Skip to content
1887

Abstract

Boid inclusion body disease (BIBD) caused by reptarenaviruses affects captive constrictor snake collections worldwide. The disease manifests by the formation of cytoplasmic inclusion bodies in various tissues. Curiously, a snake with BIBD nearly always carries a swarm of reptarenavirus small and large segments rather than a single pair, and the composition of the swarm can vary between tissues. The role of reptarenavirus coinfections in BIBD pathogenesis remains unknown, and it is unclear whether reptarenavirus infection affects the susceptibility to superinfection or to secondary infections. For mammarenaviruses, co- and/or superinfection can occur if the infecting viruses are genetically divergent enough, and we hypothesized reptarenaviruses to behave similarly. To study this hypothesis, we employed boa constrictor kidney- and brain-derived cell cultures to perform a set of co- and superinfection experiments with one hartmanivirus and five reptarenavirus isolates. While all tested viruses replicated well in the boid kidney cells, experiments on the brain-derived cells showed differences in the replication efficacy between the viruses, suggesting that reptarenaviruses could differ in their target cell spectra. The quantification of viral RNA released from infected cells as a proxy for virus replication did not reveal overt differences between mono- and coinfections. Passaging of coinfected cell cultures revealed that one of the reptarenavirus isolates requires a coinfecting reptarena- or hartmanivirus to establish a persistent infection. Superinfection experiments on persistently reptarenavirus-infected cell lines suggested some interference between genetically similar viruses. We hypothesized that such interference would be mediated by the viral Z protein (ZP) specifically locking the genetically similar viral polymerase in a catalytically inactive state. Curiously, experiments on ZP-expressing cell lines indicated ZP overexpression not to significantly affect the amount of released viral RNA. Our experiments showed very little co- or superinfection interference between genetically dissimilar reptarenaviruses, reflecting the naturally occurring reptarenavirus coinfections in snakes with BIBD.

Funding
This study was supported by the:
  • Sigrid Juséliuksen Säätiö
    • Principal Award Recipient: JussiM Hepojoki
  • Terveyden Tutkimuksen Toimikunta (Award 308613, 314119, and 335762)
    • Principal Award Recipient: JussiM Hepojoki
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.002052
2024-12-11
2025-11-07

Metrics

Loading full text...

Full text loading...

References

  1. Rohwer F, Barott K. Viral information. Biol Philos 2013; 28:283–297 [View Article] [PubMed]
    [Google Scholar]
  2. Kumar N, Sharma S, Barua S, Tripathi BN, Rouse BT. Virological and immunological outcomes of coinfections. Clin Microbiol Rev 2018; 31:e00111-17 [View Article] [PubMed]
    [Google Scholar]
  3. Griffiths EC, Pedersen AB, Fenton A, Petchey OL. The nature and consequences of coinfection in humans. J Infect 2011; 63:200–206 [View Article] [PubMed]
    [Google Scholar]
  4. Radoshitzky SR, Buchmeier MJ, Charrel RN, Gonzalez J-PJ, Günther S et al. ICTV virus taxonomy profile: arenaviridae 2023. J Gen Virol 2023; 104:109 [View Article]
    [Google Scholar]
  5. Salazar-Bravo J, Ruedas LA, Yates TL. Mammalian reservoirs of arenaviruses. In Oldstone MBA. eds Arenaviruses I: The Epidemiology, Molecular and Cell Biology of Arenaviruses Berlin, Heidelberg: Springer Berlin Heidelberg; 2002 pp 25–63 [View Article]
    [Google Scholar]
  6. Downs WG, Anderson CR, Spence L, Aitken THG, Greenhall AH. Tacaribe virus, a new agent isolated from Artibeus bats and mosquitoes in Trinidad, West Indies. Am J Trop Med Hyg 1963; 12:640–646 [View Article]
    [Google Scholar]
  7. Sayler KA, Barbet AF, Chamberlain C, Clapp WL, Alleman R et al. Isolation of Tacaribe virus, a Caribbean arenavirus, from host-seeking Amblyomma americanum ticks in Florida. PLoS One 2014; 9:e115769 [View Article]
    [Google Scholar]
  8. Bentim Góes LG, Fischer C, Almeida Campos AC, de Carvalho C, Moreira-Soto A et al. Highly diverse Arenaviruses in neotropical bats, Brazil. Emerg Infect Dis 2022; 28:2528–2533 [View Article] [PubMed]
    [Google Scholar]
  9. Radoshitzky SR, Buchmeier MJ, Charrel RN, Clegg JCS, Gonzalez J-PJ et al. ICTV Virus Taxonomy Profile: Arenaviridae. J Gen Virol 2019; 100:1200–1201 [View Article]
    [Google Scholar]
  10. Chen Y-M, Sadiq S, Tian J-H, Chen X, Lin X-D et al. RNA viromes from terrestrial sites across China expand environmental viral diversity. Nat Microbiol 2022; 7:1312–1323 [View Article] [PubMed]
    [Google Scholar]
  11. Hepojoki J, Hepojoki S, Smura T, Szirovicza L, Dervas E et al. Characterization of Haartman Institute snake virus-1 (HISV-1) and HISV-like viruses-the representatives of genus Hartmanivirus, family Arenaviridae. PLoS Pathog 2018; 14:e1007415 [View Article] [PubMed]
    [Google Scholar]
  12. Radoshitzky SR, Bào Y, Buchmeier MJ, Charrel RN, Clawson AN et al. Past, present, and future of arenavirus taxonomy. Arch Virol 2015; 160:1851–1874 [View Article]
    [Google Scholar]
  13. Hallam SJ, Koma T, Maruyama J, Paessler S. Review of mammarenavirus biology and replication. Front Microbiol 2018; 9:1751 [View Article]
    [Google Scholar]
  14. Lehmann-Grube F. Persistent infection of the mouse with the virus of lymphocytic choriomeningitis. J Clin Pathol 1972; 25:8–21 [View Article]
    [Google Scholar]
  15. Traub E. The epidemiology of lymphocytic choriomeningitis in white mice. J Exp Med 1936; 64:183–200 [View Article] [PubMed]
    [Google Scholar]
  16. Stenglein MD, Sanchez-Migallon Guzman D, Garcia VE, Layton ML, Hoon-Hanks LL et al. Differential disease susceptibilities in experimentally reptarenavirus-infected boa constrictors and ball pythons. J Virol 2017; 91:e00451-17 [View Article] [PubMed]
    [Google Scholar]
  17. Stenglein MD, Sanders C, Kistler AL, Ruby JG, Franco JY et al. Identification, characterization, and in vitro culture of highly divergent arenaviruses from boa constrictors and annulated tree boas: candidate etiological agents for snake inclusion body disease. mBio 2012; 3:e00180-12 [View Article] [PubMed]
    [Google Scholar]
  18. Bodewes R, Kik MJL, Raj VS, Schapendonk CME, Haagmans BL et al. Detection of novel divergent arenaviruses in boid snakes with inclusion body disease in The Netherlands. J Gen Virol 2013; 94:1206–1210 [View Article]
    [Google Scholar]
  19. Hetzel U, Korzyukov Y, Keller S, Szirovicza L, Pesch T et al. Experimental reptarenavirus infection of boa constrictor and python regius. J Virol 2021; 95:e01968 [View Article]
    [Google Scholar]
  20. Hetzel U, Sironen T, Laurinmäki P, Liljeroos L, Patjas A et al. Isolation, identification, and characterization of novel arenaviruses, the etiological agents of boid inclusion body disease. J Virol 2013; 87:10918–10935 [View Article] [PubMed]
    [Google Scholar]
  21. Schumacher J, Jacobson ER, Homer BL, Gaskin JM. Inclusion body disease in boid snakes. J Zoo Wildl Med 1994; 25:511
    [Google Scholar]
  22. Orós J, Tucker S, Jacobson ER. Inclusion body disease in two captive boas in the canary islands. Vet Rec 1998; 143:283–285 [View Article] [PubMed]
    [Google Scholar]
  23. Lintala A, Szirovicza L, Kipar A, Hetzel U, Hepojoki J. Persistent reptarenavirus and hartmanivirus infection in cultured boid cells. Microbiol Spectr 2022; 10:e0158522 [View Article] [PubMed]
    [Google Scholar]
  24. Keller S, Hetzel U, Sironen T, Korzyukov Y, Vapalahti O et al. Co-infecting reptarenaviruses can be vertically transmitted in boa constrictor. PLoS Pathog 2017; 13:e1006179 [View Article] [PubMed]
    [Google Scholar]
  25. Windbichler K, Michalopoulou E, Palamides P, Pesch T, Jelinek C et al. Antibody response in snakes with boid inclusion body disease. PLoS One 2019; 14:e0221863 [View Article] [PubMed]
    [Google Scholar]
  26. Chang L, Fu D, Stenglein MD, Hernandez JA, DeRisi JL et al. Detection and prevalence of boid inclusion body disease in collections of boas and pythons using immunological assays. Vet J 2016; 218:13–18 [View Article] [PubMed]
    [Google Scholar]
  27. Aqrawi T, Stohr AC, Knauf-Witzens T, Krengel A, Heckers KO et al. Identification of snake arenaviruses in live boas and pythons in a zoo in germany. Tierarztl Prax Ausg KKleintiere Heimtiere 2015; 43:239 [View Article]
    [Google Scholar]
  28. Dietz J, Kolesnik E, Heckers KO, Klingberg M-N, Marschang RE. Detection of an arenavirus in a group of captive Wagler’s pit vipers (Tropidolaemus wagleri). J Zoo Wildl Med 2020; 51:236–240 [View Article] [PubMed]
    [Google Scholar]
  29. Hyndman TH, Marschang RE, Bruce M, Clark P, Vitali SD. Reptarenaviruses in apparently healthy snakes in an Australian zoological collection. Aust Veterinary J 2019; 97:93–102 [View Article]
    [Google Scholar]
  30. Simard J, Marschang RE, Leineweber C, Hellebuyck T. Prevalence of inclusion body disease and associated comorbidity in captive collections of boid and pythonid snakes in Belgium. PLoS One 2020; 15:e0229667 [View Article] [PubMed]
    [Google Scholar]
  31. Hepojoki J, Salmenperä P, Sironen T, Hetzel U, Korzyukov Y et al. Arenavirus coinfections are common in snakes with boid inclusion body disease. J Virol 2015; 89:8657–8660 [View Article] [PubMed]
    [Google Scholar]
  32. Stenglein MD, Jacobson ER, Chang L-W, Sanders C, Hawkins MG et al. Widespread recombination, reassortment, and transmission of unbalanced compound viral genotypes in natural arenavirus infections. PLoS Pathog 2015; 11:e1004900 [View Article] [PubMed]
    [Google Scholar]
  33. Argenta FF, Hepojoki J, Smura T, Szirovicza L, Hammerschmitt ME et al. Identification of reptarenaviruses, hartmaniviruses, and a novel chuvirus in captive native Brazilian boa constrictors with boid inclusion body disease. J Virol 2020; 94:e00001-20 [View Article] [PubMed]
    [Google Scholar]
  34. Thiele T, Baggio F, Prähauser B, Ruiz Subira A, Michalopoulou E et al. Reptarenavirus S segment RNA levels correlate with the presence of inclusion bodies and the number of L segments in snakes with reptarenavirus infection-lessons learned from a large breeding colony. Microbiol Spectr 2023; 11:e0506522 [View Article] [PubMed]
    [Google Scholar]
  35. Kranzusch PJ, Whelan SPJ. Arenavirus Z protein controls viral RNA synthesis by locking a polymerase-promoter complex. Proc Natl Acad Sci U S A 2011; 108:19743–19748 [View Article] [PubMed]
    [Google Scholar]
  36. Szirovicza L, Hetzel U, Kipar A, Martinez-Sobrido L, Vapalahti O et al. Snake deltavirus utilizes envelope proteins of different viruses to generate infectious particles. mBio 2020; 11:e03250-19 [View Article] [PubMed]
    [Google Scholar]
  37. Hepojoki J, Kipar A, Korzyukov Y, Bell-Sakyi L, Vapalahti O et al. Replication of boid inclusion body disease-associated arenaviruses is temperature sensitive in both boid and mammalian cells. J Virol 2015; 89:1119–1128 [View Article] [PubMed]
    [Google Scholar]
  38. Rusanen J, Kareinen L, Levanov L, Mero S, Pakkanen SH et al. A 10-minute “Mix and Read” antibody assay for SARS-CoV-2. Viruses 2021; 13:143 [View Article] [PubMed]
    [Google Scholar]
  39. Dervas E, Hepojoki J, Laimbacher A, Romero-Palomo F, Jelinek C et al. Nidovirus-associated proliferative pneumonia in the green tree python (Morelia viridis). J Virol 2017; 91:e00718-17 [View Article] [PubMed]
    [Google Scholar]
  40. Korzyukov Y, Hetzel U, Kipar A, Vapalahti O, Hepojoki J. Generation of anti-boa immunoglobulin antibodies for serodiagnostic applications, and their use to detect anti-reptarenavirus antibodies in boa constrictor. PLoS One 2016; 11:e0158417 [View Article] [PubMed]
    [Google Scholar]
  41. Alfaro-Alarcón A, Hetzel U, Smura T, Baggio F, Morales JA et al. Boid inclusion body disease is also a disease of wild boa constrictors. Microbiol Spectr 2022; 10:e0170522 [View Article] [PubMed]
    [Google Scholar]
  42. Chang L-W, Jacobson ER. Inclusion body disease, a worldwide infectious disease of boid snakes: a review. J Exotic Pet Med 2010; 19:216–225 [View Article]
    [Google Scholar]
  43. Korzyukov Y, Iheozor-Ejiofor R, Levanov L, Smura T, Hetzel U et al. Differences in tissue and species tropism of reptarenavirus species studied by vesicular stomatitis virus pseudotypes. Viruses 2020; 12:395 [View Article] [PubMed]
    [Google Scholar]
  44. Loureiro ME, D’Antuono A, Levingston Macleod JM, López N. Uncovering viral protein-protein interactions and their role in arenavirus life cycle. Viruses 2012; 4:1651–1667 [View Article] [PubMed]
    [Google Scholar]
  45. Loureiro ME, Wilda M, Levingston Macleod JM, D’Antuono A, Foscaldi S et al. Molecular determinants of arenavirus Z protein homo-oligomerization and L polymerase binding. J Virol 2011; 85:12304–12314 [View Article] [PubMed]
    [Google Scholar]
  46. Jácamo R, López N, Wilda M, Franze-Fernández MT. Tacaribe virus Z protein interacts with the L polymerase protein to inhibit viral RNA synthesis. J Virol 2003; 77:10383–10393 [View Article] [PubMed]
    [Google Scholar]
  47. Cornu TI, de la Torre JC. RING finger Z protein of lymphocytic choriomeningitis virus (LCMV) inhibits transcription and RNA replication of an LCMV S-segment minigenome. J Virol 2001; 75:9415–9426 [View Article] [PubMed]
    [Google Scholar]
  48. Cornu TI, de la Torre JC. Characterization of the arenavirus RING finger Z protein regions required for Z-mediated inhibition of viral RNA synthesis. J Virol 2002; 76:6678–6688 [View Article] [PubMed]
    [Google Scholar]
  49. López N, Jácamo R, Franze-Fernández MaT. Transcription and RNA replication of tacaribe virus genome and antigenome analogs require N and L proteins: Z protein is an inhibitor of these processes. J Virol 2001; 75:12241–12251 [View Article] [PubMed]
    [Google Scholar]
  50. Cuypers LN, Čížková D, de Bellocq JG. Co-infection of mammarenaviruses in a wild mouse, Tanzania. Virus Evol 2022; 8:veac065 [View Article] [PubMed]
    [Google Scholar]
  51. Ellenberg P, Edreira M, Scolaro L. Resistance to superinfection of vero cells persistently infected with Junin virus. Arch Virol 2004; 149:507–522 [View Article]
    [Google Scholar]
  52. Ellenberg P, Linero FN, Scolaro LA. Superinfection exclusion in BHK-21 cells persistently infected with Junín virus. J Gen Virol 2007; 88:2730–2739 [View Article]
    [Google Scholar]
  53. Damonte EB, Mersich SE, Coto CE. Response of cells persistently infected with arenaviruses to superinfection with homotypic and heterotypic viruses. Virology 1983; 129:474–478 [View Article] [PubMed]
    [Google Scholar]
  54. Gimenez HB, Compans RW. Defective interfering Tacaribe virus and persistently infected cells. Virology 1980; 107:229–239 [View Article] [PubMed]
    [Google Scholar]
  55. Popescu M, Schaefer H, Lehmann-Grube F. Homologous interference of lymphocytic choriomeningitis virus: detection and measurement of interference focus-forming units. J Virol 1976; 20:1–8 [View Article] [PubMed]
    [Google Scholar]
  56. Staneck LD, Peau CJ. Interfering particles from a culture persistently infected with Parana virus. J Gen Virol 1974; 22:437–440 [View Article] [PubMed]
    [Google Scholar]
  57. Staneck LD, Trowbridge RS, Welsh RM, Wright EA, Pfau CJ. Arenaviruses: cellular response to long-term in vitro infection with parana and lymphocytic choriomeningitis viruses. Infect Immun 1972; 6:444–450 [View Article] [PubMed]
    [Google Scholar]
  58. Lehmann-Grube F. A carrier state of lymphocytic choriomeningitis virus in L cell cultures. Nature 1967; 213:770–773 [View Article] [PubMed]
    [Google Scholar]
  59. Dutko FJ, Wright EA, Pfau CJ. The RNAs of the defective interfering Pichinide virus. J Gen Virol 1976; 31:417–427 [View Article] [PubMed]
    [Google Scholar]
  60. Peralta LM, Bruns M, Lehmann-Grube F. Biochemical composition of lymphocytic choriomeningitis virus interfering particles. J Gen Virol 1981; 55:475–479 [View Article] [PubMed]
    [Google Scholar]
  61. Francis SJ, Southern PJ. Deleted viral RNAs and lymphocytic choriomeningitis virus persistence in vitro. J Gen Virol 1988; 69:1893–1902 [View Article] [PubMed]
    [Google Scholar]
  62. Welsh RM, O’Connell CM, Pfau CJ. Properties of defective lymphocytic choriomeningitis virus. J Gen Virol 1972; 17:355–359 [View Article] [PubMed]
    [Google Scholar]
  63. Welsh RM, Oldstone MB. Inhibition of immunologic injury of cultured cells infected with lymphocytic choriomeningitis virus: role of defective interfering virus in regulating viral antigenic expression. J Exp Med 1977; 145:1449–1468 [View Article] [PubMed]
    [Google Scholar]
  64. Welsh RM, Pfau CJ. Determinants of lymphocytic choriomeningitis interference. J Gen Virol 1972; 14:177–187 [View Article] [PubMed]
    [Google Scholar]
  65. Welsh RM Jr, Buchmeier MJ. Protein analysis of defective interfering lymphocytic choriomeningitis virus and persistently infected cells. Virology 1979; 96:503–515 [View Article]
    [Google Scholar]
  66. Jacobson S, Pfau CJ. Viral pathogenesis and resistance to defective interfering particles. Nature 1980; 283:311–313 [View Article] [PubMed]
    [Google Scholar]
  67. Jacobson S, Dutko FJ, Pfau CJ. Determinants of spontaneous recovery and persistance in MDCK cells infected with lymphocytic choriomeningitis virus. J Gen Virol 1979; 44:113–122 [View Article] [PubMed]
    [Google Scholar]
  68. Oldstone MB, Buchmeier MJ. Restricted expression of viral glycoprotein in cells of persistently infected mice. Nature 1982; 300:360–362 [View Article] [PubMed]
    [Google Scholar]
  69. Ziegler CM, Botten JW. Defective interfering particles of negative-strand RNA viruses. Trends Microbiol 2020; 28:554–565 [View Article] [PubMed]
    [Google Scholar]
/content/journal/jgv/10.1099/jgv.0.002052
Loading
/content/journal/jgv/10.1099/jgv.0.002052
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An error occurred
Approval was partially successful, following selected items could not be processed due to error