【作者】 李婕；

【导师】 奚玲；

【作者基本信息】 华中科技大学 ， 妇产科学（专业学位）， 2023， 硕士

【摘要】 目的:本研究旨于阐明T细胞免疫球蛋白粘蛋白结构域蛋白3(TIM-3)在上皮性卵巢癌(EOC)中的表达部位、程度及作用,验证TIM-3抑制剂治疗EOC的疗效。方法:利用免疫组织化学、多色免疫荧光以及流式细胞术探索TIM-3在EOC中的表达;利用流式分选技术与流式细胞术探索EOC患者腹水和肿瘤中表达TIM-3和(或)程序性死亡受体1(PD-1)的CD8+T细胞和树突状细胞(DC)的功能;利用流式分选技术与流式细胞术探索TIM-3抑制剂对EOC患者肿瘤浸润CD8+T细胞和DC的免疫调节作用;建立C57BL/6J小鼠的ID8VEGF卵巢癌皮下瘤模型,探索TIM-3抑制剂治疗上皮性卵巢癌荷瘤小鼠的疗效。结果:TIM-3与PD-1高表达且共表达于EOC患者的肿瘤和腹水中CD8+T细胞和DC上。高表达TIM-3患者肿瘤和腹水中TIM-3highPD-1+CD8+T细胞与低表达TIM-3患者肿瘤和腹水中TIM-3intPD-1+CD8+T细胞均增殖活跃且分泌功能衰竭;高表达TIM-3患者和低表达TIM-3患者肿瘤和腹水中TIM-3+PD-1+DC均增殖活跃、低表达共刺激分子且分泌功能衰竭。TIM-3抑制剂单药治疗或与PD-1抑制剂联合治疗可以改善EOC患者肿瘤浸润CD8+T细胞和DC的功能,并且可以延缓上皮性卵巢癌荷瘤小鼠的肿瘤生长速度、延长其生存期以及改善荷瘤小鼠免疫微环境中CD8+T细胞和DC的功能。结论:TIM-3与PD-1高表达且共表达于EOC患者的肿瘤和腹水中CD8+T细胞和DC上,且其表达水平与CD8+T细胞和DC的功能衰竭程度相关。TIM-3靶向治疗可改善EOC患者肿瘤浸润CD8+T细胞和DC功能,延缓上皮性卵巢癌荷瘤小鼠肿瘤生长速度以及延长其生存期,有望成为EOC免疫治疗的关键靶点。更多还原

【Abstract】 Objective:This study aims to clarify the expression site,extent,and role of T cell immunoglobulin mucin domain protein 3(TIM-3)expression in epithelial ovarian cancer(EOC),and to verify the efficacy of TIM-3 inhibitors in the treatment of EOC.Methods:Immunohistochemistry,multicolor immunofluorescence,and flow cytometry were used to explore the expression of TIM-3 in EOC;flow sorting cytometry and flow cytometry were used to explore the function of CD8+T cells and dendritic cells(DC)expressing TIM-3 and/or programmed death receptor 1(PD-1)in ascites and tumors of EOC patients;flow sorting cytometry and flow cytometry were used to explore the immunomodulatory effect of TIM-3 inhibitors on tumor-infiltrating CD8+T cells and DC from EOC patients;subcutaneous ID8VEGF ovarian tumor model in C57BL/6J mice was established to explore the efficacy of TIM-3 inhibitors in the treatment of ovarian tumorbearing mice.Results:TIM-3 and PD-1 were highly expressed and co-expressed on CD8+T cells and DC in tumors and ascites from EOC patients.TIM-3highPD-1+CD8+T cells in tumors and ascites of high TIM-3 expressers and TIM-3intPD-1+CD8+T cells in tumors and ascites of low TIM3 expressers were both active in proliferation and failure of secretory function;TIM-3+PD1+DC in tumors and ascites of high TIM-3 expressers and low TIM-3 expressers were active in proliferation,low expression of co-stimulatory molecules and failure of secretory function.TIM-3 inhibitors monotherapy or in combination with PD-1 inhibitors could improve the function of tumor-infiltrating CD8+T cells and DC of EOC patients,delay the tumor volume growth,and prolong the survival time of ovarian tumor-bearing mice and improve the function of CD8+T cells and DC in the immune microenvironment of ovarian tumor-bearing mice.Conclusions:TIM-3 and PD-1 are highly expressed and co-expressed on CD8+T cells and DC in tumors and ascites from EOC patients,and their expression levels indicate the degree of exhaustion of CD8+T cells and DC.TIM-3 targeted therapy could improve the functions of tumor-infiltrating CD8+T cells and DC of EOC patients,delay the tumor growth rate and prolong the survival of ovarian tumor-bearing mice,which is expected to be the key target for EOC immunotherapy.更多还原