J Vet Sci.  2018 Mar;19(2):200-206. 10.4142/jvs.2018.19.2.200.

Packaging of Rift Valley fever virus pseudoviruses and establishment of a neutralization assay method

• Li Y
• Zhao Y
• Wang C
• Zheng X
• Wang H
• Gai W
• Jin H
• Yan F
• Qiu B
• Gao Y
• Li N
• Yang S
• Xia X

• KMID: 2407619
• DOI: http://doi.org/10.4142/jvs.2018.19.2.200

Abstract

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV). RVF is mainly prevalent on the Arabian Peninsula, the African continent, and several islands in the Indian Ocean near southeast Africa. RVFV has been classified by the World Organisation for Animal Health (OIE) as a category A pathogen. To avoid biological safety concerns associated with use of the pathogen in RVFV neutralization assays, the present study investigated and established an RVFV pseudovirus-based neutralization assay. This study used the human immunodeficiency virus (HIV) lentiviral packaging system and RVFV structural proteins to successfully construct RVFV pseudoviruses. Electron microscopy observation and western blotting indicated that the size, structure, and shape of the packaged pseudoviruses were notably similar to those of HIV lentiviral vectors. Infection inhibition assay results showed that an antibody against RVFV inhibited the infective ability of the RVFV pseudoviruses, and an antibody neutralization assay for RVFV detection was then established. This study has successfully established a neutralization assay based on RVFV pseudoviruses and demonstrated that this method can be used to effectively evaluate antibody neutralization.

Keyword

Rift Valley fever virus; antibody neutralization assay; pseudovirus

MeSH Terms

Africa
Animals
Blotting, Western
HIV
Indian Ocean
Islands
Methods*
Microscopy, Electron
Product Packaging*
Rift Valley fever virus*
Rift Valley Fever*
Zoonoses

Figure

• 

  Fig. 1 The M gene polymerase chain reaction (PCR) product. M, marker; Lane 1, the PCR products of the Rift Valley fever virus structural M gene.

• 

  Fig. 2 Enzyme digestion of pcDNA3.1-M-rvfv. M, marker; Lane 1, polymerase chain reaction products of the restriction endonuclease digestion of recombinant plasmid pcDNA3.1-M-rvfv.

• 

  Fig. 3 Detection of pseudoviruses by transmission electron microscopy. Arrows indicate RVFV pseudoviruses. RVFV, Rift Valley fever virus. Scale bar = 200 nm.

• 

  Fig. 4 Identification of pseudoviruses by using western blotting.

• 

  Fig. 5 Determination of Rift Valley fever virus pseudovirus infectious titers. (A) Control. (B–F) Expression of fluorescence in 293T cells infected with different fold dilutions of pseudoviruses. (B) 10−1, (C) 10−2, (D) 10−3, (E) 10−4, (F) 10−5.

• 

  Fig. 6 Rift Valley fever virus (RVFV) pseudovirus infection inhibition assay. (A) Expression of fluorescence in 293T cells after incubation with RVFV pseudoviruses and negative serum reaction mixture. (B) Expression of fluorescence in 293T cells after incubation with the vesicular stomatitis virus (VSV) pseudovirus and anti-RVFV polyclonal antibody reaction mixture. (C) Expression of fluorescence in 293T cells after incubation with RVFV pseudoviruses and anti-RVFV polyclonal antibody reaction mixture.

Reference

1. Anderson GW Jr, Slone TW Jr, Peters CJ. The gerbil, Meriones unguiculatus, a model for Rift Valley fever viral encephalitis. Arch Virol. 1988; 102:187–196.

2. Ashrafi GH, Piuko K, Burden F, Yuan Z, Gault EA, Müller M, Trawford A, Reid SW, Nasir L, Campo MS. Vaccination of sarcoid-bearing donkeys with chimeric virus-like particles of bovine papillomavirus type 1. J Gen Virol. 2008; 89:148–157.

3. Bartosch B, Bukh J, Meunier JC, Granier C, Engle RE, Blackwelder WC, Emerson SU, Cosset FL, Purcell RH. In vitro assay for neutralizing antibody to hepatitis C virus: evidence for broadly conserved neutralization epitopes. Proc Natl Acad Sci U S A. 2003; 100:14199–14204.

4. Bartosch B, Dubuisson J, Cosset FL. Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J Exp Med. 2003; 197:633–642.

5. Boshra H, Lorenzo G, Busquets N, Brun A. Rift valley fever: recent insights into pathogenesis and prevention. J Virol. 2011; 85:6098–6105.

6. Brown BD, Sitia G, Annoni A, Hauben E, Sergi LS, Zingale A, Roncarolo MG, Guidotti LG, Naldini L. In vivo administration of lentiviral vectors triggers a type I interferon response that restricts hepatocyte gene transfer and promotes vector clearance. Blood. 2007; 109:2797–2805.

7. Giroglou T, Cinatl J Jr, Rabenau H, Drosten C, Schwalbe H, Doerr HW, von Laer D. Retroviral vectors pseudotyped with severe acute respiratory syndrome coronavirus S protein. J Virol. 2004; 78:9007–9015.

8. Hilgenfeld R, Peiris M. From SARS to MERS: 10 years of research on highly pathogenic human coronaviruses. Antiviral Res. 2013; 100:286–295.

9. Hofmann H, Hattermann K, Marzi A, Gramberg T, Geier M, Krumbiegel M, Kuate S, Uberla K, Niedrig M, Pöhlmann S. S protein of severe acute respiratory syndrome-associated coronavirus mediates entry into hepatoma cell lines and is targeted by neutralizing antibodies in infected patients. J Virol. 2004; 78:6134–6142.

10. Ikegami T. Molecular biology and genetic diversity of Rift Valley fever virus. Antiviral Res. 2012; 95:293–310.

11. Ikegami T, Makino S. The pathogenesis of Rift Valley fever. Viruses. 2011; 3:493–519.

12. Kim YB, Lee MK, Han DP, Cho MW. Development of a safe and rapid neutralization assay using murine leukemia virus pseudotyped with HIV type 1 envelope glycoprotein lacking the cytoplasmic domain. AIDS Res Hum Retroviruses. 2001; 17:1715–1724.

13. Li C, Bu Z, Chen H. Avian influenza vaccines against H5N1 ‘bird flu’. Trends Biotechnol. 2014; 32:147–156.

14. Li M, Gao F, Mascola JR, Stamatatos L, Polonis VR, Koutsoukos M, Voss G, Goepfert P, Gilbert P, Greene KM, Bilska M, Kothe DL, Salazar-Gonzalez JF, Wei X, Decker JM, Hahn BH, Montefiori DC. Human immunodeficiency virus type 1 env clones from acute and early subtype B infections for standardized assessments of vaccine-elicited neutralizing antibodies. J Virol. 2005; 79:10108–10125.

15. Nie Y, Wang G, Shi X, Zhang H, Qiu Y, He Z, Wang W, Lian G, Yin X, Du L, Ren L, Wang J, He X, Li T, Deng H, Ding M. Neutralizing antibodies in patients with severe acute respiratory syndrome-associated coronavirus infection. J Infect Dis. 2004; 190:1119–1126.

16. Niklasson B, Peters CJ, Grandien M, Wood O. Detection of human immunoglobulins G and M antibodies to Rift Valley fever virus by enzyme-linked immunosorbent assay. J Clin Microbiol. 1984; 19:225–229.

17. Paweska JT, Burt FJ, Anthony F, Smith SJ, Grobbelaar AA, Croft JE, Ksiazek TG, Swanepoel R. IgG-sandwich and IgM-capture enzyme-linked immunosorbent assay for the detection of antibody to Rift Valley fever virus in domestic ruminants. J Virol Methods. 2003; 113:103–112.

18. Qiu C, Huang Y, Zhang A, Tian D, Wan Y, Zhang X, Zhang W, Zhang Z, Yuan Z, Hu Y, Zhang X, Xu J. Safe pseudovirus-based assay for neutralization antibodies against influenza A(H7N9) virus. Emerg Infect Dis. 2013; 19:1685–1687.

19. Rivera A, Messaoudi I. Molecular mechanisms of Ebola pathogenesis. J Leukoc Biol. 2016; 100:889–904.

20. Simmons G, Reeves JD, Rennekamp AJ, Amberg SM, Piefer AJ, Bates P. Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry. Proc Natl Acad Sci U S A. 2004; 101:4240–4245.