IL-21 and IL-12 Inhibit Differentiation of Treg and TH17 Cells and Enhance Cytotoxicity of Peripheral Blood Mononuclear Cells in Patients With Cervical Cancer
IL-21 and IL-12 Inhibit Differentiation of Treg and TH17 Cells and Enhance Cytotoxicity of Peripheral Blood Mononuclear Cells in Patients With Cervical Cancer
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IL-21和IL-12抑制Treg和TH17细胞分化并增强宫颈癌患者外周血单个核细胞的细胞毒性
DOI：
10.1097/igc.0b013e3182358955
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发表时间：
2011-12-01
影响因子：
4.8
通讯作者：
Cui, Baoxia
中科院分区：
文献类型：
--
作者：
Tian, Yongju;Yuan, Cunzhong;Cui, Baoxia
目的白细胞介素21（IL-21）和IL-12是目前公认的有效的抗肿瘤药物。在这项研究中，我们评估了IL-21与IL-12的组合是否可以增强宫颈上皮内瘤变III和宫颈癌患者外周血单个核细胞（PBMC）的细胞毒性。方法分离宫颈上皮内瘤样病变III期患者（n = 17）和宫颈癌患者（n = 18）外周血单个核细胞。外周血单个核细胞与低浓度IL-2共同培养作为对照组。将白细胞介素2刺激的PBMC分别与抗人IL-21中和抗体、单独的IL-21、单独的IL-12和IL-21加IL-12共培养作为测试组。乳酸脱氢酶释放法检测PBMC对SiHa肿瘤细胞的杀伤活性。通过流式细胞术分析CD 4 + CD 25 + FOXP 3 + T调节（Treg）细胞和CD 4 +IL-17 A + T辅助17（TH 17）细胞。CCK-8法检测细胞增殖，流式细胞仪检测细胞凋亡。结果与对照组相比，IL-21和IL-12可显著增强PBMC对SiHa细胞的杀伤活性。此外，与单独的IL-21和单独的IL-12相比，IL-21加IL-12显著提高PBMC细胞毒性。我们还发现，与对照组相比，IL-21加IL-12显著降低了Treg和TH 17细胞比例。值得注意的是，与单独的IL-21相比，IL-21加IL-12显著降低了TH 17细胞比例。IL-21和IL-12均能显著降低PBMC的凋亡率，但对PBMC的增殖无明显影响。结论IL-21和IL-12联合应用可有效刺激PBMC产生抗SiHa细胞的细胞毒活性，其机制可能与下调Treg和TH 17细胞分化有关。
Objectives Interleukin 21 (IL-21) and IL-12 have been known to be effective antitumor agents. In this study, we evaluated whether IL-21 in combination with IL-12 could enhance the cytotoxicity of peripheral blood mononuclear cells (PBMCs) in patients with cervical intraepithelial neoplasia III and cervical cancer. Methods Peripheral blood mononuclear cells were isolated from peripheral blood of cervical intraepithelial neoplasia III patients (n = 17) and cervical cancer patients (n = 18). Peripheral blood mononuclear cells were cultured with IL-2 in low concentration as control group. Interleukin 2–stimulated PBMCs were cocultured with anti–human IL-21 neutralizing antibody, IL-21 alone, IL-12 alone, and IL-21 plus IL-12, respectively, as test groups. The cytotoxicity of PBMCs against SiHa tumor cells was examined by lactate dehydrogenase released assay. CD4+CD25+FOXP3+ T regulatory (Treg) cells and CD4+IL-17A+ T helper 17 (TH17) cells were analyzed by flow cytometry. Proliferation and apoptosis were detected by CCK-8 (cell counting kit 8) assay and flow cytometry, respectively. Results Compared with controls, IL-21 and IL-12 significantly elevated PBMC cytotoxicity against SiHa cells. Moreover, IL-21 plus IL-12 significantly elevated PBMC cytotoxicity in comparison to IL-21 alone and IL-12 alone. We also found that IL-21 plus IL-12 significantly decreased Treg and TH17 cell proportion in comparison to controls. Notably, IL-21 plus IL-12 significantly decreased TH17 cell proportion in comparison to IL-21 alone. Both IL-21 and IL-12 significantly decreased the apoptosis rate of PBMCs, whereas neither IL-21 nor IL-12 had significant effect on PBMC proliferation. Conclusions The combination of IL-21 and IL-12 could efficiently stimulate PBMCs with cytotoxicity against SiHa cells, and the possible mechanisms may be due to down-regulated Treg and TH17 cell differentiation.