Detection of enterovirus RNA in pancreas and lymphoid tissues of organ donors with type 1 diabetes

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Abstract
Aims/hypothesis: The nPOD-Virus group collaboratively applied innovative technologies to detect and sequence viral RNA in pancreas and other tissues from organ donors with type 1 diabetes. These analyses involved the largest number of pancreas samples collected to date. The aim of the current work was to examine the presence of enterovirus RNA in pancreas and lymphoid tissues of organ donors with and without type 1 diabetes. Methods: We analysed pancreas, spleen, pancreatic lymph nodes and duodenum samples from the following groups: (1) donors with type 1 diabetes (n=71) with (n=35) or without (n=36) insulin-containing islets; (2) donors with single or double islet autoantibody positivity without diabetes (n=22); and (3) autoantibody-negative donors without diabetes (control donors) (n=74). Five research laboratories participated in this collaborative effort using approaches for unbiased discovery of RNA viruses (two RNA-Seq platforms), targeted detection of Enterovirus A–D species using RT-PCR, and tests for virus growth in cell culture. Results: Direct RNA-Seq did not detect virus signal in pancreas samples, whereas RT-PCR detected enterovirus RNA confirmed by sequencing in low amounts in pancreas samples in three of the five donor groups: donors with type 1 diabetes with insulin-containing islets, 16% (5/32) being positive; donors with single islet autoantibody positivity, 53% (8/15) being positive; and non-diabetic donors, 8% (4/49) being positive. Detection of enterovirus RNA was significantly more frequent in single islet autoantibody-positive donors compared with donors with type 1 diabetes with insulin-deficient islets (pp=0.004). In some donors, pancreatic lymph nodes were also positive. RT-PCR detected enterovirus RNA also in the spleen of a small number of donors and virus enrichment in susceptible cell lines before RT-PCR resulted in much higher rate in spleen positivity, particularly in donors with type 1 diabetes. Interestingly, the enterovirus strains detected did not cause a typical lytic infection, possibly reflecting their persistence-prone nature. Conclusions/interpretation: This was the largest coordinated effort to examine the presence of enterovirus RNA in the pancreas of organ donors with type 1 diabetes, using a multitude of assays. These findings are consistent with the notion that donors with type 1 diabetes and donors with islet autoantibodies may carry a low-grade enterovirus infection in the pancreas and lymphoid tissues. Graphical
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Funding Information
  • National Institutes of Health, USA (R01-AI50237)
  • Diabetestutkimussäätiö (personal grant for JEL)
  • Yrjö Jahnssonin Säätiö (personal grants for JEL, MO)
  • Leona M. and Harry B. Helmsley Charitable Trust (Grant#2018PG-T1D053, G-2108-04793)
  • European Foundation for the Study of Diabetes (97013)
  • Global Virus Network (GVN), University of Maryland, Baltimore
  • JDRF (25-2012-770, JDRF 25-2012-516, nPOD: 5-SRA-2018-557-Q-R)
  • network for Pancreatic Organ donors with Diabetes, nPOD (RRID:SCR_014641)
  • Pirkanmaan Rahasto (personal grant for JEL)
  • Seventh Framework Programme (Persistent Virus Infection in Diabetes Network [PE)
  • Sigrid Juséliuksen Säätiö (for HH)
  • Research Council of Finland (288671)
  • Päivikki ja Sakari Sohlbergin Säätiö (personal grants for JEL, MO)
  • Tampere University

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