双类似物对重症肌无力幼鼠Foxp3＋CD4＋CD25＋ Tregs的作用机制  

Intervention mechanism of a dual analogue on Foxp3 and CD4 ＋ CD25＋ regulatory T cells in passive transferred myasthenia gravis in young mice

作　　者：魏秀丽[1,2] 黄志[1] 

机构地区：[1]重庆医科大学附属儿童医院神经内科,400014 [2]南阳市中心医院

出　　处：《中华实用儿科临床杂志》2013年第14期1108-1111,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：重庆市卫生局科研基金项目（2008-2-169）

摘　　要：目的评价鼻黏膜免疫耐受预防性双类似物Lys262-A1a207给药对重症肌无力被动转移（PTMG）幼鼠的临床疗效，探讨Foxp3对PTMG幼鼠发病的免疫干预作用和调节机制。方法应用乙酰胆碱受体单克隆抗体（mAb35）建立C57BIM6小鼠PTMG模型，将24只幼年雌性C57BL／6小鼠分为3组：耐受组（T组）、模型对照组（MC组）、正常对照组（c组），并从致敏前10d每天经鼻黏膜滴入Lys262-Ala207或PBS。检测耐受后各组小鼠的临床症状。采用流式细胞技术检测小鼠脾细胞中CD4＋ CD25＋ T淋巴细胞及Foxp3＋CD4＋ CD25＋ Tregs水平，实时荧光定量聚合酶链反应分析小鼠脾细胞中Foxp3mRNA的表达，ELISA法检测小鼠血清IL-2、IFN-1水平。结果1．T组小鼠临床症状明显轻于MC组，但未达到正常。2．T组小鼠重复频率电刺激（RNS）出现衰减的阳性率低于MC组，血清AchR—Ab水平明显低于MC组，但2组与C组相比仍明显增高，差异均有统计‘学意义（P均〈0．05）。3．T组小鼠脾细胞中CD4＋ CD25＋ T淋巴细胞／CD4＋ T 淋巴细胞、Foxp3＋CD4＋CD25＋Tregs／CD4＋ CD25＋T淋巴细胞均较MC组高，差异均有统计学意义（P均〈0．01，0．05）；且均低于C组，但差异无统计学意义（P均〉0．05）。T组小鼠脾细胞中Foxp3mRNA表达量较MC组增高，差异有统计学意义（P〈0．05）；且低于C组，差异有统计学意义（P〈0．05）。T组小鼠血清IL-2、IFN--γ水平较MC组均显著减低，较c组均显著增高，差异均有统计学意义（P〈0．01，0．05）。结论Lys262-Ala207经鼻黏膜耐受对预防幼鼠PTMG发病具有-定作用。Lys262-Ala207诱导PTMG发生免疫耐受的机制之-可能是通过上调Foxp3mRNA的表达。Foxp3有可能是MG治疗的-个靶基因。Objective To study the prophylactic effects of nasal tolerance with a dual analogue （Lys262- Ala207） ,on the mouse model of passive transferred myasthenia gravis（PTMG） and the underlying mechanism, and to explore the cell immunologic intervention function and regulatory mechanism of Foxp3 on nasal tolerance with Lys262- Ala207 on PTMG. Methods Mouse model of PTMG was established by intraperitoneal injection of mAb35. Twenty- four C57BL/6 young female mice were divided equally into 3 groups： tolerance group（ group T） , model control group （group MC）and normal control group（ group C）. Lys262-Ala207 or PBS was given nasally before 10 days of immuniza- tion for 10 days. Clinical symptoms were evaluated after immunization. Serum level of acetylcholine receptor antibody IgG was detected by using ELISA. Levels of CD4 ＋ CD25 ＋ T cells and Foxp3 ＋ CD4 ＋ CD25 ＋ Tregs in spleen cells were measured by flow cytometry. Expression of Foxp3 mRNA in spleen cells was detected by real-time fluorescent quantita- tive polymerase chain reaction. Serum levels of IL-2 and IFN-γ were detected by ELISA. Results 1. Compared with group MC, the clinical symptoms were improved in mice of group T, but which did not return to normal. 2. The positive rate of the repetitive nerve stimulation test in group T was significantly lower than that in group MC, and the serum levels of AChR-Ab IgG in group T decreased compared with that in group MC, the differences were significant. Both the test items in group T were higher than those in group C, the differences were significant（ all P 〈 0.05 ）. 3. The ratio of CD4 ＋ CD25 ＋Y cells and CD4 ＋T cells in group T increased compared with that in group MC,the differences were signi- ficant（ all P 〈0.01,0.05）;The ratio of Foxp3 ＋CD4 ＋ CD25 ＋ Tregs and CD4 ＋ CD25 ＋T cells in group T were higher than those in group MC,the differences were significant（ all P 〈 0.01 ） ;The level of Foxp3 mRNA in spleen cells in group T was higher than that in group M

关 键 词：重症肌无力 鼻黏膜耐受 双类似物 调节性T淋巴细胞 FOXP3 

分 类 号：R746.1[医药卫生—神经病学与精神病学]