Skip to main content

Advertisement

SpringerLink
Log in

Apoptosis in porcine macrophages infected in vitro with African swine fever virus (ASFV) strains with different virulence

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

African swine fever virus (ASFV) replicates in porcine macrophages. Since modulation of cell death by ASFV strains of different virulence is poorly understood, we studied the development of apoptosis in porcine macrophage cultures during in vitro infection with the high- and low-virulence isolates ASFV/L60 (L60) and ASFV/NH/P68 (NHV), respectively. In cultures inoculated with each isolate, similar numbers of cells hosting infection showed morphological signs of apoptosis, which were visible from a relatively early time of infection (8 h), although a significant proportion of the infected cell populations remained non-apoptotic until 18 h. L60 inhibited caspase-3 activation by 18 h after infection and induced less DNA internucleosomic fragmentation at 8 h than NHV. However, at the late infection time, apoptosis levels were similar in both infections and occurred, at least partially, independently of caspases, suggesting the existence of yet unknown alternative pathways committing ASFV host cells to apoptotic death.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Adrain C, Murphy BM, Martin SJ (2005) Molecular ordering of the caspase activation cascade initiated by the cytotoxic T lymphocyte/natural killer (CTL/NK) protease granzyme B. J Biol Chem 280:4663–4673

    Article  PubMed  CAS  Google Scholar 

  2. Afonso CL, Neilan JG, Kutish GF, Rock DL (1996) An African swine fever virus Bcl-2 homolog, 5-HL, suppresses apoptotic cell death. J Virol 70:4858–4863

    PubMed  CAS  Google Scholar 

  3. Aubert M, Jerome KR (2003) Apoptosis prevention as a mechanism of immune evasion. Int Rev Immunol 22:361–371

    Article  PubMed  CAS  Google Scholar 

  4. Blank M, Shiloh Y (2007) Programs for cell death: apoptosis is only one way to go. Cell Cycle 6:686–695

    PubMed  CAS  Google Scholar 

  5. Borca MV, Carrillo C, Zsak L, Laegreid WW, Kutish GF, Neilan JG, Burrage TG, Rock DL (1998) Deletion of a CD2-like gene, 8-DR, from African swine fever virus affects viral infection in domestic swine. J Virol 72:2881–2889

    PubMed  CAS  Google Scholar 

  6. Brookes SM, Dixon LK, Parkhouse RM (1996) Assembly of African Swine fever virus: quantitative ultrastructural analysis in vitro and in vivo. Virology 224:84–92

    Article  PubMed  CAS  Google Scholar 

  7. Carrasco L, de Lara FC, Martin de las Mulas J, Gomez-Villamandos JC, Perez J, Wilkinson PJ, Sierra MA (1996) Apoptosis in lymph nodes in acute African swine fever. J Comp Pathol 115:415–428

    Article  PubMed  CAS  Google Scholar 

  8. Carrascosa AL, Bustos MJ, Nogal ML, Gonzalez de Buitrago G, Revilla Y (2002) Apoptosis induced in an early step of African swine fever virus entry into Vero cells does not require virus replication. Virology 294:372–382

    Article  PubMed  CAS  Google Scholar 

  9. Chacon MR, Almazan F, Nogal ML, Vinuela E, Rodriguez JF (1995) The African swine fever virus IAP homolog is a late structural polypeptide. Virology 214:670–674

    Article  PubMed  CAS  Google Scholar 

  10. Dixon LK, Escribano JM, Martins C, Rock DL, Salas ML, Wilkinson PJ (2005) Asfarviridae. In: Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA (eds) Virus taxonomy: eighth report of the international committee on taxonomy of viruses. Elsevier/Academic Press, London, pp 135–143

    Google Scholar 

  11. Duarte M (2000) Bases moleculares da virulência e hemadsorção nos isolados nacionais Lisboa 60 e Lisboa 68 do vírus da peste suína Africana. PhD Thesis, Instituto de Tecnologia Química e Biológica. Universidade Nova de Lisboa, Oeiras, Portugal

  12. Ekert PG, Silke J, Vaux DL (1999) Caspase inhibitors. Cell Death Differ 6:1081–1086

    Article  PubMed  CAS  Google Scholar 

  13. Galindo I, Almazan F, Bustos MJ, Vinuela E, Carrascosa AL (2000) African swine fever virus EP153R open reading frame encodes a glycoprotein involved in the hemadsorption of infected cells. Virology 266:340–351

    Article  PubMed  CAS  Google Scholar 

  14. Galindo I, Hernaez B, Diaz-Gil G, Escribano JM, Alonso C (2008) A179L, a viral Bcl-2 homologue, targets the core Bcl-2 apoptotic machinery and its upstream BH3 activators with selective binding restrictions for Bid and Noxa. Virology 375:561–572

    Article  PubMed  CAS  Google Scholar 

  15. Gilmore AP (2005) Anoikis. Cell Death Differ 12(Suppl 2):1473–1477

    Article  PubMed  CAS  Google Scholar 

  16. Gupta S (2003) Molecular signaling in death receptor and mitochondrial pathways of apoptosis (Review). Int J Oncol 22:15–20

    PubMed  CAS  Google Scholar 

  17. Hernaez B, Diaz-Gil G, Garcia-Gallo M, Ignacio Quetglas J, Rodriguez-Crespo I, Dixon L, Escribano JM, Alonso C (2004) The African swine fever virus dynein-binding protein p54 induces infected cell apoptosis. FEBS Lett 569:224–228

    Article  PubMed  CAS  Google Scholar 

  18. Hernaez B, Escribano JM, Alonso C (2006) Visualization of the African swine fever virus infection in living cells by incorporation into the virus particle of green fluorescent protein-p54 membrane protein chimera. Virology 350:1–14

    Article  PubMed  CAS  Google Scholar 

  19. Hurtado C, Granja AG, Bustos MJ, Nogal ML, Gonzalez de Buitrago G, de Yebenes VG, Salas ML, Revilla Y, Carrascosa AL (2004) The C-type lectin homologue gene (EP153R) of African swine fever virus inhibits apoptosis both in virus infection and in heterologous expression. Virology 326:160–170

    Article  PubMed  CAS  Google Scholar 

  20. King KL, Jewell CM, Bortner CD, Cidlowski JA (2000) 28S ribosome degradation in lymphoid cell apoptosis: evidence for caspase and Bcl-2-dependent and -independent pathways. Cell Death Differ 7:994–1001

    Article  PubMed  CAS  Google Scholar 

  21. Leitao A, Cartaxeiro C, Coelho R, Cruz B, Parkhouse RM, Portugal F, Vigario JD, Martins CL (2001) The non-haemadsorbing African swine fever virus isolate ASFV/NH/P68 provides a model for defining the protective anti-virus immune response. J Gen Virol 82:513–523

    PubMed  CAS  Google Scholar 

  22. Martins CL, Scholl T, Mebus CA, Fisch H, Lawman MJ (1987) Modulation of porcine peripheral blood-derived macrophage functions by in vitro infection with African swine fever virus (ASFV) isolates of different virulence. Viral Immunol 1:177–190

    Article  PubMed  Google Scholar 

  23. McCullough KC, Basta S, Knotig S, Gerber H, Schaffner R, Kim YB, Saalmuller A, Summerfield A (1999) Intermediate stages in monocyte–macrophage differentiation modulate phenotype and susceptibility to virus infection. Immunology 98:203–212

    Article  PubMed  CAS  Google Scholar 

  24. Neilan JG, Lu Z, Afonso CL, Kutish GF, Sussman MD, Rock DL (1993) An African swine fever virus gene with similarity to the proto-oncogene bcl-2 and the Epstein-Barr virus gene BHRF1. J Virol 67:4391–4394

    PubMed  CAS  Google Scholar 

  25. Neilan JG, Lu Z, Kutish GF, Zsak L, Burrage TG, Borca MV, Carrillo C, Rock DL (1997) A BIR motif containing gene of African swine fever virus, 4CL, is nonessential for growth in vitro and viral virulence. Virology 230:252–264

    Article  PubMed  CAS  Google Scholar 

  26. Neilan JG, Borca MV, Lu Z, Kutish GF, Kleiboeker SB, Carrillo C, Zsak L, Rock DL (1999) An African swine fever virus ORF with similarity to C-type lectins is non-essential for growth in swine macrophages in vitro and for virus virulence in domestic swine. J Gen Virol 80(Pt 10):2693–2697

    PubMed  CAS  Google Scholar 

  27. Nogal ML, Gonzalez de Buitrago G, Rodriguez C, Cubelos B, Carrascosa AL, Salas ML, Revilla Y (2001) African swine fever virus IAP homologue inhibits caspase activation and promotes cell survival in mammalian cells. J Virol 75:2535–2543

    Article  PubMed  CAS  Google Scholar 

  28. O’Connell AR, Holohan C, Torriglia A, Lee BW, Stenson-Cox C (2006) Characterization of a serine protease-mediated cell death program activated in human leukemia cells. Exp Cell Res 312:27–39

    Article  PubMed  Google Scholar 

  29. Oura CA, Powell PP, Parkhouse RM (1998) African swine fever: a disease characterized by apoptosis. J Gen Virol 79(Pt 6):1427–1438

    PubMed  CAS  Google Scholar 

  30. Pan IC, Shimizu M, Hess WR (1980) Replication of African swine fever virus in cell cultures. Am J Vet Res 41:1357–1367

    PubMed  CAS  Google Scholar 

  31. Portugal R, Leitao A, Martins C (2009) Characterization of African swine fever virus IAP homologue expression in porcine macrophages infected with different virulence isolates. Vet Microbiol. doi:10.1016/j.vetmic.2009.04.030

  32. Ramiro-Ibanez F, Ortega A, Brun A, Escribano JM, Alonso C (1996) Apoptosis: a mechanism of cell killing and lymphoid organ impairment during acute African swine fever virus infection. J Gen Virol 77(Pt 9):2209–2219

    Article  PubMed  CAS  Google Scholar 

  33. Rodriguez JM, Yanez RJ, Almazan F, Vinuela E, Rodriguez JF (1993) African swine fever virus encodes a CD2 homolog responsible for the adhesion of erythrocytes to infected cells. J Virol 67:5312–5320

    PubMed  CAS  Google Scholar 

  34. Salguero FJ, Sanchez-Cordon PJ, Nunez A, Fernandez de Marco M, Gomez-Villamandos JC (2005) Proinflammatory cytokines induce lymphocyte apoptosis in acute African swine fever infection. J Comp Pathol 132:289–302

    Article  PubMed  CAS  Google Scholar 

  35. Trapani JA, Smyth MJ (2002) Functional significance of the perforin/granzyme cell death pathway. Nat Rev Immunol 2:735–747

    Article  PubMed  CAS  Google Scholar 

  36. Vandenabeele P, Vanden Berghe T, Festjens N (2006) Caspase inhibitors promote alternative cell death pathways. Sci STKE, 24 Oct 2006(358), pe44. doi:10.1126/stke.3582006pe44

  37. Yanez RJ, Rodriguez JM, Nogal ML, Yuste L, Enriquez C, Rodriguez JF, Vinuela E (1995) Analysis of the complete nucleotide sequence of African swine fever virus. Virology 208:249–278

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was funded by: SFRH/BD/10576/2002 PhD fellowship from Fundação para a Ciência e Tecnologia (FCT) to R. Portugal; Project POCI/CVT/59122/2004 (FCT); Project ASFRISK, GA 211691, EC, FP7-KBBE-2007-1-3-5, Seventh Framework Programme, Food, Agriculture and Fisheries; Interdisciplinary Centre of Research in Animal Health (CIISA) at Faculty of Veterinary Medicine, UTL.

Author information

Authors and Affiliations

  1. Laboratório de Doenças Infecciosas, CIISA, Faculdade de Medicina Veterinária, Technical University of Lisbon (TULisbon), Av. da Universidade Técnica, 1300-477, Lisbon, Portugal

    Raquel Portugal & Carlos Martins

  2. CVZ, CIISA, Faculdade de Medicina Veterinária, Technical University of Lisbon (TULisbon), Instituto de Investigação Científica Tropical, Av. da Universidade Técnica, 1300-477, Lisbon, Portugal

    Alexandre Leitão

Authors
  1. Raquel Portugal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexandre Leitão
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carlos Martins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carlos Martins.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Portugal, R., Leitão, A. & Martins, C. Apoptosis in porcine macrophages infected in vitro with African swine fever virus (ASFV) strains with different virulence. Arch Virol 154, 1441–1450 (2009). https://doi.org/10.1007/s00705-009-0466-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-009-0466-x

Keywords

Search

Navigation