[1]郭金东),李铁鹏),徐本玲),等.不同培养液对肾癌患者外周血T淋巴细胞体外扩增及CXC趋化因子受体3表达的影响[J].郑州大学学报(医学版),2020,(06):775-779.[doi:10.13705/j.issn.1671-6825.2019.12.190]
　GUO Jindong),LI Tiepeng),XU Benling),et al.Effects of different culture mediums on the ex vivo expansion of peripheral blood T lymphocytes from patients with renal cell carcinoma and the expression of CXCR3[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2020,(06):775-779.[doi:10.13705/j.issn.1671-6825.2019.12.190]
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不同培养液对肾癌患者外周血T淋巴细胞体外扩增及CXC趋化因子受体3表达的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2020年06期
页码:
775-779
栏目:
论著
出版日期:
2020-11-20

文章信息/Info

Title:
Effects of different culture mediums on the ex vivo expansion of peripheral blood T lymphocytes from patients with renal cell carcinoma and the expression of CXCR3
作者:
郭金东1); 李铁鹏1); 徐本玲1); 张成娟2); 杜雪相1); 王 瑶1); 高全立1)
1)郑州大学附属肿瘤医院(河南省肿瘤医院)免疫治疗科 郑州 450008 2)郑州大学附属肿瘤医院(河南省肿瘤医院)生物样本中心 郑州 450008
Author(s):
GUO Jindong1); LI Tiepeng1); XU Benling1); ZHANG Chengjuan2); DU Xuexiang1); WANG Yao1); GAO Quanli1)
1)Department of Immunotherapy,the Affiliated Cancer Hospital,Zhengzhou University(Henan Cancer Hospital),Zhengzhou 450008 2)Biological Sample Center,the Affiliated Cancer Hospital,Zhengzhou University(Henan Cancer Hospital),Zhengzhou 450008
关键词:
肾癌;  免疫细胞治疗;  T淋巴细胞;  CXCR3;  趋化因子
Keywords:
kidney cancer;  immune cell therapy;  T lymphocyte;  CXCR3;  chemokine
分类号:
R737.11
DOI:
10.13705/j.issn.1671-6825.2019.12.190
摘要:
目的:探讨不同培养液对肾癌患者外周血T淋巴细胞体外扩增及CXC趋化因子受体3(CXCR3)表达的影响。方法:密度梯度离心法分离肾癌患者PBMC,分为4组,分别用4种培养液(GT-T551、GT-T551 H3、ALyS505N、X-VIVOTM15)培养,应用CD3单克隆抗体、重组人纤维连接蛋白、白细胞介素2、干扰素γ诱导、活化PBMC。显微镜下观察细胞形态并进行细胞计数,采用流式细胞术检测T淋巴细胞亚群比例及膜表面CXCR3的表达情况。结果:4组细胞培养至第13天状态良好,第16天开始逐渐老化,组间无明显差异。4组细胞增殖倍数均于第16天达到峰值; 培养第22、25天,GT-T551 H3组细胞增殖倍数低于ALyS505N组和X-VIVOTM15组(P<0.05)。CD3+、CD3+CD8+及CD3+CD56+T淋巴细胞比例随培养时间的延长逐渐提高,CD3+CD4+T淋巴细胞的比例逐渐下降; GT-T551 H3组的CD3+T淋巴细胞比例在培养第15天低于其他3组(P=0.042),而CD3+CD56+T淋巴细胞比例在培养第11、13、15天高于其他3组(P<0.05)。与活化前相比,各组T淋巴细胞亚群中CXCR3阳性细胞比例均有显著提高,在第13天后降低,呈先高后低的变化趋势(P<0.05)。结论:4种培养液对肾癌患者PBMC形态和最高增殖倍数的影响差异无统计学意义; GT-T551 H3培养液更适用于NKT淋巴细胞的培养; 4种培养液均可以提升T淋巴细胞亚群表面CXCR3的表达水平,13 d为较好的培养周期。
Abstract:
Aim:To investigate the impact of different culture mediums on ex vivo expansion of peripheral blood monocytes(PBMC)from patients with renal cell cancer(RCC)and the expression of CXCR3.Methods:Density gradient centrifugation was used to isolate PBMC from patients with RCC.Anti-human CD3,RetroNectin,IL-2,and IFN-γ were used to activate and expand PBMC.Four culture mediums(GT-T551,GT-T551 H3,ALyS505N,and X-VIVOTM15)were used to the culture of PBMC.Cell morphology was observed regularly with a microscope.Cell number was counted after trypan blue staining.Flow cytometry was performed to determine the proportion of T lymphocyte subsets and the expression of CXCR3.Results:The cells were in good condition until day 13 after expansion with 4 different culture mediums,and gradually began to age from day 16,and there were no significant differences between 4 groups.After ex vivo expansion,cell number peaked on day 16,and the foldchange index of GT-T551 H3 group was lower than those of ALyS505N group and X-VIVOTM15 group on day 22 and 25(P<0.05).The proportion of CD3+ T cells,CD3+CD8+ T cells,and CD3+CD56+ T cells gradually increased during expansion,and the proportion of CD3+CD4+ T cells showed an opposite trend.CD3+ T cells in the GT-T551 H3 group were significantly lower than the other 3 groups on day 15(P=0.042)while CD3+CD56+ T cells were significantly higher than the other 3 groups on days 11,13,and 15(P<0.05).Compared with PBMC,the proportion of CXCR3+ cells in T lymphocyte subgroups in each group was significantly increased,which first increased and reached the peak on day 13,and then decreased(P<0.05).Conclusion:There is no significant difference in cell morphology or maximum foldchange during ex vivo expansion of PBMC from patients with RCC using 4 different mediums.GT-T551 H3 is more suitable for the expansion of NKT cells.All the 4 mediums can significantly increase the expression of CXCR3 on the surface of each T lymphocyte subgroup,and a culture period of 13 days is recommended.

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备注/Memo

备注/Memo:
【基金项目】国家自然科学基金项目(81902902,81502468); 河南省科技厅科技攻关项目(182102310387,162300410095)
【作者简介】高全立,通信作者,男,1969年4月生,博士,主任医师,研究方向:肿瘤免疫治疗,E-mail:gaoquanli2015@126.com
更新日期/Last Update: 2020-11-20