A Bivalent Human Adenovirus Type 5 Vaccine Expressing the Rabies Virus Glycoprotein and Canine Distemper Virus Hemagglutinin Protein Confers Protective Immunity in Mice and Foxes
A Bivalent Human Adenovirus Type 5 Vaccine Expressing the Rabies Virus Glycoprotein and Canine Distemper Virus Hemagglutinin Protein Confers Protective Immunity in Mice and Foxes
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表达狂犬病病毒糖蛋白和犬瘟热病毒血凝素蛋白的二价人 5 型腺病毒疫苗可赋予小鼠和狐狸保护性免疫力
DOI：
10.3389/fmicb.2020.01070
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发表时间：
2020-06-16
影响因子：
5.2
通讯作者：
Zheng, Xuexing
中科院分区：
文献类型：
--
作者：
Yan, Lina;Zhao, Zhongxin;Zheng, Xuexing
研制安全高效的多价疫苗具有广阔的应用前景。狂犬病病毒（RABV）和犬瘟热病毒（CDV）都是传染性很强的抗原，在世界范围内的家养狗和其他食肉动物中引起致命疾病。本研究构建了复制缺陷型人腺病毒5 （Ad5）载体疫苗rAd5-G-H，表达RABV糖蛋白(G)和CDV血凝素(H)蛋白。间接免疫荧光法在感染的HEK-293细胞中表达并证实了rAd5-G-H的RABV G和CDV H蛋白。电镜下rAd5-G-H保持了与rAd5-GFP相似的均匀二十面体形态。单剂量108 GFU的rAd5-G-H肌内注射小鼠，可快速、强效地产生抗RABV和CDV的中和抗体。流式细胞术检测显示，与模拟组和rAd5-GFP组相比，rad5 - g - h免疫小鼠腹股沟淋巴结的树突状细胞和B细胞明显增加。在小鼠中，rAd5-G-H还激活了Th1和th2介导的抗RABV和CDV的细胞免疫反应，这有助于在没有任何临床症状的情况下使RABV致命剂量攻击的存活率达到100%。在狐狸中，与模拟组和rAd5-GFP组相比，单剂量109 GFU的rAd5-G-H可以引起高水平的抗RABV和CDV的中和抗体。rAd5-GFP组和模拟组的狐狸全部死亡，而接种了rAd5-G-H的狐狸在被致死性野生型CDV毒株攻击后全部存活并且没有出现疾病的临床症状。这些结果表明，rAd5-G-H作为狂犬病和犬瘟热双价疫苗在高易感犬和野生动物中具有很大的潜力。
The development of a safe and efficient multivalent vaccine has great prospects for application. Both rabies virus (RABV) and canine distemper virus (CDV) are highly infectious antigens, causing lethal diseases in domestic dogs and other carnivores worldwide. In this study, a replication-deficient human adenovirus 5 (Ad5)-vectored vaccine, rAd5-G-H, expressing RABV glycoprotein (G) and CDV hemagglutinin (H) protein was constructed. The RABV G and CDV H protein of rAd5-G-H were expressed and confirmed in infected HEK-293 cells by indirect immunofluorescence assay. The rAd5-G-H retained a homogeneous icosahedral morphology similar to rAd5-GFP under an electron microscope. A single dose of 108 GFU of rAd5-G-H administered to mice by intramuscular injection elicited rapid and robust neutralizing antibodies against RABV and CDV. Flow cytometry assays indicated that the dendritic cells and B cells in inguinal lymph nodes were significantly recruited in rAd5-G-H-immunized mice in comparison with the mock and rAd5-GFP groups. rAd5-G-H also activated the Th1- and Th2-mediated cell immune responses against RABV and CDV in mice, which contributed to 100% survival of a lethal-dose RABV challenge without any clinical signs. In foxes, a single dose of 109 GFU of rAd5-G-H could elicit high levels of neutralizing antibodies against both RABV and CDV in comparison with the mock and rAd5-GFP groups. All foxes in the rAd5-GFP and mock groups died, while the foxes inoculated with rAd5-G-H all survived and showed no clinical signs of disease after being challenged with a lethal wild-type CDV strain. These results suggested that rAd5-G-H has great potential as a bivalent vaccine against rabies and canine distemper in highly susceptible dogs and wildlife animals.