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Diabetologia

, Volume 57, Issue 10, pp 2193–2200 | Cite as

Enterovirus RNA in longitudinal blood samples and risk of islet autoimmunity in children with a high genetic risk of type 1 diabetes: the MIDIA study

  • Ondrej CinekEmail author
  • Lars C. Stene
  • Lenka Kramna
  • German Tapia
  • Sami Oikarinen
  • Elisabet Witsø
  • Trond Rasmussen
  • Peter A. Torjesen
  • Heikki Hyöty
  • Kjersti S. Rønningen
Article

Abstract

Aims/hypothesis

Only a few longitudinal molecular studies of enterovirus and islet autoimmunity have been reported, and positive results seem to be limited to Finland. We aimed to investigate an association between enterovirus RNA in blood and islet autoimmunity in the MIDIA study from Norway, a country which largely shares environmental and economic features with Finland.

Methods

We analysed serial blood samples collected at ages 3, 6, and 9 months and then annually from 45 children who developed confirmed positivity for at least two autoantibodies (against insulin, GAD65 and IA-2) and 92 matched controls, all from a cohort of children with a single high-risk HLA-DQ-DR genotype. Enterovirus was tested in RNA extracted from frozen blood cell pellets, using real-time RT-PCR with stringent performance control.

Results

Out of 807 blood samples, 72 (8.9%) were positive for enterovirus. There was no association between enterovirus RNA and islet autoimmunity in samples obtained strictly before (7.6% cases, 10.0% controls, OR 0.75 [95% CI 0.36, 1.57]), or strictly after the first detection of islet autoantibodies (10.5% case, 5.8% controls, OR 2.00 [95% CI 0.64, 6.27]). However, there was a tendency towards a higher frequency of enterovirus detection in the first islet autoantibody-positive sample (15.8%) compared with the corresponding time point in matched controls (3.2%, OR 8.7 [95% CI 0.97, 77]). Neither of these results was changed by adjusting for potential confounders, restricting to various time intervals or employing various definitions of enterovirus positivity.

Conclusions/interpretation

Positivity for enterovirus RNA in blood did not predict the later induction of islet autoantibodies, but enterovirus tended to be detected more often at the islet autoantibody seroconversion stage.

Keywords

Autoimmunity Enterovirus Infancy Longitudinal Study RNA RT-PCR Virus-associated aetiology 

Abbreviation

DAISY

Diabetes Autoimmunity Study in the Young

Notes

Acknowledgements

We thank the public healthcare nurses for their recruitment efforts for the MIDIA study and the follow-up of high-risk children, and the staff at the Biobank Department at the Norwegian Institute of Public Health. In particular, we would like to express our gratitude to all of the parents for their efforts in handling their children’s type 1 diabetes risk, for allowing samples to be taken from their children and for completing questionnaires.

Funding

This study and the MIDIA project were funded by the Norwegian Organization for Health and Rehabilitation (2008/0182), the Ministry of Health of the Czech Republic (IGA MZ 11465–5), the Research Council of Norway (grants 135893/330, 155300/320, 156477/730, 205086/F20 and 166515/V50), the Norwegian Diabetes Association, the Academy of Finland (grant to HH) and the Project for the Conceptual Development of Research Organisation 00064203 (University Hospital Motol, Prague, Czech Republic). OC’s sabbatical at HH’s laboratory in 2012 was supported by an ISPAD Research Fellowship.

Duality of interest

HH is a minor shareholder (<5%) of Vactech Ltd., which develops picornavirus vaccines. All other authors declare that there is no duality of interest associated with their contribution to this manuscript.

Contribution statement

KSR and LCS conceived and designed the study. OC, LK, SO and HH collected data (performed or supervised the enterovirus testing), PAT collected data (tested islet autoantibodies), and TR managed the databases. LCS, GT, EW, TR and OC analysed and interpreted data. All authors drafted the manuscript and/or revised it for important intellectual content, and have approved the final version.

LCS is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Supplementary material

125_2014_3327_MOESM1_ESM.pdf (398 kb)
ESM Fig. 1 (PDF 397 kb)
125_2014_3327_MOESM2_ESM.pdf (32 kb)
ESM Fig. 2 (PDF 32 kb)
125_2014_3327_MOESM3_ESM.pdf (52 kb)
ESM Fig. 3 (PDF 52 kb)

References

  1. 1.
    Stene LC, Rewers M (2012) Immunology in the clinic review series; focus on type 1 diabetes and viruses: the enterovirus link to type 1 diabetes: critical review of human studies. Clin Exp Immunol 168:12–23PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Hyoty H, Taylor KW (2002) The role of viruses in human diabetes. Diabetologia 45:1353–1361PubMedCrossRefGoogle Scholar
  3. 3.
    Green J, Casabonne D, Newton R (2004) Coxsackie B virus serology and Type 1 diabetes mellitus: a systematic review of published case-control studies. Diabet Med 21:507–514PubMedCrossRefGoogle Scholar
  4. 4.
    Tauriainen S, Oikarinen S, Oikarinen M, Hyoty H (2011) Enteroviruses in the pathogenesis of type 1 diabetes. Semin Immunopathol 33:45–55PubMedCrossRefGoogle Scholar
  5. 5.
    Yeung WC, Rawlinson WD, Craig ME (2011) Enterovirus infection and type 1 diabetes mellitus: systematic review and meta-analysis of observational molecular studies. BMJ 342:d35PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Salminen KK, Vuorinen T, Oikarinen S et al (2004) Isolation of enterovirus strains from children with preclinical Type 1 diabetes. Diabet Med 21:156–164PubMedCrossRefGoogle Scholar
  7. 7.
    Salminen K, Sadeharju K, Lonnrot M et al (2003) Enterovirus infections are associated with the induction of beta-cell autoimmunity in a prospective birth cohort study. J Med Virol 69:91–98PubMedCrossRefGoogle Scholar
  8. 8.
    Oikarinen S, Martiskainen M, Tauriainen S et al (2011) Enterovirus RNA in blood is linked to the development of type 1 diabetes. Diabetes 60:276–279PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Sadeharju K, Hamalainen AM, Knip M et al (2003) Enterovirus infections as a risk factor for type I diabetes: virus analyses in a dietary intervention trial. Clin Exp Immunol 132:271–277PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Graves PM, Rotbart HA, Nix WA et al (2003) Prospective study of enteroviral infections and development of beta-cell autoimmunity. Diabetes autoimmunity study in the young (DAISY). Diabetes Res Clin Pract 59:51–61PubMedCrossRefGoogle Scholar
  11. 11.
    Tapia G, Cinek O, Rasmussen T et al (2011) Human enterovirus RNA in monthly fecal samples and islet autoimmunity in Norwegian children with high genetic risk for type 1 diabetes: the MIDIA study. Diabetes Care 34:151–155PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Stene LC, Witso E, Torjesen PA et al (2007) Islet autoantibody development during follow-up of high-risk children from the general Norwegian population from three months of age: design and early results from the MIDIA study. J Autoimmun 29:44–51PubMedCrossRefGoogle Scholar
  13. 13.
    Honkanen H, Oikarinen S, Pakkanen O et al (2013) Human enterovirus 71 strains in the background population and in hospital patients in Finland. J Clin Virol 56:348–353PubMedCrossRefGoogle Scholar
  14. 14.
    Beillard E, Pallisgaard N, van der Velden VH et al (2003) Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program. Leukemia 17:2474–2486PubMedCrossRefGoogle Scholar
  15. 15.
    Walter SD, Elwood JM (1975) A test for seasonality of events with a variable population at risk. Br J Prev Soc Med 29:18–21PubMedCentralPubMedGoogle Scholar
  16. 16.
    Hwang JY, Jun EJ, Seo I et al (2012) Characterization of infections of human leukocytes by non-polio enteroviruses. Intervirology 55:333–341PubMedCrossRefGoogle Scholar
  17. 17.
    Yin H, Berg AK, 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 51:1964–1971PubMedCrossRefGoogle Scholar
  18. 18.
    Schulte BM, Bakkers J, Lanke KH 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 23:99–104PubMedCrossRefGoogle Scholar
  19. 19.
    Perlman J, Gibson C, Pounds SB, Gu Z, Bankowski MJ, Hayden RT (2007) Quantitative real-time PCR detection of adenovirus in clinical blood specimens: a comparison of plasma, whole blood and peripheral blood mononuclear cells. J Clin Virol 40:295–300PubMedCrossRefGoogle Scholar
  20. 20.
    Ruf S, Behnke-Hall K, Gruhn B et al (2012) Comparison of six different specimen types for Epstein-Barr viral load quantification in peripheral blood of pediatric patients after heart transplantation or after allogeneic hematopoietic stem cell transplantation. J Clin Virol 53:186–194PubMedCrossRefGoogle Scholar
  21. 21.
    Sadeharju K, Knip M, Hiltunen M, Akerblom HK, Hyoty H (2003) The HLA-DR phenotype modulates the humoral immune response to enterovirus antigens. Diabetologia 46:1100–1105PubMedCrossRefGoogle Scholar
  22. 22.
    Oikarinen S, Tauriainen S, Hober D et al (2014) Virus antibody survey in different European populations indicates risk association between Coxsackievirus B1 and Type 1 diabetes. Diabetes 63:655–662PubMedCrossRefGoogle Scholar
  23. 23.
    Viskari H, Ludvigsson J, Uibo R et al (2004) Relationship between the incidence of type 1 diabetes and enterovirus infections in different European populations: results from the EPIVIR project. J Med Virol 72:610–617PubMedCrossRefGoogle Scholar
  24. 24.
    Lonnrot M, Knip M, Marciulionyte D et al (1999) Enterovirus antibodies in relation to islet cell antibodies in two populations with high and low incidence of type 1 diabetes. Diabetes Care 22:2086–2088PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Ondrej Cinek
    • 1
    Email author
  • Lars C. Stene
    • 2
  • Lenka Kramna
    • 1
  • German Tapia
    • 2
  • Sami Oikarinen
    • 3
  • Elisabet Witsø
    • 2
  • Trond Rasmussen
    • 2
  • Peter A. Torjesen
    • 4
  • Heikki Hyöty
    • 3
    • 5
  • Kjersti S. Rønningen
    • 6
  1. 1.Department of Paediatrics, 2nd Faculty of MedicineCharles University in Prague and University Hospital MotolPrague 5Czech Republic
  2. 2.Division of EpidemiologyNorwegian Institute of Public HealthOsloNorway
  3. 3.Department of VirologyUniversity of TampereTampereFinland
  4. 4.Hormone LaboratoryOslo University HospitalOsloNorway
  5. 5.Fimlab LaboratoriesPirkanmaa Hospital DistrictTampereFinland
  6. 6.Department of Pediatric ResearchOslo University HospitalOsloNorway

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