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
<p>Abstract</p> <p>Background</p> <p>The detection of insulin autoantibodies (IAA) aids in the prediction of autoimmune diabetes development. However, the long-standing, gold standard <sup>125</sup>I-insulin radiobinding assay (RBA) has low reproducibility b...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2011-11-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | http://www.translational-medicine.com/content/9/1/203 |
| _version_ | 1818158011485519872 |
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| author | Lo Bernice Swafford Austin DE 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 |
| author_facet | Lo Bernice Swafford Austin DE 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 |
| author_sort | Lo Bernice |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The detection of insulin autoantibodies (IAA) aids in the prediction of autoimmune diabetes development. However, the long-standing, gold standard <sup>125</sup>I-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.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusions</p> <p>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.</p> |
| first_indexed | 2024-12-11T15:23:18Z |
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| id | doaj.art-ceb79db15f2c46d890623dc56e99023d |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-12-11T15:23:18Z |
| publishDate | 2011-11-01 |
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| spelling | doaj.art-ceb79db15f2c46d890623dc56e99023d2022-12-22T01:00:18ZengBMCJournal of Translational Medicine1479-58762011-11-019120310.1186/1479-5876-9-203Antibodies against insulin measured by electrochemiluminescence predicts insulitis severity and disease onset in non-obese diabetic mice and can distinguish human type 1 diabetes statusLo BerniceSwafford Austin DEShafer-Weaver Kimberly AJerome Lawrence FRakhlin LubaMathern Douglas RCallahan Conor AJiang PingDavison Lucy JStevens Helen ELucas Carrie LWhite Jillvon Borstel ReidTodd John ALenardo Michael J<p>Abstract</p> <p>Background</p> <p>The detection of insulin autoantibodies (IAA) aids in the prediction of autoimmune diabetes development. However, the long-standing, gold standard <sup>125</sup>I-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.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusions</p> <p>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.</p>http://www.translational-medicine.com/content/9/1/203NOD micediabeteshuman autoantibodiesinsulinelectrochemiluminescenceIAAIAECL |
| spellingShingle | Lo Bernice Swafford Austin DE 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 Journal of Translational Medicine NOD mice diabetes human autoantibodies insulin electrochemiluminescence IAA IA ECL |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | NOD mice diabetes human autoantibodies insulin electrochemiluminescence IAA IA ECL |
| url | http://www.translational-medicine.com/content/9/1/203 |
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