دورية أكاديمية
Evaluation of the fidelity of immunolabelling obtained with clone 5D8/1, a monoclonal antibody directed against the enteroviral capsid protein, VP1, in human pancreas
| العنوان: | 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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| المؤلفون: | Richardson, Sarah J., Leete, Pia, Dhayal, S, Russell, Mark A., Oikarinen, M, Laiho, JE, Svedin, E, Lind, K, Rosenling, T, Chapman, N, Bone, AJ, Foulis, Alan K., Frisk, G, Flodström-Tullberg, M, Hober, D, Hyoty, H, Morgan, Noel G. |
| بيانات النشر: | Springer Verlag (Germany) |
| سنة النشر: | 2014 |
| المجموعة: | University of Exeter: Open Research Exeter (ORE) |
| مصطلحات موضوعية: | ATP5B, Coxsackievirus, Creatine kinase, Dako 5D8/1, Immunohistochemistry, Islets of Langerhans, Pancreatic beta cell |
| الوصف: | Journal Article ; Copyright © Springer-Verlag Berlin Heidelberg 2013 ; Aims/hypothesis: 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. Methods: 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. Results: 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. Conclusions/interpretation: 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. © 2013 Springer-Verlag Berlin Heidelberg. ; European Union’s Seventh Framework Programme PEVNET (FP7/2007-2013) ; Diabetes Research and Wellness Foundation non-clinical research fellowship ; Karolinska Institutet ... |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| تدمد: | 0012-186X |
| العلاقة: | http://link.springer.com/article/10.1007%2Fs00125-013-3094-7Test; Vol. 57, Iss. 2, pp. 392 - 401; 261441; http://hdl.handle.net/10871/18086Test; Diabetologia |
| DOI: | 10.1007/s00125-013-3094-7 |
| الإتاحة: | https://doi.org/10.1007/s00125-013-3094-7Test http://hdl.handle.net/10871/18086Test |
| حقوق: | This is the authors post print. The final publication is available at Springer via http://dx.doi.orgTest/ 10.1007/s00125-013-3094-7 |
| رقم الانضمام: | edsbas.96613E97 |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 0012186X |
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| DOI: | 10.1007/s00125-013-3094-7 |