CAJ | 학술논문

目的利用羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记大鼠肝脏NK细胞进行体内示踪,观察其在受鼠体内的存活.方法经SD大鼠门静脉转输1×106个/ml CFSE体外标记新鲜分离的SD供鼠肝脏NK细胞,于输人后1、3、7、15 d分别切取肝脏和脾脏行冰冻切片免疫荧光观察,并收集门静脉血监测淋巴细胞和中性粒细胞的比例变化.结果 CFSE对大鼠肝脏NK细胞的标记率为98.63%;转输NK细胞后第1天受鼠肝脏内CFSE荧光最强,7 d时荧光变微弱,15 d时荧光基本消失;脾脏在第1、3天时可见较弱荧光,7 d时荧光消失.转输NK细胞后相同时间点肝脏内CFSE荧光均明显强于脾脏.转输肝脏NK细胞的受鼠门静脉血中性粒细胞比例第1天时最高,随时间延长逐渐下降,第15天时基本回复正常水平,淋巴细胞比例仅一过性降低.结论转输的肝脏NK细胞在受体内的数量随时间延长而减少,在受体内的存活时间约为15 d.
목적 이용간기형광소이작산염호박선아알지(CFSE)표기대서간장NK세포진행체내시종,관찰기재수서체내적존활.방법 경SD대서문정맥전수1×106개/ml CFSE체외표기신선분리적SD공서간장NK세포,우수인후1、3、7、15 d분별절취간장화비장행빙동절편면역형광관찰,병수집문정맥혈감측림파세포화중성립세포적비례변화.결과 CFSE대대서간장NK세포적표기솔위98.63%;전수NK세포후제1천수서간장내CFSE형광최강,7 d시형광변미약,15 d시형광기본소실;비장재제1、3천시가견교약형광,7 d시형광소실.전수NK세포후상동시간점간장내CFSE형광균명현강우비장.전수간장NK세포적수서문정맥혈중성립세포비례제1천시최고,수시간연장축점하강,제15천시기본회복정상수평,림파세포비례부일과성강저.결론 전수적간장NK세포재수체내적수량수시간연장이감소,재수체내적존활시간약위15 d.
Objective To investigate the survival of donor hepatic NK cells labeled with carboxyfluorescein diacetate succiminidyl ester (CFSE) after transfusion into isogenic rats via portal vein (PV).Methods The fresh isolated hepatic NK cells from SD rats were identified by anti-rat NKR-P1 antibody staining. 1 × 106/mL NK cells labeled by CFSE in vitro were transfused into recipient SD rats via portal vein. Recipient liver and spleen tissues were harvested for detection of the fluorescence of NK cells labeled with CFSE on the day 1 to 15 after transfusion. Portal vein blood was collected for observation of the ratio of lymphocytes and neutrophilic granulocytes dynamically. Results The CFSE labeling rate of hepatic NK cells in vitro was 98.63%. The fluorescence of CFSE labeled cells in liver was the strongest on the day 1 after transfusion, while that was weak on the day 7. However, it could hardly been detected on the day 15.The fluorescence in spleen was weak on the day 1 and 3, but it could never been detected on the day 7.The ratio of lymphocytes in the portal vein blood of recipients was dropped after liver NK cells were transfused on the day 1. But the ratio in neutrophilic granulocyte field reached its peak on the day 1, dropped as time went on and returned to the normal level one the day 15. Conclusion Donor hepatic NK cells transfused via PV are decreased as time goes on in the recipients. The survival time of transfused cells in the recipient is about 15 days. The results may give us an insight into new strategy for cancer immunotherapy and transplant immunology.