摘要
目的 探讨猪囊尾蚴排泄分泌抗原(Excretory secretory antigen, ESA)富含亮氨酸重复序列结构域(Leucine-rich repeat containing 15, LRRC15)蛋白对仔猪树突状细胞(Dendritic cell, DC)成熟活化的影响。方法 收集正常仔猪DC体外诱导、培养,获得成熟与未成熟DC(immature DC, imDC)。在正常仔猪未成熟DC中,分别加入LRRC15蛋白、ESA刺激,将LRRC15组作为实验组;同时建立ESA对照组、LPS阳性对照组、LRRC15+LPS阳性对照组、LPS+ESA阳性对照组和培养基阴性对照组。流式细胞术检测DC表面标志物MHC-Ⅱ、CD80、CD86的表达量;ELISA检测DC细胞培养上清TNF-α、IL-6、IL-10、IL-12的分泌水平。结果 与未刺激DC组相比,LRRC15蛋白组诱导DC表面标志物CD80、CD86、MHC-Ⅱ表达均增加。在LPS存在情况下,LRRC15蛋白也能诱导CD80、CD86、MHC-Ⅱ分子上调;ELISA检测结果显示,与1640培养基组、LPS组和ESA组相比,LRRC15蛋白组均诱导IL-6、IL-10、IL-12、TNF-α分泌降低,LRRC15蛋白组与1640培养基组相比IL-6(t=7.529,P<0.05),IL-10(t=2.780,P<0.05),IL-12(t=4.673,P<0.05),TNF-α(t=2.894,P<0.05);LRRC15蛋白组与LPS组相比IL-6(t=26.663,P<0.05),IL-10(t=12.038,P<0.05),IL-12(t=27.594,P<0.05),TNF-α(t=29.540,P<0.05);LRRC15蛋白组与ESA组相比IL-6(t=6.343,P<0.05),IL-10(t=2.579,P<0.05),IL-12(t=8.102,P<0.05),TNF-α(t=3.890,P<0.05)。ESA组与1640培养基组相比能诱导IL-12分泌(t=3.430,P<0.05),其他细胞分子分泌水平差异均无统计学意义(P>0.05)。结论 LRRC15蛋白能诱导DC成熟活化,并抑制DC相关细胞因子的分泌,但能否通过其他途径诱导Th细胞分化还需进一步分析。
This study was aimed at investigating the effects of excretory-secretory antigen(ESA)of Cysticercus cellulosae(LRRC15)on the maturation and activation of piglet dendritic cells(DCs).Normal piglet DCs were collected and induced in vitro,and mature and immature DCs were obtained.LRRC15 protein and ESA were added to the immature DCs of normal piglets.The LRRC15 group served as the experimental group,and an ESA 1640 medium group,LPS positive 1640 medium group,LRRC15+LPS positive 1640 medium group,LPS+ESA positive 1640 medium group,and 1640 medium negative group were also analyzed.The expression of the DC surface markers MHC-Ⅱ,CD80,and CD86 was detected by flow cytometry,and the secretion levels of TNF-α,IL-6,IL-10,and IL-12 in the culture supernatants of DCs were detected by ELISA.Compared with the non-stimulated DC group,the expression of DC surface markers CD80,CD86 and MHC-Ⅱincreased in the lrrc15 protein group.In the presence of LPS,lrrc15 protein can also induce the upregulation of CD80,CD86 and MHC-Ⅱmolecules;ELISA results showed that compared with 1640 medium group,LPS group and ESA group,LRRC15 protein all induced IL-6,IL-10,IL-12,TNF-α,LRRC15 protein group compared with 1640 medium group IL-6(t=7.529,P<0.05),IL-10(t=2.780,P<0.05),IL-12(t=4.673,P<0.05),TNF-α(t=2.894,P<0.05);LRRC15 protein group and LPS group compared IL-6(t=26.663,P<0.05),IL-10(t=12.038,P<0.05),IL-12(t=27.594,P<0.05),TNF-α(t=29.540,P<0.05);LRRC15 protein group compared with ESA group IL-6(t=6.343,P<0.05),IL-10(t=2.579,P<0.05),IL-12(t=8.102,P<0.05),TNF-α(t=3.890,P<0.05).Compared with the 1640 medium group,the ESA group could induce IL-12(t=3.430,P<0.05),and there was no significant difference in other cell molecular levels(P>0.05).Thus,LRRC15 protein induces the maturation and activation of DCs,and inhibits the secretion of DC-related cytokines.However,determining whether LRRC15 induces Th cell differentiation through other routes requires further analysis.
作者
王怡
李丽竹
肖世玉
周必英
WANG Yi;LI Li-zhu;XIAO Shi-yu;ZHOU Bi-ying(The First Clinical College of Zunyi Medical University,Zunyi 563000,China;Department of Parasitology,Zunyi Medical University,Zunyi 563000,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2023年第7期675-681,共7页
Chinese Journal of Zoonoses
基金
国家自然科学基金(No.81960378)
贵州省大学生创新创业训练计划项目(No.S202210661167)
遵义医科大学大学生创新创业训练计划项目。
