人源化肿瘤-免疫微环境小鼠模型的建立  

Establishment of a humanized tumor-immune microenvironment mouse model

作　　者：陈海峰 丁一宗[2] 朱俊琳 陈倩[3] 傅于捷[2] 邓军 CHEN Haifeng;DING Yizong;ZHU Junlin;CHEN Qian;FU Yujie;DENG Jun(Shanghai Cancer Institute,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 20032,China;Department of Thoracic Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200126,China;Department of Ophthalmology,Tongren Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200336,China)

机构地区：[1]上海交通大学医学院附属仁济医院上海市肿瘤研究所,上海200032 [2]上海交通大学医学院附属仁济医院胸外科,上海200126 [3]上海交通大学医学院附属同仁医院眼科,上海200336

出　　处：《肿瘤》2024年第8期821-831,共11页Tumor

基　　金：国家自然科学基金(82101104、82071816、82271784);上海市科学技术委员会资助项目(21140903000)

摘　　要：目的:本研究旨在通过将人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)过继转移到NPSG(NOD-Prkdc^(scid)Il2rg^(null)/Shjh)小鼠,并移植人源肿瘤组织,构建h PBMC-NPSG-PDX(patient-derived xenograft)模型,以模拟人体肿瘤微环境,研究肿瘤与免疫系统的相互作用。方法:将健康志愿者外周血PBMC过继转移到NPSG小鼠,构建h PBMC-NPSG模型。将非小细胞肺癌患者的手术切除肿瘤组织移植到Nude(BALB/c-nu)小鼠,建立NudePDX模型。将传至第3代的PDX肿瘤组织移植到h PBMC-NPSG小鼠,构建h PBMC-NPSG-PDX模型。通过监测小鼠体质量、采用流式细胞术分析免疫系统重建、检测T细胞功能、评估肿瘤生长情况以及免疫组织化学法分析肿瘤组织形态学特征和免疫细胞浸润,综合评价模型的有效性。结果:h PBMC-NPSG小鼠构建28 d后,人源hu-CD45^(+)CD3^(+)T细胞比例达97%。人CD8^(+)T、CD4^(+)T、CD56^(+)自然杀伤和CD19^(+)B细胞的比例分别为64%、24%、4.6%和1.0%。CD4^(+)和CD8^(+)细胞分泌多种细胞因子(IL-2、IFN-γ和TNF-α)以及表达细胞毒性分子(Fas L、granzyme B和perforin)。在h PBMC-NPSG-PDX小鼠中,肿瘤生长初期迅速,随后趋于稳定。免疫组织化学染色结果显示,肿瘤形态学特征典型,在免疫细胞浸润的区域伴随有肿瘤细胞的凋亡。结论:本研究成功构建了h PBMC-NPSG-PDX模型,该模型可有效模拟人体肿瘤微环境,为研究肿瘤免疫学提供了理想的平台。Objective:This study aims to construct a human peripheral blood mononuclear cell(PBMC)-NPSG-PDX(patient-derived xenograft)model,by transferring PBMCs into NPSG(NOD-Prkdc^(scid)Il2rg^(null)/Shjh)mice and transplanting human tumor tissues.This model mimics the human tumor microenvironment to investigate the interactions between tumors and the immune system.Methods:PBMCs from healthy donors were transferred to NPSG mice to generate hPBMCNPSG model.Patient-derived xenografts(PDX)were established in nude(BALB/c-nu)mice.Third-generation PDX tumors were then transplanted into hPBMC-NPSG mice to establish the hPBMC-NPSG-PDX model.Mouse body weight was monitored,and flow cytometry was used to analyze immune reconstitution and T cell function.Tumor growth was evaluated,and immunohistochemistry was performed to analyze tumor morphology and immune cell infiltration.Results:Fourteen days after constructing the hPBMC-NPSG mouse model,the proportion of human hu-CD45^(+)CD3^(+)T cells reached 97%.The proportions of human CD8^(+)T,CD4^(+)T,CD56^(+)natural killer(NK),and CD19^(+)B cells were 64%,24%,4.6%,and 1.0%,respectively.Human CD4^(+)and CD8^(+)cells secreted various cytokines(IL-2,IFN-γ,and TNF-α)and expressed cytotoxic molecules(FasL,granzyme B,and perforin)28 days post reconstitution.Tumor growth in hPBMC-NPSG-PDX mice was rapid initially but then stabilized.Immunohistochemistry staining revealed typical tumor morphology,and tumor cell apoptosis was observed in areas with immune cell infiltration.Conclusion:This study successfully constructed a hPBMC-NPSG-PDX model that effectively simulates the human tumor microenvironment,providing an ideal platform for tumor immunology research.

关 键 词：人外周血单个核细胞 人源化小鼠 患者来源的异种移植瘤 肿瘤-免疫微环境 

分 类 号：R730.3[医药卫生—肿瘤]