PD-1/ICOS信号转换受体通过增强CD19-CAR-T细胞增殖能力提高其对DLBCL的杀伤活性  

PD-1/ICOS signal conversion receptor enhances the proliferation of CD19-CAR-T cells

作　　者：加尔宝·吐尔德 阿迪娜·贾库林 闻淑娟[1] JIAERBAO Tuerde;ADINA Jiakulin;WEN Shujuan(Department of Lymphoma Medicine,The Affiliated Cancer Hospital of Xinjiang Medical University,Urumqi 830011,China;Department of Breast Radiotherapy,The Cancer Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院淋巴瘤内科,新疆乌鲁木齐830011 [2]新疆医科大学附属肿瘤医院乳腺放疗科,新疆乌鲁木齐830011

出　　处：《西部医学》2022年第10期1426-1431,共6页Medical Journal of West China

基　　金：新疆维吾尔自治区自然科学基金(2020D01C268)。

摘　　要：目的探讨共表达PD-1/ICOS信号转换受体通过增强靶向CD19的嵌合抗原受体T细胞(CD19-CAR-T细胞)增殖能力提高其对弥漫大B细胞淋巴瘤(DLBCL)产生的抗肿瘤活性。方法慢病毒感染法构建CD19-CAR-T细胞(bbz)以及共表达PD-1/ICOS信号转换受体的CD19-CAR-T细胞(PD-1/ICOS-bbz);流式细胞术检测长期共孵育后T细胞CD69的表达;通过细胞计数检测CAR-T细胞的增殖情况;LDH法检测CAR-T细胞对靶细胞的细胞毒性;活体成像检测小鼠体内肿瘤细胞的生长情况;流式细胞术检测外周血中T细胞的比例。结果成功构建bbz及PD-1/ICOS-bbz;长期共孵育后,PD-1/ICOS-bbz较bbz CD69表达水平更高,增殖能力及对靶细胞的细胞毒性均显著提高(P<0.01);PD-1/ICOS-bbz可以完全清除小鼠体内的淋巴瘤细胞,且PD-1/ICOS-bbz较bbz而言可以显著延长小鼠生存期(P<0.01),同时PD-1/ICOS-bbz较bbz具有更强的体内增殖能力(P<0.05)。结论PD-1/ICOS-bbz较传统的二代CAR-T细胞具有更强的抗肿瘤活性,有望成为一种潜在的弥漫大B细胞淋巴瘤的治疗方案。Objective To investigate whether CD19-CAR T cells co-expressing PD-1/ICOS signal transforming receptor can induce stronger antitumor activity against diffuse large B-cell lymphoma.Methods CD19-CAR T cells(bbz)and CD19-CAR T cells co-expressing PD-1/ICOS signal transforming receptor(PD-1/ICOS-bbz)were constructed by lentivirus infection.The expression of CD69 on T cells after long-term co-incubation was detected by flow cytometry.The proliferation of CAR T cells was detected by cell count.LDH assay was used to detect the cytotoxicity of CAR T cells against target cells.In vivo imaging was used to detect the growth of tumor cells in mice.The proportion of T cells in peripheral blood was detected by flow cytometry.Results bbz and PD-1/ICOS-bbz were constructed successfully.After long-term co-incubation,CD69 expression level of PD-1/ICOS-bbz was higher than that of bbz(P<0.001),and proliferation ability(P<0.01)and cytotoxicity to target cells(P<0.01)were also significantly improved.PD-1/ICOS-bbz can completely eliminate lymphoma cells in mice,and PD-1/ICOS-bbz can significantly prolong the survival period of mice compared with bbz(P<0.01).PD-1/ICOS-bbz has a stronger in vivo proliferation ability than bbz.Conclusion PD-1/ICOS-bbz has stronger antitumor activity than conventional second-generation CAR T cells,and may be a promising treatment option for potential diffuse large B-cell lymphoma.

关 键 词：弥漫大B细胞淋巴瘤 嵌合抗原受体T细胞 PD-1 ICOS 免疫治疗 

分 类 号：R733[医药卫生—肿瘤]