Autoantibodies and HLA class II DR-DQ genotypes in Ugandan children and adolescents with type 1 diabetes mellitus

  • Silver Bahendeka
  • Ronald Wesonga
  • Thereza Piloya Were
  • Catherine Nyangabyaki
Original Article


The aims were to determine the prevalence of autoantibodies in type 1 diabetes mellitus (T1DM) and further to investigate the human leukocyte antigen (HLA) class II DR-DQ genotypes associated with T1DM in Ugandan children and adolescents. Cross-sectional data were collected between January and December 2015 from 85 recently detected T1DM children and adolescents and 79 age-matched healthy controls. We measured serum concentrations of C-peptide, vitamin D, insulin autoantibodies (IAA), zinc transporter family member 8 antibodies (ZnT8-Ab), and glutamic acid decarboxylase autoantibodies (GADA). HLA-DBR1 and HLA-DQB1 typing was performed on EDTA-anticoagulated blood samples. The t test, chi-square test, and univariate logistic test were performed and multivariate logistic regression model fitted to identify associated factors of T1DM. Positive IAA and ZnT8-Ab were significantly higher in T1DM than in controls. GADA showed no difference between TIDM and controls. HLA-DQB1*02, unadjusted odds ratio (UOR) 4.2 (95% CI 1.4–12.7), and HLA-DBR1*04, adjusted odds ratio (AOR) 30.6 (95% CI 5.7–161.7), were significantly associated with T1DM. IAA and ZnT8-Ab are the likely significant positive antibodies in Ugandan children and adolescents with T1DM. The T1DM was associated with HLA-DQB1*02 and HLA-DBR1*04 (HLA-DR3 and HLA-DR4) genotypes.


Type 1 diabetes Autoantibodies HLA class II Uganda 



We would like to acknowledge the contribution of Dr. William Omale, Arua Regional Hospital, for his assistance in organizing the type 1 diabetes clinic in Arua Regional Hospital, Uganda. We also like to thank MBN Clinical Laboratory and St. Francis Hospital, Nsambya, for the kind assistance in handling the laboratory samples and analysis. We thank the World Diabetes Foundation (WDF) and Novo Nordisk Changing Diabetes in Children (CDiC) for funding the type 1 diabetes clinics in Uganda. We wish to thank all the health workers and parents involved in the type 1 diabetes clinics in Uganda.

Authors contributions

All the authors, SKB, RW, TPW, and CN, contributed to the design, data collection, data analysis, and writing of the manuscript. All authors: read and approved the final manuscript.


This study was supported by a grant from the Africa Field Epidemiology Network (AFENET). The funds were towards the laboratory costs. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent

Informed consent was obtained from all individual participants included in the study: children aged 8–17 years assented to participate, with parents of children below age of 18 years giving the informed consent. Participants above the age of 18 years were eligible and gave informed consent to participate.

Ethical approval

Ethical approval was obtained from St. Francis Hospital, Nsambya and Mulago Hospital Institutional Review Boards/Review Ethical Committees; both of which are accredited by the Uganda National Council of Science and Technology (UNCST). All procedures performed in this study were in accordance with the ethical standards of UNCST and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.


  1. 1.
    American Diabetes Association. Standards of medical care in diabetes—2017 abridged for primary care providers. Clin Diabetes. 2017;35:5–26.CrossRefPubMedCentralGoogle Scholar
  2. 2.
    Bahadoran Z, Ghasemi A, Mirmiran P, Azizi F, Hadaegh F. Nitrate-nitrite-nitrosamines exposure and the risk of type 1 diabetes: a review of current data. World J Diabetes. 2016;7:433–40.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Rewers M, Ludvigsson J. Environmental risk factors for type 1 diabetes. Lancet. 2016;387:2340–8.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    DIAMOND Project Group. Incidence and trends of childhood type 1 diabetes worldwide 1990-1999. Diabet Med. 2006;23:857–66.CrossRefGoogle Scholar
  5. 5.
    International Diabetes Federation. IDF diabetes atlas, Seventh edn (Brussels 2015).Google Scholar
  6. 6.
    Borchers AT, Uibo R, Gershwin ME. The geoepidemiology of type 1 diabetes. Autoimmun Rev. 2010;9:A355–65.CrossRefPubMedGoogle Scholar
  7. 7.
    Kalk WJ, Huddle KRL, Raal FJ. The age of onset and sex distribution of insulin-dependent diabetes mellitus in Africans in South Africa. Postgrad Med J. 1993;69:552–6.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    You W-P, Henneberg M. Type 1 diabetes prevalence increasing globally and regionally: the role of natural selection and life expectancy at birth. BMJ Open Diabetes Res Care. 2016;4(1):e000161. Scholar
  9. 9.
    Cook AR. Notes on the diseases met with in Uganda, Central Africa. J Trop Med. 1901;4:175–8.Google Scholar
  10. 10.
    Piloya-Were T, Sunni M, Ogle GD, Moran A. Childhood diabetes in Africa. Curr Opin Endocrinol Diabetes Obes. 2016;23:306–11.CrossRefPubMedGoogle Scholar
  11. 11.
    World Health Organization. Global report on diabetes. Geneva: World Health Organization; 2016.Google Scholar
  12. 12.
    Padoa C. The epidemiology and pathogenesis of type 1 diabetes mellitus in Africa. J Endocrinol Metab Diabetes S Afr. 2011;16:130–6.CrossRefGoogle Scholar
  13. 13.
    Steck AK, Rewers MJ. Genetics of type 1 diabetes. Clin Chem. 2011;57:176–85.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Noble JA, Erlich HA. Genetics of type 1 diabetes. Cold Spring Harb Perspect Med. 2012;2:a007732. Scholar
  15. 15.
    Nguyen C, Varney MD, Harrison LC, Morahan G. Definition of high-risk type 1 diabetes HLA-DR and HLA-DQ types using only three single nucleotide polymorphisms. Diabetes. 2013;62:2135–40.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Howson JMM, Stevens H, Smyth DJ, Walker NM, Chandler KA, Bingley PJ, et al. Evidence that HLA class I and II associations with type 1 diabetes, autoantibodies to GAD and autoantibodies to IA-2, are distinct. Diabetes. 2011;60:2635–44.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Roark CL, Anderson KM, Simon LJ, Schuyler RP, Aubrey MT, Freed BM. Multiple HLA epitopes contribute to type 1diabetes susceptibility. Diabetes. 2014;63:323–31.CrossRefPubMedGoogle Scholar
  18. 18.
    Noble JA, Valdes AM, Varney MD, Carlson JA, Moonsamy P, Fear AL, et al. HLA class I and genetic susceptibility to type 1 diabetes. Results from the type 1 diabetes genetics consortium. Diabetes. 2010;59:2972–9.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Barrett JC, et al. Genome-wide association study and meta-analysis find that over 40 loci affect risk of type 1 diabetes. Nat Genet. 2009;41:703–7.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Kantarova D, Vrlik M, Buc M. Genetic determination and immunopathogenesis of type 1 diabetes mellitus in humans. Acta Med Mart. 2012;12:19–31.Google Scholar
  21. 21.
    Christie MR, Vohra G, Champagne P, Daneman D, Delovitch TL. Distinct antibody specificities to a 64-kD islet cell antigen in type 1 diabetes as revealed by trypsin treatment. J Exp Med. 1990;172:789–94.CrossRefPubMedGoogle Scholar
  22. 22.
    Biekkeskov S, et al. Antibodies to a 64,000 Mr human islet cell antigen precede the clinical onset of insulin-dependent diabetes. J Clin Invest. 1987;79:926–34.CrossRefGoogle Scholar
  23. 23.
    Ludvigsson J, Carlsson A, Forsander G, Ivarsson S, Kockum I, Lernmark Å, et al. C-peptide in the classification of diabetes in children and adolescents. Pediatr Diabetes. 2012;13:45–50.CrossRefPubMedGoogle Scholar
  24. 24.
    Kuhtreiber WM, Washer SLL, Hsu E, Zhao M, Reinhold P III, Burger D, et al. Low levels of C-peptide have clinical significance for established type 1 diabetes. Diabet Med. 2015;32:1346–53.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Jones AG, Hattersley AT. The clinical utility of C-peptide measurement in the care of patients with diabetes. Diabet Med. 2013;30:803–17.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Davis AK, DuBose S, Haller MJ, Miller KM, DiMeglio L, Bethin KE, et al. Prevalence of detectable C-peptide according to age at diagnosis and duration of type 1 diabetes. Diabete Care. 2015;38:476–81.CrossRefGoogle Scholar
  27. 27.
    Nokoff N, Rewers M. Pathogenesis of type 1 diabetes: lessons from natural history studies of high-risk individuals. Ann N Y Acad Sci. 2013;1281:1–15.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Atkinson MA. The pathogenesis and natural history of type 1 diabetes. Cold Spring Harb Perspect Med. 2012;2:a007641. Scholar
  29. 29.
    The Expert Committee On the Diagnosis and Classification of Diabetes Mellitus. Report of the expert committee on the diagnosis and classification of diabetes mellitus. Diabetes Care. 2003;26:S5–20.CrossRefGoogle Scholar
  30. 30.
    Acharjee S, Ghosh B, Al-Dhubiab BE, Nair AB. Understanding type 1 diabetes: etiology and models. Can J Diabetes. 2013;37:269–76.CrossRefPubMedGoogle Scholar
  31. 31.
    Rubio-Cabezas O, Hattersley AT, Njølstad PR, Mlynarski W, Ellard S, White N, et al. ISPAD clinical practice consensus guidelines 2014. The diagnosis and management of monogenic diabetes in children and adolescents. Pediatr Diabetes. 2014;15(Suppl 20):47–64.CrossRefPubMedGoogle Scholar
  32. 32.
    Asanghanwa M, et al. Clinical and biological characteristics of diabetic patients under age 40 in Cameroon: relation to autoantibody status and comparison with Belgian patients. DRCP. 2014;103:97–105.Google Scholar
  33. 33.
    Holick MF. High prevalence of vitamin D inadequacy and implications for health. Mayo Clin Proc. 2006;81:353–73.CrossRefPubMedGoogle Scholar
  34. 34.
    Mack SJ, Cano P, Hollenbach JA, He J, Hurley CK, Middleton D, et al. Common and well-documented HLA alleles: 2012 update to the CWD catalogue. Tissue Antigens. 2013;81:194–203.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Mayor NP, Robinson J, McWhinnie AJM, Ranade S, Eng K, Midwinter W, et al. HLA typing for the next generation. PLoS One. 2015;10:e0127153. Scholar
  36. 36.
    Xie M, Li J, Jiang T. Accurate HLA type inference using a weighted similarity graph. BMC Bioinformatics. 2010;11(Suppl 11):S10. Scholar
  37. 37.
    Morran MP, Vonberg A, Khadra A, Pietropaolo M. Immunogenetics of type 1 diabetes mellitus. Mol Asp Med. 2015;42:42–60.CrossRefGoogle Scholar
  38. 38.
    Patrick R. A pocket-calculator algorithm for the Shapiro-Francia test for non-normality: an application to medicine. Stat Med. 1993;12:181–4.CrossRefGoogle Scholar
  39. 39.
    Berry KJ, Mielke PW Jr, Mielke HW. The Fisher-Pitman permutation test: an attractive alternative to the F test. Psychol Rep. 2002;90:495–502.CrossRefPubMedGoogle Scholar
  40. 40.
    Pundziute-Lycka A, et al. The incidence of type I diabetes has not increased but shifted to a younger age at diagnosis in the 0-34 years group in Sweden 1983-1998. Diabetologia. 2002;45:783–91.CrossRefPubMedGoogle Scholar
  41. 41.
    Del Gobbo LC, Song Y, Dannenbaum DA, Dewailly E, Egeland GM. Serum 25-hydroxyvitamin D is not associated with insulin resistance or beta cell function in Canadian Cree. J Nutr. 2011;141:290–5.CrossRefPubMedGoogle Scholar
  42. 42.
    Svoren BM, Volkening LK, Wood JR, Laffel LM. Significant vitamin D deficiency in youth with type 1 diabetes mellitus. J Pediatr. 2009;154:132–4.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    The NS, Crandell JL, Lawrence JM, King IB, Dabelea D, Marcovina SM, et al. Vitamin D in youth with type 1 diabetes: prevalence of insufficiency and association with insulin resistance in the SEARCH Nutrition Ancillary Study. Diabet Med. 2013;30:1324–32.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Regnell SE, Lernmark Å. Early prediction of autoimmune (type 1) diabetes. Diabetologia. 2017;60:1370–81. Scholar
  45. 45.
    Kanatsuna N, Papadopoulos GK, Moustakas AK, Lenmark A. Etiopathogenesis of insulin autoimmunity. Anat Res Int. 2012;2012:457546.PubMedPubMedCentralGoogle Scholar
  46. 46.
    Ziegler AG, Rewers M, Simell O, Simell T, Lempainen J, Steck A, et al. Seroconversion to multiple islet autoantibodies and risk of progression to diabetes in children. JAMA. 2013;309:2473–9.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Mendivil CO, Toloza FJK, Ricardo-Silgado ML, Morales-Alvarez MC, Mantilla-Rivas JO, Pinzón-Cortés JA, et al. Antibodies against glutamic acid decarboxylase and indices of insulin resistance and insulin secretion in nondiabetic adults: a cross-sectional study. Diabetes Metab Syndr Obes. 2017;10:179–85.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Miao D, Steck AK, Zhang L, Guyer KM, Jiang L, Armstrong T, et al. Electrochemiluminescence assays for insulin and glutamic acid decarboxylase autoantibodies improve prediction of type 1 diabetes risk. Diabetes Technol Ther. 2015;17:119–27.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Lutale, J.J., Thordarson, H., Holm, P.I., Eide, G.E. & Vetvik, K. Islet cell autoantibodies in African patients with type 1 and type 2 diabetes in Dar es Salaam Tanzania: a cross sectional study. J Autoimmune Dis 4, 4 (2007).Google Scholar
  50. 50.
    Ekpebegh CO, Longo-Mbenza B. Clinical, immunologic and insulin secretory characteristics of young black South African patients with diabetes: hospital based single centre study. Diabetes Res Clin Pract. 2013;99:380–4.CrossRefPubMedGoogle Scholar
  51. 51.
    Pociot F, Lernmark Å. Genetic risk factors for type 1 diabetes. Lancet. 2016;387:2331–9.CrossRefPubMedGoogle Scholar

Copyright information

© Research Society for Study of Diabetes in India 2018

Authors and Affiliations

  • Silver Bahendeka
    • 1
    • 2
  • Ronald Wesonga
    • 3
  • Thereza Piloya Were
    • 4
  • Catherine Nyangabyaki
    • 2
  1. 1.Mother Kevin Postgraduate Medical School, Uganda Martyrs UniversityKampalaUganda
  2. 2.St. Francis HospitalKampalaUganda
  3. 3.School of Statistics and PlanningMakerere UniversityKampalaUganda
  4. 4.School of Medicine, College of Health SciencesMakerere UniversityKampalaUganda

Personalised recommendations