【摘要】 目的探究溶瘤单纯疱疹病毒（oHSV）和CD19 CAR-T细胞联合治疗弥漫大B细胞淋巴瘤的疗效以及CD19 CAR-T细胞是否为oHSV的良好载体。方法将oHSV与CD19 CAR-T细胞共培养,制备携带oHSV的CD19 CAR-T细胞(CD19 CAR-T^oHSV细胞)。通过细胞增殖实验、细胞杀伤实验以及重要指标如细胞因子、细胞亚群、耗竭表型的测定,评估体外CD19CAR-T^oHSV细胞相较于CD19 CAR-T细胞的抗肿瘤效果和功能。构建弥漫大B肿瘤细胞系小鼠模型,比较未转导T细胞组、CD19 CAR-T细胞组和CD19 CAR-T^oHSV细胞组小鼠瘤体变化和重要脏器指标探究CD19 CAR-T^oHSV细胞在体内的安全性和有效性。结果成功制备出CD19 CAR-T^oHSV细胞。体外实验发现,与CD19 CAR-T细胞相比,CD19 CAR-T^oHSV细胞表现出更强的抗肿瘤效果(t_24h=2.989 P<0.05;t_48h=8.525,P<0.01)。联合治疗促进了干扰素（IFN）-γ的分泌（t=4.578,P<0.05）,并降低CD19 CAR-T细胞表面PD-1的表达（t=2.946,P<0.05）。在小鼠模型中,CD19CAR-T^oHSV只感染肿瘤,不影响其他脏器功能显示出安全性。同时,CART^oHSV细胞组瘤体质量显著小于CD19 CAR-T细胞组（t=6.010,P<0.01）。免疫组化也发现,CD19 CAR-T细胞和oHSV的联合治疗促进了CAR-T细胞向肿瘤的浸润（t=12.68,P<0.001）。结论:CD19 CAR-T细胞可以作为oHSV的良好载体。CD19 CAR-T^oHSV细胞治疗可安全且有效提升弥漫大B细胞淋巴瘤治疗效果。更多还原

【Abstract】 Objective:To explore the efficacy of combined treatment with oncolytic herpes simplex virus（oHSV） and CD 19 CAR-T cells in diffuse large B-cell lymphoma（DLBCL）,and determine whether CD19 CAR-T cells can serve as a good carrier for oHSV.Methods:CD19 CAR-T cells were co-cultured with oHSV to prepare CD19 CAR-T cells carrying oHSV(CD19 CAR-T^oHSV cells).The anti-tumor effect and function of CD19 CAR-T^oHSV cells were evaluated in vitro compared to CD19 CAR-T cells through cell proliferation assays,cytotoxicity assays,and the determination of important indicators such as cytokines,cell subsets,and exhaustion phenotypes.A mouse model of DLBCL was established to compare the tumor growth and important organ function in mice treated with non-transduced T cells,CD 19 CAR-T cells,and CD 19 CAR-T^oHSV cells, and to investigate the safety and efficacy of CD 19 CAR-T^oHSV cells in vivo.Results:CD 19 CAR-T^oHSV cells were successfully prepared.In vitro experiments showed that CD 19CAR-T^oHSV cells exhibited stronger anti-tumor effects compared with CD19 CAR-T cells(t_24h=2.989,P<0.05;t_48h=8.525,P<0.01).The combined treatment promoted the secretion of interferon（IFN）-γ（t=4.578,P<0.05） and reduced the expression of PD-1 on the surface of CD19 CAR-T cells（t=2.946,P<0.05）.In the mouse model,CD19 CAR-T^oHSV cells showed,safety by only infecting tumors but not affecting the function of other organs,demonstrating safety.Additionally,the tumor weight in the CAR-T^oHSV cells group was significantly lower than that in the CD19 CAR-T cells group（t=6.010,P<0.01）.Immunohistochemistry also revealed that the combined treatment of CD 19 CAR-T cells and oHSV promoted the infiltration of CAR-T cells into tumors（t=12.68,P<0.001）.Conclusion:CD19 CAR-T cells can serve as a good carrier for oHSV.The treatment with CD19 CAR-T^oHSV cells can safely and effectively enhance the therapeutic effect on diffuse large B-cell lymphoma.更多还原