肝癌荷瘤小鼠调节性T细胞数量的改变及其与肿瘤生长的关系  

Changes of regulatory T cell number in hepatocellular carcinoma-bearing mice and its relationship with tumor growth

作　　者：张蘋[1] 张利宁[1] 朱法良[1] 王群[1] 王晓燕[1] 李海艳[1] 刘春梅[1] 高飞[1] 刘成虎[1] 

机构地区：[1]山东大学医学院免疫学研究所,济南250012

出　　处：《中华肿瘤杂志》2007年第5期342-345,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金资助项目（30271247）

摘　　要：目的研究肝癌荷瘤小鼠调节性T细胞数量的改变及其与肿瘤生长的关系。方法采用小鼠肝癌细胞系H22接种BALB／c小鼠,建立肝癌模型;采用流式细胞术方法检测CD4^＋ CD25^＋T／CD4^＋T细胞的比例;以RT-PCR和流式细胞术检测Foxp3基因的表达。以免疫磁珠分选法纯化CD4^＋CD25^＋T和CD4^＋CD25^-T细胞;在体外,用3H-TdR掺入法检测T细胞的增殖情况;在体内,观察荷瘤小鼠来源的CD4^＋CD25^＋T细胞对肿瘤生长的作用。结果（1）荷瘤小鼠在引流淋巴结中,CD4^＋CD25^＋T细胞占CD4＋T细胞（18．80％±0．06％）比例增高,与对照组（9．50％±0．03％）相比,差异有统计学意义（P〈0．01）;在非引流淋巴结（LN）和脾脏（SP）中,荷瘤小鼠CD4^＋CD25^＋T／CD4^＋T比例分别为16．28％±0．02％和17．28％±0．06％,与对照组9．50％±0．03％和11．08％±0．04％相比,差异有统计学意义（P〈0．01,P〈0．05）;同时,调节性T细胞特异性标志Foxp3 mRNA的表达也升高。在同一只荷瘤小鼠中,引流淋巴结中CD4^＋CD25^＋T细胞数量（18．8％±0．06％）较对侧非引流淋巴结（16．28％±0．02％）略有升高,但差异无统计学意义（P〉0．05）。（2）从荷瘤小鼠中纯化的CD4^＋CD25^＋T细胞,在体外对抗CD3单抗的刺激无反应,但能抑制CD4^＋CD25^-T细胞的增殖。（3）CD4^＋CD25^＋T／CD4＋T比例与肿瘤大小呈正相关,并且可以抑制CD4^＋CD25^-T细胞的抗肿瘤效应。结论肝癌细胞在小鼠体内的生长可以提高调节性T细胞的数量,其数量的高低与肿瘤的大小呈正相关,提示清除调节性T细胞将是肿瘤免疫治疗的策略之一。Objective To study the relationship between the change of regulatory T cell number in CD4^＋ T subset and the growth of tumor in H22 hepatocellular carcinoma-beating mice. Methods Tumor-beating mice were established by subcutaneous inoculation of H22 hepatocelluler carcinoma cells. Flow cytometry was used to detect the expression of CD4 and CD25 molecules of the T cells which came from the tumor-beating mice. The Foxp3 gene expression was detected by RT-PCR and flow cytometry. CD4^＋ CD25^＋ T cells and CD4^＋ CD25^- T cells were separated and purified by immuno-maguetic beads. The proliferation and suppressive function of the CD4^＋ CD25^＋ T cells coming from tumor-beating mice was measured by [^3H]-thymidines incorporation experiment in vitro, and then effect of CD4^＋ CD25^＋ T cells originated from hepatocellular carcinoma-bearing mice on tumor growth was observed in vivo. Results （ 1 ） Compared with mice of the control group, the percentage of CD4^＋ CD25^＋ T cells of CD4^＋ T cells in tumor-bearing mice is not only higher in draining lymph nodes（18.80%±0.06% ） vs. （9.50%±0.03% ）, （P 〈0.01 ）, but also higher in non-draining lymph nodes （LN） and spleen （SP）, LN： （ 16.28 %±0.02% ） vs. （ 9.50%±0.03%）,P〈0.01; SP：（17.28%±0.06%） vs. （11.08%±0.04%）,（P〈0.05）. The expression of regulatory T cell specific marker Foxp3 gene was also increased. In the same tumor-beating mice, the number of CD4^＋ CD25^＋ T cells in draining lymph node was relatively higher than the contralateral nondraining lymph node, but the difference was statistically not significant（ 18.8%±0.06% ） vs. （16.28%±0.02% ）, （ P 〉 0. 05 ）. （ 2 ） The CD4^＋ CD25^＋ T cells purified from tumor-beating mice- like naturally occurring regulatory T cells - were anergic to anti-CD3 monoclonal antibody stimulation in vitro, but it could Suppress CD4^＋ CD25^- T cells proliferation. （ 3 ） The percentage of CD4^＋ CD25^＋ T cells was positively related to tumor s

关 键 词：CD4^+CD25^+调节性T细胞 FOXP3 肝癌荷瘤小鼠 

分 类 号：R686[医药卫生—骨科学]