Shahsavandi Shahla, Ebrahimi Mohammad Majid, Nazari Ali, Khalili Iraj
Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran.
Vet Res Forum. 2024;15(1):49-55. doi: 10.30466/vrf.2023.2009350.3978. Epub 2024 Jan 15.
Purification is an important step in the production of viral vaccines that strongly affects product recovery and subsequent immune responses. The present study was carried out with the aim of improving the purification of infectious bursal disease virus (IBDV) by the tangential flow filtration (TFF) method. Then, the effect of the purified virus on the induction of immune responses against IBDV in specific pathogen free (SPF) chickens was investigated. The IBD07IR strain was propagated in embryonated SPF eggs. The virus was purified using a 100 kDa cassette. The quality of the recovered viruses was evaluated by titration. A total number of 60 SPF chickens were randomly divided into three groups (n = 20) and received the concentrated viral antigen, commercial live IBDV vaccine and phosphate-buffered saline at the age of 3 weeks by eye drop method. The bursa of Fabricius was examined histopathologically for possible changes. Sera were collected at 1-week intervals from day 0 until the end of 6 weeks after vaccination. The IBDV-specific antibody levels, induction of cell-mediated immunity and mRNA expression levels of cytokines were evaluated. The results showed that despite a relative raise in virus titer from 7.66 to 8.17 embryo infectious dose (EID) mL following purification, both the purified IBDV and commercial vaccine are able to induce strong immune responses against the virus. Within a context of egg-based IBDV vaccine production, a single-step TFF can be applied for the relatively purification. This platform requires a further study in the selection of multiple membranes to optimize the operating conditions and final product.
纯化是病毒疫苗生产中的重要步骤,对产品回收率和后续免疫反应有很大影响。本研究旨在通过切向流过滤(TFF)方法改进传染性法氏囊病病毒(IBDV)的纯化。然后,研究了纯化病毒对无特定病原体(SPF)鸡诱导针对IBDV免疫反应的影响。IBD07IR毒株在SPF胚胎蛋中增殖。使用100 kDa滤器纯化病毒。通过滴定评估回收病毒的质量。将60只SPF鸡随机分为三组(n = 20),在3周龄时通过滴眼法分别给予浓缩病毒抗原、商业活IBDV疫苗和磷酸盐缓冲盐水。对法氏囊进行组织病理学检查以观察可能的变化。从接种后第0天开始,每隔1周收集血清,直至6周结束。评估IBDV特异性抗体水平、细胞介导免疫的诱导情况以及细胞因子的mRNA表达水平。结果表明,尽管纯化后病毒滴度从7.66提高到8.17胚感染剂量(EID)/mL,但纯化的IBDV和商业疫苗均能诱导针对该病毒的强烈免疫反应。在基于鸡蛋的IBDV疫苗生产中,单步TFF可用于相对纯化。该平台需要进一步研究以选择多种膜来优化操作条件和最终产品。