冻存的亲属活体供肾者的骨髓源性树突状细胞介导受者淋巴细胞反应  

Donor dendritic cells-mediated recipient lymphocyte reaction after living related kidney transplantation

作　　者：黄赤兵[1] 许晓婷[1] 冯嘉瑜[1] 范明齐[1] 张艮甫[1] 王平贤[1] 肖亚[1] 

机构地区：[1]第三军医大学附属新桥医院肾移植中心,重庆400037

出　　处：《中华器官移植杂志》2011年第1期39-42,共4页Chinese Journal of Organ Transplantation

摘　　要：目的 探讨冻存的供者骨髓源性树突状细胞（DC）介导肾移植受者淋巴细胞反应的可行性.方法 肾移植前取供者骨髓,分离单个核细胞,液氮冻存备用.术后1、3、6、9个月,取冻存的供者骨髓细胞分离CD34＋细胞,并培养DC,检测各时期DC活细胞的比率;同时,分别以供者DC和外周血淋巴细胞（PBL）作为刺激细胞,以术前未冻存过的供者骨髓源性DC作为对照,观察健康志愿者以及受者淋巴细胞对冻存不同时间的供者DC和PBL刺激的反应.结果术前每10 ml骨髓中可分离骨髓单个核细胞（BMMC）的数量平均为6.8 × 107个,免疫磁珠分选获得CD34＋细胞数为（4.10±0.58）×105个.冻存不同时间的BMMC复苏后,所分离到的CD34＋细胞的比率并无明显差异.术后1个月时,BMMC、CD34＋细胞存活率均大幅明显下降,以后呈缓慢进行性下降,BMMC的下降幅度大于CD34＋细胞.DC的活细胞比率维持在93.2%～94.8%.肾移植术后,各组DC对健康志愿者和受者淋巴细胞的刺激能力均显著强于PBL（P＜0.05）;在术后各时间观察点,健康志愿者淋巴细胞对DC刺激的反应波动在12 067±1190至14 238±1220（P＞0.05）,受者淋巴细胞对DC刺激的反应波动较大,在9490±1386～15 233±1098.结论 供者骨髓源性DC具有稳定的细胞活力和刺激能力,其作为术后长时间内介导肾移植受者淋巴细胞反应的刺激原,明显优于供者外周淋巴细胞,术前仅需一次性取少量骨髓冻存,术后可随时且多次用于监测.Objective To explore the feasibility of mediating recipient lymphocyte reaction with donor dendritic cells （DCs） in renal allograft recipients. Methods Donor bone marrow monocytes （BMMCs） were isolated and cryopreserved in liquid nitrogen before kidney transplantation. At 0 day, 1month,3 month, 6 month and 9 month post-operation, CD34＋ cells which were isolated from frozen BMMCs and cultured into DCs as well as the peripheral blood lymphocytes （PBLs） of donors were used as the stimulating cells to the PBLs of recipients and healthy volunteers. The number of viable DCs from frozen- and room temperature-preserved BMMCs was counted and the reactions of recipients＇and healthy volunteers＇ lymphocytes to DCs and donor PBLs were measured. Results 6. 8 × 107BMMCs were isolated from each 10 ml of donor bone marrow on average while （4. 10 ± 0. 58） × 105CD34＋ cells were isolated by magnetic active cell sorting （MACS）. There was no significant difference in the isolating rate of recovered CD34＋ cells at each observation point postoperatively. The percentage of viable BMMCs and CD34＋ was decreased significantly at 1 month after surgery, then, decreased slowly and progressively. The decreasing rate of BMMCs was higher than CD34＋. The rate of viable DCs was maintained stable （93. 2%-94. 8% ） in each group. The reactions of recipients＇ and healthy volunteers＇ lymphocytes to DCs were stronger than those to donor PBLs （P〈0. 05）. The reactions of healthy volunteers＇ lymphocytes to DCs were maintained stable while those of recipients＇ were fluctuating. Conclusion Bone marrow-derived DCs are superior to PBLs in mediating long-term lymphocyte reaction after kidney transplantation due to their stable viability and stimulating ability to lymphocytes. Only once collection of a small quality of bone marrow of donors is needed to meet the demand of immune monitoring at any time after transplantation.

关 键 词：肾移植 树突细胞 低温保存 淋巴细胞培养试验 混合 

分 类 号：R699.2[医药卫生—泌尿科学]