嵌合犬瘟热病毒H基因的重组狂犬病病毒的构建及免疫原性研究

doi:10.16431/j.cnki.1671-7236.2022.12.026

Abstract

Abstract: 【Objective】 The aim of this experiment was to construct recombinant Rabies virus (RABV) expressing the hemagglutinin H gene of Canine distemper virus (CDV), and study its biological characteristics and immunogenicity.【Method】 H gene of CDV was inserted between G and L genes of RABV HEP-dG genome and the full-length cDNA plasmid pHEP-dG(H) was constructed.The plasmid pHEP-dG(H) and help plasmid were co-transfected into BHK cell and the recombinant RABV carrying double G and H genes was rescued.The recombinant virus was inoculated into BHK cells to analyze the diffusion ability of the virus.Mice were inoculated with the recombinant virus, and the pathogenicity and immunogenicity of the virus were analyzed.【Result】 RT-PCR and direct immunofluorescence showed that H gene could still be detected in the third generation virus, indicating that the CDV H gene had been successfully inserted into the RABV genome and stably inherited and correctly expressed in HEP-dG(H).The growth curves of HEP-dG(H) virus and HEP-dG strain maintained similar and the titers of two viruses both reached the peak at 96 h.But the titer of recombinant virus HEP-dG(H) was slightly lower than that of HEP-dG at each time point.Spread and infection experiments showed when the multiplicity of infection (MOI) was 0.005, proliferation ability of HEP-dG(H) was lower than HEP-dG strain.HEP-dG(H) and HEP-dG were not lethal to 6 weeks adult mice, however, HEP-dG(H) had a weaker effect on the body weight of adult mice than HEP-dG.HEP-dG (H) and HEP-dG were both able to induce mice to produce antibody, On the 7th day post infection, antibody reached the level of protection (0.5 EU/mL), and the recombinant virus HEP-dG(H) could also induce neutralizing antibody against CDV.Virus challenge test indicated that antibody induced by HEP-dG(H) and HEP-dG could resist the attack of RABV CVS-24 after immunization for three weeks.【Conclusion】 In this study, the recombinant RABV HEP-dG(H) was successfully constructed, which had good immunogenicity and safety, and could be used as a new dual genetic engineering candidate vaccine of RABV-CDV.

Key words: Rabies virus (RABV); Canine distemper virus (CDV); H gene; rabies inactivated vaccine(dG); immunogenicity

CLC Number:

S852.65^＋5

ZHAO Jing, CHU Ying, LUO Jun, GUO Xiaofeng. Construction and Immunogenicity of Recombinant Rabies Virus with Canine Distemper Virus H Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4776-4785.

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Construction and Immunogenicity of Recombinant Rabies Virus with Canine Distemper Virus H Gene

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