程序性死亡配体1单克隆抗体增强DC疫苗致敏的B细胞靶向肿瘤干细胞的体液免疫  

Programmed death-ligand 1 monoclonal antibody enhances humoral immunity elicited by sensitized B cells with DC vaccine targeting CSCs

作　　者：胡阳阳 汪毅 HU Yang-yang;WANG Yi(Department of Rheumatology and Immunology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China;Department of Oncology,The Third Affiliated Hospital of Anhui Medical University,Anhui Hefei 230022,China)

机构地区：[1]华中科技大学同济医学院附属同济医院风湿免疫内科,湖北武汉430030 [2]安徽医科大学第三附属医院肿瘤科,安徽合肥230022

出　　处：《内科急危重症杂志》2024年第2期155-159,192,共6页Journal of Critical Care In Internal Medicine

基　　金：国家自然科学基金(81703058)。

摘　　要：目的:探讨程序性死亡配体1(PD-L1)单克隆抗体是否可以增强ALDH high CSC-DC疫苗致敏的B细胞靶向ALDH high肿瘤干细胞(CSCs)的体液免疫作用。方法:建立B16-F10黑色素瘤小鼠模型,各组小鼠分别接受PBS、ALDH high CSC-DC+IgG、ALDH high CSC-DC疫苗、PD-L1单克隆抗体、ALDH high CSC-DC联合PD-L1单克隆抗体的治疗,记录小鼠的生存时间及肿瘤的体积。实验结束时收集各组小鼠的肿瘤,单个肿瘤细胞悬液进行ALDEFLUOR染色检测CSCs的比例。流式细胞术检测各组小鼠脾脏B细胞上PD-1的表达量。同时进一步行抗体结合试验和补体依赖的细胞毒性作用(CDC)试验检测B细胞培养上清中的抗体结合和裂解CSCs的能力。结果:相较于单独治疗组,PD-L1单克隆抗体与ALDH high CSC-DC疫苗的联合治疗可以更加显著地抑制肿瘤生长,延长小鼠生存时间。联合治疗组小鼠活化的B淋巴细胞上PD-1的表达水平显著降低,仅为6.5%。抗体结合试验提示,与ALDH high CSC-DC疫苗单独治疗组11.3%的结合率相比,联合治疗组小鼠B细胞培养上清中的抗体可以特异地结合15.7%ALDH high CSCs。同时CDC试验结果显示,联合治疗组的B细胞培养上清特异性地裂解ALDH high CSCs。结论:PD-L1单克隆抗体可以显著增强ALDH high CSC-DC疫苗致敏的B细胞产生靶向ALDH high CSCs的体液免疫反应。Objective:To explore whether the programmed death-ligand 1(PD-L1)monoclonal antibody can enhance humoral immunity elicited by sensitized B cells with DC vaccine targeting CSCs.Methods:A B16-F10 melanoma mouse model was established.Mice were treated with PBS,ALDH high CSC-DC vaccine with IgG,ALDH high CSC-DC vaccine,PD-L1 monoclonal antibody,or ALDH high CSC-DC vaccine plus PD-L1 monoclonal antibody,respectively.The survival time and tumor volume of the mice were recorded.At the end of the experiment,tumors from each group of mice were collected,and a single tumor cell suspension was subjected to ALDEFLUOR staining to detect the proportion of CSCs.Flow cytometry was used to detect the expression of PD-1 on B cells in the spleens of mice from each group.Additionally,antibody binding assays and complement-dependent cytotoxicity(CDC)assays were conducted to assess the ability of antibodies in the cultured supernatant of B cells to bind and lyse ALDH high CSCs.Results:Compared to the solo treatment group,the combined treatment of PD-L1 monoclonal antibody with ALDH high CSC-DC vaccine significantly inhibited tumor growth and prolonged the survival time of mice.The expression level of PD-1 in activated B lymphocytes in the combined treatment group was significantly decreased to 6.5%.Antibody binding assays suggested that antibodies in the cultured supernatant of B cells from the combined treatment group could bind more specifically to ALDH high CSCs(15.7%)compared to the ALDH high CSC-DC vaccine solo treatment(11.3%).Furthermore,CDC assay results showed that the B cell cultured supernatant from the combined treatment group specifically lysed ALDH high CSCs.Conclusion:The PD-L1 monoclonal antibody can significantly enhance the humoral immune response elicited by sensitized B cells with DC vaccine targeting ALDH high CSCs.

关 键 词：黑色素瘤 肿瘤干细胞 B细胞 树突状细胞 程序性死亡配体1 

分 类 号：R392[医药卫生—免疫学]