CFSE标记抗原特异性CD4+CD25+T细胞在胰岛移植体内归巢探究.docVIP

CFSE标记抗原特异性CD4+CD25+T细胞在胰岛移植体内归巢探究 　 作者：张梅,　徐书杭,　徐瑜,　刘翠萍,　茅晓东,　徐宽枫,　刘超 【摘要】 　　目的: 观察抗原特异性CD4+CD25+Treg细胞在小鼠同种异体胰岛移植后体内的分布特点, 探讨抗原特异性CD4+CD25+Treg细胞免疫调节可能机制。方法:　构建小鼠同种异体胰岛移植模型。CFSE对抗原特异性CD4+CD25+Treg细胞标记, 经尾静脉输注。用组织冰冻切片和流式细胞术动态了解CFSE标记的抗原特异性CD4+CD25+Treg细胞在受体内的分布和增殖变化。结果:　抗原特异性CD4+CD25+ Treg细胞联合胰岛移植组胰岛移植物平均生存期为(34.57±17.15) d, 较胰岛移植组生存时间(10.6±1.82) d, 差异有统计学意义(Plt;0.01)。抗原特异性CD4+CD25+ Treg 细胞在各级淋巴结均有能定居, 但是胰腺淋巴结定居的细胞数量明显多于其他淋巴结。结论:　抗原特异性Treg细胞抑制STZ诱导的糖尿病小鼠的急性移植排斥反应, 细胞抑制功能与细胞在体内淋巴结的归巢有关。 　　AIM: To observe the distribution and proliferation of the antigenspecific CD4+CD25+ regulatory T (Treg ) cells and investigate the potential mechanism of antigenspecific CD4+CD25+ regulatory T cells on the suppression of rejection for allogenetic islet transplantation in vivo. METHODS: Antigenspecific CD4+CD25+ Treg cells were generated by the addition of multiple intravenous injections of ICR mice splenocytes in vivo. After labeled by CFSE, 8×105 antigenspecific Treg cells were injected via tail vein with islets transplantation. Homing and distribution of antigenspecific CD4+CD25+ Treg cells were investigated by flow cytometric analysis and immunofluorescence. RESULTS: In vivo, the mean survival time of recipients with islets and antigenspecific CD4+CD25+ Treg cells were (34.57±17.15) days, whereas transplanted islets without Treg treatment survived (10.6±1.82) days in control mice. The transferred antigenspecific CD4+CD25+ Treg cells were mainly resident in pancreatic node and mesenteric node. CONCLUSION: Allogeneic antigenspecific CD4+CD25+ Treg cells prolong islet graft survival, and draining lymphoid tissue play a key role in the immune response. 【关键词】 CD4+CD25+调节性T细胞; 胰岛/移植; CFSE 　　胰岛移植是治疗1型糖尿病理想的方法。近年来, 较多研究发现CD4+CD25+调节性T细胞(regulatory T cell, Treg)在调节移植免疫中起重要作用［1］。Treg细胞主动免疫调节作用为移植排斥反应的治疗开辟了一条新途径。选择合适的细胞标记物跟踪观察, 对于研究移植后的细胞在体内作用机制具有重要作用。在构建小鼠胰岛移植模型的基础上, 我们将标记荧光染料羧基荧光素二醋酸盐琥珀酰亚胺酯(5, 6carboxyfluorescein diacetate succinimidyl ester, CFSE) 抗原特异性CD4+CD25+ Tre