【摘要】 目的研究细粒棘球蚴囊液对健康人外周血单个核细胞(PBMC)及JurkatT淋巴细胞等的影响。方法PBMC及JurkatT淋巴细胞用含不同浓度(1000、100和10μg/ml)囊液的DMEM培养基培养5d后,台盼蓝染色检测细胞活力。用含植物血凝素(PHA)的DMEM培养基培养PBMC,以MTT法观察PBMC增殖情况,用流式细胞仪检测CD4+、CD8+细胞数量及CD4+/CD8+值,用原位末端标记法和Hoechst33258荧光染色法检测PBMC细胞凋亡。结果各浓度囊液对PBMC和JurkatT淋巴细胞未显示出毒性作用。囊液不影响PHA对PBMC的增殖功能。在1000μg/ml囊液组CD4+、CD8+细胞数量与空白对照组(CD4+72.08±4.10,CD8+29.32±2.62)相比,CD4+(49.32±8.32)显著减少(P<0.05),CD8+数量(32.02±6.57)略增加(P>0.05),CD4+/CD8+比值减少。未见细胞凋亡现象。结论细粒棘球蚴囊液对外周血单个核细胞的生长无抑制作用,但可以使CD4+、CD8+细胞的数量和比例改变,向Th2免疫抑制方向转变。更多还原

【Abstract】 Objective To study the effect of cyst fluid of the parasite Echinococcus granulosus on healthy human peripheral blood mononuclear cells (PBMC) and lymphocytoma cell line Jurkat T growth. Methods After PBMC and Jurkat T cell were incubated with cyst fluid (the protein concentration was 1.65 mg/ml) at different concentration(1 000 μg/ml, 100 μg/ml, 10 μg/ml) in DMEM medium for 5 days, the cell viability was checked by trypan blue. PBMC proliferation uptaked by cyst fluid on stimulation by mitogen phytohemagglutinin A was assayed with MTT method. The number of CD4+, CD8+ and CD4+/CD8+ ratio of the PBMCs were also inspected with flow cytometer (FCM). The PBMCs of culture stained by method of Hoechst No.33258 were inspected under fluorescence microscope. And cell apoptosis was examined using TUNEL assay. Results No apparent inhibition was found compared with control in both PBMCs and Jurkat T cell. The MTT assay showed no significance compared with control. CD4+ T cell analyzed by FCM showed significantly decrease (49.32±8.32) under high concentration (1 000 μg/ml) of cyst fluid and CD8+ T cell increased slightly (32.02±6.57,P>0.05) compared with control(CD4+: 72.08±4.10, CD8+:29.32±2.62). The median and low concentration(100 μg/ml and 10 μg/ml) showed no apparent inhibition on PBMCs growth. The typical apoptosis phenomenon could not be found in both methods. Conclusion The cyst fluid had no inhibition on PBMCs and could reduce CD4+ T cell numbers and CD4+/CD8+ ratio, and turn to Th2 type immune inhibition.更多还原