Virologica Sinica

, Volume 27, Issue 4, pp 221–227 | Cite as

Replication kinetics of coxsackievirus A16 in human rhabdomyosarcoma cells

  • Jun Jin
  • Mingming Han
  • Lin Xu
  • Dong An
  • Wei KongEmail author
  • Chunlai JiangEmail author
Research Article


Coxsackievirus A16 (CVA16), together with enterovirus type 71 (EV71), is responsible for most cases of hand, foot and mouth disease (HFMD) worldwide. Recent findings suggest that the recombination between CVA16 and EV71, and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years. It is therefore important to further understand the virology, epidemiology, virus-host interactions and host pathogenesis of CVA16. In this study, we describe the viral kinetics of CVA16 in human rhabdomyosarcoma (RD) cells by analyzing the cytopathic effect (CPE), viral RNA replication, viral protein expression, viral RNA package and viral particle secretion in RD cells. We show that CVA16 appears to first attach, uncoat and enter into the host cell after adsorption for 1 h. Later on, CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1. At MOI 0.1, CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i. CPE was observed after 12 h p.i., and viral antigen was first detected at 6 h p.i. at MOI 1 and at 9 h p.i. at MOI 0.1. Thus, our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.


Coxsackievirus A16 (CVA16) Hand, foot and mouth disease (HFMD) Viral kinetics qRT-PCR Western blotting 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Ang L W, Koh B K, Chan K P, et al. 2009. Epidemiology and control of hand, foot and mouth disease in Singapore, 2001–2007. Ann Acad Med Singapore, 38(2):106–112.PubMedGoogle Scholar
  2. 2.
    Baccam P, Beauchemin C, Macken C A, et al. 2006. Kinetics of influenza A virus infection in humans. J Virol, 80(15):7590–7599.PubMedCrossRefGoogle Scholar
  3. 3.
    Cardosa M J, Perera D, Brown B A, et al. 2003. Molecular epidemiology of human enterovirus 71 strains and recent outbreaks in the Asia-Pacific region: comparative analysis of the VP1 and VP4 genes. Emerg Infect Dis, 9(4):461–468.PubMedGoogle Scholar
  4. 4.
    Chun T W, Carruth L, Finzi D, et al. 1997. Quantification of latent tissue reservoirs and total body viral load in HIV-1 infection. Nature, 387(6629):183–188.PubMedCrossRefGoogle Scholar
  5. 5.
    Cromeans T, Fields H A, Sobsey M D. 1989. Replication kinetics and cytopathic effect of hepatitis A virus. J Gen Virol, 70( Pt 8):2051–2062.PubMedCrossRefGoogle Scholar
  6. 6.
    Egger D, Bienz K. 2005. Intracellular location and translocation of silent and active poliovirus replication complexes. J Gen Virol, 86(Pt 3):707–718.PubMedCrossRefGoogle Scholar
  7. 7.
    Garcia P M, Kalish L A, Pitt J, et al. 1999. Maternal levels of plasma human immunodeficiency virus type 1 RNA and the risk of perinatal transmission. Women and Infants Transmission Study Group. N Engl J Med, 341(6):394–402.PubMedCrossRefGoogle Scholar
  8. 8.
    Hadinoto V, Shapiro M, Sun C C, et al. 2009. The dynamics of EBV shedding implicate a central role for epithelial cells in amplifying viral output. PLoS Pathog, 5(7):e1000496.PubMedCrossRefGoogle Scholar
  9. 9.
    Hollingsworth T D, Laeyendecker O, Shirreff G, et al. HIV-1 transmitting couples have similar viral load set-points in Rakai, Uganda. PLoS Pathog, 6(5):e1000876.Google Scholar
  10. 10.
    Hosoya M, Kawasaki Y, Sato M, et al. 2007. Genetic diversity of coxsackievirus A16 associated with hand, foot, and mouth disease epidemics in Japan from 1983 to 2003. J Clin Microbiol, 45(1):112–120.PubMedCrossRefGoogle Scholar
  11. 11.
    Jabs D A, Gilpin A M, Min Y I, et al. 2002. HIV and cytomegalovirus viral load and clinical outcomes in AIDS and cytomegalovirus retinitis patients: Monoclonal Antibody Cytomegalovirus Retinitis Trial. AIDS, 16(6):877–887.PubMedCrossRefGoogle Scholar
  12. 12.
    Kung Y H, Huang S W, Kuo P H, et al. Introduction of a strong temperature-sensitive phenotype into enterovirus 71 by altering an amino acid of virus 3D polymerase. Virology, 396(1):1–9.Google Scholar
  13. 13.
    Lin K H, Hwang K P, Ke G M, et al. 2006. Evolution of EV71 genogroup in Taiwan from 1998 to 2005: an emerging of subgenogroup C4 of EV71. J Med Virol, 78(2):254–262.PubMedCrossRefGoogle Scholar
  14. 14.
    Lu J, He M L, Wang L, et al. MiR-26a inhibits cell growth and tumorigenesis of nasopharyngeal carcinoma through repression of EZH2. Cancer Res, 71(1):225–233.Google Scholar
  15. 15.
    Lu J, He Y Q, Yi L N, et al. Viral kinetics of enterovirus 71 in human abdomyosarcoma cells. World J Gastroenterol, 17(36):4135–4142.Google Scholar
  16. 16.
    Lyles R H, Munoz A, Yamashita T E, et al. 2000. Natural history of human immunodeficiency virus type 1 viremia after seroconversion and proximal to AIDS in a large cohort of homosexual men. Multicenter AIDS Cohort Study. J Infect Dis, 181(3):872–880.Google Scholar
  17. 17.
    Ma Y, Yu J, Chan H L, et al. 2009. Glucose-regulated protein 78 is an intracellular antiviral factor against hepatitis B virus. Mol Cell Proteomics, 8(11):2582–2594.PubMedCrossRefGoogle Scholar
  18. 18.
    McMinn P C. 2002. An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev, 26(1):91–107.PubMedCrossRefGoogle Scholar
  19. 19.
    Mellors J W, Munoz A, Giorgi J V, et al. 1997. Plasma viral load and CD4+ lymphocytes as prognostic markers of HIV-1 infection. Ann Intern Med, 126(12):946–954.PubMedGoogle Scholar
  20. 20.
    Muir P, Kammerer U, Korn K, et al. 1998. Molecular typing of enteroviruses: current status and future requirements. The European Union Concerted Action on Virus Meningitis and Encephalitis. Clin Microbiol Rev, 11(1):202–227.PubMedGoogle Scholar
  21. 21.
    Neumann A U, Lam N P, Dahari H, et al. 1998. Hepatitis C viral dynamics in vivo and the antiviral efficacy of interferon-alpha therapy. Science, 282(5386):103–107.PubMedCrossRefGoogle Scholar
  22. 22.
    Nijhuis M, van Maarseveen N, Schuurman R, et al. 2002. Rapid and sensitive routine detection of all members of the genus enterovirus in different clinical specimens by real-time PCR. J Clin Microbiol, 40(10):3666–3670.PubMedCrossRefGoogle Scholar
  23. 23.
    Nishimura Y, Shimojima M, Tano Y, et al. 2009. Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71. Nat Med, 15(7):794–797.PubMedCrossRefGoogle Scholar
  24. 24.
    Perelson A S, Neumann A U, Markowitz M, et al. 1996. HIV-1 dynamics in vivo: virion clearance rate, infected cell life-span, and viral generation time. Science, 271(5255):1582–1586.PubMedCrossRefGoogle Scholar
  25. 25.
    Podin Y, Gias E L, Ong F, et al. 2006. Sentinel surveillance for human enterovirus 71 in Sarawak, Malaysia: lessons from the first 7 years. BMC Public Health, 6:180.PubMedCrossRefGoogle Scholar
  26. 26.
    Rabenau H F, Richter M, Doerr H W. Hand, foot and mouth disease: seroprevalence of Coxsackie A16 and Enterovirus 71 in Germany. Med Microbiol Immunol, 199(1):45–51.Google Scholar
  27. 27.
    Ribeiro R M, Lo A, Perelson A S. 2002. Dynamics of hepatitis B virus infection. Microbes Infect, 4(8):829–835.PubMedCrossRefGoogle Scholar
  28. 28.
    Solomon T, Lewthwaite P, Perera D, et al. Virology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect Dis, (11):778–790.Google Scholar
  29. 29.
    Yamayoshi S, Yamashita Y, Li J, et al. 2009. Scavenger receptor B2 is a cellular receptor for enterovirus 71. Nat Med, 15(7):798–801.PubMedCrossRefGoogle Scholar
  30. 30.
    Yi L, He Y, Chen Y, et al. Potent inhibition of human enterovirus 71 replication by type I interferon subtypes. Antivir Ther, 16(1):51–58.Google Scholar
  31. 31.
    Thomas D L, Villano S A, Riester K A, et al. 1998. Perinatal transmission of hepatitis C virus from human immunodeficiency virus type 1-infected mothers. Women and Infants Transmission Study. J Infect Dis, 177(6):1480–1488.PubMedCrossRefGoogle Scholar
  32. 32.
    Yip C C, Lau S K, Zhou B, et al. Emergence of enterovirus 71 “double-recombinant” strains belonging to a novel genotype D originating from southern China: first evidence for combination of intratypic and intertypic recombination events in EV71. Arch Virol, 155(9):1413–1424.Google Scholar
  33. 33.
    Zhang H M, Li C R, Liu Y J, et al. 2009. To investigate pathogen of hand, foot and mouth disease in Shenzhen in 2008. Chin J Exp Clin Virol, 23(5):334–336.(in Chinese)Google Scholar
  34. 34.
    Zhang Y, Zhu Z, Yang W, et al. An emerging recombinant human enterovirus 71 responsible for the 2008 outbreak of hand foot and mouth disease in Fuyang city of China. Virol J, 7:94.Google Scholar
  35. 35.
    Zoll J, Heus H A, van Kuppeveld F J, et al. 2009. The structure-function relationship of the enterovirus 3′-UTR. Virus Res, 139(2):209–216.PubMedCrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  1. 1.National Engineering Laboratory for AIDS Vaccine, College of Life ScienceJilin UniversityChangchunChina

Personalised recommendations