负载人蛔虫和猪蛔虫抗原树突状细胞影响T细胞亚群活性研究  

Comparative analysis of T cell subsets response induced by DC fused with Ag of Ascaris lumbricoides and Ascaris suum

作　　者：祝伟[1,2] 黄川[1,2] 黄艳琴[1,2] 胡银英[1,2] 戴志芳[1,2] 袁芳[1,2] 袁铿[1,2] 

机构地区：[1]江西省医学研究院,南昌330006 [2]南昌大学医学院

出　　处：《中国人兽共患病学报》2013年第8期797-802,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.30960151)资助~~

摘　　要：目的探讨人蛔虫和猪蛔虫抗原致敏树突状细胞(Dendritic Cell,DC)对T细胞亚群活性的调节作用差异。方法用IL-4和GM-CSF联合体外诱导小鼠骨髓源性DC,分别负载两种不同基因型蛔虫抗原,由低到高各分3组,并设置空白对照进行培养。负载了蛔虫抗原的DC,按1∶25的比例分别与同源性脾淋巴细胞共培养。用流式细胞术、ELISA和qPCR分别检测比较负载后DC表面分子表达变化及其混合淋巴细胞反应Th1/Th2类细胞因子表达的变化和FoxP3mRNA的表达水平。结果骨髓细胞诱导所得初始DC高表达表面分子CD86(85.5%)、CD83(38.24%)和CD80(94%),在负载蛔虫抗原后,各组仍高表达,但CD83和CD80较初始DC有所下降,且猪蛔虫抗原组和人蛔虫抗原组分别低和高于空白对照组。ELISA检测显示,各时间段实验组TNF-α的表达量都低于对照组,且差异都有统计学意义(P<0.05);IL-10在各组的表达量随时间延长而增加,各时间段的相同浓度除24h外,人蛔虫抗原组高于猪蛔虫抗原组(P<0.05)。qPCR结果显示,相对于人蛔虫抗原组Foxp3mRNA的表达量,空白对照组和住蛔虫抗原组各个时间段都较低,且都与前者存在差异性(P<0.05);人蛔虫抗原组的表达量在短期内增加明显,但又随时间而减少;猪蛔虫抗原组则先增后减,但都与空白对照组间无差异性(P>0.05)。结论负载了人蛔虫抗原的DC能更强促进淋巴细胞Th2类反应和Foxp3表达,影响调节性T细胞功能。To investigate comparatively the different responses of T cell subsets induced by dendritic cell （DC） fused respectively with three different concentrations of A. lumbricoides and A. suum antigens fAg）, the levels of the expression of DC surface moleculars, Th1/Th2 cytokines and Foxp3 mRNA in different MLR groups, which were antigen-loaded DC co-cultured with C57BL/6 mouse-derived splenic lymphocytes at the effect-target ratio of 1 ： 25, were detected respeetively by flow cytometry （FCM）, ELISA and real time PCR. And the DCs were isolated from bone marrow cells cultured with granulocyte/ macrophage colony-stimulating factor （GM-CSF） and IL-4 in vitro. The FCM results showed that DCs isolated from bone mar row cells were with high expression of kinds of surface molecules （CD86, 85.5%; CD83 38.24%; CD80, 94%）. CD80 and CD83 declined after being cultured with ascaris extract for 2 days and that of the groups fused separately with Ag Of A. lumbricoides and A. suum was respectively higher and lower than the control group. ELISA showed the level of expression of TNF-α in each period of each experimental group was lower than the control group （P〈0.05）; IL- 10 in each group increased over time, and the groups with ascaris antigen were higher than groups with same concentration of A. suum antigen （P〈0.05） in each time except the time point of 24 hours. The qPCR results showed that the levels of the groups with A. lumbricoides Ag were declined over time but significantly higher than the control groups and the groups with A. suum Ag in different time points （P〈0.05）. The differences between the groups with A. suurn Ag and the control groups were not statistically significant （P〈0.05）. In conclusion, DC fused with the Ag of A. lumbricoides promoted a stronger Th2 type response and the expression of Foccp3 mRNA in vitro.

关 键 词：树突状细胞 T细胞 人蛔虫 猪蛔虫 抗原 

分 类 号：R383[医药卫生—医学寄生虫学]