Muniraju Murali, Mahapatra Mana, Buczkowski Hubert, Batten Carrie, Banyard Ashley C, Parida Satya
The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK.
Animal and Plant Health Agency, Weybridge, Surrey KT15 3NB, UK.
Vaccine. 2015 Jan 9;33(3):465-71. doi: 10.1016/j.vaccine.2014.10.050. Epub 2014 Nov 1.
Across the developing world peste des petits ruminants virus places a huge disease burden on agriculture, primarily affecting the production of small ruminant. The disease is most effectively controlled by vaccinating sheep and goats with live attenuated vaccines that provide lifelong immunity. However, the current vaccines and serological tests are unable to enable Differentiation between naturally Infected and Vaccinated Animals (DIVA). This factor precludes meaningful assessment of vaccine coverage and epidemiological surveillance based on serology, in turn reducing the efficiency of control programmes. The availability of a recombinant PPRV vaccine with a proven functionality is a prerequisite for the development of novel vaccines that may enable the development of DIVA tools for PPRV diagnostics. In this study, we have established an efficient reverse genetics system for PPRV Nigeria 75/1 vaccine strain and, further rescued a version of PPRV Nigeria 75/1 vaccine strain that expresses eGFP as a novel transcription cassette and a version of PPRV Nigeria 75/1 vaccine strain with mutations in the haemagglutinin (H) gene to enable DIVA through disruption of binding to H by the C77 monoclonal antibody used in the competitive (c) H-ELISA. All three rescued viruses showed similar growth characteristics in vitro in comparison to parent vaccine strain and, following in vivo assessment the H mutant provided full protection in goats. Although the C77 monoclonal antibody used in the cH-ELISA was unable to bind to the mutated form of H in vitro, the mutation was not sufficient to enable DIVA in vivo.
在整个发展中世界,小反刍兽疫病毒给农业带来了巨大的疾病负担,主要影响小型反刍动物的生产。通过给绵羊和山羊接种能提供终身免疫力的减毒活疫苗,可以最有效地控制这种疾病。然而,目前的疫苗和血清学检测无法区分自然感染动物和接种疫苗动物(DIVA)。这一因素妨碍了基于血清学对疫苗接种覆盖率进行有意义的评估以及流行病学监测,进而降低了防控计划的效率。拥有一种经证实具有功能的重组小反刍兽疫病毒疫苗,是开发新型疫苗的先决条件,这些新型疫苗可能有助于开发用于小反刍兽疫病毒诊断的DIVA工具。在本研究中,我们为小反刍兽疫病毒尼日利亚75/1疫苗株建立了一个高效的反向遗传学系统,并进一步拯救出了一个表达绿色荧光蛋白(eGFP)作为新型转录盒的小反刍兽疫病毒尼日利亚75/1疫苗株版本,以及一个血凝素(H)基因发生突变的小反刍兽疫病毒尼日利亚75/1疫苗株版本,通过竞争性(c)H-ELISA中使用的C77单克隆抗体破坏与H的结合来实现DIVA。与亲本疫苗株相比,所有三种拯救出的病毒在体外均表现出相似的生长特性,并且在体内评估后,H突变株在山羊中提供了完全保护。尽管cH-ELISA中使用的C77单克隆抗体在体外无法与突变形式的H结合,但该突变不足以在体内实现DIVA。