Meng Ming, Li Chunxiao, Wang Zerui, Hua Han, Shi Jianhong, Chen Hua, Li Xiaoliu, Chen Dongzhi
School of Medicine, College of Chemistry and Environmental Science, Hebei University, Baoding 071000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Apr;29(4):345-9.
To explore the immunostimulatory effect of immunostimulant CH2a bearing thiazolidin-4-one on human invariant natural killer T (iNKT) cell function in vitro.
Peripheral blood mononuclear cells (PBMCs) were obtained from healthy adults, and then amplified with α-Galcer and IL-2 in vitro. The iNKT cells were isolated from these proliferating cells by magnetic-activated cell sorting (MACS) method. The purified iNKT cells were labled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then incubated with CH2a for functional analysis, including cell proliferation, expressions of IFN-γ and IL-4 by flow cytometry, proliferation rate and cytotoxicity by MTT assay. In additon, ELISA was performed to determine the levels of IFN-γ and IL-4 in cell culture media.
CH2a significantly promoted the proliferation of iNKT cells induced by IL-2 in vitro, stimulated the release of both IFN-r and IL-4, and led to the increase in IFN-γ/IL-4 ratio. More importantly, the cytotoxicity of iNKT was also markedly elevated under the stimulation of CH2a.
Immunostimulant CH2a probably promote the production of Th1-like cytokines and the differentiation of Th0 to Th1, so as to improve cellular immune function. In addition, CH2a could significantly enhance the cytotoxicity of iNKT cells.
探讨含噻唑烷-4-酮的免疫刺激剂CH2a对人不变自然杀伤T(iNKT)细胞体外功能的免疫刺激作用。
从健康成年人获取外周血单个核细胞(PBMCs),然后在体外用α-半乳糖神经酰胺(α-Galcer)和白细胞介素-2(IL-2)进行扩增。通过磁珠激活细胞分选(MACS)方法从这些增殖细胞中分离出iNKT细胞。将纯化的iNKT细胞用5-(和6)-羧基荧光素二乙酸琥珀酰亚胺酯(CFDA-SE)标记,然后与CH2a孵育进行功能分析,包括通过流式细胞术检测细胞增殖、IFN-γ和IL-4的表达,通过MTT法检测增殖率和细胞毒性。此外,采用酶联免疫吸附测定(ELISA)法测定细胞培养基中IFN-γ和IL-4的水平。
CH2a显著促进体外IL-2诱导的iNKT细胞增殖,刺激IFN-γ和IL-4的释放,并导致IFN-γ/IL-4比值升高。更重要的是,在CH2a刺激下iNKT的细胞毒性也明显升高。
免疫刺激剂CH2a可能促进Th1样细胞因子的产生以及Th0向Th1的分化,从而改善细胞免疫功能。此外,CH2a可显著增强iNKT细胞的细胞毒性。