Skip to main content
NCBI home page
Virologica Sinica logoLink to Virologica Sinica
. 2015 Jun 19;30(3):190–199. doi: 10.1007/s12250-015-3597-0

A respiratory syncytial virus persistent-infected cell line system reveals the involvement of SOCS1 in the innate antiviral response

Junwen Zheng 1, Pu Yang 1, Yan Tang 2, Dongchi Zhao 1,✉
PMCID: PMC8200879  PMID: 26122642

Abstract

HEp-2 cells persistently infected with respiratory syncytial virus (RSV) are a heterogeneous mixture of viral antigen-positive and -negative variants; however, the mechanism through which viral replication becomes latent remains unclear. In this study, we investigated the potential mechanism by which RSV escapes from innate immune surveillance. Persistent-infected RSV HEp-2 cells were isolated and cell clones were passaged. The RSV-persistent cells produced viruses at a lower titer, resisted wild-type RSV re-infection, and secreted high levels of interferon-ß (IFN-ß), macrophage inflammatory protein-1α (Mip-1α), interleukin-8 (IL-8), and Rantes. Toll-like receptor 3 (TLR3), retinoic acid inducible gene-I (RIG-I), and suppressor of cytokine signaling 1 (SOCS1) levels were upregulated in these cells. The silencing of TLR3 mRNA decreased the expression of SOCS1 protein and the secretion of cytokines. RSV-persistent cells are in an inflammatory state; upregulation of SOCS1 is related to the TLR3 signaling pathway, which could be associated with the mechanism of viral persistence.

Keywords: respiratory syncytial virus (RSV), suppressor of cytokine signaling 1 (SOCS1), interferon, toll-like receptor 3 (TLR3), RSV-persistent cell line

Footnotes

ORCID: 0000-0002-9454-6041

References

  1. Baetz A., Frey M., Heeg K., Dalpke A.H. Suppressor of cytokine signaling (SOCS) proteins indirectly regulate toll-like receptor signaling in innate immune cells. J Biol Chem. 2004;279:54708–54715. doi: 10.1074/jbc.M410992200. [DOI] [PubMed] [Google Scholar]
  2. Bangham C.R.M., McMichael A.J. Specific human cytotoxic T cells recognize B cell lines persistently infected with respiratory syncytial virus. Proc Natl Acad Sci USA. 1986;83:9183–9187. doi: 10.1073/pnas.83.23.9183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bode J.G., Ludwig S., Ehrhardt C., Albrecht U., Erhardt A., Schaper F., Heinrich P.C., Häussinger D. IFN-alpha antagonistic activity of HCV core protein involves induction of suppressor of cytokine signaling-3. Faseb J. 2003;17:488–490. doi: 10.1096/fj.02-0664fje. [DOI] [PubMed] [Google Scholar]
  4. Borg I., Rohde G., Löseke S., Bittscheidt J., Schultze-Werninghaus G., Stephan V., Bufe A. Evaluation of quantitative real-time PCR for the detection of respiratory syncytial virus in pulmonary diseases. Eur Respir J. 2003;21:944–951. doi: 10.1183/09031936.03.00088102. [DOI] [PubMed] [Google Scholar]
  5. Dalpke A., Heeg K., Bartz H., Baetz A. Regulation of innate immunity by suppressor of cytokine signaling (SOCS) proteins. Immunobiology. 2008;213:225–235. doi: 10.1016/j.imbio.2007.10.008. [DOI] [PubMed] [Google Scholar]
  6. Falsey A.R., Formica M.A., Hennessey P.A., Criddle M.M., Sullender W.M., Walsh E.E. Detection of respiratory syncytial virus in adults with chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2006;173:639–643. doi: 10.1164/rccm.200510-1681OC. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Groskreutz D.J., Monick M.M., Powers L.S., Yarovinsky T.O., Look D.C., Hunninghake G.W. Respiratory syncytial virus induces TLR3 protein and protein kinase R, leading to increased double-stranded RNA responsiveness in airway epithelial cells. J Immunol. 2006;176:1733–1740. doi: 10.4049/jimmunol.176.3.1733. [DOI] [PubMed] [Google Scholar]
  8. Hashimoto K., Ishibashi K., Ishioka K., Zhao D., Sato M., Ohara S., Abe Y., Kawasaki Y., Sato Y., Yokota S., Fujii N., Peebles R. J., Hosoya M., Suzutani T. RSV replication is attenuated by counteracting expression of the suppressor of cytokine signaling (SOCS) molecules. Virology. 2009;391:162–170. doi: 10.1016/j.virol.2009.06.026. [DOI] [PubMed] [Google Scholar]
  9. Herranz C., Melero J.A., Martínez I. Reduced innate immune response, apoptosis, and virus release in cells cured of respiratory syncytial virus persistent infection. Virology. 2011;410:56–63. doi: 10.1016/j.virol.2010.10.035. [DOI] [PubMed] [Google Scholar]
  10. Hobson L., Everard M.L. Persistent of respiratory syncytial virus in human dendritic cells and influence of nitric oxide. Clin Exp Immun. 2008;151:359–366. doi: 10.1111/j.1365-2249.2007.03560.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jeanette I., Webster M., Jacqueline C. Poly I:C and respiratory syncytial virus (RSV) inhibit glucocorticoid receptor (GR)-mediated transactivation in lung epithelial, but not monocytic, cell lines. Virus Res. 2013;176:303–306. doi: 10.1016/j.virusres.2013.06.011. [DOI] [PubMed] [Google Scholar]
  12. Le Goffic R., Pothlichet J., Vitour D., Fujita T., Meurs E., Chignard M., Si-Tahar M. Cutting Edge: Influenza A virus activates TLR3-dependent inflammatory and RIG-I-dependent antiviral responses in human lung epithelial cells. J Immunol. 2007;178:3368–3372. doi: 10.4049/jimmunol.178.6.3368. [DOI] [PubMed] [Google Scholar]
  13. Liu P., Jamaluddin M., Li K., Garofalo R.P., Casola A., Brasier A.R. Retinoic acid inducible gene-I mediates early anti-viral response and toll like receptor 3 expression in respiratory syncytial virus-infected airway epithelial cells. J Virol. 2007;81:1401–1411. doi: 10.1128/JVI.01740-06. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Martínez I., Lombardía L., Herranz C., García-Barreno B., Domínguez O., Melero J.A. Cultures of HEp-2 cells persistently infected by human respiratory syncytial virus differ in chemokine expression and resistance to apoptosis as compared to lytic infections of the same cell type. Virology. 2009;388:31–41. doi: 10.1016/j.virol.2009.03.008. [DOI] [PubMed] [Google Scholar]
  15. Matsuse H., Behera A.K., Kumar M., Rabb H., Lockey R.F., Mohapatra S.S. Recurrent respiratory syncytial virus infections in allergen-sensitized mice lead to persistent airway inflammation and hyperresponsiveness. J Immunol. 2000;164:6583–6592. doi: 10.4049/jimmunol.164.12.6583. [DOI] [PubMed] [Google Scholar]
  16. Oshansky C.M., Krunkosky T.M., Barber J., Jones L.P., Tripp R.A. Respiratory syncytial virus proteins modulate suppressors of cytokine signaling 1 and 3 and the type I interferon response to infection by a toll-like receptor pathway. Viral Immunol. 2009;22:147–161. doi: 10.1089/vim.2008.0098. [DOI] [PubMed] [Google Scholar]
  17. Pothlichet J., Chignard M., Si-Tahar M. Cutting Edge: Innate immune response triggered by influenza A virus is negatively regulated by SOCS1 and SOCS3 through a RIG-I/IFNAR1-dependent pathway. J Immunol. 2008;80:2034–2038. doi: 10.4049/jimmunol.180.4.2034. [DOI] [PubMed] [Google Scholar]
  18. Prêle C.M., Woodward E.A., Bisley J., Keith-Magee A., Nicholson S.E., Hart P.H. SOCS1 regulates the IFN but not NFκB pathway in TLR-stimulated human monocytes and macrophages. J Immunol. 2008;181:8018–8026. doi: 10.4049/jimmunol.181.11.8018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Psarras S., Papadopoulos N.G., Johnston S.L. Paediatr Respir Rev. 2004. Pathogenesis of respiratory syncytial virus bronchiolitis-related wheezing; pp. 179–184. [DOI] [PubMed] [Google Scholar]
  20. Riedel F., Oberdieck B., Streckert H.J., Philippou S., Krusat T., Marek W. Persistence of airway hyperresponsiveness and viral antigen following respiratory syncytial virus bronchiolitis in young guinea-pigs. Eur Respir J. 1997;10:639–645. [PubMed] [Google Scholar]
  21. Schwarze J., O’Donnell D.R., Rohwedder A., Openshaw P.J. Latency and persistence of respiratory syncytial virus despite T cell immunity. Am J Respir Crit Care Med. 2004;169:801–805. doi: 10.1164/rccm.200308-1203OC. [DOI] [PubMed] [Google Scholar]
  22. Seemungal T., Harper-Owen R., Bhowmik A., Moric I., Sanderson G., Message S., Maccallum P., Meade T.W., Jeffries D.J., Johnston S.L., Wedzicha J.A. Respiratory viruses, symptoms, and inflammatory markers in acute exacerbations and stable chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2001;164:1618–1623. doi: 10.1164/ajrccm.164.9.2105011. [DOI] [PubMed] [Google Scholar]
  23. Sigurs N., Bjarnason R., Sigurbergsson F., Kjellman B. Respiratory syncytial virus bronchiolitis in infancy is an important risk factor for asthma and allergy at age 7. Am J Respir Crit. Care Med. 2000;161:1501–1507. doi: 10.1164/ajrccm.161.5.9906076. [DOI] [PubMed] [Google Scholar]
  24. Sikkel M.B., Quint J.K., Mallia P., Wedzicha J.A., Johnston S.L. Respiratory syncytial virus persistence in chronic obstructive pulmonary disease. Pediatr Infect Dis J. 2008;27:63–70. doi: 10.1097/INF.0b013e3181684d67. [DOI] [PubMed] [Google Scholar]
  25. Stowell N.C., Seideman J., Raymond H.A., Smalley K.A., Lamb R.J., Egenolf D.D., Bugelski P.J., Murray L.A., Marsters P.A., Bunting R.A., Flavell R.A., Alexopoulou L., San Mateo L.R., Griswold D.E., Sarisky R.T., Mbow M.L., Das A.M. Long-term activation of TLR3 by Poly (I: C) induces inflammation and impairs lung function in mice. Respir Res. 2009;10:43. doi: 10.1186/1465-9921-10-43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tripp R.A. The brume surrounding respiratory syncytial virus persistence. Am. J Respir Crit Care Med. 2004;169:778–779. doi: 10.1164/rccm.2401013. [DOI] [PubMed] [Google Scholar]
  27. Valdovinos M.R., Gomez B. Establishment of respiratory syncytial virus persistence in cell lines: association with defective interfering particles. Intervirology. 2003;46:190–198. doi: 10.1159/000071461. [DOI] [PubMed] [Google Scholar]
  28. Vlotides G., Sörensen A.S., Kopp F., Zitzmann K., Cengic N., Brand S., Zachoval R., Auernhammer C.J. SOCS-1 and SOCS-3 inhibit IFN-alpha-induced expression of the antiviral proteins 2,5-OAS and MxA. Biochem Biophys Res Commun. 2004;320:1007–1014. doi: 10.1016/j.bbrc.2004.06.051. [DOI] [PubMed] [Google Scholar]
  29. Xu X., Zheng J., Zheng K., Hou Y., Zhao F., Zhao D. Respiratory syncytial virus NS1 protein degrades STAT2 by inducing SOCS1 expression. Intervirology. 2014;57:65–73. doi: 10.1159/000357327. [DOI] [PubMed] [Google Scholar]
  30. Yasukawa H., Yajima T., Duplain H., Iwatate M., Kido M., Hoshijima M., Weitzman M.D., Nakamura T., Woodard S., Xiong D., Yoshimura A., Chien K.R., Knowlton K.U. The suppressor of cytokine signaling-1 (SOCS1) is a novel therapeutic target for enterovirus-induced cardiac injury. J Clin Invest. 2003;111:469–478. doi: 10.1172/JCI16491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zhao D.C., Tang Y., Li L., You S.Y., Zhang C.Y. Respiratory syncytial virus inhibits interferon-a-inducible signaling in macrophage- like U937 cells. J Infect. 2007;54:393–398. doi: 10.1016/j.jinf.2006.06.005. [DOI] [PubMed] [Google Scholar]

Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

RESOURCES