Journal of Clinical Immunology

, Volume 32, Issue 4, pp 778–785

Autoantibodies and High-Risk HLA Susceptibility Markers in First-Degree Relatives of Brazilian Patients with Type 1 Diabetes Mellitus: A Progression to Disease Based Study

  • L. I. Alves
  • E. Davini
  • M. R. Correia
  • R. T. Fukui
  • R. F. Santos
  • M. R. Cunha
  • D. M. Rocha
  • W. M. G. Volpini
  • M. E. R. Silva



The objective of this study was to determine the frequencies of autoantibodies to heterogeneous islet-cell cytoplasmic antigens (ICA), glutamic acid decarboxylase65 (GAD65A), insulinoma-associated antigen-2 (IA-2A) and insulin (IAA)–and human leukocyte antigen (HLA) class II markers (HLA-DR and -DQ) in first degree relatives of heterogeneous Brazilian patients with type I diabetes(T1DM). A major focus of this study was to determine the influence of age, gender, proband characteristics and ancestry on the prevalence of autoantibodies and HLA-DR and -DQ alleles on disease progression and genetic predisposition to T1DM among the first-degree relatives.


IAA, ICA, GAD65A, IA-2A and HLA- class II alleles were determined in 546 first-degree-relatives, 244 siblings, 55 offspring and 233 parents of 178 Brazilian patients with T1DM.


Overall, 8.9% of the relatives were positive for one or more autoantibodies. IAA was the only antibody detected in parents. GAD65 was the most prevalent antibody in offspring and siblings as compared to parents and it was the sole antibody detected in offspring. Five siblings were positive for the IA-2 antibody. A significant number (62.1%) of siblings had 1 or 2 high risk HLA haplotypes. During a 4-year follow-up study, 5 siblings (expressing HLA-DR3 or –DR4 alleles) and 1 offspring positive for GAD65A progressed to diabetes.


The data indicated that the GAD65 and IA-2 antibodies were the strongest predictors of T1DM in our study population. The high risk HLA haplotypes alone were not predictive of progression to overt diabetes.


Type I diabetes autoantibodies HLA-DR/DQ GAD65A IAA IA2-A 



Anti-glutamic acid decarboxylase 65 autoantibodies


Anti-insulin autoantibodies


Anti-Islet cell autoantibodies


Anti-protein tyrosine phosphatase-like IA-2 (IA-2A) autoantibodies


Human leukocyte antigen


Type 1 diabetes mellitus


  1. 1.
    Eisenbarth GS. Diabetes: a chronic autoimmune disease. N Engl J Med. 1986;314:1360–8.PubMedCrossRefGoogle Scholar
  2. 2.
    Atkinson MA, Maclaren NK. The pathogenesis of insulin-dependent diabetes mellitus. N Engl J Med. 1994;331:1428–36.PubMedCrossRefGoogle Scholar
  3. 3.
    Gillespie KM. Type 1 pathogenesis and prevention. CMAJ. 2006;175:165–70.PubMedCrossRefGoogle Scholar
  4. 4.
    Zhang L, Gianani R, Nakayama M, Liu E, Kobayashi M, Baschal E, Yu L, Babu S, Dawson A, Johnson K, Jahromi M, Aly T, Fain P, Barker J, Rewers M, Eisenbarth GS. Type 1 diabetes: chronic progressive autoimmune disease. Novartis Found Symp. 2008;292:85–94.PubMedCrossRefGoogle Scholar
  5. 5.
    Spielman RS, Baker L, Zmijewski CM. Gene dosage and susceptibility to insulin-dependent diabetes. Ann Hum Genet. 1980;2:135–50.CrossRefGoogle Scholar
  6. 6.
    Kumar D, Gemayel NS, Deapen D, Kapadia D, Yamashita PH, Lee M, Dwyer JH, Roy-Burman P, Bray GA, Mack TM. North-American twins with IDDM, genetic, etiological, and clinical significance of disease concordance according to age, zygosity, and the interval after diagnosis in first twin. Diabetes. 1993;42:1351–63.PubMedCrossRefGoogle Scholar
  7. 7.
    Sheehly MJ. Prospective in diabetes. HLA and insulin-dependent diabetes. A protective perspective. Diabetes. 1992;41:123–9.CrossRefGoogle Scholar
  8. 8.
    Davies JL, Kawaguchi Y, Bennett ST, Copeman JB, Cordell HJ, Pritchard LE, Reed PW, Gough SC, Jenkins SC, Palmer SM, et al. A genome-wide search for human type I diabetes susceptibility genes. Nature. 1994;371:130.PubMedCrossRefGoogle Scholar
  9. 9.
    Todd JA, Bell JJ, McDevitt HO. HLA-DQ/β gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature. 1987;329:599–604.PubMedCrossRefGoogle Scholar
  10. 10.
    Erlich H, Valdes AM, Noble J, Carlson JA, Varney M, Concannon P, Mychaleckyj JC, Todd JA, Bonella P, Fear AL, Lavant E, Louey A, Moonsamy P. HLA DR-DQ halotypes and genotypes and type 1 diabetes risk: analysis of the type 1 diabetes genetics consortium families. Diabetes. 2008;57:1084–92.PubMedCrossRefGoogle Scholar
  11. 11.
    Zhang L, Eisenbarth GS. Prediction and prevention of Type 1 diabetes mellitus. J Diabetes. 2011;3:48–57.PubMedCrossRefGoogle Scholar
  12. 12.
    Bonifacio E, Bingley PJ, Shattock M, Dean BM, Dunger D, Gale EA, Bottazzo GF. Quantification of islet-cell antibodies and prediction of insulin dependent diabetes. Lancet. 1990;335:147–9.PubMedCrossRefGoogle Scholar
  13. 13.
    Greenbaum CJ, Seras KL, Kahn SE, Palmer JP. Relationship of b-cells function and autoantibodies to progression and nonprogression of subclinical type 1 diabetes. Diabetes. 1999;48:170–5.PubMedCrossRefGoogle Scholar
  14. 14.
    Maclaren N, Lan M, Coutant R, Schatz D, Silverstein J, Muir A, Clare-Salzer M, She JX, Malone J, Crockett S, Schwartz S, Quattrin T, DeSilva M, Vander Vegt P, Notkins A, Krischer J. Only multiple autoantibodies to islet cells (ICA), insulin, GAD65, IA-2, and IA-2b predict immune-mediated (type 1) diabetes relatives. J Autoimmun. 1999;12:279–87.PubMedCrossRefGoogle Scholar
  15. 15.
    Achenbach P, Bonifacio E, Koczwara K, Ziegler AG. Natural history of type I diabetes. Diabetes. 2005;54 Suppl 2:S25–31.PubMedCrossRefGoogle Scholar
  16. 16.
    Bottazo GF, Florin-Christensen A, Doniach D. Islet cell antibodies in diabetes mellitus with autoimmune polyendocrine deficiencies. Lancet. 1974;2:1279–82.CrossRefGoogle Scholar
  17. 17.
    Lendrum R, Walker G, Cudworth AG, Theophanides C, Pyke DA, Bloom A, Gamble DR, et al. Islet-cell antibodies in diabetes mellitus. Lancet. 1976;2:1273–6.PubMedCrossRefGoogle Scholar
  18. 18.
    Baekkeskov S, Aanstoot H-J, Christgau S, Reetz A, Solimena M, Cascalho M, Folli F, Richter-Olesen H, De Camilli P. et al. Identification of the 64 K autoantigen in insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic acid decorboxylase [published erratum appears in (1990) Nature. 1990;347:782]. Nature.1990; 347151–156.Google Scholar
  19. 19.
    Payton MA, Hawkes CJ, Christie MR. Relationship of 37,000 and 40,000-Mr tryptic fragment of islet antigens in insulin dependent diabetes to the protein tyrosine phosphatase-like molecule IA-2 (ICA 512). J Clin Invest. 1995;96:1506–11.PubMedCrossRefGoogle Scholar
  20. 20.
    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;222:1337–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Pihoker C, Gilliam LK, Hampe CS, Lenmark Å. Autoantibodies in diabetes. Diabetes. 2005;54 Suppl 2:S52–61.PubMedCrossRefGoogle Scholar
  22. 22.
    Serrano-Rios M, Gutierrez-López MD, Pérez-Bravo F, Martínez MT, Antona J, Rowley M, Mackay I, Zimmet P. HLA-DR, DQ and anti-GAD antibodies in first degree relatives of type 1 diabetes mellitus. Diabetes Res Clin Pract. 1994;34 Suppl 34:S133–9.Google Scholar
  23. 23.
    Zeidler A, Raffel LJ, Costin G, Shaw SJ, Buchanan TA, Noble J, Rotter JI, Palmer J, Krischer JP, Wait C, Maclaren NK. Autoantibodies and human leucocyte antigen class II in first-degree family members of Mexican-American type 1 diabetic patients. J Clin Endocrinol Metab. 2001;86:4957–62.PubMedCrossRefGoogle Scholar
  24. 24.
    Wang J-P, Zhou Z-G, Lin J, Li X, Huang G, Zhou WD, Lin J, Zhou ZG, et al. Islet autoantibodies are associated with HLA-DQ genotypes in Han Chinese patients with type 1 diabetes and their relatives. Tissue Antigens. 2007;70:369–75.PubMedCrossRefGoogle Scholar
  25. 25.
    Williams AJ, Aitken RJ, Chandler MA, Gillespie KM, Lampsona V, Bingley PJ. Autoantibodies to islet antigen-2 are associated with KLA-DRBI*07, and DRBI*09 haplotypes as well as DRBI*04 at onset of type 1 diabetes: the possible role of HLA-DQA in autoimmunity to IA-2. Diabetologia. 2008;51:1444–8.PubMedCrossRefGoogle Scholar
  26. 26.
    Qu HQ, Polychronakos C. The effect of the MHC locus on autoantibodies in type 1 diabetes. J Med Genet. 2009;46:469–71.PubMedCrossRefGoogle Scholar
  27. 27.
    Howson JM, Stevens H, Smyth DJ, et al. Evidence that HLA class I and II associations with type 1 diabetes, autoantibodies to GAD and autoantibodies to 1A-2 are distinct. Diabetes. 2011;60:2635–44.PubMedCrossRefGoogle Scholar
  28. 28.
    van Belle TL, Coppieters KT, von Herrath MG. Type 1 diabetes: etiology, immunology, and therapeutic strategies. Physiol Rev. 2011;91:79–118.PubMedCrossRefGoogle Scholar
  29. 29.
    Lins TC, Vieira RG, Abreu BS, Grattapaglia D, Pereira RW. Genetic composition of Brazilian population samples based on a set of twenty-eight ancestry informative SNPs. Am J Hum Biol. 2010;22:187–92.PubMedGoogle Scholar
  30. 30.
    Wild S, Roglic G, Green A, Sicree R, King H. Global prevalence of diabetes. Estimates for the year 2000 and projections for 2030. Diabetes Care. 2004;27:1047–53.PubMedCrossRefGoogle Scholar
  31. 31.
    Ferreira SR, Franco LJ, Vivolo MA, Negrato CA, Simoes AC, Ventureli CR. Population based incidence of IDDM in the state of Sao Paulo, Brazil. Diabetes Care. 1993;16:701–4.PubMedCrossRefGoogle Scholar
  32. 32.
    Lisboa HR, Graebin R, Butzke L, Rodrigues CS. Incidence of type 1 diabetes mellitus in Passo Fundo, RS Brazil. Braz J Med Biol Res. 1998;31:553–6.CrossRefGoogle Scholar
  33. 33.
    Alves C, Meyer I, Vieira N, Toralles MBP, LeMaire D. Distribuiçã e freqüência de alelos e haplptipos HLA em Brasileiros com diabetes melito tipo 1. Arq Bras Endocrinol Metab. 2006;50:436–44.Google Scholar
  34. 34.
    Davini E, Silva MER, Alves LI, Correia MRS, Fukui RT, Latronico AC. O genótipo I/I do locus VNTR do gene da insulina confere risco independente para diabetes mellitus tipo 1 nos pacientes sem os genótipos DRB1 e DQB1 de susceptibilidade. Arq Bras Endocrinol Metab. 2005;49 Suppl 2:S876.Google Scholar
  35. 35.
    Bingley PJ, Colman P, Eisenbarth GS, Jackson RA, McCulloch DK, Riley WJ, Gale EA. Standardization of IVGTT to predict IDDM. Diabetes Care. 1992;15:1313–6.PubMedCrossRefGoogle Scholar
  36. 36.
    American Diabetes Association. Diagnosis and classification of diabetes. Diabetes Care. 2008;31 Suppl 1:S55–60.CrossRefGoogle Scholar
  37. 37.
    Vardi P, Ziegler AG, Mattews JH, Dib S, Keller RJ, Richer AT. Concentration of insulin autoantibodies at onset of type 1 diabetes. Inverse log-linear correlation with age. Diabetes Care. 1998;11:736–9.Google Scholar
  38. 38.
    Dib SA, Colman PG, Dotta F, Tautkus M, Rabizadeh A, Eisenbarth GS. Expression of cytoplasmic islet cell antigens by rat pancreas. Diabetes. 1987;36:982–5.PubMedCrossRefGoogle Scholar
  39. 39.
    Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16:1215.PubMedCrossRefGoogle Scholar
  40. 40.
    Eisenbarth GS, Lafferty K. Type 1 diabetes: cellular, molecular and clinical immunology. 2009.
  41. 41.
    Park Y. Why is type 1 diabetes uncommon in Asia? Ann NY Acad Sci. 2006;1079:31–40.PubMedCrossRefGoogle Scholar
  42. 42.
    Zeidler A, Frasier SD, Penny R, Stein RB, Nicoloff JT. Pancreatic islet cell and thyroid antibodies, and islet cell function in diabetic patients of Mexican-American origin. J Clin Endocrinol Metab. 1982;54:949–54.PubMedCrossRefGoogle Scholar
  43. 43.
    Shaw SJ, Vagheim CM, Rotter JI, Costin G, Beshkov Y, Zeidler A. Ancestral origin of insulin-dependent diabetes in Mexican-Americans. Intern J Epidemiol. 1992;21:725–9.CrossRefGoogle Scholar
  44. 44.
    Baker JM. Clinical review: type 1 diabetes-associated autoimmunity: natural history, genetic associations, and screening. J Clin Endocrinol Metab. 2006;91:1210–7.CrossRefGoogle Scholar
  45. 45.
    Bingley PJ. Interaction of age, islet cell antibodies, insulin antibodies, and first-phase insulin response in predicting risk of progression to IDDM in ICA + relatives. The ICARUS data. Islet Cell Antibody Register Users Study. Diabetes. 1996;45:1720–8.PubMedCrossRefGoogle Scholar
  46. 46.
    Verge CF, Gianani R, Kawasaki E, Yu L, Pietropolo M, Jackson RA, Chase HP, Eisenbarth GS. Prediction of type I diabetes in first-degree relatives using a combination of insulin, GAD, ICA512bdc/IA-2 autoantibodies. Diabetes. 1996;45:926–33.PubMedCrossRefGoogle Scholar
  47. 47.
    Cantor AB, Krischer JP, Cuthbertson DD, Schatz DA, Riley WJ, Malone J, Schwartz S, Quattrin T, Maclaren NK. Age and family relationship accentuate the risk of insulin-dependent diabetes mellitus (IDDM) in relatives of patients with IDDM. J Clin Endocrinol Metab. 1995;80:3739–43.PubMedCrossRefGoogle Scholar
  48. 48.
    Vandewalle CL, Falorni A, Svanholm S, Lernmark A, Pipeleers DG, Gorus FK. High diagnostic sensitivity of glutamate decarboxylase autoantibodies in insulin-dependent diabetes mellitus with clinical onset between age 20 and 40 years. The Belgian Diabetes Registry. J Clin Endocrinol Metab. 1995;80:846–51.PubMedCrossRefGoogle Scholar
  49. 49.
    Pugliese A, Gianani R, Moromissato R, Awdeh ZL, Alper CA, Erlich HA, Jackson RA, Eisenbath GS. HLA-DQB1*0602 is associated with dominant protection from diabetes even among islet cell antibody-positive first-degree relatives of patients with IDDM. Diabetes. 1995;44:608–13.PubMedCrossRefGoogle Scholar
  50. 50.
    Redondo MJ, Babu S, Zeidler A, Orban T, Yu L, Greenbaum C, Palmer JP, Cuthbertson D, Eisenbarth GS, Krischer JP, Schatz D. Diabetes Prevention Trial Type 1 StudyGroup. Specific human leukocyte antigen DQ influence on expression of antiislet autoantibodies and progression to type 1 diabetes. J Clin Endocrinol Metab. 2006;91:1705–13.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • L. I. Alves
    • 1
  • E. Davini
    • 1
  • M. R. Correia
    • 1
  • R. T. Fukui
    • 1
  • R. F. Santos
    • 1
  • M. R. Cunha
    • 1
  • D. M. Rocha
    • 1
  • W. M. G. Volpini
    • 2
  • M. E. R. Silva
    • 1
    • 3
  1. 1.Laboratory of Medical Investigation LIM-18Ambulatory of Endocrinology of Hospital das Clinicas of University São Paulo Medical SchoolSão PauloBrazil
  2. 2.Laboratory of ImmunologyUniversity of CampinasCampinasBrazil
  3. 3.Laboratory of Medical Investigation—LIM-18University of São Paulo Medical SchoolCerqueira CesarBrazil

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