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

Bernice Lo, Austin D E Swafford, Kimberly A. Shafer-Weaver, Lawrence F. Jerome, Luba Rakhlin, Douglas R. Mathern, Conor A. Callahan, Ping Jiang, Lucy J. Davison, Helen E. Stevens, Carrie L. Lucas, Jill White, Reid von Borstel, John A. Todd, Michael J. Lenardo

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Article number203
JournalJournal of Translational Medicine
Volume9
Issue number1
DOIs
Publication statusPublished - 28 Nov 2011
Externally publishedYes

Fingerprint

Insulin Antibodies
Inbred NOD Mouse
Medical problems
Type 1 Diabetes Mellitus
Assays
Autoantibodies
Insulin
Antibodies
Serum
Plasmas
Blood Glucose
Anti-Idiotypic Antibodies

Keywords

  • Diabetes
  • ECL
  • Electrochemiluminescence
  • Human autoantibodies
  • IA
  • IAA
  • Insulin
  • NOD mice

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

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. / Lo, Bernice; Swafford, Austin D E; Shafer-Weaver, Kimberly A.; Jerome, Lawrence F.; Rakhlin, Luba; Mathern, Douglas R.; Callahan, Conor A.; Jiang, Ping; Davison, Lucy J.; Stevens, Helen E.; Lucas, Carrie L.; White, Jill; von Borstel, Reid; Todd, John A.; Lenardo, Michael J.

In: Journal of Translational Medicine, Vol. 9, No. 1, 203, 28.11.2011.

Research output: Contribution to journalArticle

Lo, B, Swafford, ADE, Shafer-Weaver, KA, Jerome, LF, Rakhlin, L, Mathern, DR, Callahan, CA, Jiang, P, Davison, LJ, Stevens, HE, Lucas, CL, White, J, von Borstel, R, Todd, JA & Lenardo, MJ 2011, '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, vol. 9, no. 1, 203. https://doi.org/10.1186/1479-5876-9-203
Lo, Bernice ; Swafford, Austin D E ; Shafer-Weaver, Kimberly A. ; Jerome, Lawrence F. ; Rakhlin, Luba ; Mathern, Douglas R. ; Callahan, Conor A. ; Jiang, Ping ; Davison, Lucy J. ; Stevens, Helen E. ; Lucas, Carrie L. ; White, Jill ; von Borstel, Reid ; Todd, John A. ; Lenardo, Michael J. / 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. In: Journal of Translational Medicine. 2011 ; Vol. 9, No. 1.
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abstract = "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.",
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AU - Lo, Bernice

AU - Swafford, Austin D E

AU - Shafer-Weaver, Kimberly A.

AU - Jerome, Lawrence F.

AU - Rakhlin, Luba

AU - Mathern, Douglas R.

AU - Callahan, Conor A.

AU - Jiang, Ping

AU - Davison, Lucy J.

AU - Stevens, Helen E.

AU - Lucas, Carrie L.

AU - White, Jill

AU - von Borstel, Reid

AU - Todd, John A.

AU - Lenardo, Michael J.

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N2 - 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.

AB - 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.

KW - Diabetes

KW - ECL

KW - Electrochemiluminescence

KW - Human autoantibodies

KW - IA

KW - IAA

KW - Insulin

KW - NOD mice

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