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Evaluation of the f...
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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Richardson, Sarah J. (författare)
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Leete, Pia (författare)
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Dhayal, Shalinee (författare)
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Russell, Mark A. (författare)
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Oikarinen, Maarit (författare)
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Laiho, Jutta E. (författare)
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Svedin, Emma (författare)
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- Lind, Katharina (författare)
- Karolinska Institutet
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- Rosenling, Therese (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi
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Chapman, Nora (författare)
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Bone, Adrian J. (författare)
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Foulis, Alan K. (författare)
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- Frisk, Gun (författare)
- Uppsala universitet,Klinisk immunologi
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- Flodstrom-Tullberg, Malin (författare)
- Karolinska Institutet
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Hober, Didier (författare)
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Hyoty, Heikki (författare)
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Morgan, Noel G. (författare)
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(creator_code:org_t)
- 2013-11-05
- 2014
- Engelska.
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Ingår i: Diabetologia. - : Springer Science and Business Media LLC. - 0012-186X .- 1432-0428. ; 57:2, s. 392-401
- Relaterad länk:
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https://hdl.handle.n...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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http://kipublication...
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Abstract
Ämnesord
Stäng
- Aims/hypothesisEnteroviral 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.MethodsEnteroviral 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.ResultsClone 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.Conclusions/interpretationWhen 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.
Nyckelord
- ATP5B
- Coxsackievirus
- Creatine kinase
- Dako 5D8/1
- Immunohistochemistry
- Islets of Langerhans
- Pancreatic beta cell
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- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Richardson, Sara ...
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Leete, Pia
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Dhayal, Shalinee
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Russell, Mark A.
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Oikarinen, Maari ...
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Laiho, Jutta E.
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visa fler...
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Svedin, Emma
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Lind, Katharina
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Rosenling, There ...
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Chapman, Nora
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Bone, Adrian J.
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Foulis, Alan K.
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Frisk, Gun
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Flodstrom-Tullbe ...
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Hober, Didier
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Hyoty, Heikki
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Morgan, Noel G.
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visa färre...
- Artiklar i publikationen
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Diabetologia
- Av lärosätet
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Uppsala universitet
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Karolinska Institutet