CAJ | 학술논문

目的：探讨4-1BBL基因转染对Raji细胞功能及利妥昔单抗活性的影响。方法采用脂质体法将4-1BBL基因转染Raji细胞，实验分为转染组（4-1BBL组）、转染空质粒组（ Mock组）及未转染组（对照组）。 Western blotting鉴定转染48h后各组4-1BBL蛋白水平，采用CCK-8法检测不同浓度（20?0、10?0、5?0、2?5、1?25、0?625mg/ml）利妥昔单抗处理48h后Raji细胞的增殖率，分别采用CCK-8及ELISA法检测与不同效/靶比（5∶1和10∶1）活化外周血淋巴细胞共培养48h各组淋巴细胞增殖率及培养上清液的白介素（ IL）-2浓度。结果4-1BBL组中4-1BBL蛋白水平均高于对照组和Mock组（ P＜0?05），4-1BBL组Raji细胞增殖率随利妥昔单抗浓度升高而降低，且当浓度＞2?5mg/ml，增殖率均低于对照组和Mock组，差异有统计学意义（ P＜0?05）。在5∶1和10∶1两种效/靶比下，4-1BBL组淋巴细胞的增殖率及上清液IL-2水平均高于对照组和Mock组，差异有统计学意义（ P＜0?05）。对照组和Mock组中上述指标的差异均无统计学意义（ P＞0?05）。结论4-1BBL基因转染Raji细胞可增强利妥昔单抗活性，提示细胞免疫治疗结合抗体靶向治疗将成为肿瘤免疫治疗的重要策略。
목적：탐토4-1BBL기인전염대Raji세포공능급리타석단항활성적영향。방법채용지질체법장4-1BBL기인전염Raji세포，실험분위전염조（4-1BBL조）、전염공질립조（ Mock조）급미전염조（대조조）。 Western blotting감정전염48h후각조4-1BBL단백수평，채용CCK-8법검측불동농도（20?0、10?0、5?0、2?5、1?25、0?625mg/ml）리타석단항처리48h후Raji세포적증식솔，분별채용CCK-8급ELISA법검측여불동효/파비（5∶1화10∶1）활화외주혈림파세포공배양48h각조림파세포증식솔급배양상청액적백개소（ IL）-2농도。결과4-1BBL조중4-1BBL단백수평균고우대조조화Mock조（ P＜0?05），4-1BBL조Raji세포증식솔수리타석단항농도승고이강저，차당농도＞2?5mg/ml，증식솔균저우대조조화Mock조，차이유통계학의의（ P＜0?05）。재5∶1화10∶1량충효/파비하，4-1BBL조림파세포적증식솔급상청액IL-2수평균고우대조조화Mock조，차이유통계학의의（ P＜0?05）。대조조화Mock조중상술지표적차이균무통계학의의（ P＞0?05）。결론4-1BBL기인전염Raji세포가증강리타석단항활성，제시세포면역치료결합항체파향치료장성위종류면역치료적중요책략。
Objective To explore the influence on function and activity of rituximab of Raji cells via 4-1BBL gene transfec-tion. Methods Stable expression of 4-1BBL in Raji cells was carried out through transfection mediated by liposome. The experiments contained 4-1BBL group, Mock group and control group. The 4-1BBL expression was identified by Western blotting analysis at 48h after transfection. CCK-8 method was used to detect the proliferation at 48h after treatment with different concentrations (20?0, 10?0, 5?0, 2?5, 1?25, 0?625 mg/ml) of rituximab. The proliferation rates of lymphocyte and supernatant IL-2 levels at 48h after co-culture with the activated peripheral blood lymphocytes under the effector/target ratio of 5∶1 and 10∶1 by CCK-8 and enzyme-linked immunosorbent assay. Results The protein level of 4-1BBL in 4-1BBL group was higher than those in Mock group and control group ( P<0?05) . The proliferative rates decreased with the increasing concentrations of rituximab in 4-1BBL group. When the concentration was over 2?5mg/ml, there were lower proliferative rate in 4-1BBL group versus Mock group and control group ( P<0?05) . Under the effector/target ratio of 5∶1 and 10∶1, the proliferative rate of lymphocyte and supernatant IL-2 level in 4-1BBL group was higher than those in Mock group and control group ( P<0?05) . Conclusion The 4-1BBL gene transfection can enhance the activity of rituximab on Raji cells, sugges-ting that combined activation of 4-1BBL and targeting-antibody may be a new strategy for cancer immunotherapy.