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Evaluation of the fidelity of immunolabelling obtained with clone 5D8/1, a monoclonal antibody directed against the enteroviral capsid protein, VP1, in human pancreas
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2014 (English)In: Diabetologia, ISSN 0012-186X, E-ISSN 1432-0428, Vol. 57, no 2, 392-401 p.Article in journal (Refereed) Published
Abstract [en]


Enteroviral infection has been implicated in the development of islet autoimmunity in type 1 diabetes and enteroviral antigen expression has been detected by immunohistochemistry in the pancreatic beta cells of patients with recent-onset type 1 diabetes. However, the immunohistochemical evidence relies heavily on the use of a monoclonal antibody, clone 5D8/1, raised against an enteroviral capsid protein, VP1. Recent data suggest that the clone 5D8/1 may also recognise non-viral antigens; in particular, a component of the mitochondrial ATP synthase (ATP5B) and an isoform of creatine kinase (CKB). Therefore, we evaluated the fidelity of immunolabelling by clone 5D8/1 in the islets of patients with type 1 diabetes.


Enteroviral VP1, CKB and ATP5B expression were analysed by western blotting, RT-PCR and immunocytochemistry in a range of cultured cell lines, isolated human islets and human tissue.


Clone 5D8/1 labelled CKB, but not ATP5B, on western blots performed under denaturing conditions. In cultured human cell lines, isolated human islets and pancreas sections from patients with type 1 diabetes, the immunolabelling of ATP5B, CKB and VP1 by 5D8/1 was readily distinguishable. Moreover, in a human tissue microarray displaying more than 80 different cells and tissues, only two (stomach and colon; both of which are potential sites of enterovirus infection) were immunopositive when stained with clone 5D8/1.


When used under carefully optimised conditions, the immunolabelling pattern detected in sections of human pancreas with clone 5D8/1 did not reflect cross-reactivity with either ATP5B or CKB. Rather, 5D8/1 is likely to be representative of enteroviral antigen expression.

Place, publisher, year, edition, pages
2014. Vol. 57, no 2, 392-401 p.
Keyword [en]
ATP5B, Coxsackievirus, Creatine kinase, Dako 5D8/1, Immunohistochemistry, Islets of Langerhans, Pancreatic beta cell
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-218545DOI: 10.1007/s00125-013-3094-7ISI: 000329615100017OAI: oai:DiVA.org:uu-218545DiVA: diva2:696042
Available from: 2014-02-12 Created: 2014-02-12 Last updated: 2014-02-12Bibliographically approved

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Frisk, Gun
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