Skip to main content

Advertisement

Log in

Suppression of lytic replication of Kaposi’s sarcoma-associated herpesvirus by autophagy during initial infection in NIH 3T3 fibroblasts

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

Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the infectious cause of the angioproliferative neoplasm Kaposi’s sarcoma (KS). We first confirmed the susceptibility of NIH 3T3 fibroblasts to KSHV by infecting them with BCP-1-derived KSHV. Lytic replication of KSHV was confirmed by PCR amplification of viral DNA isolated from culture supernatants of KSHV-infected cells. The template from KSHV-infected NIH 3T3 cells resulted in an intense viral DNA PCR product. A time course experiment revealed the disappearance of KSHV-specific DNA in culture supernatant of NIH 3T3 cells during a period between 48 h and 72 h postinfection. Furthermore, 3 days postinfection, infected NIH 3T3 cells showed no evidence of latent or lytic transcripts, including LANA, vFLIP, vCyclin, and vIL-6. These results imply that KSHV infection in NIH 3T3 cells is unstable and is rapidly lost on subsequent culturing. Additionally, we detected an enhancement of autophagy early in infection with KSHV. More interestingly, inhibition of autophagy by Beclin 1 siRNA or 3-methyladenine significantly increased the amount of KSHV-specific DNA in the culture supernatant of NIH 3T3 cells when compared to the group treated with KSHV infection alone, implying that autophagy prevents lytic replication of KSHV. Taken together, our data suggest that autophagy could be one of the cellular mechanisms utilized by host cells to promote viral clearance.

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

Access this article

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

Similar content being viewed by others

References

  1. Chang Y, Cesarman E, Pessin MS, Lee F, Culpepper J, Knowles DM, Moore PS (1994) Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi’s sarcoma. Science 266(5192):1865–1869

    Article  CAS  PubMed  Google Scholar 

  2. Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM (1995) Kaposi’s sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. N Engl J Med 332(18):1186–1191

    Article  CAS  PubMed  Google Scholar 

  3. Soulier J, Grollet L, Oksenhendler E, Cacoub P, Cazals-Hatem D, Babinet P, d’Agay MF, Clauvel JP, Raphael M, Degos L, Sigaux F (1995) Kaposi’s sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman’s disease. Blood 86(4):1276–1280

    CAS  PubMed  Google Scholar 

  4. Veettil MV, Bandyopadhyay C, Dutta D, Chandran B (2014) Interaction of KSHV with host cell surface receptors and cell entry. Viruses 6(10):4024–4046

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Deng H, Liang Y, Sun R (2007) Regulation of KSHV lytic gene expression. Curr Top Microbiol Immunol 312:157–183

    CAS  PubMed  Google Scholar 

  6. Yang Z, Yan Z, Wood C (2008) Kaposi’s sarcoma-associated herpesvirus transactivator RTA promotes degradation of the repressors to regulate viral lytic replication. J Virol 82(7):3590–3603

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Ye F, Lei X, Gao SJ (2011) Mechanisms of Kaposi’s sarcoma-associated herpesvirus latency and reactivation. Adv Virol 2011:193860

    Article  PubMed Central  PubMed  Google Scholar 

  8. Tempera I, Lieberman PM (2010) Chromatin organization of gammaherpesvirus latent genomes. Biochim Biophys Acta 1799(3–4):236–245

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Dittmer D, Lagunoff M, Renne R, Staskus K, Haase A, Ganem D (1998) A cluster of latently expressed genes in Kaposi’s sarcoma-associated herpesvirus. J Virol 72(10):8309–8315

    PubMed Central  CAS  PubMed  Google Scholar 

  10. Rainbow L, Platt GM, Simpson GR, Sarid R, Gao SJ, Stoiber H, Herrington CS, Moore PS, Schulz TF (1997) The 222- to 234-kilodalton latent nuclear protein (LNA) of Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8) is encoded by orf73 and is a component of the latency-associated nuclear antigen. J Virol 71(8):5915–5921

    PubMed Central  CAS  PubMed  Google Scholar 

  11. Sarid R, Flore O, Bohenzky RA, Chang Y, Moore PS (1998) Transcription mapping of the Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8) genome in a body cavity-based lymphoma cell line (BC-1). J Virol 72(2):1005–1012

    PubMed Central  CAS  PubMed  Google Scholar 

  12. Bechtel JT, Liang Y, Hvidding J, Ganem D (2003) Host range of Kaposi’s sarcoma-associated herpesvirus in cultured cells. J Virol 77(11):6474–6481

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Akula SM, Naranatt PP, Walia NS, Wang FZ, Fegley B, Chandran B (2003) Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8) infection of human fibroblast cells occurs through endocytosis. J Virol 77(14):7978–7990

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Pertel PE (2002) Human herpesvirus 8 glycoprotein B (gB), gH, and gL can mediate cell fusion. J Virol 76(9):4390–4400

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Chaumorcel M, Souquère S, Pierron G, Codogno P, Esclatine A (2007) Human cytomegalovirus controls a new autophagy-dependent cellular antiviral defense mechanism. Autophagy 4(1):46–53

    Article  PubMed  Google Scholar 

  16. Orvedahl A, Alexander D, Tallóczy Z, Sun Q, Wei Y, Zhang W, Burns D, Leib DA, Levine B (2007) HSV-1 ICP34.5 confers neurovirulence by targeting the Beclin 1 autophagy protein. Cell Host Microbe 1(1):23–35

    Article  CAS  PubMed  Google Scholar 

  17. Orvedahl A, Levine B (2008) Viral evasion of autophagy. Autophagy 4(3):280–285

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Kudchodkar SB, Levine B (2009) Viruses and autophagy. Rev Med Virol 19(6):359–378

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Leidal AM, Cyr DP, Hill RJ, Lee PWK, McCormick C (2012) Subversion of autophagy by Kaposi’s sarcoma-associated herpesvirus impairs oncogene-induced senescence. Cell Host Microbe 11(2):167–180

    Article  CAS  PubMed  Google Scholar 

  20. Lee DY, Sugden B (2008) The latent membrane protein 1 oncogene modifies B-cell physiology by regulating autophagy. Oncogene 27(20):2833–2842

    Article  CAS  PubMed  Google Scholar 

  21. Wen HJ, Yang Z, Zhou Y, Wood C (2010) Enhancement of autophagy during lytic replication by the Kaposi’s sarcoma-associated herpesvirus replication and transcription activator. J Virol 84(15):7448–7458

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Boshoff C, Gao SJ, Healy LE, Matthews S, Thomas AJ, Coignet L, Warnke RA, Strauchen JA, Matutes E, Kamel OW, Moore PS, Weiss RA, Chang Y (1998) Establishing a KSHV+ cell line (BCP-1) from peripheral blood and characterizing its growth in Nod/SCID mice. Blood 91(5):1671–1679

    CAS  PubMed  Google Scholar 

  23. McGeoch DJ, Davison AJ (1999) The descent of human herpesvirus 8. Semin Cancer Biol 9(3):201–209

    Article  CAS  PubMed  Google Scholar 

  24. Poole LJ, Zong JC, Ciufo DM, Alcendor DJ, Cannon JS, Ambinder R, Orenstein JM, Reitz MS, Hayward GS (1999) Comparison of genetic variability at multiple loci across the genomes of the major subtypes of Kaposi’s sarcoma-associated herpesvirus reveals evidence for recombination and for two distinct types of open reading frame K15 alleles at the right-hand end. J Virol 73(8):6646–6660

    PubMed Central  CAS  PubMed  Google Scholar 

  25. Miller G, Rigsby MO, Heston L, Grogan E, Sun R, Metroka C, Levy JA, Gao SJ, Chang Y, Moore P (1996) Antibodies to butyrate inducible antigens of Kaposi’s sarcoma-associated herpesvirus in patients with HIV-1 infection. N Engl J Med 334(20):1292–1297

    Article  CAS  PubMed  Google Scholar 

  26. Berenson JR, Vescio RA (1999) HHV-8 is present in multiple myeloma patients. Blood 93(10):3157–3159

    CAS  PubMed  Google Scholar 

  27. Pan L, Milligan L, Michaeli J, Cesarman E, Knowles DM (2001) Polymerase chain reaction detection of Kaposi’s sarcoma-associated herpesvirus-optimized protocols and their application to myeloma. J Mol Diagn 3(1):32–38

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Greene W, Kuhne K, Ye F, Chen J, Zhou F, Lei X, Gao SJ (2007) Molecular biology of KSHV in relation to AIDS-associated oncogenesis. Cancer Treat Res 133:69–127

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Zhu FX, Cusano T, Yuan Y (1999) Identification of the immediate-early transcripts of Kaposi’s sarcoma-associated herpesvirus. J Virol 73(7):5556–5567

    PubMed Central  CAS  PubMed  Google Scholar 

  30. Sun Q, Fan W, Zhong Q (2009) Regulation of Beclin 1 in autophagy. Autophagy 5(5):713–716

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Hartford CM, Ratain MJ (2007) Rapamycin: something old, something new, sometimes borrowed and now renewed. Clin Pharmacol Ther 82:381–388

    Article  CAS  PubMed  Google Scholar 

  32. Renne R, Blackbourn D, Whitby D, Levy J, Ganem D (1998) Limited transmission of Kaposi’s sarcoma-associated herpesvirus in cultured cells. J Virol 72(6):5182–5188

    PubMed Central  CAS  PubMed  Google Scholar 

  33. Hahn A, Birkmann A, Wies E, Dorer D, Mahr K, Stürzl M, Titgemeyer F, Neipel F (2009) Kaposi’s sarcoma-associated herpesvirus gH/gL: glycoprotein export and interaction with cellular receptors. J Virol 83(1):396–407

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Nicholas J, Ruvolo VR, Burns WH, Sandford G, Wan X, Ciufo D, Hendrickson SB, Guo HG, Hayward GS, Reitz MS (1997) Kaposi’s sarcoma-associated human herpesvirus-8 encodes homologues of macrophage inflammatory protein-1 and interleukin-6. Nat Med 3(3):287–292

    Article  CAS  PubMed  Google Scholar 

  35. Mesri EA, Cesarman E, Boshoff C (2010) Kaposi’s sarcoma and its associated herpesvirus. Nat Rev Cancer 10(10):707–719

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  36. Lee JS, Li Q, Lee JY, Lee SH, Jeong JH, Lee HR, Chang H, Zhou FC, Gao SJ, Liang C, Jung JU (2009) FLIP-mediated autophagy regulation in cell death control. Nat Cell Biol 11(11):1355–1362

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Mijaljica D, Devenish RJ (2013) Nucleophagy at a glance. J Cell Sci 126(Pt 19):4325–4330

    Article  CAS  PubMed  Google Scholar 

  38. English L, Chemali M, Duron J, Rondeau C, Laplante A, Gingras D, Alexander D, Leib D, Norbury C, Lippé R, Desjardins M (2009) Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection. Nat Immunol 10(5):480–487

    Article  CAS  PubMed  Google Scholar 

  39. Pankiv S, Lamark T, Bruun JA, Øvervatn A, Bjørkøy G, Johansen T (2010) Nucleocytoplasmic shuttling of p62/SQSTM1 and its role in recruitment of nuclear polyubiquitinated proteins to promyelocytic leukemia bodies. J Biol Chem 285(8):5941–5953

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  40. Kirkegaard K, Taylor MP, Jackson WT (2004) Cellular autophagy: surrender, avoidance and subversion by microorganisms. Nat Rev Microbiol 2(4):301–304

    Article  CAS  PubMed  Google Scholar 

  41. Levine B (2005) Eating oneself and uninvited guests: autophagy-related pathways in cellular defense. Cell 120(2):159–162

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Hara Kang or Michael Lee.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jang, GH., Lee, J., Kim, NY. et al. Suppression of lytic replication of Kaposi’s sarcoma-associated herpesvirus by autophagy during initial infection in NIH 3T3 fibroblasts. Arch Virol 161, 595–604 (2016). https://doi.org/10.1007/s00705-015-2698-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-015-2698-2

Keywords

Navigation