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鞭毛蛋白的截短形式(tFlic)使二维帽亚基疫苗在小鼠中具有更好的免疫原性和保护作用。

The truncated form of flagellin (tFlic) provides the 2dCap subunit vaccine with better immunogenicity and protective effects in mice.

作者信息

Lu Ying, Liu Zehui, Li Yingxiang, Deng Zhuofan, Fang Weihuan, He Fang

机构信息

Department of Veterinary Medicine, Institute of Preventive Veterinary Medicine & Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, 866 Yuhangtang road, Hangzhou, 310058 China.

出版信息

Anim Dis. 2022;2(1):11. doi: 10.1186/s44149-022-00043-x. Epub 2022 Jun 2.

Abstract

Porcine circovirus type 2 (PCV2) is the main causative agent of porcine circovirus-associated diseases, and it causes substantial economic losses in the swine industry each year. It is crucial to develop an effective vaccine against the circulating strain PCV2d, which is prone to substantial degrees of mutation. In this study, a truncated form of flagellin (tFlic: 85-111 aa) was inserted into the C-terminal sequence of 2dCap, and Western blotting results showed that recombinant Cap-tFlic VLPs were successfully expressed. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) data indicated that purified recombinant Cap-tFlic fusion proteins existed in the form of polymers and that tFlic could not affect the formation and internalization of VLPs. Integrated Cap-tFlic VLPs induced the expression of antigen presentation-related factors (MHC-II and CD86) by bone marrow-derived dendritic cells (BM-DCs), and the expression of TLR5-related factors (TNF-α) was dramatically elevated. Mice intramuscularly immunized with Cap-tFlic VLPs exhibited significantly higher levels of Cap-specific antibodies and neutralizing antibodies than mice immunized with wild-type Cap VLPs. The data obtained in the current study indicate that Cap-tFlic may be a candidate for a subunit vaccine against PCV2 in the future.

摘要

猪圆环病毒2型(PCV2)是猪圆环病毒相关疾病的主要病原体,每年给养猪业造成巨大经济损失。开发一种针对流行毒株PCV2d的有效疫苗至关重要,因为该毒株极易发生大量突变。在本研究中,将截短形式的鞭毛蛋白(tFlic:85-111氨基酸)插入2dCap的C末端序列,蛋白质免疫印迹结果表明重组Cap-tFlic病毒样颗粒(VLPs)成功表达。透射电子显微镜(TEM)和动态光散射(DLS)数据表明,纯化的重组Cap-tFlic融合蛋白以聚合物形式存在,且tFlic不影响VLPs的形成和内化。整合的Cap-tFlic VLPs诱导骨髓来源的树突状细胞(BM-DCs)表达抗原呈递相关因子(MHC-II和CD86),且TLR5相关因子(TNF-α)的表达显著升高。用Cap-tFlic VLPs肌肉注射免疫的小鼠比用野生型Cap VLPs免疫的小鼠表现出显著更高水平的Cap特异性抗体和中和抗体。本研究获得的数据表明,Cap-tFlic未来可能成为一种针对PCV2的亚单位疫苗候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/690a/9160859/1fddacce2281/44149_2022_43_Fig1_HTML.jpg

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