Advertisement

Pancreatic beta cells persistently infected with coxsackievirus B4 are targets of NK cell-mediated cytolytic activity

  • Magloire Pandoua Nekoua
  • Antoine Bertin
  • Famara Sane
  • Enagnon Kazali Alidjinou
  • Delphine Lobert
  • Jacques Trauet
  • Christine Hober
  • Ilka Engelmann
  • Kabirou Moutairou
  • Akadiri Yessoufou
  • Didier HoberEmail author
Original Article

Abstract

It has been suggested that the persistence of coxsackieviruses-B (CV-B) in pancreatic beta cells plays a role in the pathogenesis of type 1 diabetes (T1D). Yet, immunological effectors, especially natural killer (NK) cells, are supposed to clear virus-infected cells. Therefore, an evaluation of the response of NK cells to pancreatic beta cells persistently infected with CV-B4 was conducted. A persistent CV-B4 infection was established in 1.1B4 pancreatic beta cells. Infectious particles were found in supernatants throughout the culture period. The proportion of cells containing viral protein VP1 was low (< 5%), although a large proportion of cells harbored viral RNA (around 50%), whilst cell viability was preserved. HLA class I cell surface expression was downregulated in persistently infected cultures, but HLA class I mRNA levels were unchanged in comparison with mock-infected cells. The cytolytic activities of IL-2-activated non-adherent peripheral blood mononuclear cells (PBMCs) and of NK cells were higher towards persistently infected cells than towards mock-infected cells, as assessed by an LDH release assay. Impaired cytolytic activity of IL-2-activated non-adherent PBMCs from patients with T1D towards infected beta cells was observed. In conclusion, pancreatic beta cells persistently infected with CV-B4 can be lysed by NK cells, implying that impaired cytolytic activity of these effector cells may play a role in the persistence of CV-B in the host and thus in the viral pathogenesis of T1D.

Keywords

Enterovirus Persistence HLA class I Type 1 diabetes LDH assay 

Notes

Acknowledgements

This work was supported by Ministere de l’Education Nationale de la Recherche et de la Technologie, Universite Lille 2 (Equipe d’accueil 3610), Centre Hospitalier Regional et Universitaire de Lille, and by EU FP7 (GA-261441-PEVNET: Persistent virus infection as a cause of pathogenic inlammation in type 1 diabetes—an innovative research program of biobanks and expertise). M. P. N was supported by a “CABRI 2016” scholarship of Universite Lille 2 and a “Programme Eifel 2017” scholarship of Ministere des Afaires etrangeres et du Developpement International de la Republique Francaise. Funding was supported by Campus France (EIFFELDOCTORAT 2017/n°P714914K). The authors thank Dr Sarah Richardson (Exeter, UK) for helpful discussion. The authors thank Dr Adrian J. F. Luty for reading the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Zell R, Delwart E, Gorbalenya AE, Hovi T, King AMQ, Knowles NJ, Lindberg AM, Pallansch MA, Palmenberg AC, Reuter G, Simmonds P, Skern T, Stanway G, Yamashita TIRC (2017) ICTV virus taxonomy profile: Picornaviridae. J Gen Virol 98:2421–2422CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Hober D, Alidjinou EK (2013) Enteroviral pathogenesis of type 1 diabetes: queries and answers. Curr Opin Infect Dis 26:263–269CrossRefPubMedGoogle Scholar
  3. 3.
    Hober D, Sauter P (2010) Pathogenesis of type 1 diabetes mellitus: interplay between enterovirus and host. Nat Rev Endocrinol 6:279–289CrossRefPubMedGoogle Scholar
  4. 4.
    Oikarinen S, Martiskainen M, Tauriainen S et al (2011) Enterovirus RNA in blood is linked to the development of type 1 diabetes. Diabetes 60:276–279CrossRefPubMedGoogle Scholar
  5. 5.
    Oikarinen M, Tauriainen S, Oikarinen S et al (2012) Type 1 diabetes is associated with enterovirus infection in gut mucosa. Diabetes 61:687–691CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Yeung W-CG, Rawlinson WD, Craig ME (2011) Enterovirus infection and type 1 diabetes mellitus: systematic review and meta-analysis of observational molecular studies. BMJ 342:d35–d35CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Alidjinou EK, Sané F, Engelmann I et al (2014) Enterovirus persistence as a mechanism in the pathogenesis of type 1 diabetes. Discov Med 18:273–282PubMedGoogle Scholar
  8. 8.
    Krogvold L, Edwin B, Buanes T et al (2015) Detection of a low-grade enteroviral infection in the islets of langerhans of living patients newly diagnosed with type 1 diabetes. Diabetes 64:1682–1687CrossRefPubMedGoogle Scholar
  9. 9.
    Alidjinou EK, Chehadeh W, Weill J et al (2015) Monocytes of patients with type 1 diabetes harbour enterovirus RNA. Eur J Clin Investig 45:918–924CrossRefGoogle Scholar
  10. 10.
    Yin H, Berg A-K, Tuvemo T, Frisk G (2002) Enterovirus RNA is found in peripheral blood mononuclear cells in a majority of type 1 diabetic children at onset. Diabetes 51:1964–1971CrossRefPubMedGoogle Scholar
  11. 11.
    Sane F, Caloone D, Gmyr V et al (2013) Coxsackievirus B4 can infect human pancreas ductal cells and persist in ductal-like cell cultures which results in inhibition of Pdx1 expression and disturbed formation of islet-like cell aggregates. Cell Mol Life Sci 70:4169–4180CrossRefPubMedGoogle Scholar
  12. 12.
    Alidjinou EK, Engelmann I, Bossu J et al (2017) Persistence of Coxsackievirus B4 in pancreatic ductal-like cells results in cellular and viral changes. Virulence 8:1229–1244CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Chehadeh W, Kerr-Conte J, Pattou F et al (2000) Persistent infection of human pancreatic islets by coxsackievirus B is associated with alpha interferon synthesis in beta cells. J Virol 74:10153–10164CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Richardson SJ, Morgan NG, Foulis AK (2014) Pancreatic pathology in type 1 diabetes mellitus. Endocr Pathol 25:80–92CrossRefPubMedGoogle Scholar
  15. 15.
    Warren H, Smyth M (1999) NK cells and apoptosis. Immunol Cell Biol 77:64–75CrossRefPubMedGoogle Scholar
  16. 16.
    Seliger B, Ritz U, Ferrone S (2006) Molecular mechanisms of HLA class I antigen abnormalities following viral infection and transformation. Int J Cancer 118:129–138CrossRefPubMedGoogle Scholar
  17. 17.
    Smyth MJ, Cretney E, Kelly JM et al (2005) Activation of NK cell cytotoxicity. Mol Immunol 42:501–510CrossRefPubMedGoogle Scholar
  18. 18.
    Vitale C, Chiossone L, Morreale G et al (2005) Human natural killer cells undergoing in vivo differentiation after allogeneic bone marrow transplantation: analysis of the surface expression and function of activating NK receptors. Mol Immunol 42:405–411CrossRefPubMedGoogle Scholar
  19. 19.
    Dotta F, Censini S, van Halteren AGS et al (2007) Coxsackie B4 virus infection of beta cells and natural killer cell insulitis in recent-onset type 1 diabetic patients. Proc Natl Acad Sci USA 104:5115–5120CrossRefPubMedGoogle Scholar
  20. 20.
    Baba M, Hasegawa H, Nakayabu M et al (1993) Cytolytic activity of natural killer cells and lymphokine activated killer cells against hepatitis A virus infected fibroblasts. J Clin Lab Immunol 40:47–60PubMedGoogle Scholar
  21. 21.
    Biron CA, Nguyen KB, Pien GC et al (1999) Natural killer cells in antiviral defense: function and regulation by innate cytokines. Annu Rev Immunol 17:189–220CrossRefPubMedGoogle Scholar
  22. 22.
    Godeny EK, Gauntt CJ (1986) Involvement of natural killer cells in coxsackievirus B3-induced murine myocarditis. J Immunol 137:1695–1702PubMedGoogle Scholar
  23. 23.
    Godeny EK, Gauntt CJ (1987) Murine natural killer cells limit coxsackievirus B3 replication. J Immunol 139:913–918PubMedGoogle Scholar
  24. 24.
    Hühn MH, Hultcrantz M, Lind K, Ljunggren HG, Malmberg KJF-TM (2008) IFN-γ production dominates the early human natural killer cell response to Coxsackievirus infection. Cell Microbiol 10:426–436PubMedGoogle Scholar
  25. 25.
    Alidjinou EK, Sané F, Engelmann I, Hober D (2013) Serum-dependent enhancement of Coxsackievirus B4-induced production of IFNα, IL-6 and TNFα by peripheral blood mononuclear cells. J Mol Biol 425:5020–5031CrossRefPubMedGoogle Scholar
  26. 26.
    Alidjinou EK, Sané F, Trauet J et al (2015) Coxsackievirus B4 can infect human peripheral blood-derived macrophages. Viruses 7:6067–6079CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Marca V, Gianchecchi E, Fierabracci A (2018) Type 1 diabetes and its multi-factorial pathogenesis: the putative role of NK cells. Int J Mol Sci.  https://doi.org/10.3390/ijms19030794 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Cornell CT, Kiosses WB, Harkins S, Whitton JL (2007) Coxsackievirus B3 proteins directionally complement each other to downregulate surface major histocompatibility complex class I. J Virol 81:6785–6797CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Campbell IL, Bizilj K, Colman PG et al (1986) Interferon-gamma induces the expression of HLA-A, B, C but not HLA-DR on human pancreatic beta-cells. J Clin Endocr Metab 62:1101–1109CrossRefPubMedGoogle Scholar
  31. 31.
    Rodríguez T, Méndez R, Del Campo A et al (2007) Distinct mechanisms of loss of IFN-gamma mediated HLA class I inducibility in two melanoma cell lines. BMC Cancer 7:34.  https://doi.org/10.1186/1471-2407-7-34 CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Cao J, Brouwer NJ, Jordanova ES et al (2018) HLA class I antigen expression in conjunctival melanoma is not associated with PD-L1/PD-1 status. Investig Ophthalmol Vis Sci 59:1005–1015CrossRefGoogle Scholar
  33. 33.
    Roivainen M, Rasilainen S, Ylipaasto P et al (2000) Mechanisms of coxsackievirus-induced damage to human pancreatic beta-cells. J Clin Endocr Metab 85:432–440PubMedGoogle Scholar
  34. 34.
    Vuorinen T, Nikolakaros G, Simell O et al (1992) Mumps and Coxsackie B3 virus infection of human fetal pancreatic islet-like cell clusters. Pancreas 7:460–464CrossRefPubMedGoogle Scholar
  35. 35.
    Yoon JW, Onodera T, Jenson AB, Notkins AL (1978) Virus-induced diabetes mellitus. XI. Replication of coxsackie B3 virus in human pancreatic beta cell cultures. Diabetes 27:778–781CrossRefPubMedGoogle Scholar
  36. 36.
    McCluskey JT, Hamid M, Guo-Parke H et al (2011) Development and functional characterization of insulin-releasing human pancreatic beta cell lines produced by electrofusion. J Biol Chem 286:21982–21992CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Benkahla MA, Alidjinou EK, Sane F et al (2018) Fluoxetine can inhibit coxsackievirus-B4 E2 in vitro and in vivo. Antiviral Res 159:130–133CrossRefPubMedGoogle Scholar
  38. 38.
    Alidjinou EK, Sané F, Bertin A et al (2015) Persistent infection of human pancreatic cells with Coxsackievirus B4 is cured by fluoxetine. Antiviral Res 116:51–54CrossRefPubMedGoogle Scholar
  39. 39.
    Heim A, Canu A, Kirschner P et al (1992) Synergistic interaction of interferon-beta and interferon-gamma in coxsackievirus B3-infected carrier cultures of human myocardial fibroblasts. J Infect Dis 166:958–965CrossRefPubMedGoogle Scholar
  40. 40.
    Heim A, Brehm C, Stille-Siegener M et al (1995) Cultured human myocardial fibroblasts of pediatric origin: natural human interferon-alpha is more effective than recombinant interferon-alpha 2a in carrier-state coxsackievirus B3 replication. J Mol Cell Cardiol 27:2199–2208CrossRefPubMedGoogle Scholar
  41. 41.
    Pinkert S, Klingel K, Lindig V et al (2011) Virus-host coevolution in a persistently coxsackievirus B3-infected cardiomyocyte cell line. J Virol 85:13409–13419CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Richardson SJ, Willcox A, Bone AJ et al (2009) The prevalence of enteroviral capsid protein vp1 immunostaining in pancreatic islets in human type 1 diabetes. Diabetologia 52:1143–1151CrossRefPubMedGoogle Scholar
  43. 43.
    Pujol-Borrell R, Todd I, Doshi M et al (1986) Differential expression and regulation of MHC products in the endocrine and exocrine cells of the human pancreas. Clin Exp Immunol 65:128–139PubMedPubMedCentralGoogle Scholar
  44. 44.
    Deitz SB, Dodd DA, Cooper S et al (2000) MHC I-dependent antigen presentation is inhibited by poliovirus protein 3A. Proc Natl Acad Sci USA 97:13790–13795CrossRefPubMedGoogle Scholar
  45. 45.
    Moffat K, Howell G, Knox C et al (2005) Effects of foot-and-mouth disease virus nonstructural proteins on the structure and function of the early secretory pathway: 2BC but not 3A blocks endoplasmic reticulum-to-Golgi transport. J Virol 79:4382–4395CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Kirkegaard K, Taylor MP, Jackson WT (2004) Cellular autophagy: surrender, avoidance and subversion by microorganisms. Nat Rev Microbiol 2:301–314CrossRefPubMedGoogle Scholar
  47. 47.
    de Jong AS, Visch H-J, de Mattia F et al (2006) The coxsackievirus 2B protein increases efflux of ions from the endoplasmic reticulum and Golgi, thereby inhibiting protein trafficking through the Golgi. J Biol Chem 281:14144–14150CrossRefPubMedGoogle Scholar
  48. 48.
    Cornell CT, Kiosses WB, Harkins S, Whitton JL (2006) Inhibition of protein trafficking by coxsackievirus b3: multiple viral proteins target a single organelle. J Virol 80:6637–6647CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Salvesen GS, Dixit VM (1997) Caspases: intracellular signaling by proteolysis. Cell 91:443–446CrossRefPubMedGoogle Scholar
  50. 50.
    Li ZM, Liu ZC, Guan ZZ et al (2004) Inhibition of DNA primase and induction of apoptosis by 3,3′-diethyl-9-methylthia-carbocyanine iodide in hepatocellular carcinoma BEL-7402 cells. World J Gastroenterol 10:514–520CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Vives-Pi M, Rodríguez-Fernández S, Pujol-Autonell I (2015) How apoptotic β-cells direct immune response to tolerance or to autoimmune diabetes: a review. Apoptosis 20:263–272CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Mathis D, Vence L, Benoist C (2001) Beta-cell death during progression to diabetes. Nature 414:792–798CrossRefPubMedGoogle Scholar
  53. 53.
    O’Brien BA, Geng X, Orteu CH et al (2006) A deficiency in the in vivo clearance of apoptotic cells is a feature of the NOD mouse. J Autoimmun 26:104–115CrossRefPubMedGoogle Scholar
  54. 54.
    Eizirik DL, Grieco FA (2012) On the immense variety and complexity of circumstances conditioning pancreatic-cell apoptosis in type 1 diabetes. Diabetes 61:1661–1663CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Wilson RG, Anderson J, Shenton BK et al (1986) Natural killer cells in insulin dependent diabetes mellitus. Br Med J 293:244CrossRefGoogle Scholar
  56. 56.
    Hussain MJ, Alviggi L, Millward BA et al (1987) Evidence that the reduced number of natural killer cells in type 1 (insulin-dependent) diabetes may be genetically determined. Diabetologia 30:907–911CrossRefPubMedGoogle Scholar
  57. 57.
    Negishi K, Waldeck N, Chandy G et al (1986) Natural killer cell and islet killer cell activities in type 1 (insulin-dependent) diabetes. Diabetologia 29:352–357CrossRefPubMedGoogle Scholar
  58. 58.
    Lorini R, Moretta A, Valtorta A et al (1994) Cytotoxic activity in children with insulin-dependent diabetes mellitus. Diabetes Res Clin Pract 23:37–42CrossRefPubMedGoogle Scholar
  59. 59.
    Qin H, Lee IF, Panagiotopoulos C et al (2011) Natural killer cells from children with type 1 diabetes have defects in NKG2D-dependent function and signaling. Diabetes 60:857–866CrossRefPubMedPubMedCentralGoogle Scholar
  60. 60.
    Rodacki M, Svoren B, Butty V et al (2007) Altered natural killer cells in type 1 diabetic patients. Diabetes 56:177–185CrossRefPubMedGoogle Scholar
  61. 61.
    Hofmann P, Schmidtke M, Stelzner A, Gemsa D (2001) Suppression of proinflammatory cytokines and induction of IL-10 in human monocytes after coxsackievirus B3 infection. J Med Virol 64:487–498CrossRefPubMedGoogle Scholar
  62. 62.
    Ylipaasto P, Klingel K, Lindberg AM et al (2004) Enterovirus infection in human pancreatic islet cells, islet tropism in vivo and receptor involvement in cultured islet beta cells. Diabetologia 47:225–239CrossRefPubMedGoogle Scholar
  63. 63.
    Schulte BM, Bakkers J, Lanke KHW et al (2010) Detection of enterovirus RNA in peripheral blood mononuclear cells of type 1 diabetic patients beyond the stage of acute infection. Viral Immunol 23:99–104CrossRefPubMedGoogle Scholar
  64. 64.
    Willcox A, Richardson SJ, Bone AJ et al (2011) Immunohistochemical analysis of the relationship between islet cell proliferation and the production of the enteroviral capsid protein, VP1, in the islets of patients with recent-onset type 1 diabetes. Diabetologia 54:2417–2420CrossRefPubMedGoogle Scholar
  65. 65.
    Lima JF, Oliveira LMS, Pereira NZ et al (2017) Polyfunctional natural killer cells with a low activation profile in response to Toll-like receptor 3 activation in HIV-1-exposed seronegative subjects. Sci Rep 7:524.  https://doi.org/10.1038/s41598-017-00637-3 CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Flodström M, Maday A, Balakrishna D et al (2002) Target cell defense prevents the development of diabetes after viral infection. Nat Immunol 3:373–382CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Magloire Pandoua Nekoua
    • 1
    • 2
  • Antoine Bertin
    • 1
  • Famara Sane
    • 1
  • Enagnon Kazali Alidjinou
    • 1
  • Delphine Lobert
    • 1
  • Jacques Trauet
    • 3
  • Christine Hober
    • 4
  • Ilka Engelmann
    • 1
  • Kabirou Moutairou
    • 2
  • Akadiri Yessoufou
    • 2
  • Didier Hober
    • 1
    • 5
    Email author
  1. 1.Université de Lille, Faculté de Médecine, CHU de Lille, Laboratoire de Virologie EA3610LilleFrance
  2. 2.Université d’Abomey-Calavi, Faculté des Sciences et Techniques, Institut des Sciences Biomédicales Appliquées (ISBA), Laboratoire de Biologie et Physiologie CellulairesCotonouBenin
  3. 3.Université de Lille, INSERM U995, LIRIC-Lille, CHU de Lille, Institut d’ImmunologieLilleFrance
  4. 4.Polyclinique, Service de Médecine ProgramméeHenin-BeaumontFrance
  5. 5.Laboratoire de Virologie EA3610, Centre Paul Boulanger, Hôpital A Calmette, CHRULille CedexFrance

Personalised recommendations