双阴性T细胞促进Babl/C小鼠B16F10种植瘤生长的机制研究  

DNT promotes the growth of B16F10 melanoma and its possible mechanism

作　　者：蓝弘文 王于昌 徐利军[1] 周鸿敏[1] 李军[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院心胸外科,湖北武汉430030

出　　处：《现代肿瘤医学》2015年第20期2879-2883,共5页Journal of Modern Oncology

基　　金：国家自然基金资助项目(编号:30901364)

摘　　要：目的:探讨双阴性T细胞(DNT)促进B16F10黑色素瘤生长的可能机制。方法:以B16F10-luc-G5细胞系接种Babl/C小鼠建立种植瘤模型,IVIS监测生长动态。流式细胞仪检测外周血中T淋巴细胞的比例。流式分选正常小鼠的DNT及CD4+细胞。在体内,以从正常小鼠体内获得的外源性DNT干预种植瘤,IVIS监测肿瘤的生长动态,病理切片分析肿瘤组织病变。在体外,建立混合培养系统,观察DNT对淋巴细胞的杀伤功能及分化的影响,以及对CD4+细胞杀伤功能的影响。结果:荷瘤鼠DNT含量高于正常鼠(P<0.05);CD4+细胞含量与肿瘤大小高度负相关(R2=-0.99)。外源性DNT干预后种植瘤生长速度加快(P<0.05),病理显示肿瘤组织中淋巴细胞浸润较对照组少。体外实验中,DNT抑制总淋巴细胞的杀伤功能;DNT细胞直接抑制CD4+细胞杀伤功能。结论:DNT促进B16F10种植瘤生长主要机制之一可能为DNT直接抑制CD4+细胞杀伤功能,从而减少了CD4+细胞对肿瘤的杀伤作用。Objective:To study the possible mechanism that DNT promotes the growth of B16F10 melanoma. Methods:Tumor -bearing mice were established by subcutaneous inoculation of B16F10 -luc -G5 cells.Flow cy-tometry was used to detect the composition changes of lymphocyte in peripheral blood from tumor -bearing mice and normal mice.DNT and CD4 + cells were separated by Aria Cell Sorter.DNT was injected in tumor -bearing mice and the effect on tumor growth was observed by IVIS in vivo and pathological changes.Mixture culture system (MLC)in vitro were established to reveal the mechanism of DNT negative -effect on tumor growth:DNT influence lymphocytes, killing function and differentiation,respectively were monitored by IVIS and flow cytometry after MLC.Effects of DNT direct influence on lymphoid killing function of CD4 + cells were detected by IVIS after MLC.Results:Luminescence between DNT group and tumor control group:there was no significant difference (P >0.05)of mean photon count [photon/(cm2 .ser.s)]on day 0,but significant difference (P <0.05)on day 3 ~9.Differentiation of T cells in pe-ripheral blood:In tumor control group,CD4 + increased first to peak on day 5 and then decreased,with a strong nega-tive correlation to tumor cells (bioluminescence)(R2 =-0.99).Contrast to normal mice,in tumor -bearing mice, DNT increased (P <0.05)but CD4 +CD25 +T decreased (P <0.05),there was strong correlation between two cells (R2 =-0.99).Also a strong correlation between number of CD8 + cells and DNT cells (R2 =-0.97)was also demonstrated in tumor control group.Pathology analysis of tumor:With less lymphocytes infiltration,the tumor was bigger in DNT effect group than in tumor control group.But in following day 5 ~9,the differences decreased gradual-ly.DNT influenced lymphoid killing function:)DNT influenced lymphoid killing function of total lymphocytes:24 ~72h,there was significant difference (P <0.05)of mean photon count and contents of total lymphocytes between any two groups of three.DNT also influenced lymphoid killing function

关 键 词：T淋巴细胞 调节性 双阴性T细胞 黑色素瘤 小鼠 近交系Babl/C 机制 

分 类 号：R73-362[医药卫生—肿瘤] R738.6[医药卫生—临床医学]