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Establishment of persistent foot-and-mouth disease virus (FMDV) infection in MDBK cells

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In addition to acute infection and disease, foot-and-mouth disease virus (FMDV) can cause persistent infection in ruminants. Such “carrier” animals represent a potential risk for FMDV transmission to susceptible animals. However, the mechanisms and the factors that determine FMDV persistence remain unknown. We describe here the establishment of FMDV type O persistent infection in a bovine epithelial cell line (Madin-Darby bovine kidney; MDBK). Preliminary experiments to assess the permissivity of MDBK cells to FMDV O infection revealed an unusual pattern of infection: after the initial phase of acute cell lysis, new monolayers formed within 48-72 h post-infection. We found that some cells survived cytolytic infection and subsequently regrew, thereby demonstrating that this bovine cell line can be persistently infected with FMDV type O. Further evidence that MDBK cells were persistently infected with FMDV includes: (i) detection of viral RNA in cells as well as in cell culture supernatants, (ii) detection of viral antigens in the cells by immunofluorescence analysis, and (iii) production of infectious viral particles for up to 36 cell passages. Furthermore, preliminary sequence analysis of persistent virus revealed a single nucleotide substitution within the VP1 coding region, resulting in the V50A amino acid substitution. This bovine model of FMDV persistence holds promise for the investigation of the viral and cellular molecular determinants that promote FMDV persistence.

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  1. 1.

    Alexandersen S, Zhang Z, Donaldson AI (2002) Aspects of the persistence of foot-and-mouth disease virus in animals-the carrier problem. Microbes Infect 4:1099–1110

  2. 2.

    Alexandersen S, Zhang Z, Donaldson AI, Garland AJ (2003) The pathogenesis and diagnosis of foot-and-mouth disease. J Comp Pathol 129:1–36

  3. 3.

    Arzt J, Baxt B, Grubman MJ, Jackson T, Juleff N, Rhyan J, Rieder E, Waters R, Rodriguez LL (2011) The pathogenesis of foot-and-mouth disease II: viral pathways in swine, small ruminants, and wildlife; myotropism, chronic syndromes, and molecular virus-host interactions. Transbound Emerg Dis 58:305–326

  4. 4.

    Bao HF, Li D, Sun P, Zhou Q, Hu J, Bai XW, Fu YF, Lu ZJ, Liu ZX (2011) The infectivity and pathogenicity of a foot-and-mouth disease virus persistent infection strain from oesophageal-pharyngeal fluid of a Chinese cattle in 2010. Virol J 8:536

  5. 5.

    Barreca C, O’Hare P (2004) Suppression of herpes simplex virus 1 in MDBK cells via the interferon pathway. J Virol 78:8641–8653

  6. 6.

    Barreca C, O’Hare P (2006) Characterization of a potent refractory state and persistence of herpes simplex virus 1 in cell culture. J Virol 80:9171–9180

  7. 7.

    Barros JJ, Malirat V, Rebello MA, Costa EV, Bergmann IE (2007) Genetic variation of foot-and-mouth disease virus isolates recovered from persistently infected water buffalo (Bubalus bubalis). Vet Microbiol 120:50–62

  8. 8.

    Bastos AD, Boshoff CI, Keet DF, Bengis RG, Thomson GR (2000) Natural transmission of foot-and-mouth disease virus between African buffalo (Syncerus caffer) and impala (Aepyceros melampus) in the Kruger National Park, South Africa. Epidemiol Infect 124:591–598

  9. 9.

    Bossert B, Conzelmann KK (2002) Respiratory syncytial virus (RSV) nonstructural (NS) proteins as host range determinants: a chimeric bovine RSV with NS genes from human RSV is attenuated in interferon-competent bovine cells. J Virol 76:4287–4293

  10. 10.

    Cai KJ, Meng QL, Qiao J, Huang J, Zhang ZC, Wang GC, Wang JW, Chen CF (2013) Expression of bovine Mx1 protein inhibits the replication of foot-and-mouth disease virus in BHK-21 cells. Acta Virol 57:429–434

  11. 11.

    Chang H, Ma Y, Lin T, Cong G, Du J, Ma J (2013) Foot-and-mouth disease virus carrier status in Bos grunniens yaks. Virol J 10:81

  12. 12.

    Chauveau E, Doceul V, Lara E, Adam M, Breard E, Sailleau C, Viarouge C, Desprat A, Meyer G, Schwartz-Cornil I, Ruscanu S, Charley B, Zientara S, Vitour D (2012) Sensing and control of bluetongue virus infection in epithelial cells via RIG-I and MDA5 helicases. J Virol 86:11789–11799

  13. 13.

    Dawe PS, Flanagan FO, Madekurozwa RL, Sorensen KJ, Anderson EC, Foggin CM, Ferris NP, Knowles NJ (1994) Natural transmission of foot-and-mouth disease virus from African buffalo (Syncerus caffer) to cattle in a wildlife area of Zimbabwe. Vet Rec 134:230–232

  14. 14.

    Dawe PS, Sorensen K, Ferris NP, Barnett IT, Armstrong RM, Knowles NJ (1994) Experimental transmission of foot-and-mouth disease virus from carrier African buffalo (Syncerus caffer) to cattle in Zimbabwe. Vet Rec 134:211–215

  15. 15.

    de la Torre JC, Davila M, Sobrino F, Ortin J, Domingo E (1985) Establishment of cell lines persistently infected with foot-and-mouth disease virus. Virology 145:24–35

  16. 16.

    Diez J, Davila M, Escarmis C, Mateu MG, Dominguez J, Perez JJ, Giralt E, Melero JA, Domingo E (1990) Unique amino acid substitutions in the capsid proteins of foot-and-mouth disease virus from a persistent infection in cell culture. J Virol 64:5519–5528

  17. 17.

    Donn A, Castagnaro M, Donaldson AI (1995) Ultrastructural and replicative features of foot-and-mouth disease virus in persistently infected BHK-21 cells. Arch Virol 140:13–25

  18. 18.

    Fry EE, Newman JW, Curry S, Najjam S, Jackson T, Blakemore W, Lea SM, Miller L, Burman A, King AM, Stuart DI (2005) Structure of Foot-and-mouth disease virus serotype A10 61 alone and complexed with oligosaccharide receptor: receptor conservation in the face of antigenic variation. J Gen Virol 86:1909–1920

  19. 19.

    Garland AJ, de Clercq K (2011) Cattle, sheep and pigs vaccinated against foot and mouth disease: does trade in these animals and their products present a risk of transmitting the disease? Rev Sci Tech 30:189–206

  20. 20.

    Gebauer F, de la Torre JC, Gomes I, Mateu MG, Barahona H, Tiraboschi B, Bergmann I, de Mello PA, Domingo E (1988) Rapid selection of genetic and antigenic variants of foot-and-mouth disease virus during persistence in cattle. J Virol 62:2041–2049

  21. 21.

    Goris N, Vandenbussche F, Herr C, Villers J, Van der Stede Y, De Clercq K (2009) Validation of two real-time RT-PCR methods for foot-and-mouth disease diagnosis: RNA-extraction, matrix effect, uncertainty of measurement and precision. J Virol Methods 160:157–162

  22. 22.

    Gorna K, Houndje E, Romey A, Relmy A, Blaise-Boisseau S, Kpodekon M, Saegerman C, Moutou F, Zientara S, Bakkali Kassimi L (2014) First isolation and molecular characterization of foot-and-mouth disease virus in Benin. Vet Microbiol 171:175–181

  23. 23.

    Grubman MJ, Baxt B (2004) Foot-and-mouth disease. Clin Microbiol Rev 17:465–493

  24. 24.

    Herrera M, Grande-Perez A, Perales C, Domingo E (2008) Persistence of foot-and-mouth disease virus in cell culture revisited: implications for contingency in evolution. J Gen Virol 89:232–244

  25. 25.

    Huang X, Li Y, Fang H, Zheng C (2011) Establishment of persistent infection with foot-and-mouth disease virus in BHK-21 cells. Virol J 8:169

  26. 26.

    Jamal SM, Belsham GJ (2013) Foot-and-mouth disease: past, present and future. Vet Res 44:116

  27. 27.

    Juleff N, Windsor M, Reid E, Seago J, Zhang Z, Monaghan P, Morrison IW, Charleston B (2008) Foot-and-mouth disease virus persists in the light zone of germinal centres. PLoS ONE 3:e3434

  28. 28.

    Juleff ND, Maree FF, Waters R, Bengis RG, Charleston B (2012) The importance of FMDV localisation in lymphoid tissue. Vet Immunol Immunopathol 148:145–148

  29. 29.

    Klein J (2009) Understanding the molecular epidemiology of foot-and-mouth-disease virus. Infect Genet Evol 9:153–161

  30. 30.

    Lee CD, Yan YP, Liang SM, Wang TF (2009) Production of FMDV virus-like particles by a SUMO fusion protein approach in Escherichia coli. J Biomed Sci 16:69

  31. 31.

    Li X, Wang J, Liu J, Li Z, Wang Y, Xue Y, Cao H, Zheng SJ (2013) Engagement of soluble resistance-related calcium binding protein (sorcin) with foot-and-mouth disease virus (FMDV) VP1 inhibits type I interferon response in cells. Vet Microbiol 166:35–46

  32. 32.

    Luna VE, Luk AD, Tyring SK, Hellman JM, Lefkowitz SS (1984) Properties of bovine interferons. Experientia 40:1410–1412

  33. 33.

    Maddur MS, Kishore S, Gopalakrishna S, Singh N, Suryanarayana VV, Gajendragad MR (2009) Immune response and viral persistence in Indian buffaloes (Bubalus bubalis) infected with foot-and-mouth disease virus serotype Asia 1. Clin Vaccine Immunol 16:1832–1836

  34. 34.

    Mannini-Palenzona A, Bartoletti AM, Foa’-Tomasi L, Baserga M, Tognon M, Manservigi R (1985) Establishment and characterization of a persistent infection of MDBK cells with herpes simplex virus. Microbiologica 8:165–180

  35. 35.

    Martin-Acebes MA, Herrera M, Armas-Portela R, Domingo E, Sobrino F (2010) Cell density-dependent expression of viral antigens during persistence of foot-and-mouth disease virus in cell culture. Virology 403:47–55

  36. 36.

    Martin Hernandez AM, Carrillo EC, Sevilla N, Domingo E (1994) Rapid cell variation can determine the establishment of a persistent viral infection. Proc Natl Acad Sci USA 91:3705–3709

  37. 37.

    Martinez-Salas E, Saiz JC, Davila M, Belsham GJ, Domingo E (1993) A single nucleotide substitution in the internal ribosome entry site of foot-and-mouth disease virus leads to enhanced cap-independent translation in vivo. J Virol 67:3748–3755

  38. 38.

    Mattoscio D, Segre CV, Chiocca S (2013) Viral manipulation of cellular protein conjugation pathways: the SUMO lesson. World J Virol 2:79–90

  39. 39.

    Nobiron I, Remond M, Kaiser C, Lebreton F, Zientara S, Delmas B (2005) The nucleotide sequence of foot-and-mouth disease virus O/FRA/1/2001 and comparison with its British parental strain O/UKG/35/2001. Virus Res 108:225–229

  40. 40.

    O’Donnell V, Pacheco JM, Larocco M, Gladue DP, Pauszek SJ, Smoliga G, Krug PW, Baxt B, Borca MV, Rodriguez L (2014) Virus-host interactions in persistently FMDV-infected cells derived from bovine pharynx. Virology 468–470:185–196

  41. 41.

    OIE (2012) Foot and mouth disease. Manual of diagnostic tests and vaccines for terrestrial animals

  42. 42.

    Opperman PA, Rotherham LS, Esterhuysen J, Charleston B, Juleff N, Capozzo AV, Theron J, Maree FF (2014) Determining the epitope dominance on the capsid of a serotype SAT2 foot-and-mouth disease virus by mutational analyses. J Virol 88:8307–8318

  43. 43.

    Otsuki K, Maeda J, Yamamoto H, Tsubokura M (1979) Studies on avian infectious bronchitis virus (IBV). III. Interferon induction by and sensitivity to interferon of IBV. Arch Virol 60:249–255

  44. 44.

    Reed LJ, Muench LH (1938) A simple method of estimating fifty percent endpoints. Am J Hyg 27:493–497

  45. 45.

    Salt JS (1993) The carrier state in foot and mouth disease–an immunological review. Br Vet J 149:207–223

  46. 46.

    Sarangi LN, Mohapatra JK, Subramaniam S, Sanyal A, Pattnaik B (2013) Antigenic site variation in foot-and-mouth disease virus serotype O grown under vaccinal serum antibodies in vitro. Virus Res 176:273–279

  47. 47.

    Stenfeldt C, Heegaard PM, Stockmarr A, Belsham GJ (2012) Modulation of cytokine mRNA expression in pharyngeal epithelial samples obtained from cattle infected with foot-and-mouth disease virus. J Comp Pathol 146:243–252

  48. 48.

    Stenfeldt C, Pacheco JM, Smoliga GR, Bishop E, Pauszek SJ, Hartwig EJ, Rodriguez LL, Arzt J (2014) Detection of foot-and-mouth disease virus RNA and capsid protein in lymphoid tissues of convalescent pigs does not indicate existence of a carrier state. Transbound Emerg Dis

  49. 49.

    Sutmoller P, McVicar JW, Cottral GE (1968) The epizootiological importance of foot-and-mouth disease carriers. I. Experimentally produced foot-and-mouth disease carriers in susceptible and immune cattle. Arch Gesamte Virusforsch 23:227–235

  50. 50.

    Sutmoller P, Barteling SS, Olascoaga RC, Sumption KJ (2003) Control and eradication of foot-and-mouth disease. Virus Res 91:101–144

  51. 51.

    Tenzin Dekker A, Vernooij H, Bouma A, Stegeman A (2008) Rate of foot-and-mouth disease virus transmission by carriers quantified from experimental data. Risk Anal 28:303–309

  52. 52.

    Woodbury EL (1995) A review of the possible mechanisms for the persistence of foot-and-mouth disease virus. Epidemiol Infect 114:1–13

  53. 53.

    Zhang H, Li Y, Huang X, Zheng C (2013) Global transcriptional analysis of model of persistent FMDV infection reveals critical role of host cells in persistence. Vet Microbiol 162:321–329

  54. 54.

    Zhang ZD, Kitching RP (2001) The localization of persistent foot and mouth disease virus in the epithelial cells of the soft palate and pharynx. J Comp Pathol 124:89–94

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This work was funded by ANSES, INRA, and ENVA. Lela Kopliku received a PhD grant from ANSES. We thank Damien Vitour for providing reagents, critical reading of the manuscript, and useful discussions.

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Correspondence to Sandra Blaise-Boisseau.

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Kopliku, L., Relmy, A., Romey, A. et al. Establishment of persistent foot-and-mouth disease virus (FMDV) infection in MDBK cells. Arch Virol 160, 2503–2516 (2015). https://doi.org/10.1007/s00705-015-2526-8

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  • MDBK Cell
  • Persistent Virus
  • African Buffalo
  • Cytolytic Infection
  • Cold Minimum Essential Medium