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Virologica Sinica

, Volume 30, Issue 3, pp 208–213 | Cite as

Monoclonal neutralizing antibodies against EV71 screened from mice immunized with yeast-produced virus-like particles

  • Tao Lin
  • Lingzhi Xianyu
  • Songya Lyu
Research Article

Abstract

Periodic outbreaks of hand, foot and mouth disease (HFMD) occur in children under 5 years old, and can cause death in some cases. The C4 strain of enterovirus 71 (EV71) is the main pathogen that causes HFMD in China. Although no drugs against EV71 are available, some studies have shown that candidate vaccines or viral capsid proteins can produce anti-EV71 immunity. In this study, female BABL/c mice (6–8 weeks old) were immunized with virus-like particles (VLPs) of EV71 produced in yeast to screen for anti-EV71 antibodies. Two hybridomas that could produce neutralizing antibodies against EV71 were obtained. Both neutralizing mAbs (D4 and G12) were confirmed to bind the VP1 capsid protein of EV71, and could protect >95% cells from 100 TCID50 EV71 infection at 25 µg/mL solution (lowest concentration). Those two neutralizing mAbs identified in the study may be promising candidates in development for mAbs to treat EV71 infection, and utilized as suitable reagents for use in diagnostic tests and biological studies.

Keywords

hand, foot and mouth disease enterovirus 71 (EV71) virus-like particles (VLPs) hybridoma neutralizing antibodies 

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References

  1. Baek K, Park K, Jung E, Chung E, Park J, Choi H, Baek S, Jee Y, Cheon D, Ahn G. 2009. Molecular and epidemiological characterization of enteroviruses isolated in Chungnam, Korea from 2005 to 2006. J Microbiol Biotechnol, 19: 1055–1064.PubMedCrossRefGoogle Scholar
  2. Chang GH, Luo YJ, Wu XY, Si BY, Lin L, Zhu QY. 2010. Monoclonal antibody induced with inactived EV71-Hn2 virus protects mice against lethal EV71-Hn2 virus infection. Virol J, 7: 106.PubMedCentralPubMedCrossRefGoogle Scholar
  3. Chang HW, Liu CC, Lin MH, Ho HM, Yang YT, Chow YH, Chong P, Sia C. 2011. Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates. J Virol Methods, 173: 189–195.PubMedCrossRefGoogle Scholar
  4. Chatproedprai S, Theanboonlers A, Korkong S, Thongmee C, Wananukul S, Poovorawan Y. 2010. Clinical and molecular characterization of hand-foot-and-mouth disease in Thailand, 2008–2009. Jpn J Infect Dis, 63: 229–233.PubMedGoogle Scholar
  5. Chen HF, Chang MH, Chiang BL, Jeng ST. 2006. Oral immunization of mice using transgenic tomato fruit expressing VP1 protein from enterovirus 71. Vaccine, 24: 2944–2951.PubMedCrossRefGoogle Scholar
  6. Chen HL, Huang JY, Chu TW, Tsai TC, Hung CM, Lin CC, Liu FC, Wang LC, Chen YJ, Lin MF, Chen CM. 2008. Expression of VP1 protein in the milk of transgenic mice: a potential oral vaccine protects against enterovirus 71 infection. Vaccine, 26: 2882–2889.PubMedCrossRefGoogle Scholar
  7. Chonmaitree T, Menegus MA, Schervish-Swierkosz EM, Schwalenstocker E. 1981. Enterovirus 71 infection: report of an outbreak with two cases of paralysis and a review of the literature. Pediatrics, 67: 489–493.PubMedGoogle Scholar
  8. Fujimoto T, Chikahira M, Yoshida S, Ebira H, Hasegawa A, Totsuka A, Nishio O. 2002. Outbreak of central nervous system disease associated with hand, foot, and mouth disease in Japan during the summer of 2000. detection and molecular epidemiology of enterovirus 71. Microbiol Immunol, 46: 621–627.PubMedCrossRefGoogle Scholar
  9. Ho M, Chen ER, Hsu KH, Twu SJ, Chen KT, Tsai SF, Wang JR, Shih SR. 1999. An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group. N Engl J Med, 341: 929–935.PubMedCrossRefGoogle Scholar
  10. Lee TC, Guo HR, Su HJ, Yang YC, Chang HL, Chen KT. 2009. Diseases caused by enterovirus 71 infection. Pediatr Infect Dis J, 28: 904–910.PubMedCrossRefGoogle Scholar
  11. Li HY, Han JF, Qin CF, Chen R. 2013. Virus-like particles for enterovirus 71 produced from Saccharomyces cerevisiae potently elicits protective immune responses in mice. Vaccine, 31: 3281–3287.PubMedCrossRefGoogle Scholar
  12. Li X, Mao C, Ma S, Wang X, Sun Z, Yi Y, Guo M, Shen X, Sun L, Bi S. 2009. Generation of neutralizing monoclonal antibodies against Enterovirus 71 using synthetic peptides. Biochem Biophys Res Commun, 390: 1126–1128.PubMedCrossRefGoogle Scholar
  13. Liu CC, Guo MS, Lin FH, Hsiao KN, Chang KH, Chou AH, Wang YC, Chen YC, Yang CS, Chong PC. 2011. Purification and characterization of enterovirus 71 viral particles produced from vero cells grown in a serum-free microcarrier bioreactor system. PLoS One, 6: e20005.PubMedCentralPubMedCrossRefGoogle Scholar
  14. Roldao A, Mellado MC, Castilho LR, Carrondo MJ, Alves PM. 2010. Virus-like particles in vaccine development. Expert Rev Vaccines, 9: 1149–1176.PubMedCrossRefGoogle Scholar
  15. Singh S, Chow VT, Phoon MC, Chan KP, Poh CL. 2002. Direct detection of enterovirus 71 (EV71) in clinical specimens from a hand, foot, and mouth disease outbreak in Singapore by reverse transcription-PCR with universal enterovirus and EV71-specific primers. J Clin Microbiol, 40: 2823–2827.PubMedCentralPubMedCrossRefGoogle Scholar
  16. Sivasamugham LA, Cardosa MJ, Tan WS, Yusoff K. 2006. Recombinant Newcastle Disease virus capsids displaying enterovirus 71 VP1 fragment induce a strong immune response in rabbits. J Med Virol, 78: 1096–1104.PubMedCrossRefGoogle Scholar
  17. Wang X, Peng W, Ren J, Hu Z, Xu J, Lou Z, Li X, Yin W, Shen X, Porta C, Walter TS, Evans G, Axford D, Owen R, Rowlands DJ, Wang J, Stuart DI, Fry EE, Rao Z. 2012. A sensor-adaptor mechanism for enterovirus uncoating from structures of EV71. Nat Struct Mol Biol, 19: 424–429.PubMedCentralPubMedCrossRefGoogle Scholar
  18. Wong SS, Yip CC, Lau SK, Yuen KY. 2010. Human enterovirus 71 and hand, foot and mouth disease. Epidemiol Infect, 138: 1071–1089.PubMedCrossRefGoogle Scholar
  19. Wu CN, Lin YC, Fann C, Liao NS, Shih SR, Ho MS. 2001. Protection against lethal enterovirus 71 infection in newborn mice by passive immunization with subunit VP1 vaccines and inactivated virus. Vaccine, 20: 895–904.PubMedCrossRefGoogle Scholar
  20. Zhang Y, Tan XJ, Wang HY, Yan DM, Zhu SL, Wang DY, Ji F, Wang XJ, Gao YJ, Chen L, An HQ, Li DX, Wang SW, Xu AQ, Wang ZJ, Xu WB. 2009. An outbreak of hand, foot, and mouth disease associated with subgenotype C4 of human enterovirus 71 in Shandong, China. J Clin Virol, 44: 262–267.PubMedCrossRefGoogle Scholar
  21. Zhou SL, Ying XL, Han X, Sun XX, Jin Q, Yang F. 2015. Characterization of the enterovirus 71 VP1 protein as a vaccine candidate. J Med Virol, 87: 256–262.PubMedCrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.State Key Laboratory of Virology, College of Life SciencesWuhan UniversityWuhanChina

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