Induction of Hantaan virus-specific immune responses in C57BL/6 mice by immunization with a modified recombinant adenovirus containing the chimeric gene, GcS0.7

Li,Kai; Li,Pu-Yuan; Wu,Xing-An; Zhang,Liang; Liu,Zi-Yu; Yu,Lan; Zhang,Lei; Cheng,Lin-Feng; Bai,Wen-Tao; Zhang,Fang-Lin; Xu,Zhi-Kai

doi:10.3892/ijmm.2013.1421

Induction of Hantaan virus-specific immune responses in C57BL/6 mice by immunization with a modified recombinant adenovirus containing the chimeric gene, GcS0.7

Authors:
- Kai Li
- Pu-Yuan Li
- Xing-An Wu
- Liang Zhang
- Zi-Yu Liu
- Lan Yu
- Lei Zhang
- Lin-Feng Cheng
- Wen-Tao Bai
- Fang-Lin Zhang
- Zhi-Kai Xu
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Affiliations: Department of Microbiology, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Minimally Invasive Surgery of the Military General Surgery Center, General Hospital of People's Liberation Army Chengdu Military Region, Chengdu, Sichuan 610083, P.R. China
Published online on: June 20, 2013 https://doi.org/10.3892/ijmm.2013.1421
Pages: 709-716

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Abstract

Hantavirus glycoprotein Gc is one of the main components that contribute to the generation of humoral immune responses, while the nucleocapsid protein (NP) is involved in cellular immune responses through the induction of antibody-dependent cytotoxic T cells. In this study, a chimeric gene, GcS0.7, which encodes a fusion protein containing Gc and truncated NP, was constructed as a candidate for Hantaan virus (HTNV) vaccine development. The chimeric gene was cloned into an adenoviral vector in conjunction with the powerful hybrid cytomegalovirus (CMV) enhancer/chicken β-actin (CAG) promoter or the woodchuck hepatitis virus (WHV) post-transcriptional regulatory element (WPRE), or both. Both elements increased the expression level of the fusion protein. The rAd-GcS0.7-pCAG group demonstrated the highest fusion protein expression level, with a 2.3‑fold increase compared with the unmodified adenoviral vector. To further evaluate the humoral and cellular immunity induced by the recombinant adenovirus, the antibody titers, interferon (IFN)-γ secretion level and cytotoxic T cell ratio were detected in immunized mice. The strongest HTNV‑specific humoral and cellular immune responses were detected in the rAd-GcS0.7‑pCAG group. The immunogenicity of these recombinant adenoviruses was compared with that of the inactivated vaccine through a series of immunological assays. In terms of the cellular immune responses, the rAd-GcS0.7-pCAG group even exceeded those induced by the vaccine control. The CAG hybrid promoter improved not only the expression level, but also the immunogenicity of the fusion protein, and may thus provide a promising strategy for HTNV vaccine research.

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1	Hepojoki J, Strandin T, Lankinen H and Vaheri A: Hantavirus structure - molecular interactions behind the scene. J Gen Virol. 93:1631–1644. 2012. View Article : Google Scholar : PubMed/NCBI
2	Plyusnin A, Vapalahti O and Vaheri A: Hantaviruses: genome structure, expression and evolution. J Gen Virol. 77:2677–2687. 1996. View Article : Google Scholar
3	Charrel RN, Coutard B, Baronti C, et al: Arenaviruses and hantaviruses: from epidemiology and genomics to antivirals. Antiviral Res. 90:102–114. 2011. View Article : Google Scholar : PubMed/NCBI
4	Hart CA and Bennett M: Hantavirus infections: epidemiology and pathogenesis. Microbes Infect. 1:1229–1237. 1999. View Article : Google Scholar : PubMed/NCBI
5	Clement JP: Hantavirus. Antiviral Res. 57:121–127. 2003. View Article : Google Scholar : PubMed/NCBI
6	Kruger DH, Schonrich G and Klempa B: Human pathogenic hantaviruses and prevention of infection. Hum Vaccin. 7:685–693. 2011. View Article : Google Scholar : PubMed/NCBI
7	Simmons JH and Riley LK: Hantaviruses: an overview. Comp Med. 52:97–110. 2002.
8	Kariwa H, Yoshimatsu K and Arikawa J: Hantavirus infection in East Asia. Comp Immunol Microbiol Infect Dis. 30:341–356. 2007. View Article : Google Scholar
9	Schmaljohn C: Vaccines for hantaviruses. Vaccine. 27(Suppl 4): D61–D64. 2009. View Article : Google Scholar
10	Khaiboullina SF and St Jeor SC: Hantavirus immunology. Viral Immunol. 15:609–625. 2002. View Article : Google Scholar : PubMed/NCBI
11	Van Epps HL, Schmaljohn CS and Ennis FA: Human memory cytotoxic T-lymphocyte (CTL) responses to Hantaan virus infection: identification of virus-specific and cross-reactive CD8(+) CTL epitopes on nucleocapsid protein. J Virol. 73:5301–5308. 1999.PubMed/NCBI
12	Yoshimatsu K, Yoo YC, Yoshida R, Ishihara C, Azuma I and Arikawa J: Protective immunity of Hantaan virus nucleocapsid and envelope protein studied using baculovirus-expressed proteins. Arch Virol. 130:365–376. 1993. View Article : Google Scholar
13	Xue X, Xu Z and Ma W: Expression of truncated HTNV nucleoprotein and analysis of antigenic epitope. Virol Sin. 15:220–225. 2000.
14	Zhang FL, Wu XA, Luo W, et al: The expression and genetic immunization of chimeric fragment of Hantaan virus M and S segments. Biochem Biophys Res Commun. 354:858–863. 2007. View Article : Google Scholar : PubMed/NCBI
15	Luo W, Zhang F, Yan Y, et al: Immunological properties of a fusion protein containing nucleocapsid protein and glycoprotein Gn of Hantaan virus. Acta Virol. 52:243–249. 2008.PubMed/NCBI
16	Gribaudo G, Ravaglia S, Caliendo A, et al: Interferons inhibit onset of murine cytomegalovirus immediate-early gene transcription. Virology. 197:303–311. 1993. View Article : Google Scholar : PubMed/NCBI
17	Donello JE, Loeb JE and Hope TJ: Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element. J Virol. 72:5085–5092. 1998.PubMed/NCBI
18	Real G, Monteiro F, Burger C and Alves PM: Improvement of lentiviral transfer vectors using cis-acting regulatory elements for increased gene expression. Appl Microbiol Biotechnol. 91:1581–1591. 2011. View Article : Google Scholar : PubMed/NCBI
19	Xu ZL, Mizuguchi H, Mayumi T and Hayakawa T: Woodchuck hepatitis virus post-transcriptional regulation element enhances transgene expression from adenovirus vectors. Biochim Biophys Acta. 1621:266–271. 2003. View Article : Google Scholar : PubMed/NCBI
20	Zufferey R, Donello JE, Trono D and Hope TJ: Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J Virol. 73:2886–2892. 1999.
21	Garg S, Oran AE, Hon H and Jacob J: The hybrid cytomegalovirus enhancer/chicken beta-actin promoter along with woodchuck hepatitis virus posttranscriptional regulatory element enhances the protective efficacy of DNA vaccines. J Immunol. 173:550–558. 2004. View Article : Google Scholar
22	Li PY, Yu L, Wu XA, et al: Modification of the adenoviral transfer vector enhances expression of the Hantavirus fusion protein GnS0.7 and induces a strong immune response in C57BL/6 mice. J Virol Methods. 179:90–96. 2012. View Article : Google Scholar : PubMed/NCBI
23	Xu Z, Wei L, Wang L, Wang H and Jiang S: The in vitro and in vivo protective activity of monoclonal antibodies directed against Hantaan virus: potential application for immunotherapy and passive immunization. Biochem Biophys Res Commun. 298:552–558. 2002. View Article : Google Scholar
24	Zhang F, Liu Y, Yu L, et al: Construction and identification of recombinant adenovirus containing chimeric gene G2S0.7 of Hantaan virus. J Fourth Mil Med Univ. 25:1057–1060. 2004.
25	Kruger DH, Ulrich R and Lundkvist AA: Hantavirus infections and their prevention. Microbes Infect. 3:1129–1144. 2001. View Article : Google Scholar : PubMed/NCBI
26	Hooper JW and Li D: Vaccines against hantaviruses. Curr Top Microbiol Immunol. 256:171–191. 2001.PubMed/NCBI
27	Schonrich G, Rang A, Lutteke N, Raftery MJ, Charbonnel N and Ulrich RG: Hantavirus-induced immunity in rodent reservoirs and humans. Immunol Rev. 225:163–189. 2008. View Article : Google Scholar : PubMed/NCBI
28	Tischler ND, Gonzalez A, Perez-Acle T, Rosemblatt M and Valenzuela PD: Hantavirus Gc glycoprotein: evidence for a class II fusion protein. J Gen Virol. 86:2937–2947. 2005. View Article : Google Scholar : PubMed/NCBI
29	Klein R, Ruttkowski B, Knapp E, Salmons B, Gunzburg WH and Hohenadl C: WPRE-mediated enhancement of gene expression is promoter and cell line specific. Gene. 372:153–161. 2006. View Article : Google Scholar : PubMed/NCBI
30	Richardson JS, Yao MK, Tran KN, et al: Enhanced protection against Ebola virus mediated by an improved adenovirus-based vaccine. PLoS One. 4:e53082009. View Article : Google Scholar : PubMed/NCBI
31	Lehmann PV and Zhang W: Unique strengths of ELISPOT for T cell diagnostics. Methods Mol Biol. 792:3–23. 2012. View Article : Google Scholar : PubMed/NCBI