METHODS: Peripheral blood samples were collected from PTB patients and healthy controls. The cytometric bead array was used to detect plasma IL-33, TGF-β, IL-4, IL-5, IL-6, IL-10, IL-13, and soluble ST2 (sST2). ILC2s, Th2, and Treg cells were detected with flow cytometry. Quantitative real-time PCR was used to measure mRNA levels. ILC2s were co-cultured with peripheral blood mononuclear cells and then intervened with IL-33 or anti-ST2 antibody + IL-33 in vitro. IL-4, IL-6, IL-5, IL-10, IL-13, and TGF-β levels were measured by enzyme-linked immunosorbent assay.
RESULTS: Compared with healthy controls, the levels of IL-33, sST2, TGF-β, IL-10, and IL-6 in the plasma of PTB patients were significantly higher. No significant difference was found in the plasma IL-4, IL-5, and IL-13 levels. Patients with PTB had significantly increased ILC2s proportion and mRNA levels of RAR-related orphan receptor α and GATA binding protein 3. After 48 h of IL-33 stimulation in vitro, Treg cell proportion significantly increased and the IL-10 level was significantly elevated. Treatment with anti-ST2 abolished these effects. No significant difference was found in cytokines of IL-4, IL-6, IL-5, IL-13, and TGF-β, or Th2 cells before and after IL-33 treatment. ILC2s proportion in peripheral blood was increased and plasma IL-33 was upregulated in PTB patients.
CONCLUSIONS: IL-33 may promote the growth of ILC2s and the production of Treg-related cell cytokines, but not Th2-related cell cytokines, to participate in immune response to PTB.
方法:从PTB患者和健康对照者收集外周血样本。细胞计数珠阵列用于检测血浆IL-33,TGF-β,IL-4、IL-5、IL-6、IL-10、IL-13和可溶性ST2(sST2)。ILC2s,流式细胞仪检测Th2、Treg细胞。定量实时PCR用于测量mRNA水平。将ILC2s与外周血单个核细胞共培养,然后用IL-33或抗ST2抗体+IL-33进行体外干预。采用酶联免疫吸附法检测IL-4、IL-6、IL-5、IL-10、IL-13和TGF-β水平。
结果:与健康对照组相比,IL-33,sST2,TGF-β,IL-10、IL-6在PTB患者血浆中显著增高。血浆IL-4、IL-5和IL-13水平无显著差异。PTB患者的ILC2s比例和RAR相关孤儿受体α和GATA结合蛋白3的mRNA水平显着增加。在体外IL-33刺激48小时后,Treg细胞比例显著增加,IL-10水平显著升高。用抗ST2治疗消除了这些作用。细胞因子IL-4、IL-6、IL-5、IL-13和TGF-β无显著差异,或IL-33治疗前后的Th2细胞。PTB患者外周血中ILC2s比例增加,血浆IL-33上调。
结论:IL-33可能促进ILC2s的生长和Treg相关细胞因子的产生,但不是Th2相关的细胞细胞因子,参与对PTB的免疫应答。