摘要
目的探讨猪囊尾蚴排泄分泌抗原(excretory secretory antigen,ESA)对仔猪体外培养的树突状细胞(dendritic cell,DC)成熟活化的影响。方法建立囊虫病猪动物模型,收集囊尾蚴体外培养并制备其ESA。实验分为control组、LPS阳性对照组、ESA及LPS+ESA实验组,体外用囊尾蚴ESA分别刺激仔猪DC。流式细胞术检测DC表面标志物MHC-II、CD80、CD86的表达量;ELISA检测细胞培养上清IL-6、IL-10、IL-12、TNF-α分泌水平。结果与control组相比,LPS及LPS+ESA组DC表面标志物表达量及细胞因子分泌水平均显著升高,差异具有统计学意义(P<0.05);ESA组MHC-II、CD80、CD86表达量及IL-6、TNF-α分泌水平均显著升高,差异具有统计学意义(P<0.05),IL-10及IL-12分泌水平差异无统计学意义(P>0.05)。与LPS组相比,ESA组CD86、MHC-II表达及IL-6、IL-10、IL-12、TNF-ɑ分泌均较低,差异具有统计学意义(P<0.05);LPS+ESA组CD80、CD86、MHC-II表达及IL-6、IL-10、IL-12、TNF-α分泌水平均显著升高,差异具有统计学意义(P<0.05),且细胞因子分泌水平均高于LPS组及ESA组,差异具有统计学意义(P<0.05)。结论猪囊尾蚴ESA可刺激DC成熟活化并分泌IL-6及TNF-α,本研究为进一步探讨DC参与的囊虫病免疫发病机制提供了实验基础。
Objective To investigate the effect of an excretory secretory antigen(ESA)of Cysticercus cellulosae on the maturation and activation of dendritic cells(DCs)of piglets cultured in vitro.Methods An animal model of cysticercosis was created by feeding healthy piglets with the gravid proglottids of Taenia solium.C.cellulosae was collected to prepare ESA by dissecting the infected piglets after 2-3 months.DCs derived from the bone marrow of piglets were isolated and cultured.Cells were divided into a control group,an LPS positive control group,an ESA group,and an LPS+ESA experimental group.The concentration of ESA was 40μg/ml and that of LPS was 100 ng/ml.Under suitable conditions,treated cells were co-cultured with immature DCs and then centrifuged.Flow cytometry was used to detect the expression of DC surface markers CD80,CD86,and MHC-Ⅱand ELISA was used to analyze the levels of IL-6,IL-10,IL-12,and TNF-ɑexpression in the culture supernatant.Results In this study,immature DCs(with a purity of more than 80%)of piglets were stimulated with C.cellulosae to successfully prepare an ESA of C.cellulosae.The expression of DC surface markers and the secretion of cytokines in LPS group and LPS+ESA group were significantly higher than those in the control group(P<0.05).The expression of DC surface markers CD80,CD86 and MHC-II in ESA group was higher than those in the control group(t=68.38,19.46 and 16.50,P>0.05).The secretion of the cytokines IL-6 and TNF-ɑin the ESA group was higher than that in the control group(t=33.62 and 52.94,P<0.05).There were no significant differences in the levels of IL-10 and IL-12 secretion in the ESA group and the control group(t=0.39 and 2.19,P>0.05).The expression of MHC-II and CD86 and the secretion of IL-6,IL-10,IL-12,and TNF-α in ESA group were significantly lower than those in the LPS group(P<0.05),but there were no significant differences in the expression of CD80(t=1.98,P>0.05).The expression of surface markers and the level of cytokine secretion in the LPS+ESA group were significant
作者
张雷
范贤敏
马琴
马敏梅
周必英
ZHANG Lei;FAN Xian-min;MA Qin;MA Min-mei;ZHOU Bi-ying(Department of Parasitology,Zunyi Medical University,Zunyi 563000,China)
出处
《中国病原生物学杂志》
CSCD
北大核心
2021年第11期1249-1253,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.81960378)
遵义医科大学大学生创新创业训练计划项目(No.ZYDC2019039)
关键词
猪带绦虫
猪囊尾蚴
排泄分泌抗原
树突状细胞
Taenia solium
cysticercus cellulosae
excretory secretory antigen
dendritic cell
