燕麦来源β-葡聚糖通过Dectin-1增强粗抗原对旋毛虫感染C57BL/6J小鼠的免疫保护作用  

Oat derivedβ-glucan enhances immune protection of crude antigen against C57BL/6Jmice infected by Trichinella spiralis via Dectin-1

作　　者：毛瀚海 李承瑶 吕庆博 丁静 刘晓雷 刘明远 杨姝 金雪敏 MAO Hanhai;LI Chengyao;LYU Qingbo;DING Jing;LIU Xiaolei;LIU Mingyuan;YANG Shu;JIN Xuemin(State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases/Ministry of Education Key Laboratory for Zoonoses Research,College of Veterinary Medicine,Jilin University,Changchun 130062,China;Qinghe Clinic of Jingbei Medical District,PLA General Hospital,Beijing100000,China)

机构地区：[1]吉林大学动物医学学院人兽共患病研究教育部重点实验室/人畜共患传染病重症诊治全国重点实验室,吉林长春130062 [2]解放军总医院京北医疗区清河门诊部,北京100000

出　　处：《中国兽医学报》2024年第4期706-711,共6页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2023YFD1802404);吉林省科技发展计划资助项目(20220101294JC);吉林省自然科学基金人才专项-青年成长科技计划资助项目(20220508052RC)。

摘　　要：将48只雌性C57BL/6J小鼠随机分为4组,即PBS对照组、粗抗原(crude antigen,CA)处理组、β-葡聚糖(BG)+CA处理组和BG+CA+Dectin-1中和抗体(2A11)处理组,每组各12只。PBS对照组仅皮下注射PBS作为对照,CA组、BG+CA组和BG+CA+2A11组每间隔2周进行皮下注射,共计免疫3次,最后1次免疫2周后,所有组经口灌胃等量旋毛虫。感染后7d,统计PBS对照组、CA处理组、BG+CA处理组和BG+CA+2A11处理组成虫荷虫量及感染后35d肌幼虫荷虫量。取感染后7d小鼠小肠组织,qPCR法检测肠道Dectin-1基因表达量;通过间接酶联免疫吸附法测定感染后0~7周IgG滴度变化;使用ELISA检测IFN-γ和IL-4的表达量。结果显示,与CA处理组相比,BG+CA联合免疫小鼠成虫和肌幼虫荷虫量极显著降低(P<0.0001),BG+CA诱导更高水平的IgG免疫反应以及更多的Dectin-1、IFN-γ和IL-4表达;而给予2A11抑制剂小鼠与CA处理组相比荷虫量、IgG滴度以及IFN-γ、IL-4表达量均无显著差异(P>0.05),靶向2A11抗体后显著降低了BG引起的免疫反应。结果表明,Dectin-1与BG触发CA对旋毛虫的保护型免疫反应有关,BG可能是一种有效的免疫佐剂,为旋毛虫疫苗的开发提供参考。Studies have shown that Dectin-1is one of the important ways for BG to activate innate immunity and is involved in the infection process generated by various pathogens.It is a typical pattern recognition receptor,but whether BG enhances the immune response induced by crude antigens of Trichinellathrough dectin-1pathway remains unclear.Forty-eight female C57BL/6Jmice were randomly divided into four groups:the PBS control group;crude antigen(CA)treatment group;BG and CA treatment group;BG,CA and Dectin-1neutralizing antibody(2A11)treatment group,with 12animals in each group.The PBS control group was only immunized subcutaneously,and the CA group,BG+CA group and BG+CA+2A11groups were immunized subcutaneously three times every two weeks.Two weeks after the last immunization,the same amount of T.spiralis was given orally in all groups.On the 7th day after infection,the parasite load of PBS control group,CA treatment group,BG+CA treatment group and BG+CA+2A11treatment group and the parasite load of muscle larvae at the 35th day were counted.The expression of Dectin-1gene was detected by qPCR on the 7th day after infection.IgG titers were measured 2,4,6,7weeks after infection by indirect enzyme-linked immunosorbent assay.The expression levels of IFN-γand IL-4 were detected by ELISA.The results showed that compared with the CA-treated group,the burden of adult and muscle larvae in mice immunized with BG+CA was significantly reduced(P<0.0001),and BG+CA induced higher levels of IgG immune response and more expression of Dectin-1,IFN-γ,and IL-4.There were no significant differences in parasite load,IgG titer,IFN-γand IL-4expression between 2A11inhibitor mice and CA treatment group(P>0.05),targeting 2A11 antibody significantly reduced the immune response caused by BG.The results show that Dectin-1 is related to the protective immune response of CA triggered by BG against T.spiralis,and BG may be an effective immune adjuvant,providing a reference for the development of T.spiralis vaccine.

关 键 词：β-葡聚糖(BG) 粗抗原(CA) 旋毛虫 DECTIN-1 IgG IFN-γ IL-4 

分 类 号：S852.73[农业科学—基础兽医学] R535[农业科学—兽医学]