目的:研究脂氧素对Jurkat T细胞增殖和白细胞介素-2表达的影响。方法:体外培养Jurkat T细胞,anti—CD3(2ms/L)和anti—CD28(2ms/L)单抗刺激Jurkat细胞活化,加入不同浓度脂氧素(0.1nmol/L-100nmol/L)共同孵育24h后,加入[^3H]-TdR继续孵育6h,液闪仪测放射性活度;或收集培养上清,ELISA测IL-2水平,收集细胞,流式细胞仪检测细胞表面IL-2受体α亚单位CD25表达,PI染色后经流式细胞仪进行细胞周期分析。结果:脂氧素剂量依赖性抑制anti—CD3和anti—CD28活化的Jurkat T细胞增殖(P〈0.05);细胞周期分析发现脂氧素处理组S期细胞比例减少;脂氧素显著降低培养上清中IL-2含量(P〈0.05)但对CD25表达无明显影响(P〉0.05)。结论:脂氧素能抑制活化Jurkat T细胞增殖和白细胞介素-2表达;脂氧素可通过影响T淋巴细胞的活化增殖进而发挥免疫负性调节作用。
AIM: To investigate the effect of lipoxin A4 on the proliferation and IL -2 production in Jurkat T cells. METHODS: Jurkat T cells were activated in vitro with anti - CD3 (2 rag/L) and anti - CD28 (2 rag/L) antibodies in the absence or presence of lipoxin A4 (0. 1 nmol/L- 100 nmol/L) for 24 h, then [^3H] -TdR was added into the medium and radioactivities were measured by scintillation counting. The concentrations of IL -2 in the supemants were determined by ELISA. Cells were harvested and the expression of CD25 was assessed by FCM. For analysing the cell cycle, the cells were stained with PI and DNA contents were detected by FCM. RESULTS: Lipoxin A4 suppressed the proliferation of anti - CD3 and anti - CD28 antibodies activated Jurkat ceils in a dose - dependent manner, which was associated with reduced proportion of S phase cells. Furthemore, lipoxin A4 significantly inhibited the production of IL - 2 but had no obvious effect on CD25 expression. CONCLUSION: Lipoxin A4 can suppress proliferation of activate Jurkat cells and IL -2 production, through which lipoxin A4 might negatively regulate immune response.