Antibodies against insulin measured by electrochemiluminescence predicts insulitis severity and disease onset in non-obese diabetic mice and can distinguish human type 1 diabetes status

Journal of Translational Medicine

Research

John A Todd¹  Helen E Stevens¹  Lucy J Davison¹  Douglas R Mathern²  Kimberly A Shafer-Weaver²  Carrie L Lucas²  Ping Jiang²  Conor A Callahan²  Bernice Lo²  Michael J Lenardo²  Luba Rakhlin²  Austin DE Swafford³  Lawrence F Jerome⁴  Jill White⁴  Reid von Borstel⁵ 

[1] Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Biomedical Research Centre, University of Cambridge, Addenbrooke's Hospital, CB2 0XY, Cambridge, UK;Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892, Bethesda, MD, USA;Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 20892, Bethesda, MD, USA;Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Biomedical Research Centre, University of Cambridge, Addenbrooke's Hospital, CB2 0XY, Cambridge, UK;Wellstat Diagnostics, LLC, 930 Clopper Rd, 20878, Gaithersburg, MD, USA;Wellstat Therapeutics Corporation, 930 Clopper Rd, 20878, Gaithersburg, MD, USA;

关键词: NOD mice;    diabetes;    human autoantibodies;    insulin;    electrochemiluminescence;    IAA;    IA;    ECL;   

DOI  :  10.1186/1479-5876-9-203

 received in 2011-11-23, accepted in 2011-11-28,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe detection of insulin autoantibodies (IAA) aids in the prediction of autoimmune diabetes development. However, the long-standing, gold standard 125I-insulin radiobinding assay (RBA) has low reproducibility between laboratories, long sample processing times and requires the use of newly synthesized radiolabeled insulin for each set of assays. Therefore, a rapid, non-radioactive, and reproducible assay is highly desirable.MethodsWe have developed electrochemiluminescence (ECL)-based assays that fulfill these criteria in the measurement of IAA and anti-insulin antibodies (IA) in non-obese diabetic (NOD) mice and in type 1 diabetic individuals, respectively. Using the murine IAA ECL assay, we examined the correlation between IAA, histopathological insulitis, and blood glucose in a cohort of female NOD mice from 4 up to 36 weeks of age. We developed a human IA ECL assay that we compared to conventional RBA and validated using samples from 34 diabetic and 59 non-diabetic individuals in three independent laboratories.ResultsOur ECL assays were rapid and sensitive with a broad dynamic range and low background. In the NOD mouse model, IAA levels measured by ECL were positively correlated with insulitis severity, and the values measured at 8-10 weeks of age were predictive of diabetes onset. Using human serum and plasma samples, our IA ECL assay yielded reproducible and accurate results with an average sensitivity of 84% at 95% specificity with no statistically significant difference between laboratories.ConclusionsThese novel, non-radioactive ECL-based assays should facilitate reliable and fast detection of antibodies to insulin and its precursors sera and plasma in a standardized manner between laboratories in both research and clinical settings. Our next step is to evaluate the human IA assay in the detection of IAA in prediabetic subjects or those at risk of type 1 diabetes and to develop similar assays for other autoantibodies that together are predictive for the diagnosis of this common disorder, in order to improve prediction and facilitate future therapeutic trials.

【 授权许可】

Unknown   
© Lo et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.