Wang Yan, Gao Yi, Sun Er-wei, Chen Zhong-hua
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Zhonghua Gan Zang Bing Za Zhi. 2006 May;14(5):370-4.
To track the location of the transfused apoptotic allogeneic lymphocytes and asses the process of their accumulation and phagocytosis removal as consequences on allograft tolerance in recipient mice.
Donor spleen lymphocytes were labeled by CFSE and induced to apoptosis by dexamethasone incubation. After purification by anti-annexin V-conjugated magnetic beads isolation, apoptotic lymphocytes were transfused into recipient mice through the tail veins. Tissue samples from various organs were taken at various time points to analyze the fates of the apoptotic allogeneic lymphocytes and the phagocytosis of them by organ resident APCs.
Using fluorescent microscopy and flow cytometry, after the apoptotic cells were recognized and uptaken, the largest amount of labeled cells were accumulated in the livers and disappeared within not more than 12 hours. Recipient liver APCs were highly efficacious in phagocytosis of apoptotic allogeneic lymphocytes; the removal was completed within 15 minutes after incubation. LSEC, KC and LDC all phagocytosized the apoptotic allogeneic lymphocytes but with significantly different rates. Considering the numbers of those cells in a normal liver, it could be calculated that LSEC and KC had greater effects in this activity.
The liver deserves foremost attention for study of the mechanism of allografts tolerance induced by pre-transfusion of apoptotic donor spleen lymphocytes. LSEC and KC are the main functional APCs to the alloantigens.
追踪输注的凋亡同种异体淋巴细胞的位置,并评估其积累和吞噬清除过程,以及这些过程对受体小鼠同种异体移植耐受的影响。
用CFSE标记供体脾淋巴细胞,并通过地塞米松孵育诱导其凋亡。经抗膜联蛋白V偶联磁珠分离纯化后,将凋亡淋巴细胞经尾静脉输注到受体小鼠体内。在不同时间点采集各器官的组织样本,分析凋亡同种异体淋巴细胞的命运及其被器官驻留抗原呈递细胞吞噬的情况。
利用荧光显微镜和流式细胞术观察到,凋亡细胞被识别并摄取后,标记细胞大量积聚在肝脏中,且在不超过12小时内消失。受体肝脏抗原呈递细胞对凋亡同种异体淋巴细胞的吞噬效率很高;孵育后15分钟内清除完成。肝血窦内皮细胞、枯否细胞和肝树突状细胞均能吞噬凋亡同种异体淋巴细胞,但吞噬速率差异显著。根据正常肝脏中这些细胞的数量计算,肝血窦内皮细胞和枯否细胞在该活动中作用更大。
在研究凋亡供体脾淋巴细胞预输注诱导同种异体移植耐受的机制时,肝脏应得到首要关注。肝血窦内皮细胞和枯否细胞是针对同种异体抗原的主要功能性抗原呈递细胞。