CAJ | 학술논문

目的：研究并探讨重组人纤维连接蛋白（ RetroNectin ， RN ）对细胞因子诱导的杀伤细胞（ CIK）增殖、免疫特性及对人类K562、LOVO细胞杀伤活性的影响。方法提取患者外周血单个核细胞，标本分为四组，对照组（ CIK组）：按常规方法诱导CIK增殖；C－CIK组：提前通过CD3 Ab包被诱导；R－CIK组：提前通过RetroNectin包被诱导；R＋C－CIK组：提前通过RetroNectin及CD3 Ab包被诱导，记录各组免疫效应细胞不同时间段的增殖情况。流式细胞术检测各组细胞不同时间段的免疫表型变化；并用LDH法检测各组细胞对肿瘤细胞K562、LOVO杀伤活性。结果 R＋C－CIK组细胞增殖倍数高于其他三组，差异有统计学意义（ P＜0．05）；R＋C－CIK组及R－CIK组的CD3＋CD56＋双阳性细胞所占百分比较C－CIK及CIK组高，d12和d15时有统计学差异（P＜0．05）；在效靶比20∶1和40∶1时，R＋C－CIK及R－CIK组对K562的杀伤活性高于C－CIK及CIK组，杀伤活性有统计学差异（P＜0．05），R＋C－CIK组、R－CIK组及C－CIK组细胞对结肠癌细胞系LOVO的杀伤活性高于CIK 组，有统计学差异（ P＜0．05）。结论 RetroNectin联合抗CD3MAb包被技术应用于CIK体外培养可以获得增殖率和活性更高的免疫活性细胞，是值得临床推荐的一种有效的培养方法。
목적：연구병탐토중조인섬유련접단백（ RetroNectin ， RN ）대세포인자유도적살상세포（ CIK）증식、면역특성급대인류K562、LOVO세포살상활성적영향。방법제취환자외주혈단개핵세포，표본분위사조，대조조（ CIK조）：안상규방법유도CIK증식；C－CIK조：제전통과CD3 Ab포피유도；R－CIK조：제전통과RetroNectin포피유도；R＋C－CIK조：제전통과RetroNectin급CD3 Ab포피유도，기록각조면역효응세포불동시간단적증식정황。류식세포술검측각조세포불동시간단적면역표형변화；병용LDH법검측각조세포대종류세포K562、LOVO살상활성。결과 R＋C－CIK조세포증식배수고우기타삼조，차이유통계학의의（ P＜0．05）；R＋C－CIK조급R－CIK조적CD3＋CD56＋쌍양성세포소점백분비교C－CIK급CIK조고，d12화d15시유통계학차이（P＜0．05）；재효파비20∶1화40∶1시，R＋C－CIK급R－CIK조대K562적살상활성고우C－CIK급CIK조，살상활성유통계학차이（P＜0．05），R＋C－CIK조、R－CIK조급C－CIK조세포대결장암세포계LOVO적살상활성고우CIK 조，유통계학차이（ P＜0．05）。결론 RetroNectin연합항CD3MAb포피기술응용우CIK체외배양가이획득증식솔화활성경고적면역활성세포，시치득림상추천적일충유효적배양방법。
Objective To investigate the recombinant human fibronectin (RetroNectin,RN)on prolifera-tion,immunological characteristics and cytotoxicity of cytokine -induced killer cells ( CIK) .Methods Peripheral blood mononuclear cells(PBMCs)were cultured in vitro by precoating with RetroNectin (R-CIK group),CD3Ab (C-CIK group),RetroNectin combined with CD3Ab(R+C-CIK group)and traditional method(CIK group) were to generate CIK.The changes of growth rate,characterization,cytotoxicity and apoptosis of CIK were deter-mined by cell counting ,LDH assay and flow cytometry respectively .Results R+C-CIK cell showed a higher proliferation rate than other three groups .The difference was statistically significant (P<0.05);The percentage of CD3/CD56 double positive cells in R +C-CIK and R-CIK group had a higher proportion than those in C -CIK and CIK group at day12 and day15(P<0.05);Cytotoxicity of CIK towards K562 cells in R+C-CIK group and R-CIK group were significantly higher than that in C -CIK and CIK group ,when effector-to-target ratio was 20∶1 and 40∶1(P<0.05).We also found that the cytotoxicity of CIK towards LOVO cells in R +C-CIK group and R-CIK group were higher than that in C -CIK group and CIK group(P<0.05).Conclusion RetroNectin and Anti-CD3-MAb synergistically promote antitumor efficiency of CIKs by increasing proliferation and cyto-toxicity.