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Journal of Zhejiang University SCIENCE B 2017 Vol.18 No.8 P.653-661

10.1631/jzus.B1600106


Adjuvant effects mediated by the carbohydrate recognition domain of Agrocybe aegerita lectin interacting with avian influenza H9N2 viral surface glycosylated proteins


Author(s):  Li-bao Ma, Bao-yang Xu, Min Huang, Lv-hui Sun, Qing Yang, Yi-jie Chen, Ya-lin Yin, Qi-gai He, Hui Sun

Affiliation(s):  College of Animal Sciences and Technology, Huazhong Agricultural University, Wuhan 430070, China; more

Corresponding email(s):   heqigai@yahoo.com, sunhui@whu.edu.cn

Key Words:  Adjuvant, Agrocybe aegerita lectin, Carbohydrate recognition domain, Glycosylated protein, Avian influenza H9N2 virus


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Li-bao Ma, Bao-yang Xu, Min Huang, Lv-hui Sun, Qing Yang, Yi-jie Chen, Ya-lin Yin, Qi-gai He, Hui Sun. Adjuvant effects mediated by the carbohydrate recognition domain of Agrocybe aegerita lectin interacting with avian influenza H9N2 viral surface glycosylated proteins[J]. Journal of Zhejiang University Science B, 2017, 18(1): 653-661.

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author="Li-bao Ma, Bao-yang Xu, Min Huang, Lv-hui Sun, Qing Yang, Yi-jie Chen, Ya-lin Yin, Qi-gai He, Hui Sun",
journal="Journal of Zhejiang University Science B",
volume="18",
number="8",
pages="653-661",
year="2017",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1600106"
}

%0 Journal Article
%T Adjuvant effects mediated by the carbohydrate recognition domain of Agrocybe aegerita lectin interacting with avian influenza H9N2 viral surface glycosylated proteins
%A Li-bao Ma
%A Bao-yang Xu
%A Min Huang
%A Lv-hui Sun
%A Qing Yang
%A Yi-jie Chen
%A Ya-lin Yin
%A Qi-gai He
%A Hui Sun
%J Journal of Zhejiang University SCIENCE B
%V 18
%N 8
%P 653-661
%@ 1673-1581
%D 2017
%I Zhejiang University Press & Springer

TY - JOUR
T1 - Adjuvant effects mediated by the carbohydrate recognition domain of Agrocybe aegerita lectin interacting with avian influenza H9N2 viral surface glycosylated proteins
A1 - Li-bao Ma
A1 - Bao-yang Xu
A1 - Min Huang
A1 - Lv-hui Sun
A1 - Qing Yang
A1 - Yi-jie Chen
A1 - Ya-lin Yin
A1 - Qi-gai He
A1 - Hui Sun
J0 - Journal of Zhejiang University Science B
VL - 18
IS - 8
SP - 653
EP - 661
%@ 1673-1581
Y1 - 2017
PB - Zhejiang University Press & Springer
ER -


Abstract: 
Objective: To evaluate the potential adjuvant effect of Agrocybe aegerita lectin (AAL), which was isolated from mushroom, against a virulent H9N2 strain in vivo and in vitro. Methods: In trial 1, 50 BALB/c male mice (8 weeks old) were divided into five groups (n=10 each group) which received a subcutaneous injection of inactivated H9N2 (control), inactivated H9N2+0.2% (w/w) alum, inactivated H9N2+0.5 mg recombinant AAL/kg body weight (BW), inactivated H9N2+1.0 mg AAL/kg BW, and inactivated H9N2+2.5 mg AAL/kg BW, respectively, four times at 7-d intervals. In trial 2, 30 BALB/c male mice (8 weeks old) were divided into three groups (n=10 each group) which received a subcutaneous injection of inactivated H9N2 (control), inactivated H9N2+2.5 mg recombinant wild-type AAL (AAL-wt)/kg BW, and inactivated H9N2+2.5 mg carbohydrate recognition domain (CRD) mutant AAL (AAL-mutR63H)/kg BW, respectively, four times at 7-d intervals. Seven days after the final immunization, serum samples were collected from each group for analysis. Hemagglutination assay, immunogold electron microscope, lectin blotting, and co-immunoprecipitation were used to study the interaction between AAL and H9N2 in vitro. Results: IgG, IgG1, and IgG2a antibody levels were significantly increased in the sera of mice co-immunized with inactivated H9N2 and AAL when compared to mice immunized with inactivated H9N2 alone. No significant increase of the IgG antibody level was detected in the sera of the mice co-immunized with inactivated H9N2 and AAL-mutR63H. Moreover, AAL-wt, but not mutant AAL-mutR63H, adhered to the surface of H9N2 virus. The interaction between AAL and the H9N2 virus was further demonstrated to be associated with the CRD of AAL binding to the surface glycosylated proteins, hemagglutinin and neuraminidase. Conclusions: Our findings indicated that AAL could be a safe and effective adjuvant capable of boosting humoral immunity against H9N2 viruses in mice through its interaction with the viral surface glycosylated proteins, hemagglutinin and neuraminidase.

杨树菇血凝素糖识别域与禽流感病毒H9N2表面糖蛋白结合介导的佐剂效果

目的:探讨杨树菇血凝素(AAL)作为疫苗佐剂的作用机理,为有囊膜病毒疫苗佐剂的研究提供新思路和数据。
创新点:首次研究AAL作为禽流感疫苗佐剂的作用机理。
方法:大肠杆菌重组表达、纯化野生型的AAL(AAL-wt)和糖识别结构域(CRD)突变型的AAL(AAL-mutR63H)(图1);动物试验证实AAL-wt具有免疫佐剂活性,AAL-mutR63H失去免疫佐剂活性(图2);免疫胶体金电镜(immunogold electron microscopy,IEM)试验发现AAL-wt能吸附H9N2病毒粒子,而AAL-mutR63H不能吸附病毒粒子(图4);凝集素印迹(lectin blot)、免疫共沉淀(co-immunoprecipitation,Co-IP)试验证明了CRD区与H9N2病毒中血球凝集素(HA)和神经氨酸酶(NA)这两种糖蛋白的相互作用(图5)。
结论:AAL的CRD区是发挥佐剂作用的关键区域,CRD区与禽流感病毒表面的HA糖蛋白结合,可能暴露病毒的免疫识别位点,或者将病毒粒子连接在一起,形成大的病毒复合体,增加病毒的抗原性(图6)。

关键词:杨树菇凝集素;禽流感病毒H9N2;糖识别结构域(CRD);血球凝集素(HA);神经氨酸酶(NA)

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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