表达PEDVRBD结构域的重组仙台病毒构建及其免疫原性评价
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中图分类号：S852.65 文献标志码：A
Construction of a recombinant Sendai virus expressing the PEDV RBD domain and its immunogenicity evaluation
WEI Liping1，LI Jiarui1，PAN Shuonan1，SHI Kaichuang²，CHEN Zhenhai1’³，MOU Chunxiao 1，3
（1. College of Veterinary Medicine， Yangzhou University， Yangzhou 2250o9， China;2. Guangxi Zhuang Autonomous Region Animal Disease Prevention and Control Center， Nanning 530028， China;3. Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Diseases and Zoonosis，Yangzhou 225oo9，China）
ABSTRACT：To construct a recombinant Sendai virus（SeV）expressing the receptor-binding domain（RBD）of porcine epidemic diarea virus （PEDV） and evaluate its immunogenicity，we designed a sequence-optimized PEDV spike（S） gene RBD domain （amino acids 507-757 ）engineered to contain four pairs of intramolecular disulfide bonds，referred to as RBD（8C）. The optimized gene fragment was inserted into a SeV vector to generate the recombinant plasmid pBeloBAC-SeV-PE-RBD（8C）-3Flag.Folowing virus rescue andcharacterizationof its biological properties，therecombinant virus was used to immunize mice. PEDV-specific IgG and IgA antibody levels in serum and bronchoalveolar lavage fluid （BALF），serum neutralizing antibody titers，as well as ΔT lymphocyte proliferation and T cell subset activation in lymphoid tisses，were evaluated.The results demonstrated that the recombinant SeV specifically expressed the RBD（8C）proteinand exhibiteda growth curve similar tothatof wild-type SeV.Immunization withtherecombinantvirus efectively induced PEDV-specific IgG ， IgA，and neutralizing antibodies in mice，significantly promoted PEDV-specific T cell proliferation，and increased the proportions of CD4+ and CD8+T cells in spleen and lymph nodes. In conclusion，the recombinant virus induced robust humoral，cellular，and mucosal immune responses in mice，providing experimental evidence to sup port the development and optimization of PEDV vaccines.
KEY WORDS： PEDV; RBD protein; Sendai virus vector; immunogenicity evaluation
猪流行性腹泻病毒（porcine epidemic diarrhea virus，PEDV）是一种引起猪流行性腹泻的接触性肠道病毒，该病毒可感染各生长阶段的猪群，尤其是哺乳仔猪，引起以急性肠炎和水样腹泻为主的临床症状，新生仔猪发病率和死亡率高达 100%[1] 。（剩余15738字）
