Zhang Pin, Zhang Li-ning, Zhu Fa-liang, Wang Qun, Wang Xiao-yan, Li Hai-yan, Liu Chun-mei, Gao Fei, Liu Cheng-hu
Institute of Immunology, Medical School of Shandong University, Jinan 250012, China.
Zhonghua Zhong Liu Za Zhi. 2007 May;29(5):342-5.
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-bearing mice.
Tumor-bearing 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-bearing 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-magnetic beads. The proliferation and suppressive function of the CD4+ CD25+ T cells coming from tumor-bearing 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.
(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% < or = 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-bearing 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-bearing 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 size. It could also suppress the anti-tumor effect of CD4+ CD25- T cells in vivo. Conclusion The growth of hepatocellular carcinoma in mice can boost the amount of regulatory T cells. The amount of regulatory T cells is positively related to tumor size, indicating that attack on regulatory T cells could be used as one of modalities in cancer treatment in the future.
研究荷H22肝癌小鼠CD4+T亚群中调节性T细胞数量变化与肿瘤生长的关系。
通过皮下接种H22肝癌细胞建立荷瘤小鼠模型。采用流式细胞术检测荷瘤小鼠T细胞中CD4和CD25分子的表达。通过RT-PCR和流式细胞术检测Foxp3基因表达。用免疫磁珠分离纯化CD4+CD25+T细胞和CD4+CD25-T细胞。体外采用[3H]-胸腺嘧啶核苷掺入实验检测荷瘤小鼠CD4+CD25+T细胞的增殖及抑制功能，然后在体内观察肝癌荷瘤小鼠来源的CD4+CD25+T细胞对肿瘤生长的影响。
（1）与对照组小鼠相比，荷瘤小鼠CD4+T细胞中CD4+CD25+T细胞百分比不仅在引流淋巴结中更高（18.80%±0.06% vs. 9.50%±0.03%，P<0.01），在非引流淋巴结和脾脏中也更高，非引流淋巴结：（16.28%±0.02% vs. 9.50%±0.03%，P<0.01）；脾脏：（17.28%±0.06% vs. 11.08%±0.04%，P<0.05）。调节性T细胞特异性标志物Foxp3基因表达也增加。在同一荷瘤小鼠中，引流淋巴结中CD4+CD25+T细胞数量相对高于对侧非引流淋巴结，但差异无统计学意义（18.8%±0.06% vs. 16.28%±0.02%，P>0.05）。（2）从荷瘤小鼠中纯化的CD4+CD
