表达VII型新城疫病毒F蛋白和传染性法氏囊病病毒VP2蛋白的重组复制缺陷型人腺病毒5型的免疫原性及保护效力构建与评价
Construction and Evaluation of the Immunogenicity and Protective Efficacy of Recombinant Replication-Deficient Human Adenovirus-5 Expressing Genotype VII Newcastle Disease Virus F Protein and Infectious Bursal Disease Virus VP2 Protein.
作者信息
Xu Ting, Xiong Ting, Xie Wenting, Wu Jing, Liu Xiao, Li Guimin, Lv Yadi, Li Linyu, Yang Zekun, Wang Han, Liu Dingxiang, Chen Ruiai
机构信息
Zhaoqing Branch of Guangdong Laboratory of Lingnan Modern Agricultural Science and Technology, Zhaoqing 526238, China.
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510640, China.
出版信息
Vaccines (Basel). 2023 May 31;11(6):1051. doi: 10.3390/vaccines11061051.
Newcastle disease (ND) and infectious bursal disease (IBD) are two key infectious diseases that significantly threaten the health of the poultry industry. Although existing vaccinations can effectively prevent and treat these two diseases through multiple immunizations, frequent immunization stresses significantly impact chicken growth. In this study, three recombinant adenoviruses, rAd5-F expressing the NDV (genotype VII) F protein, rAd5-VP2 expressing the IBDV VP2 protein, and rAd5-VP2-F2A-F co-expressing F and VP2 proteins, were constructed using the AdEasy system. The F and VP2 genes of the recombinant adenoviruses could be transcribed and expressed normally in HEK293A cells as verified by RT-PCR and Western blot. The three recombinant viruses were shown to have similar growth kinetics as rAd5-EGFP. Compared with the PBS and rAd5-EGFP groups, SPF chickens immunized with recombinant adenoviruses produced higher antibody levels, more significant lymphocyte proliferation, and significantly higher CD4+/CD3+ and CD8+/CD3+ cells in peripheral blood. The survival rate of SPF chickens immunized with rAd5-F and rAd5-VP2-F2A-F after the challenge with DHN3 was 100%, and 86% of SPF chickens showed no viral shedding at 7 dpc. The survival rate of SPF chickens immunized with rAd5-VP2 and rAd5-VP2-F2A-F after the challenge with BC6/85 was 86%. rAd5-VP2 and rAd5-VP2-F2A-F significantly inhibited bursal atrophy and pathological changes compared to the rAd5-EGFP and PBS groups. This study provides evidence that these recombinant adenoviruses have the potential to be developed into safe and effective vaccine candidates for the prevention and control of ND and IBD.
新城疫(ND)和传染性法氏囊病(IBD)是严重威胁家禽业健康的两种关键传染病。尽管现有疫苗接种可通过多次免疫有效预防和治疗这两种疾病,但频繁的免疫应激会显著影响鸡的生长。在本研究中,使用AdEasy系统构建了三种重组腺病毒,即表达新城疫病毒(基因型VII)F蛋白的rAd5 - F、表达传染性法氏囊病病毒VP2蛋白的rAd5 - VP2以及共表达F和VP2蛋白的rAd5 - VP2 - F2A - F。经逆转录聚合酶链反应(RT - PCR)和蛋白质免疫印迹法(Western blot)验证,重组腺病毒的F和VP2基因可在人胚肾细胞系293A(HEK293A)细胞中正常转录和表达。这三种重组病毒显示出与rAd5 - 增强绿色荧光蛋白(rAd5 - EGFP)相似的生长动力学。与磷酸盐缓冲盐水(PBS)组和rAd5 - EGFP组相比,用重组腺病毒免疫的无特定病原体(SPF)鸡产生了更高的抗体水平、更显著的淋巴细胞增殖,并且外周血中CD4⁺/CD3⁺和CD8⁺/CD3⁺细胞显著增多。用rAd5 - F和rAd5 - VP2 - F2A - F免疫的SPF鸡在受到DHN3攻击后的存活率为100%,并且86%的SPF鸡在攻毒后7天未出现病毒脱落。用rAd5 - VP2和rAd5 - VP2 - F2A - F免疫的SPF鸡在受到BC6/85攻击后的存活率为86%。与rAd5 - EGFP和PBS组相比,rAd5 - VP2和rAd5 - VP2 - F2A - F显著抑制了法氏囊萎缩和病理变化。本研究提供了证据表明这些重组腺病毒有潜力被开发成用于预防和控制新城疫和传染性法氏囊病的安全有效的候选疫苗。