Summary
Type 1 (insulin-dependent) diabetes mellitus is associated with abnormalities of circulating lymphocyte subsets and autoantibodies. To investigate the prevalence of these in non-diabetic siblings and nondiabetic parents of children with Type 1 diabetes, we analysed T-cell subsets of function and activation in 31 families with an index case of Type 1 diabetes and related these to autoantibodies and HLA DR type. Using two and three colour cytofluorimetry, we studied total and activated (HLA-DR + ) CD3 +, CD4 +, CD8 + lymphocytes and on CD4 + lymphocytes the CD45RA/RO “naive” and “memory” cell phenotypes. Diabetic children (mean duration of disease 3.1 years) had a reduced total lymphocyte count (p < 0.05), their non-diabetic siblings a reduced CD4+ T-helper cell count (p < 0.05), and their parents a reduced percentage and number of CD3 + T cells (p < 0.01 and p < 0.05) compared with age-matched control subjects. Diabetic children, their siblings and parents all had significantly increased levels of activated CD4 + T-helper cells (p < 0.01, p < 0.05 and p < 0.01). In diabetic children and their siblings there was a significant over-expression of the CD45RO “memory” cell marker and significant under-expression of the CD45RA “naive” cell marker, whilst these were normal in the parents. Islet cell antibody positive diabetic children had significantly higher levels of CD45RO-expressing CD4 + lymphocytes than those who were islet cell antibody negative (p < 0.05). Amongst the siblings and parents, possession of HLA-DR4 was associated with lower percentages of CD4 + and higher percentages of CD8 + T cells. These findings extend current knowledge about the role of immunoregulatory CD45RA/ RO cells in Type 1 diabetes. In addition, they demonstrate lymphocyte subset abnormalities in unaffected family members, some of which may be influenced by HLA DR alleles.
Article PDF
Similar content being viewed by others
References
Bottazzo GF, Dean BM, McNally JM, Mackay EH, Swift PGF, Gamble R (1985) In situ characterization of autoimmune phenomena and expression of HLA molecules in the pancreas in diabetic insulitis. N Engl J Med 313: 353–360
Sibley RK, Sutherland DER, Goetz F, Michael AF (1985) Recurrent diabetes mellitus in the pancreas iso and allograft. A light and electron microscopic and immunohistochemical analysis of four cases. Lab Invest 53: 132–144
Leslie RDG, Lazarus NR, Vergani D (1989) Aetiology of insulin dependent diabetes. Br Med Bull 45: 58–72
Drell DW, Notkins AL (1987) Multiple immunological abnormalities in patients with type 1 (insulin-dependent) diabetes mellitus. Diabetologia 30: 132–143
Alviggi L, Johnston C, Hoskins PJ et al. (1984) Pathogenesis of insulin dependent diabetes: a role for activated T lymphocytes. Lancet II: 4–5
Muñoz A, Gallart T, Viñas O, Gomis R (1991) Increased CD5-positive B lymphocytes in type I diabetes. Clin Exp Immunol 83: 304–308
Smerdon RA, Peakman M, Hussain MJ et al. (1993) Increase in simultaneous co-expression of naive and memory markers at diagnosis of insulin dependent diabetes mellitus. Diabetes 42: 127–133
Peakman M, Vergani D (1992) Cell mediated immunity and type 1 diabetes. Diabetes Rev 1: 5–8
Todd JA (1990) Genetic control of autoimmunity in type 1 diabetes. Immunol Today 11: 122–128
Todd JA, Bell JI, McDevitt HO (1987) HLA-DQ beta gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature 329: 599–604
Khalil I, D’ Auriol L, Gobet M et al. (1990) A combination of HLA-DQβ Asp57-negative and HLA DQα Arg52 confers susceptibility to insulin-dependent diabetes mellitus. J Clin Invest 85: 1315–1319
Wen L, Peakman M, Lobo-Yeo A et al. (1991) T-cell-directed hepatocyte damage in autoimmune chronic active hepatitis. Lancet 336: 1527–1530
Van Vliet E, Roep VO, Meulenbroek L, Bruining GJ, De Vries RRP (1989) Human T cell clones with specificity for insulinoma cell antigens. Eur J Immunol 19: 213–216
Roep BO, Aarden SD, De Vries RRP, Hutton JC (1990) Tcell clones from a type-1 diabetes patient respond to insulin secretory granule proteins. Nature 345: 632–634
Altmann DM, Sansom D, Marsh SGE (1991) What is the basis for HLA-DQ associations with autoimmune disease? Immunol Today 12: 267–270
National Diabetes Data Group (1979) Classification and diagnosis of diabetes mellitus and other categories of glucose intolerance. Diabetes 28: 1039–1057
Peakman M, Tredger JM, Davies ET, Davenport M, Williams R, Vergani D (1993) Analysis of peripheral blood mononuclear cells in rodents by 3-colour flow cytometry using a small-volume lysed whole blood technique. J Immunol Methods 158: 87–94
Peakman M, Buggins A, Nicolaides K, Layton DM, Vergani D (1992) Analysis of lymphocyte phenotypes in cord blood from early gestation foetuses. Clin Exp Immunol 90: 345–350
Doherty DG, Donaldson PT (1991) HLA-DRB and DQB typing by a combination of serology, retriction fragment length polymorphism analysis and oligonucleotide probing. Eur J Immunogenet 18: 111–124
Bidwell JL, Jarrold EA (1986) HLA-DR allogenotyping using exon-specific cDNA probes and application of rapid minigel methods. Mol Immunol 23: 1111–1116
Bidwell JL, Bidwell EA, Savage DA, Middleton D, Klouda PT, Bradley BA (1988) A DNA-RFLP typing system that positively identifies serologically well-defined and ill-defined HLA-DR and DQ alleles, including DRw10. Transplantation 45: 640–646
Bidwell JL (1988) DNA-RFLP analysis and genotyping of HLA-DR and DQ antigens. Immunol Today 9: 18–23
Millward BA, Hussain MJ, Peakman M, Pyke DA, Leslie RDG, Vergani D (1988) Characterization of islet cell antibody in insulin dependent diabetes: evidence for IgGl subclass restriction and polyclonality. Clin Exp Immunol 71: 353–356
Peakman M, Lobo-Yeo A, Mieli-Vergani G, Davies ET, Mowat AP, Vergani D (1987) Characterization of anti-liver kidney microsomal antibody in childhood autoimmune chronic active hepatitis: evidence for IgGl subclass restriction, polyclonality and non-cross reactivity with hepatocyte surface antigens. Clin Exp Immunol 69: 543–549
Pontesilli O, Chase HP, Carotenuto P, Herberger MJ, Hayward AR (1982) T-lymphocyte subpopulations in insulin-dependent diabetes mellitus. Clin Exp Immunol 63: 68–72
Johnston C, Alviggi L, Millward BA, Leslie RDG, Pyke DA, Vergani D (1988) Alterations in T-lymphocyte subpopulations in type 1 diabetes: exploration of genetic influence in identical twins. Diabetes 37: 1484–1488
Galluzzo A, Giordano C, Rubino G, Bompiani GD (1984) Immunoregulatory T-lymphocyte subset deficiency in newly diagnosed type 1 (insulin-dependent) diabetes mellitus. Diabetologia 26: 426–430
Quiniou-Debrie MC, Debray-Sachs M, Dardenne M, Czernichow P, Assan R, Bach JF (1985) Anti-islet cellular and humoral autoimmunity, T-cell subsets, and thymic function in type 1 diabetes. Diabetes 34: 373–379
Richards SJ, Scott CS (1990) Immunophenotypic dissection of normal peripheral blood NK-associated (NKa) subpopulations by flow cytometry: morphological features and relationships between NKa (CD11b, CD16, CD56 and CD57) and T-cell (CD2, CD3, TCR, CD5, CD7, CD8 and CD38) associated determinant expression. Leukemia Lymphoma 2: 111–126
Selam JL, Clot J, Andary M, Mirouze J (1979) Circulating lymphocyte subpopulations in juvenile onset diabetes: a correction of abnormalities by adequate metabolic control. Diabetologia 16: 35–40
Crosti JL, Secchi A, Ferrero E et al. (1986) Impairment of lymphocyte-suppressive system in recent-onset insulin dependent diabetes: correlation with metabolic control. Diabetes 35: 1053–1057
Pozzilli P, Zuccarini O, Iavicoli M et al. (1983) Monoclonal antibodies defined abnormalities of T-lymphocytes in type 1 (insulin-dependent) diabetes. Diabetes 32: 91–94
Peakman M, Hussain MJ, Millward BA, Leslie RDG, Vergani D (1990) Effect of initiation of insulin therapy on T-lymphocyte activation in type 1 diabetes. Diabetic Med 7: 327–330
Jackson RA, Morris MA, Haynes BF, Eisenbarth GS (1982) Increased circulating la-antigen bearing T cells in type I diabetes mellitus. N Engl J Med 306: 785–788
Gilbey SG, Hussain MJ, Watkins PJ, Vergani D (1988) Cellmediated immunity and symptomatic diabetic autonomic neuropathy. Diabetic Med 5: 845–848
Bending JJ, Lobo-Yeo A, Vergani D, Viberti GC (1988) Proteinuria and activated T lymphocytes in diabetic neuropathy. Diabetes 37: 507–511
Allen C, Palta M, D’ Allessio DJ (1991) Risk of diabetes in siblings and other relatives of IDDM subjects. Diabetes 40: 831–836
Lawley TJ, Hall RP, Fauci AS, Katz SI, Hamburger MI, Frank MM (1981) Defective Fc-receptor functions associated with the HLA-B8/DRw3 haplotype: studies on patients with dermatitis herpetiformis and normal subjects. N Engl J Med 304: 185–192
Jaworski MA, Colle L, Guttmann RD (1983) Abnormal immunoregulation in patients with insulin dependent diabetes mellitus and their healthy first degree relatives. Hum Immunol 7: 25–34
Peakman M, Buggins AGS, Nicolaides KH, Layton DM, Vergani D (1992) Analysis of lymphocyte populations in cord blood from early gestation fetuses. Clin Exp Immunol 90: 345–350
Beverley PCL (1990) Is T-cell memory maintained by crossreactive stimulation? Immunol Today 11: 203–205
Morimoto C, Hafler DA, Weiner HL et al. (1987) Selective loss of the suppressor-inducer T cell subset in progressive multiple sclerosis: analysis with anti-2H4 monoclonal antibody. N Engl J Med 316: 67–73
Morimoto C, Steinberg AD, Letvin NL et al. (1987) A defect of immunoregulatory T cell subsets in systemic lupus erythematosus patients demonstrated with anti-2H4 antibody. J Clin Invest 79: 762–763
Faustman D, Eisenbarth GS, Daley J, Breitmeyer J (1989) Abnormal T-lymphocyte subsets in type 1 diabetes. Diabetes 38: 1462–1468
Schatz DA, Riley WJ, Maclaren NK, Barrett DJ (1991) Defective inducer T-cell function before the onset of insulin-dependent diabetes mellitus. J Autoimmunity 4: 125–136
Faustman D, Schoenfeld D, Ziegler R (1991) T-lymphocyte changes linked to autoantibodies: association of insulin autoantibodies with CD4 + CD45R + lymphocyte subpopulation in prediabetic subjects. Diabetes 40: 590–597
Sanders ME, Makgoba MW, Shaw S (1988) Human naive and memory T cells: reinterpretation of helper-inducer and suppressor-inducer subsets. Immunol Today 9: 195–199
Suzuki T, Yamada Y, Fujimura T et al. (1987) Diabetogenic effects of lymphocyte transfusion on the NOD or NOD nude mouse. In: Rygaard J, Brunner N, Graem N, Spang-Thomsen M (eds) Immune-deficient animals in biomedical research. Karger, Basel, pp 112–116
Like AA, Guberski DL, Butler L (1991) Influence of environmental viral agents on frequency and tempo of diabetes mellitus in BB/Wor rats. Diabetes 40: 259–262
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Peakman, M., Warnock, T., Vats, A. et al. Lymphocyte subset abnormalities, autoantibodies and their relationship with HLA DR types in children with Type 1 (insulin-dependent) diabetes and their first degree relatives. Diabetologia 37, 155–165 (1994). https://doi.org/10.1007/s001250050087
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s001250050087