Yang Yupeng, Qi Ruibin, Chen Mengru, Feng Kexin, Liu Zhe, Kang Hongtao, Jiang Qian, Qu Liandong, Liu Jiasen
State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
Vaccines (Basel). 2024 Oct 24;12(11):1205. doi: 10.3390/vaccines12111205.
Feline calicivirus (FCV), a pathogen that causes upper respiratory tract diseases in felids, primarily leads to oral ulcers and various respiratory symptoms, which can be fatal in severe cases. Currently, FCV prevention and control rely primarily on vaccination; however, the existing vaccine types in China are mainly inactivated vaccines, leading to a single prevention and control method with suboptimal outcomes.
This study commences with a genetic evolution analysis of Chinese FCV isolates, confirming the presence of two major genotypes, GI and GII with GI emerging as the dominant form. We subsequently selected the broadly neutralizing vaccine candidate strain DL39 as the template for the truncation and expression of multiple recombinant proteins. Through serological assays, we successfully confirmed the optimal protective antigen region, which is designated CE (CDE). Further investigation revealed the location of the optimal protective antigen region within the CE region for both the GI and GII genotype strains. Capitalizing on this discovery, a bivalent recombinant protein, designated CE-CE, was generated. Cat antisera generated against CE and CE-CE proteins were used in cross-neutralization against various strains of different genotypes, yielding high neutralization titers ranging from 1:45 to 1:15 and from 1:48 to 1:29, respectively, which surpassed those induced by antisera from cats vaccinated with Mi-aosanduo (commercial vaccine, strain 255). Ultimately, in vivo challenge experiments were per-formed after immunizing cats with the CE and CE-CE proteins, utilizing Miaosanduo as a control for comparison. The results demonstrated that immunization with both proteins effectively made cats less susceptible to FCV GI, GII, and VSD strains infection, resulting in superior immune efficacy compared with that in the Miaosanduo group.
These results indicate that this study successfully identified the antigen CE, which has broad-spectrum antigenicity, through in vivo and in vitro experiments. These findings pre-liminarily demonstrate that the optimal protective antigen region of FCV strains is the CE region, laying a theoretical foundation for the development of novel broad-spectrum vaccines against FCV disease.
猫杯状病毒(FCV)是一种可导致猫科动物上呼吸道疾病的病原体,主要引发口腔溃疡和各种呼吸道症状,严重时可能致命。目前,FCV的防控主要依靠疫苗接种;然而,中国现有的疫苗类型主要是灭活疫苗,导致防控方法单一,效果欠佳。
本研究首先对中国FCV分离株进行遗传进化分析,确定存在两种主要基因型,即GI和GII,其中GI为优势型。随后,我们选择具有广泛中和作用的候选疫苗株DL39作为模板,进行多种重组蛋白的截短和表达。通过血清学检测,我们成功确定了最佳保护性抗原区域,命名为CE(CDE)。进一步研究揭示了GI和GII基因型毒株的最佳保护性抗原区域在CE区域内的位置。基于这一发现,构建了一种二价重组蛋白,命名为CE-CE。用针对CE和CE-CE蛋白产生的猫抗血清对不同基因型的各种毒株进行交叉中和试验,中和效价分别高达1:45至1:15和1:48至1:29,超过了用妙三多(商业疫苗,毒株255)免疫的猫所产生的抗血清诱导的中和效价。最终,在用CE和CE-CE蛋白免疫猫后进行体内攻毒实验,并以妙三多作为对照进行比较。结果表明,用这两种蛋白免疫可有效降低猫对FCV GI、GII和VSD毒株感染的易感性,免疫效果优于妙三多组。
这些结果表明,本研究通过体内和体外实验成功鉴定出具有广谱抗原性的抗原CE。这些发现初步证明FCV毒株的最佳保护性抗原区域是CE区域,为开发新型抗FCV疾病的广谱疫苗奠定了理论基础。