B7-H4对宫颈癌患者外周血活化T细胞增殖、凋亡和分泌细胞因子的影响

Influence of B7-H4 on cell proliferation,apoptosis and cytokine secretion of peripheral blood activated T cells in cervical cancer patients

目的观察重组人B7-H4蛋白对宫颈癌患者外周血T细胞增殖周期、凋亡及分泌细胞因子的影响。方法 B7-H4分别与15例宫颈癌患者外周血活化淋巴细胞混合培养48h后,采用流式细胞术分析T细胞增殖、细胞周期、凋亡及各亚群T细胞的变化,酶联免疫吸附试验(ELISA)芯片检测培养上清液细胞因子含量。结果宫颈癌患者外周血T细胞与B7-H4混合培养48h后,G1、G2和S期的T细胞分别占90.59%、8.55%和0.87%,空白对照组T细胞各期占92.83%、6.09%和1.13%;B7-H4组CD4+T和CD8+T细胞的Ki67阳性率分别为2.13%±0.13%和1.03%±1.33%,空白对照组为2.74%±0.98%和1.71%±1.32%。B7-H4组较空白对照组CD8+T和CD4+T细胞占T细胞的比例下降,但CD4+T/CD8+T比值增高,CD4+CD25+Foxp3+T细胞增多;B7-H4组混合培养上清液中TGF-β1含量为(259.25±32.78)pg/mL,空白对照组为(202.75±20.17)pg/mL。B7-H4对宫颈癌患者外周血活化T细胞凋亡无明显影响。结论 B7-H4使宫颈癌患者外周血活化T细胞被阻滞于G2期,S期细胞明显减少;B7-H4抑制CD4+T和CD8+T细胞增殖,但对Foxp3+T细胞增殖和分泌TGF-β1可能有促进作用;B7-H4对T细胞凋亡无明显影响。B7-H4在抑制宫颈癌抗肿瘤细胞免疫中发挥作用,它可能成为宫颈癌免疫治疗的潜在靶点。

Objective To observe the in vitro influence of recombinant human B7-H4 protein on the cell proliferation cycle,apoptosis and cytokine secretion of peripheral blood activated T lymphocytes in cervical cancer patients.Methods After 48 hcoculture of peripheral blood T lymphocytes in 15 cases of cervical cancer with B7-H4 48 h,T lymphocytes′cell proliferation cycle,apoptosis and T lymphocytes subtypes changes were detected by FCM;the cytokines concentration in the culture supernatant was tested by ELISA array.Results After 48hco-culture of peripheral blood T lymphocytes with B7-H4 48 hs,G1,G2 and S phase of T cells accounted for 90.59%,8.55%and 0.87%respectively,which of the blank group were 92.83%,6.09%and 1.13%respectively;the Ki67 positive rates of CD4＋T and CD8＋T cells in the B7-H4 group were 2.13%±0.13% and 1.03%±1.33% respectively,which of the blank group were 2.74% ±0.98% and 1.71% ±1.32% respectively;the proportion of CD4＋T and CD8＋T cells accounting for T cells in the B7-H4 group was decreased compared with the blank group,but the ratio of CD4＋T/CD8＋T and the proportion of CD4＋CD25＋Foxp3＋T cells were increased,in addition,the TGF-β1secretion;concentration in the co-culture supernatant in the B7-H4 group was（259.25±32.78）pg/mL,which was higher than（202.75±20.17）pg/mL in the blank group.B7-H4 had no significant influence on the peripheral blood activated T cells apoptosis.Conclusion B7-H4 block the peripheral blood activated T cells at G2 phase,the S phase cells are obviously decreased;B7-H4 inhibits the cellular proliferation of CD4＋T and CD8＋cells,but may have the promoting effect on Foxp3＋T proliferation and TGF-β1secretion;B7-H4 has no significant influence on T cell apoptosis.B7-H4 plays a role in depressing anti-tumor T cell immune response of cervical cancer and may become a potential target of cervical cancer immunotherapy.