 ( WT control) or TLR2 knock-out ( KO control) were respectively involved as negative controls. The edema and transparency were observed twice per week under slit lamp nucroscope after surgery and the rejection times were recorded according to Sonoda’ s criteria. At 14 days after transplantation , the frequencies of CD4 + T cells in ipsilateral cervical drairung lymph nodes were analyzed by flow cytometry, and the expression of IFN-7 , TLR2 and MyD88 were detected by immunohistochemical staining and qPCR. Results The median survival time ( MST) in KO group was ( 35. 0 +3. 8) days , while in WT group was ( 21. 0 + 1. 5 ) days, there was significant difference between WT and KO group (P < 0. 05 ) . Lymphoid CD4 + T cells percentage of WT group was significantly higher than that of WT control group ( P < 0. 05 ) . There was no statistical difference between ISO and WT control group , as well as KO and KO control group ( all P > 0. 05 ) . Immunohistochemistry staining revealed that IFN-7 and MyD88 expressed almost alike in ISO and KO group, but there was a significant decreasing in ISO and KO group compared with WT group. No TLR2 could be detected in KO group, while expressed weakly in ISO group, and increased in WT group. The mRNA expression of IFN-7 and MyD88 in ISO and KO group were nearly the same ( all P > 0. 05 ) ,but there was a marked decrease in ISO and KO group while compared with WT group ( P < 0. 05 ) . The MyD88 mRNA expression level in KO group was much higher than that in ISO group (P < 0. 05 ) , and both of them were lower than that in WT group ( all P < 0. 05 ) . Conclusion TLR2 knock-out may attenuate immune rejection after allogeneic corneal transplantation.

相似文献/References:

[1]苏亚茹,白浪,刘晶,等.Toll样受体2调节Th细胞极化对小鼠角膜移植排斥反应发生的影响[J].眼科新进展,2016,36(7):601.[doi:10.13389/j.cnki.rao.2016.0159]
 SU Ya-Ru,BAI Lang,LIU Jing,et al.Effects of Th cell polarization regulated by TLR2 0n immune rejection after corneal transplantation in mouse[J].Recent Advances in Ophthalmology,2016,36(11):601.[doi:10.13389/j.cnki.rao.2016.0159]

备注/Memo

备注/Memo:
国家自然科学基金资助(编号:81170887);南方医科大学南方医院横向课题匹配基金资助(编号:G201202)
更新日期/Last Update: 2016-11-29