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

Michael J Lenardo¹  John A Todd⁴  Reid von Borstel²  Jill White³  Carrie L Lucas¹  Helen E Stevens⁴  Lucy J Davison⁴  Ping Jiang¹  Conor A Callahan¹  Douglas R Mathern¹  Luba Rakhlin¹  Lawrence F Jerome³  Kimberly A Shafer-Weaver¹  Austin DE Swafford⁴  Bernice Lo¹ 

[1] Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA;Wellstat Therapeutics Corporation, 930 Clopper Rd, Gaithersburg, MD 20878, USA;Wellstat Diagnostics, LLC, 930 Clopper Rd, Gaithersburg, MD 20878, 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, Cambridge, CB2 0XY, UK

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

Others  :  1207815
DOI  :  10.1186/1479-5876-9-203

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

【 摘 要 】

Background

The 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.

Methods

We 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.

Results

Our 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.

Conclusions

These 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】

   
2011 Lo et al; licensee BioMed Central Ltd.

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