摘要
目的在体外诱导形成三级淋巴器官(tertiary lymphoid organs,TLO),并评价其在抗肿瘤免疫中的功能。方法利用慢病毒系统在NIH3T3细胞上过表达淋巴毒素-β受体(lymphotoxin-beta receptor,LTβR),并检测LTβRNIH3T3细胞中LTβR的过表达效率;通过免疫印迹实验和qPCR探究过表达LTβR的NIH3T3细胞内非经典核因子(nuclear factor,NF)-κB信号通路的情况。构建B16-OVA黑色素瘤小鼠荷瘤模型,探究LTβR-NIH3T3细胞诱导TLO形成的能力及抗肿瘤效果。结果利用慢病毒感染在NIH3T3细胞中过表达LTβR,流式检测发现GFP+细胞比例达99%。过表达LTβR能在NIH3T3细胞内激活非经典NF-κB信号通路。小鼠肿瘤模型结果表明,注射LTβR-NIH3T3细胞能在肿瘤附近诱导形成淋巴样组织,并促进了T细胞和MHCⅡ+巨噬细胞的浸润,明显抑制荷瘤小鼠的肿瘤生长,并延长小鼠生存期。结论LTβRNIH3T3细胞通过诱导TLO的形成,促进抗肿瘤免疫,为肿瘤免疫治疗提供新的思路。
Objective To induce the development of tertiary lymphoid organs(TLO)in a mouse model of melanoma and to evaluate TLO’s functions in antitumor immunity.Methods Lymphotoxin-beta receptor(LTβR)was overexpressed in NIH3T3 cells through the lentivirus system and the overexpression efficiency of LTβR in LTβR-NIH3T3 cells was examined.Western blot and qPCR were used to examine the non-canonical nuclear factor(NF)-κB signaling pathway in NIH3T3 cells overexpressing LTβR.B16-OVA melanoma mouse model was constructed to explore the induction of TLO and anti-tumor functions of TLO in LTβR-NIH3T3 cells.Results LTβR was overexpressed in NIH3T3 cells through the lentivirus system,and flow cytometry showed that the proportion of GFP+cells reached 99%.The overexpression of LTβR activated the non-canonical NF-κB signaling pathway in NIH3T3 cells.Findings from the mouse tumor model suggest that the injection of LTβR-NIH3T3 cells successfully induced the development of lymphoid tissue around the tumor and enhanced the tumor infiltration of T cells and MHCⅡ+macrophages,significantly inhibiting tumor growth and prolonging the survival of tumor-bearing mice.Conclusion LTβR-NIH3T3 cells promoted antitumor immunity by inducing TLO development,which may provide new perspectives for tumor immunotherapy.
作者
陈红
胡翔
张惠媛
胡洪波
CHEN Hong;HU Xiang;ZHANG Hui-yuan;HU Hong-bo(State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu 610041,China;Center for Immunology and Hematology Research,West China Hospital,Sichuan University,Chengdu 610041,China)
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2022年第1期35-42,共8页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.82025002、No.81871232)
科技部重点研发专项(No.2019YFA0110200)资助。
