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Protective immunity of E. coli-synthesized NS1 protein of Japanese encephalitis virus

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Abstract

Immunogenicity and protective efficacy of recombinant Japanese encephalitis virus (JEV) NS1 proteins generated using DNA vaccines and recombinant viruses have been demonstrated to induce protection in mice against a challenge of JEV at a lethal dose. The West Nile virus NS1 region expressed in E. coli is recognized by these protective monoclonal antibodies and, in this study, we compare immunogenicity and protective immunity of the E. coli-synthesized NS1 protein with another protective immunogen, the envelope domain III (ED3). Pre-challenge, detectable titers of JEV-specific neutralizing antibody were detected in the immunized mice with E. coli-synthesized ED3 protein (PRNT50 = 1:28) and the attenuated JEV strain T1P1 (PRNT50 = 1:53), but neutralizing antibodies were undetectable in the immunized mice with E. coli-synthesized NS1 protein (PRNT50 < 1:10). However, the survival rate of the NS1-immunized mice against the JEV challenge was 87.5% (7/8), showing significantly higher levels of protection than the ED3-immunized mice, 62.5% (5/8) (P = 0.041). In addition, E. coli-synthesized NS1 protein induced a significant increase of anti-NS1 IgG1 antibodies, resulting in an ELISA titer of 100,1000 in the immunized sera before lethal JEV challenge. Surviving mice challenged with the virulent JEV strain Beijing-1 showed a ten-fold or greater rise in IgG1 and IgG2b titers of anti-NS1 antibodies, implying that the Th2 cell activation might be predominantly responsible for antibody responses and mice protection.

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References

  • Abbas AK, Murphy KM, Sher A (1996) Functional diversity of helper T lymphocytes. Nature 383:787–793

    Article  PubMed  CAS  Google Scholar 

  • Burke DS, Monath TP (2001) Flaviviridae. In: Knipe DM, Howley PM (eds) Fields virology, 4th ed., Lippincott-Williams and Wilkins, New York, pp 1043–1125

    Google Scholar 

  • Chia SC, Leung PS, Liao CP, Huang JH, Lee ST (2001) Fragment of Japanese encephalitis virus envelope protein produced in Escherichia coli protects mice from virus challenge. Microb Pathog 31:9–19

    Article  PubMed  CAS  Google Scholar 

  • Chiou SS, Chen WJ (2001) Mutations in the NS3 gene and 3′-NCR of Japanese encephalitis virus isolated from an unconventional ecosystem and implications for natural attenuation of the virus. Virology 289:129–136

    Article  PubMed  CAS  Google Scholar 

  • Chung KM, Nybakken GE, Thompson BS, Engle MJ, Marri A, Fremont DH, Diamond MS (2006) Antibodies against West Nile Virus nonstructural protein NS1 prevent lethal infection through Fc gamma receptor-dependent and -independent mechanisms. J Virol 80:1340–1351

    Article  PubMed  CAS  Google Scholar 

  • Fan WF, Mason PW (1990) Membrane accociation and secretion of the Japanese encephalitis virus NS1 protein from cells expressing NS1 cDNA. Virology 177:470–476

    Article  PubMed  CAS  Google Scholar 

  • Flamand M, Chevalier M, Henchal E, Girard M, Deubel V (1995) Purification and renaturation of Japanese encephalitis virus nonstructural glycoprotein NS1 overproduced by insect cells. Protein Expr Purif 6:519–527

    Article  PubMed  CAS  Google Scholar 

  • Kar NJ, Saxena VK (1998) Some epidemiological characteristics of Japanese encephalitis in Haryana state of northern India. J Commun Dis 30:129–131

    PubMed  CAS  Google Scholar 

  • Konishi E, Pincus S, Paoletti E, Shope RE, Mason PW (1994) Avipox virus-vectored Japanese encephalitis virus vaccines: use as vaccine candidates in combination with purified subunit immunogens. Vaccine 12:633–638

    Article  PubMed  CAS  Google Scholar 

  • Konishi E, Kurane I, Mason PW, Shope RE, Ennis FA (1997) Poxvirus-based Japanese encephalitis virus vaccine candidates induce JE virus-specific CD8+ cytotoxic T lymphocytes in mice. Virology 227:353–360

    Article  PubMed  CAS  Google Scholar 

  • Lieberman MM, Clements DE, Ogata S, Wang G, Corpuz G, Wong T, Martyak T, Gilson L, Coller BA, Leung J, Watts DM, Tesh RB, Siirin M, Travassos da Rosa A, Humphreys T, Weeks-Levy C (2007) Preparation and immunogenic properties of a recombinant West Nile subunit vaccine. Vaccine 25:414–423

    Article  PubMed  CAS  Google Scholar 

  • Lin CW, Wu SC (2003) A functional epitope determinant on domain III of the Japanese encephalitis virus envelope protein interactions with neutralizing-antibody combining sites. J Virol 77:2600–2606

    Article  PubMed  CAS  Google Scholar 

  • Lin YL, Chen LK, Liao CL, Yeh CT, Ma SH, Chen JL, Huang YL, Chen SS, Chiang HY (1998) DNA immunization with Japanese encephalitis virus nonstructural protein NS1 elicits protective immunity in mice. J Virol 72:191–200

    PubMed  CAS  Google Scholar 

  • Lindenbach BD, Rice MC (2001) Flaviviridae: the viruses and their replication. In: Knipe DM, Howley PM (eds), Fields virology, 4th ed, Lippincott-Williams and Wilkins, New York, pp 991–1041

    Google Scholar 

  • Mason PW, McAda PC, Dalrymple JM, Fournier MJ, Mason TL (1987) Expression of Japanese encephalitis virus antigens in Escherichia coli. Virology 158:361–72

    Article  PubMed  CAS  Google Scholar 

  • Mason PW, Pincus S, Fournier MJ, Mason TL, Shope RE, Paoletti E (1991) Japanese encephalitis virus-vaccinia recombinants produce particulate forms of the structural membrane proteins and induce high levels of protection against lethal JEV infection. Virology 180:294–305

    Article  PubMed  CAS  Google Scholar 

  • McCown J, Cochran M, Putnak R (1990) Protection of mice against lethal Japanese encephalitis with a recombinant baculovirus vaccine. Am J Trop Med Hyg 42:491–499

    PubMed  CAS  Google Scholar 

  • McMinn PC (1997) The molecular basis of virulence of the encephalitogenic flaviviruses. J Gen Virol 78:2711–2722

    PubMed  CAS  Google Scholar 

  • Plesner AM, Ronne T (1997) Allergic mucocutaneous reactions to Japanese encephalitis vaccine. Vaccine 15:1239–1243

    Article  PubMed  CAS  Google Scholar 

  • Poland JD, Cropp CB, Craven RB, Monath TP (1990) Evaluationof the potency and safety of inactivated Japanese encephalitis vaccine in US inhabitants. J Infect Dis 161:817–882

    Google Scholar 

  • Pugachev KV, Mason PW, Shope RE, Frey TK (1995) Double subgenomic sindbis virus recombinants expressing immunogenic proteins of Japanese encephalitis virus induce significant protection against lethal JEV infection. Virology 212:587–594

    Article  PubMed  CAS  Google Scholar 

  • Rey FA, Heinz FX, Mandl C, Kunz C, Harrison SC (1995) The envelope glycoprotein from tick-borne encephalitis virus at 2 A resolution. Nature 375:291–298

    Article  PubMed  CAS  Google Scholar 

  • Sakaguchi M, Yoshida M, Kuroda W, Harayama O, Matsunaga Y, Inouye S (2000) Systemic immediate-type reactions to gelatin included in Japanese encephalitis vaccines. Vaccine 15:121–122

    Article  Google Scholar 

  • Solomon T, Dung NM, Kneen R, Gainsborough M, Vaughn DW, Khanh VT (2000) Japanese encephalitis. J Neurol Neurosurg Psychiatry 68:405–415

    Article  PubMed  CAS  Google Scholar 

  • Volpina OM, Volkova TD, Koroev DO, Ivanov VT, Ozherelkov SV, Khoretonenko MV, Vorovitch MF, Stephenson JR, Timofeev AV (2005) A synthetic peptide based on the NS1 non-structural protein of tick-borne encephalitis virus induces a protective immune response against fatal encephalitis in an experimental animal model. Virus Res 112:95–99

    Article  PubMed  CAS  Google Scholar 

  • WHO (2005) Global Advisory Committee on Vaccine Safety, 9–10 June 2005, Wkly. Epidemiol Rec 80:242–243

    Google Scholar 

  • Wu HH, Chen CT, Lin YL, Lee ST (2004) Sub-fragments of the envelope gene are highly protective against the Japanese encephalitis virus lethal infection in DNA priming–protein boosting immunization strategies. Vaccine 22:793–800

    PubMed  Google Scholar 

  • Wu SC, Lian WC, Hsu LC, Liau MY (1997) Japanese encephalitis virus antigenic variants with characteristic differences in neutralization resistance and mouse virulence. Virus Res 51:173–181

    Article  PubMed  CAS  Google Scholar 

  • Wu SC, Yu CH, Lin CW, Chu IM (2003) The domain III fragment of Japanese encephalitis virus envelope protein: mouse immunogenicity and liposome adjuvanticity. Vaccine 21:2516–2252

    Article  PubMed  CAS  Google Scholar 

  • Xu G, Xu X, Li Z, He Q, Wu B, Sun S, Chen H (2004) Construction of recombinant pseudorabies virus expressing NS1 protein of Japanese encephalitis (SA14-14-2) virus and its safety and immunogenicity. Vaccine 22:1846–1853

    Article  PubMed  CAS  Google Scholar 

  • Yang DK, Kweon CH, Kim BH, Lim SI, Kwon JH, Kim SH, Song JY, Han HR (2005) Immunogenicity of baculovirus expressed recombinant proteins of Japanese encephalitis virus in mice. J Vet Sci 6:125–133

    PubMed  Google Scholar 

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Acknowledgements

We would like to thank the China Medical University and National Science Council, Taiwan for financial supports (CMU94-086, NSC94-2320-B-039-010 and NSC94-2745-B-039-003-URD).

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Authors and Affiliations

  1. Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 404, Taiwan, ROC

    Cheng-Wen Lin, Kuang-Ting Liu & Hong-Da Huang

  2. Clinical Virology Laboratory, Department of Laboratory Medicine, China Medical University Hospital, Taichung, 404, Taiwan, ROC

    Cheng-Wen Lin

  3. Department of Public Health and Parasitology, College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan, ROC

    Wei-June Chen

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  1. Cheng-Wen Lin
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  2. Kuang-Ting Liu
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Correspondence to Cheng-Wen Lin.

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Lin, CW., Liu, KT., Huang, HD. et al. Protective immunity of E. coli-synthesized NS1 protein of Japanese encephalitis virus. Biotechnol Lett 30, 205–214 (2008). https://doi.org/10.1007/s10529-007-9529-9

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  • DOI: https://doi.org/10.1007/s10529-007-9529-9

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