天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2013年
7期
632-635
,共4页
李青%邓琦%刘鹏江%白雪%穆娟%李玉明
李青%鄧琦%劉鵬江%白雪%穆娟%李玉明
리청%산기%류붕강%백설%목연%리옥명
重组融合蛋白质类%纤连蛋白类%抗原, CD3%抗体,单克隆%细胞因子类%杀伤细胞
重組融閤蛋白質類%纖連蛋白類%抗原, CD3%抗體,單剋隆%細胞因子類%殺傷細胞
중조융합단백질류%섬련단백류%항원, CD3%항체,단극륭%세포인자류%살상세포
recombinant fusion proteins%fibronectins%antigens,CD3%antibodies,monoclonal%cytokines%killer cells
目的探讨重组人纤维连接片段（RetroNectin）与抗CD3单抗（CD3Ab）联合包被培养对细胞因子诱导的杀伤细胞（CIK）增殖和杀伤活性的影响。方法取完全缓解期急性白血病(AL)患者外周血单个核细胞，采用Retro-Nectin包被（RN组）、CD3Ab包被（CD3Ab组）及RetroNectin联合CD3Ab包被（RN+CD3Ab组）与传统方法培养（对照组）获得CIK细胞，比较各组增殖、表达及杀伤活性。结果 CIK扩增：各实验组CIK扩增倍数高于对照组，且RN+CD3Ab组高于RN组和CD3Ab组（P<0.05）。收获细胞CD25+表达：RN组和RN+CD3Ab组高于对照组和CD3Ab组（P<0.05）。各期细胞比例：RN组和RN+CD3Ab组的G1期细胞比例低于CD3Ab组和对照组，而S期细胞高于CD3Ab组和对照组（P<0.05）。CIK对自体AL细胞的杀伤活性：效靶比10∶1、20∶1和40∶1时，RN组和RN+CD3Ab组高于对照组和CD3Ab组（P<0.05）。收获细胞凋亡率：RN组和RN+CD3Ab组较CD3Ab组和对照组下降（P<0.05）。结论RetroNectin联合CD3Ab包被用于CIK体外培养可获得增殖率和活性更高的免疫活性细胞，是一种值得推荐的方法。
目的探討重組人纖維連接片段（RetroNectin）與抗CD3單抗（CD3Ab）聯閤包被培養對細胞因子誘導的殺傷細胞（CIK）增殖和殺傷活性的影響。方法取完全緩解期急性白血病(AL)患者外週血單箇覈細胞，採用Retro-Nectin包被（RN組）、CD3Ab包被（CD3Ab組）及RetroNectin聯閤CD3Ab包被（RN+CD3Ab組）與傳統方法培養（對照組）穫得CIK細胞，比較各組增殖、錶達及殺傷活性。結果 CIK擴增：各實驗組CIK擴增倍數高于對照組，且RN+CD3Ab組高于RN組和CD3Ab組（P<0.05）。收穫細胞CD25+錶達：RN組和RN+CD3Ab組高于對照組和CD3Ab組（P<0.05）。各期細胞比例：RN組和RN+CD3Ab組的G1期細胞比例低于CD3Ab組和對照組，而S期細胞高于CD3Ab組和對照組（P<0.05）。CIK對自體AL細胞的殺傷活性：效靶比10∶1、20∶1和40∶1時，RN組和RN+CD3Ab組高于對照組和CD3Ab組（P<0.05）。收穫細胞凋亡率：RN組和RN+CD3Ab組較CD3Ab組和對照組下降（P<0.05）。結論RetroNectin聯閤CD3Ab包被用于CIK體外培養可穫得增殖率和活性更高的免疫活性細胞，是一種值得推薦的方法。
목적탐토중조인섬유련접편단（RetroNectin）여항CD3단항（CD3Ab）연합포피배양대세포인자유도적살상세포（CIK）증식화살상활성적영향。방법취완전완해기급성백혈병(AL)환자외주혈단개핵세포，채용Retro-Nectin포피（RN조）、CD3Ab포피（CD3Ab조）급RetroNectin연합CD3Ab포피（RN+CD3Ab조）여전통방법배양（대조조）획득CIK세포，비교각조증식、표체급살상활성。결과 CIK확증：각실험조CIK확증배수고우대조조，차RN+CD3Ab조고우RN조화CD3Ab조（P<0.05）。수획세포CD25+표체：RN조화RN+CD3Ab조고우대조조화CD3Ab조（P<0.05）。각기세포비례：RN조화RN+CD3Ab조적G1기세포비례저우CD3Ab조화대조조，이S기세포고우CD3Ab조화대조조（P<0.05）。CIK대자체AL세포적살상활성：효파비10∶1、20∶1화40∶1시，RN조화RN+CD3Ab조고우대조조화CD3Ab조（P<0.05）。수획세포조망솔：RN조화RN+CD3Ab조교CD3Ab조화대조조하강（P<0.05）。결론RetroNectin연합CD3Ab포피용우CIK체외배양가획득증식솔화활성경고적면역활성세포，시일충치득추천적방법。
Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P<0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P<0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P<0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P<0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P < 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.
