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凋亡抑制蛋白Survivin在移植物内T淋巴细胞中的表达及意义

Expression of Survivin in T lymphocytes from allografts and its clinical significance

摘要:

目的 观察T淋巴细胞在体内和体外接受同种抗原刺激后凋亡抑制蛋白Survivin的表达规律并探讨其意义.方法 实验分三个部分.(1)体外实验中以刀豆素A(ConA)10 mg/L刺激C57BL/6小鼠脾细胞并进行培养,然后加入抗白细胞介素2(IL-2)受体的单克隆抗体阻断细胞增殖,最后检测脾细胞中Survivin表达的情况.(2)通过对Balb/c×C57BL/6杂交F1代小鼠输注C57BL/6小鼠脾细胞建立移植物抗宿主反应(GVHR)模型;并于不同时间点检测供者淋巴细胞中Survivin的表达.(3)选取临床移植肾穿刺活检标本73例,按移植肾Banff 97活检病理学诊断和分级标准对标本进行病理诊断和分级,并检测Survivin表达.结果 ConA刺激后脾细胞中CD3阳性细胞表达Survivin,第3天阳性细胞数目达到峰值,随后逐渐减少.GVHR小鼠于输注脾细胞后第4天,肝脏门静脉周围和汇管区出现淋巴细胞浸润并表达Survivin,可维持到第12天;第14天即不能检测到浸润淋巴细胞表达Survivin.对移植肾穿刺活检标本的检测发现,未诊断为急性细胞性排斥反应的13例标本淋巴细胞Survivin表达均呈阴性;而诊断为急性细胞性排斥反应的60例标本中有44例Survivin表达阳性(73%),两者比较,差异有统计学意义(P<0.01).结论 T淋巴细胞激活后可表达Survivin,且表达具有时间依赖性.鉴于T淋巴细胞表达Survivin的特性,移植物内T淋巴细胞表达Survivin可用于鉴别是否存在排斥反应,以及排斥反应所处的阶段.

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abstracts:

Objective To investigate the expression of survivin in T lymphocytes that were stimulated by Con A and alloantigens in renal grafts in vitro and in vivo. Methods According to the different treatments, the experiment was divided into three parts. (1) The C57BL/6 mice splenocytes stimulated by Con A (10 mg/L) were cultured in the RPMI-1640 medium. Following the proliferation blockade or not, the expression of Survivin in the splenocytes was detected. (2) The GVHR model was established by transfusing the C57BL/6 mice splenocytes into Balb/c× C57BL/6 F1 mice, and the expression of Survivin in the donor splenocytes was detected at the different time points. (3) Seventythree cases of clinical renal allograft biopsy specimens were collected, pathologically diagnosed and classified according to the Banff 97 classification, and then the expression of Survivin was detected.Results Survivin was expressed in the CD3+ splenocytes that received Con A stimulation. The positive cell count reached the peak on the day 3, and subsequently declined. In the GVHR model, the lymphocytes infiltration and Survivin expression were detected around the portal vein and portal area on the post-splenocytes-transfused day (PSTD) 4 to 12. But on the PSTD 14, the Survivin expression could not be detected in the infiltrated lymphocytes. In the renal allograft biopsy specimens,lymphocytes did not express Survivin in 13 specimens of the group without acute cellular rejection.was difference between the two groups (P<0. 01 ). Conclusion The activated T cells possess the capacity to express Survivin, and the expression is time-dependent. For the characteristics of Survivin expression of T cells, it may be applied as an approach to diagnose the acute cellular rejection and judge its degree and stage in the clinical allograft biopsy specimens.

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