Kebede Wakjira, Bitew Molalegne, Bari Fufa Dawo, Edao Bedaso Mammo, Mohammed Hawa, Yami Martha, Getachew Belayneh, Abayneh Takele, Gelaye Esayas
National Veterinary Institute, Bishoftu, Ethiopia.
Ethiopian Biotechnology Institute, Addis Ababa, Ethiopia.
Vet Med (Auckl). 2021 Oct 1;12:261-270. doi: 10.2147/VMRR.S326479. eCollection 2021.
Infectious bursal disease virus (IBDV) is an avian viral pathogen that causes infectious bursal disease (IBD) of chickens. The disease has been endemic in Ethiopia since 2002, and vaccination has been practiced as the major means of disease prevention and control. An IBD vaccine is produced in Ethiopia using primary chicken embryo fibroblast (CEF) cell, which is time-consuming, laborious, and uneconomical. The present study was carried out to develop cell-based IBDV LC-75 vaccine using Vero cells and to evaluate the safety, immunogenicity and protection level.
Identity of the vaccine seed was confirmed with gene-specific primers using reverse transcription polymerase chain reaction (RT-PCR). Confluent monolayer of Vero cells was infected with vaccine virus and serial passage continued till passage 10. A characteristic virus-induced cytopathic effect (CPE) was observed starting from passage 2 on the third day post-infection. The infectious titer of adapted virus showed a linear increment along the passage level. The virus-induced specific antibody was determined using indirect ELISA after vaccination of chicks through ocular route.
The antibody titer measured from Vero cells vaccinated chicks revealed similar level with the currently available CEF cell-based vaccine, hence no significant difference. Chicks vaccinated with Vero cell adapted virus showed complete protection against very virulent IBDV, while unvaccinated group had 60% morbidity and 25% mortality.
It is concluded that the IBDV vaccine strain well adapted on Vero cells and found to be immunogenic induces antibody development and successfully protects chicks against challenge with the circulating field IBDV isolate. Hence, it is recommended to produce IBD vaccine using Vero cell culture at the industrial scale to conquer the limitations caused by using CEF cells and thus to vaccinate chicks population to protect against the circulating IBDV infection.
传染性法氏囊病病毒(IBDV)是一种引起鸡传染性法氏囊病(IBD)的禽类病毒病原体。自2002年以来,该病在埃塞俄比亚一直呈地方流行状态,疫苗接种一直是疾病预防和控制的主要手段。埃塞俄比亚使用原代鸡胚成纤维细胞(CEF)生产IBD疫苗,这既耗时、费力又不经济。本研究旨在利用Vero细胞开发基于细胞的IBDV LC - 75疫苗,并评估其安全性、免疫原性和保护水平。
使用逆转录聚合酶链反应(RT - PCR),通过基因特异性引物确认疫苗种子的身份。用疫苗病毒感染汇合的Vero细胞单层,并连续传代直至第10代。从感染后第三天的第2代开始观察到特征性的病毒诱导细胞病变效应(CPE)。适应病毒的感染滴度随传代水平呈线性增加。通过眼内途径给雏鸡接种疫苗后,使用间接ELISA测定病毒诱导的特异性抗体。
从接种Vero细胞的雏鸡测得的抗体滴度与目前可用的基于CEF细胞的疫苗相似,因此无显著差异。接种Vero细胞适应病毒的雏鸡对超强毒IBDV表现出完全保护,而未接种疫苗的组发病率为60%,死亡率为25%。
得出结论,IBDV疫苗株能很好地适应Vero细胞,具有免疫原性,可诱导抗体产生,并成功保护雏鸡免受循环野毒株IBDV的攻击。因此,建议在工业规模上使用Vero细胞培养生产IBD疫苗,以克服使用CEF细胞带来的局限性,从而为雏鸡群体接种疫苗以预防循环IBDV感染。