Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on sleep in brain health

LIVE: Thursday 2nd May 2024, 18:00-19:30 (CEST)

Quality sleep is essential for health. But what happens to our brains when sleep patterns are disturbed? Join our experts to explore the interplay between sleep disruption and neurological diseases, and the questions that you need to be asking your patients to help you prevent the harmful effects of sleep deprivation.
ADVANCED SEARCH
Log in
Top
Journal of Translational Medicine
Published in: Journal of Translational Medicine 1/2011

Open Access 01-12-2011 | Research

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

Authors: Bernice Lo, Austin DE 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

Published in: Journal of Translational Medicine | Issue 1/2011

Login to get access

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.
Appendix
Available only for authorised users
Literature
1.
Wucherpfennig KW, Sethi D: T cell receptor recognition of self and foreign antigens in the induction of autoimmunity. Semin Immunol. 2011, 23: 84-91. 10.1016/j.smim.2011.01.007.PubMedCentralCrossRefPubMed
2.
Lleo A, Invernizzi P, Gao B, Podda M, Gershwin ME: Definition of human autoimmunity--autoantibodies versus autoimmune disease. Autoimmun Rev. 2010, 9: A259-266. 10.1016/j.autrev.2009.12.002.CrossRefPubMed
3.
Melendez-Ramirez LY, Richards RJ, Cefalu WT: Complications of type 1 diabetes. Endocrinol Metab Clin North Am. 2010, 39: 625-640. 10.1016/j.ecl.2010.05.009.CrossRefPubMed
4.
van Belle TL, Coppieters KT, von Herrath MG: Type 1 diabetes: etiology, immunology, and therapeutic strategies. Physiol Rev. 2011, 91: 79-118. 10.1152/physrev.00003.2010.CrossRefPubMed
5.
6.
7.
Verge CF, Gianani R, Kawasaki E, Yu L, Pietropaolo M, Jackson RA, Chase HP, Eisenbarth GS: Prediction of type I diabetes in first-degree relatives using a combination of insulin, GAD, and ICA512bdc/IA-2 autoantibodies. Diabetes. 1996, 45: 926-933. 10.2337/diabetes.45.7.926.CrossRefPubMed
8.
Verge CF, Gianani R, Kawasaki E, Yu L, Pietropaolo M, Chase HP, Eisenbarth GS: Number of autoantibodies (against insulin, GAD or ICA512/IA2) rather than particular autoantibody specificities determines risk of type I diabetes. J Autoimmun. 1996, 9: 379-383. 10.1006/jaut.1996.0051.CrossRefPubMed
9.
Siljander HT, Veijola R, Reunanen A, Virtanen SM, Akerblom HK, Knip M: Prediction of type 1 diabetes among siblings of affected children and in the general population. Diabetologia. 2007, 50: 2272-2275. 10.1007/s00125-007-0799-5.CrossRefPubMed
10.
Korhonen S, Knip MM, Kulmala P, Savola K, Akerblom HK, Knip M: Autoantibodies to GAD, IA-2 and insulin in ICA-positive first-degree relatives of children with type 1 diabetes: a comparison between parents and siblings. Diabetes Metab Res Rev. 2002, 18: 43-48. 10.1002/dmrr.258.CrossRefPubMed
11.
Wenzlau JM, Liu Y, Yu L, Moua O, Fowler KT, Rangasamy S, Walters J, Eisenbarth GS, Davidson HW, Hutton JC: A common nonsynonymous single nucleotide polymorphism in the SLC30A8 gene determines ZnT8 autoantibody specificity in type 1 diabetes. Diabetes. 2008, 57: 2693-2697. 10.2337/db08-0522.PubMedCentralCrossRefPubMed
12.
Verge CF, Stenger D, Bonifacio E, Colman PG, Pilcher C, Bingley PJ, Eisenbarth GS: Combined use of autoantibodies (IA-2 autoantibody, GAD autoantibody, insulin autoantibody, cytoplasmic islet cell antibodies) in type 1 diabetes: Combinatorial Islet Autoantibody Workshop. Diabetes. 1998, 47: 1857-1866. 10.2337/diabetes.47.12.1857.CrossRefPubMed
13.
Achenbach P, Warncke K, Reiter J, Naserke HE, Williams AJ, Bingley PJ, Bonifacio E, Ziegler AG: Stratification of type 1 diabetes risk on the basis of islet autoantibody characteristics. Diabetes. 2004, 53: 384-392. 10.2337/diabetes.53.2.384.CrossRefPubMed
14.
Kimpimaki T, Kulmala P, Savola K, Kupila A, Korhonen S, Simell T, Ilonen J, Simell O, Knip M: Natural history of beta-cell autoimmunity in young children with increased genetic susceptibility to type 1 diabetes recruited from the general population. J Clin Endocrinol Metab. 2002, 87: 4572-4579. 10.1210/jc.2002-020018.CrossRefPubMed
15.
Yu L, Robles DT, Abiru N, Kaur P, Rewers M, Kelemen K, Eisenbarth GS: Early expression of antiinsulin autoantibodies of humans and the NOD mouse: evidence for early determination of subsequent diabetes. Proc Natl Acad Sci USA. 2000, 97: 1701-1706. 10.1073/pnas.040556697.PubMedCentralCrossRefPubMed
16.
Colman PG, Steele C, Couper JJ, Beresford SJ, Powell T, Kewming K, Pollard A, Gellert S, Tait B, Honeyman M, Harrison LC: Islet autoimmunity in infants with a Type I diabetic relative is common but is frequently restricted to one autoantibody. Diabetologia. 2000, 43: 203-209. 10.1007/s001250050030.CrossRefPubMed
17.
Mrena S, Virtanen SM, Laippala P, Kulmala P, Hannila ML, Akerblom HK, Knip M: Models for predicting type 1 diabetes in siblings of affected children. Diabetes Care. 2006, 29: 662-667. 10.2337/diacare.29.03.06.dc05-0774.CrossRefPubMed
18.
Siljander H, Harkonen T, Hermann R, Simell S, Hekkala A, Salonsaari RT, Simell T, Simell O, Ilonen J, Veijola R, Knip M: Role of insulin autoantibody affinity as a predictive marker for type 1 diabetes in young children with HLA-conferred disease susceptibility. Diabetes Metab Res Rev. 2009, 25: 615-622. 10.1002/dmrr.998.CrossRefPubMed
19.
Steck AK, Johnson K, Barriga KJ, Miao D, Yu L, Hutton JC, Eisenbarth GS, Rewers MJ: Age of islet autoantibody appearance and mean levels of insulin, but not GAD or IA-2 autoantibodies, predict age of diagnosis of type 1 diabetes: diabetes autoimmunity study in the young. Diabetes Care. 2011, 34: 1397-1399. 10.2337/dc10-2088.PubMedCentralCrossRefPubMed
20.
Melanitou E, Devendra D, Liu E, Miao D, Eisenbarth GS: Early and quantal (by litter) expression of insulin autoantibodies in the nonobese diabetic mice predict early diabetes onset. J Immunol. 2004, 173: 6603-6610.CrossRefPubMed
21.
Robles DT, Eisenbarth GS, Dailey NJ, Peterson LB, Wicker LS: Insulin autoantibodies are associated with islet inflammation but not always related to diabetes progression in NOD congenic mice. Diabetes. 2003, 52: 882-886. 10.2337/diabetes.52.3.882.CrossRefPubMed
22.
Thayer TC, Wilson SB, Mathews CE: Use of nonobese diabetic mice to understand human type 1 diabetes. Endocrinol Metab Clin North Am. 2010, 39: 541-561. 10.1016/j.ecl.2010.05.001.PubMedCentralCrossRefPubMed
23.
Delovitch TL, Singh B: The nonobese diabetic mouse as a model of autoimmune diabetes: immune dysregulation gets the NOD. Immunity. 1997, 7: 727-738. 10.1016/S1074-7613(00)80392-1.CrossRefPubMed
24.
Anderson MS, Bluestone JA: The NOD mouse: a model of immune dysregulation. Annu Rev Immunol. 2005, 23: 447-485. 10.1146/annurev.immunol.23.021704.115643.CrossRefPubMed
25.
Greenbaum CJ, Palmer JP, Kuglin B, Kolb H: Insulin autoantibodies measured by radioimmunoassay methodology are more related to insulin-dependent diabetes mellitus than those measured by enzyme-linked immunosorbent assay: results of the Fourth International Workshop on the Standardization of Insulin Autoantibody Measurement. J Clin Endocrinol Metab. 1992, 74: 1040-1044. 10.1210/jc.74.5.1040.PubMed
26.
Schlosser M, Mueller PW, Torn C, Bonifacio E, Bingley PJ: Diabetes Antibody Standardization Program: evaluation of assays for insulin autoantibodies. Diabetologia. 2010, 53: 2611-2620. 10.1007/s00125-010-1915-5.CrossRefPubMed
27.
Bonifacio E, Atkinson M, Eisenbarth G, Serreze D, Kay TW, Lee-Chan E, Singh B: International Workshop on Lessons From Animal Models for Human Type 1 Diabetes: identification of insulin but not glutamic acid decarboxylase or IA-2 as specific autoantigens of humoral autoimmunity in nonobese diabetic mice. Diabetes. 2001, 50: 2451-2458. 10.2337/diabetes.50.11.2451.CrossRefPubMed
28.
Bingley PJ: Clinical applications of diabetes antibody testing. J Clin Endocrinol Metab. 2010, 95: 25-33. 10.1210/jc.2009-1365.CrossRefPubMed
29.
Williams AJ, Curnock R, Reed CR, Easton P, Rokni S, Bingley PJ: Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays. J Immunol Methods. 2010, 362: 199-203. 10.1016/j.jim.2010.09.009.CrossRefPubMed
30.
Williams AJ, Bingley PJ, Bonifacio E, Palmer JP, Gale EA: A novel micro-assay for insulin autoantibodies. J Autoimmun. 1997, 10: 473-478. 10.1006/jaut.1997.0154.CrossRefPubMed
31.
Zancheta R, Russo V, Presotto F, Magrin L, Pedini B, Betterle C: Detection of insulin autoantibodies using an ELISA technique in first-degree relatives of IDDM patients and in autoimmune patients. Diabetes Res. 1987, 6: 189-194.PubMed
32.
Forster RJ, Bertoncello P, Keyes TE: Electrogenerated chemiluminescence. Annu Rev Anal Chem (Palo Alto Calif). 2009, 2: 359-385. 10.1146/annurev-anchem-060908-155305.CrossRef
33.
Bertoncello P, Forster RJ: Nanostructured materials for electrochemiluminescence (ECL)-based detection methods: recent advances and future perspectives. Biosens Bioelectron. 2009, 24: 3191-3200. 10.1016/j.bios.2009.02.013.CrossRefPubMed
34.
Blackburn GF, Shah HP, Kenten JH, Leland J, Kamin RA, Link J, Peterman J, Powell MJ, Shah A, Talley DB: Electrochemiluminescence detection for development of immunoassays and DNA probe assays for clinical diagnostics. Clin Chem. 1991, 37: 1534-1539.PubMed
35.
Lewczuk P, Kamrowski-Kruck H, Peters O, Heuser I, Jessen F, Popp J, Burger K, Hampel H, Frolich L, Wolf S: Soluble amyloid precursor proteins in the cerebrospinal fluid as novel potential biomarkers of Alzheimer's disease: a multicenter study. Mol Psychiatry. 2010, 15: 138-145. 10.1038/mp.2008.84.CrossRefPubMed
36.
Jesse S, Steinacker P, Lehnert S, Sdzuj M, Cepek L, Tumani H, Jahn O, Schmidt H, Otto M: A proteomic approach for the diagnosis of bacterial meningitis. PLoS One. 2010, 5: e10079-10.1371/journal.pone.0010079.PubMedCentralCrossRefPubMed
37.
Gisslen M, Krut J, Andreasson U, Blennow K, Cinque P, Brew BJ, Spudich S, Hagberg L, Rosengren L, Price RW, Zetterberg H: Amyloid and tau cerebrospinal fluid biomarkers in HIV infection. BMC Neurol. 2009, 9: 63-10.1186/1471-2377-9-63.PubMedCentralCrossRefPubMed
38.
Guglielmo-Viret V, Attree O, Blanco-Gros V, Thullier P: Comparison of electrochemiluminescence assay and ELISA for the detection of Clostridium botulinum type B neurotoxin. J Immunol Methods. 2005, 301: 164-172. 10.1016/j.jim.2005.04.003.CrossRefPubMed
39.
Guglielmo-Viret V, Thullier P: Comparison of an electrochemiluminescence assay in plate format over a colorimetric ELISA, for the detection of ricin B chain (RCA-B). J Immunol Methods. 2007, 328: 70-78. 10.1016/j.jim.2007.08.003.CrossRefPubMed
40.
McInerney MF, Pek SB, Thomas DW: Prevention of insulitis and diabetes onset by treatment with complete Freund's adjuvant in NOD mice. Diabetes. 1991, 40: 715-725. 10.2337/diabetes.40.6.715.CrossRefPubMed
41.
Leiter EH: The NOD mouse: a model for insulin-dependent diabetes mellitus. Curr Protoc Immunol. 2001, Chapter 15 (Unit 15): 19-
42.
Hanley JA, McNeil BJ: A method of comparing the areas under receiver operating characteristic curves derived from the same cases. Radiology. 1983, 148: 839-843.CrossRefPubMed
43.
Deaver DR: A new non-isotopic detection system for immunoassays. Nature. 1995, 377: 758-760. 10.1038/377758a0.CrossRefPubMed
44.
Kimpimaki T, Kupila A, Hamalainen AM, Kukko M, Kulmala P, Savola K, Simell T, Keskinen P, Ilonen J, Simell O, Knip M: The first signs of beta-cell autoimmunity appear in infancy in genetically susceptible children from the general population: the Finnish Type 1 Diabetes Prediction and Prevention Study. J Clin Endocrinol Metab. 2001, 86: 4782-4788. 10.1210/jc.86.10.4782.PubMed
45.
Polychronakos C, Li Q: Understanding type 1 diabetes through genetics: advances and prospects. Nat Rev Genet. 2011, 12: 781-792. 10.1038/nrg3069.CrossRefPubMed
46.
Nakayama M, Abiru N, Moriyama H, Babaya N, Liu E, Miao D, Yu L, Wegmann DR, Hutton JC, Elliott JF, Eisenbarth GS: Prime role for an insulin epitope in the development of type 1 diabetes in NOD mice. Nature. 2005, 435: 220-223. 10.1038/nature03523.PubMedCentralCrossRefPubMed
47.
Butler JE: Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays. Methods. 2000, 22: 4-23. 10.1006/meth.2000.1031.CrossRefPubMed
48.
Ohsugi T, Kurosawa T: Increased incidence of diabetes mellitus in specific pathogen-eliminated offspring produced by embryo transfer in NOD mice with low incidence of the disease. Lab Anim Sci. 1994, 44: 386-388.PubMed
49.
Baxter AG, Koulmanda M, Mandel TE: High and low diabetes incidence nonobese diabetic (NOD) mice: origins and characterisation. Autoimmunity. 1991, 9: 61-67. 10.3109/08916939108997125.CrossRefPubMed
50.
Palmer JP, Asplin CM, Clemons P, Lyen K, Tatpati O, Raghu PK, Paquette TL: Insulin antibodies in insulin-dependent diabetics before insulin treatment. Science. 1983, 222: 1337-1339. 10.1126/science.6362005.CrossRefPubMed
51.
Thomas JW, Virta VJ, Nell LJ: Heterogeneity and specificity of human anti-insulin antibodies determined by isoelectric focusing. J Immunol. 1985, 134: 1048-1052.PubMed
52.
Thomas JW, Beckwith M, Nell LJ: Spectrotypic analysis of antibodies to insulin A and B chains. Mol Immunol. 1988, 25: 173-179. 10.1016/0161-5890(88)90065-X.CrossRefPubMed
53.
Nell LJ, Hulbert C, Thomas JW: Human insulin autoantibody fine specificity and H and L chain use. J Immunol. 1989, 142: 3063-3069.PubMed
54.
Greenbaum CJ, Palmer JP: Insulin antibodies and insulin autoantibodies. Diabet Med. 1991, 8: 97-105. 10.1111/j.1464-5491.1991.tb01553.x.CrossRefPubMed
55.
Wenzlau JM, Juhl K, Yu L, Moua O, Sarkar SA, Gottlieb P, Rewers M, Eisenbarth GS, Jensen J, Davidson HW, Hutton JC: The cation efflux transporter ZnT8 (Slc30A8) is a major autoantigen in human type 1 diabetes. Proc Natl Acad Sci USA. 2007, 104: 17040-17045. 10.1073/pnas.0705894104.PubMedCentralCrossRefPubMed
56.
LaGasse JM, Brantley MS, Leech NJ, Rowe RE, Monks S, Palmer JP, Nepom GT, McCulloch DK, Hagopian WA: Successful prospective prediction of type 1 diabetes in schoolchildren through multiple defined autoantibodies: an 8-year follow-up of the Washington State Diabetes Prediction Study. Diabetes Care. 2002, 25: 505-511. 10.2337/diacare.25.3.505.CrossRefPubMed
Metadata
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
Authors
Bernice Lo
Austin DE 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
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Journal of Translational Medicine / Issue 1/2011
Electronic ISSN: 1479-5876
DOI
https://doi.org/10.1186/1479-5876-9-203

Other articles of this Issue 1/2011

Journal of Translational Medicine 1/2011 Go to the issue

Research

Cathepsin B: a potential prognostic marker for inflammatory breast cancer

Research

Periostin: a promising target of therapeutical intervention for prostate cancer

Research

Buffy coat specimens remain viable as a DNA source for highly multiplexed genome-wide genetic tests after long term storage

Research

Safety and pharmacokinetics of recombinant human hepatocyte growth factor (rh-HGF) in patients with fulminant hepatitis: a phase I/II clinical trial, following preclinical studies to ensure safety

Research

Bioactivities of berberine metabolites after transformation through CYP450 isoenzymes

Research

Therapeutic effects of pyrrolidine dithiocarbamate on acute lung injury in rabbits

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24 to hear Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits