溶瘤病毒诱导下过继细胞疗法用于骨肿瘤治疗可行性研究

Feasibility study of adoptive cell therapy induced by oncolytic virus for the treatment of bone tumors

目的探讨溶瘤病毒诱导下过继细胞疗法用于骨肿瘤治疗可行性。方法选取45只成年健康雄性大鼠建立骨肿瘤模型。随机分组后分别给予生理盐水(对照组)及溶瘤病毒诱导下过继细胞(VVDDCCL5、VVDD-CxC11、VVDD-IL15Rα、VVDD-IL2)干预,选取抗肿瘤免疫反应最优一组作为研究对象。比较对照组与VVDD干预组大鼠生存期,比较不同组别CD3^(+)、CD4^(+)、CD8^(+)表达及CTLA-4、TIM-3、PD-1^(high)TIM-3^(+)、Foxp3^(+)CD4^(+)表达,初始T细胞、TCM、TEM、4-1BB表达。进行体外实验,检测VVDD干预下不同细胞IFN-γ及4-1BB表达。结果干预10 d后可见VVDD-IL 2治疗的肿瘤具有较高的肿瘤反应性CD8^(+)TILs频率,VVDDIL 2组大鼠IFN-γ阳性斑点表达高于其他组别(P<0.05);VVDD-IL 2组CD3^(+)、CD4^(+)、CD8^(+)细胞表达及生存时间高于对照组(P<0.05);VVDD-IL 2组CTLA-4、TIM-3表达高于对照组,PD-1^(high)TIM-3^(+)、Foxp3^(+)CD4^(+)表达低于对照组(P<0.05);VVDD-IL 2组TCM、TEM、4-1BB表达高于对照组,初始T细胞表达低于对照组(P<0.05);体外实验结果显示,肿瘤细胞中VVDD-IL 2干预后IFN-γ、4-1BB表达高于未干预(P<0.05),正常细胞中两组差异无统计学意义(P>0.05)。结论溶瘤病毒诱导下过继细胞疗法用于骨肿瘤治疗可延长大鼠生存时间,IL2武装溶瘤病毒可促进T细胞浸润,并提高肿瘤反应性TILS的数量。促进肿瘤反应性TILS在低或低免疫原性肿瘤中的生成和浸润,并扩大这种TILS用于细胞过继治疗可能是骨肿瘤治疗的新策略。

ObjectiveeTo explore the feasibility of oncolytic virus-induced adoptive cell therapy for the treatment of bone tumors.Methods 45 adult healthy male rats were selected to establish bone tumor model.After randomization,normal saline(control group)and oncolytic virus-induced adoptive cells(VVDD-CCL5,VVDDCxC11,VVDD-IL15Rα,VVDD-IL2)were respectively administered,and the group with the best anti-tumor immune response was selected as the study object.The survival period of rats in control group and VVDD intervention group was compared,and the expressions of CD3^(+),CD4^(+),CD8^(+),CTLA-4,TIM-3,PD-1^(high)TIM-3^(+),Foxp3^(+)CD4,initial T cells,TCM,TEM,4-1BB were compared in different groups.The expression of IFN-and 4-1BB in different cells under VVDD intervention was detected in vitro.Results After 10 days of intervention,VVDD-IL-2 treated tumors showed a higher frequency of tumor reactivity CD8 TILs,and the expression of IFN-positive spots in VVDD-IL-2 group was higher than that in other groups(P<0.05).The expression and survival time of CD3^(+),CD4^(+)and CD8^(+)cells in VVDD-IL 2 group were higher than those in control group(P<0.05).The expressions of CTLA-4 and TIM-3 in VVDD-IL 2 group were higher than those in control group,while the expressions of PD-1^(high)TIM-3t and Foxp3^(+)CD4^(+)in VVDD-IL 2 group were lower than those in control group(P<0.05).The expression of TCM,TEM and 4-1BB in VVDD-IL 2 group was higher than that in control group,and the initial T cell expression was lower than that in control group(P<0.05).The results of in vitro experiment showed that the expressions of IFN-and 4-1BB in tumor cells after VVDD-IL 2 intervention were higher than those without intervention(P<0.05),and there was no statistical significance between the two groups in normal cells(P>0.05).Conclusion Adoptive cell therapy induced by oncolytic virus can prolong the survival time of rats in the treatment of bone tumor.Il2-armed oncolytic virus can promote T cell invasion and increase the number of tumor-reactive TILS.Promoting the formation and infiltration of tumor-reactive TILS in low or low immunogenic tumors and expanding such TILS for cellular adoptive therapy may be a new strategy for bone tumor treatment.