The CD4+ regulatory T-cells is decreased in adults with latent autoimmune diabetes

https://doi.org/10.1016/j.diabres.2006.08.013Get rights and content

Abstract

The latent autoimmune diabetes in adults (LADA) is a subgroup of type 1 diabetes, which procession of autoimmune destruction of β-cells was slower than classic type 1 diabetes. To investigate the pathogenesis of LADA, we examined the lymphocyte subsets including the CD4+CD25+ T-cells in 60 LADA patients and 30 patients of type 2 diabetes and 30 healthy individuals by FACS. And we compared the expression of FOXP3 mRNA in CD4+ T-cell between 10 patients of LADA and 10 matched healthy individuals by real time PCR. The percent of CD4+CD25+ T-cells were higher (11.89 ± 4.96% versus 8.16 ± 3.65%, P < 0.01), and the percent CD8+ T-cells elevated (24.58 ± 6.80% versus 19.39 ± 7.12, P < 0.01) in LADA patients than healthy individuals. While the expression of FOXP3 mRNA in CD4+ T-cell was markedly decreased in LADA patients (0.52-fold, n = 10, P = 0.004) compared with normal subjects. In addition, the percent of CD8+ T-cells related with GAD-Ab titers in LADA patients (r = 0.292, P = 0.03). Our results showed that there were cellular immune disorder and decreased CD4+ regulatory T-cells in LADA patients. The adoptive transfer regulatory T-cells seem to be a potential therapeutics for LADA.

Introduction

Type 1 diabetes is a T-cell-mediated autoimmune disease that results in the destruction of the insulin-secreting β-cells [1]. Autoreactive T-cells that recognize islet autoantigens have been identified and are thought to play a direct role in type 1 diabetes immunopathogenesis [2], [3]. But the mechanisms that lead to disease development are not known with certainty.

Recent interest has focused on a feature of tolerance that seems to bridge the central and peripheral processes, namely the CD4+ regulatory T-cell [4]. The CD4+regulatory T-cells that exert their suppressive function in vitro in a contact-dependent manner and preferentially express high levels of CD25 and the forkhead and winged-helix family transcription factor forkhead box P3 (FOXP3). FOXP3 plays crucial role in the development and function of CD4+ regulatory T-cells [5], [6]. The regulatory T-cells have been shown to be defective in patients with type 1 diabetes, multiple sclerosis, rheumatoid arthritis and other autoimmune diseases [7], [8], [9], [10].

The latent autoimmune diabetes in adults (LADA) is a subgroup of type 1 diabetes who, showed preserved β-cell function, initially misclassified as type 2 diabetes [11], [12]. The development of symptoms is insidious and the clinical diabetes becomes overt at the age of 30s or older, and often is maintained in good metabolic control on diet or oral hypoglycemic therapy for up to several years before insulin dependency [13], [14]. So it has been suggested that LADA deviates from classic type 1 diabetes [15]. Until now, LADA remains poorly understood at both a clinical and research level [16].

Our approach was to investigate the pathogenesis of LADA. Thus, we examined the lymphocyte subsets including the CD4+CD25+ T fells and the expression of FOXP3 mRNA in CD4+ T-cell of the patients with LADA.

Section snippets

Patients and healthy control subjects

We enrolled 60 patients in this study with definite LADA and 30 patients with type 2 diabetes upon informed consent. This work was approved by the Institutional Review Board at the Second Xiangya Hospital. The range of diabetes duration in LADA patients were from 1 month to 8 years, eight recent onset LADA patients included. All patients had no other autoimmune diseases and did not receive any immunomodulatory drugs. Thirty normal healthy donors with no history of autoimmune diseases and

The general characteristics

Table 1 showed general characteristics of all groups. There were matched in age, sex, BMI in three groups. After statistical analysis, LADA patients had higher FBS and HbA1c compared with patients of type 2 diabetes. No significant differences were present in duration of diabetes and FCP levels in the two groups.

The lymphocyte subsets in peripheral blood

The percentage of CD4+CD25+ T-cells was evaluated as a proportion of CD4 T lymphocyte population. They were matched in CD3 T lymphocyte and no CD4 lymphopenia noted in three groups.

In

Discussion

Immune responses need to be constantly regulated—a balancing act that ensures appropriate reactivity to pathogens while preventing the development of an unwanted autoimmune reaction. Recently, this has been shown to be mediated by the CD4+ regulatory T-cells, which are characterized by expression of CD25 (the interleukin-2 (IL-2) receptor α-chain). These cells comprise ∼5–10% of the total population of CD4+ T-cells in mice and are thought to be crucial in the repression of autoimmune disorders

Acknowledgments

This study was supported, in part, by the National Natural Science Foundation of China (39370343) and Hunan Health Bureau Key Research Funds (9736, 2001-Z04). We thank Dr. Bill Hagopian for providing the recombinant hGAD65 plasmid.

We are grateful to all the patients and control subjects who donated blood samples.

References (36)

  • M.A. Atkinson et al.

    Response of peripheral-blood mononuclear cells to glutamate decarboxylase in insulin-dependent diabetes

    Lancet

    (1992)
  • S. Sakaguchi

    Regulatory T-cells: key controllers of immunologic self-tolerance

    Cell

    (2000)
  • L. Castano et al.

    Type-I diabetes: a chronic autoimmune disease of human, mouse, and rat

    Annu. Rev. Immunol.

    (1990)
  • B.O. Roep

    The role of T-cells in the pathogenesis of type 1 diabetes: from cause to cure

    Diabetologia

    (2003)
  • H. Yagi et al.

    Crucial role of FOXP3 in the development and function of human CD25+CD4+ regulatory T cells

    Int. Immunol.

    (2004)
  • S. Hori et al.

    Control of regulatory T cell development by the transcription factor Foxp3

    Science

    (20031057)
  • S. Lindley et al.

    Defective suppressor function in CD4+CD25+ T-cells from patients with type 1 diabetes

    Diabetes

    (2005)
  • T.M. Brusko et al.

    Functional defects and the influence of age on the frequency of CD4+CD25+ T-cells in type 1 diabetes

    Diabetes

    (2005)
  • M.R. Ehrenstein et al.

    Compromised function of regulatory T cells in rheumatoid arthritis and reversal by anti-TNFα therapy

    J. Exp. Med.

    (2004)
  • V. Viglietta et al.

    Loss of functional suppression by CD4+CD25+ regulatory T cells in patients with multiple sclerosis

    J. Exp. Med.

    (2004)
  • B. Littorin et al.

    Islet cell and glutamic acid decarboxylase antibodies present at diagnosis of diabetes predict the need for insulin treatment. A cohort study in young adults whose disease was initially labeled as type 2 or unclassifiable diabetes

    Diabetes Care

    (1999)
  • P. Pozzilli et al.

    Autoimmune diabetes not requiring insulin at diagnosis (latent autoimmune diabetes of adults): definition, characterization, and potential prevention

    Diabetes Care

    (2001)
  • G. Stenström et al.

    Latent autoimmune diabetes in adults: definition, prevalence, β-cell function, and treatment

    Diabetes

    (2005)
  • T. Kobayashi et al.

    Immunogenetic and clinical characterization of slowly progressive IDDM

    Diabetes Care

    (1993)
  • J.P. Palmer et al.

    Is latent autoimmune diabetes in adults distinct from type 1 diabetes or just type 1 diabetes at an older age?

    Diabetes

    (2005)
  • S. Fourlanos et al.

    Latent autoimmune diabetes in adults (LADA) should be less latent

    Diabetologia

    (2005)
  • World Health Organization, Definition, Diagnosis, and Classification of Diabetes Mellitus and its Complications: Report...
  • J.S. Petersen et al.

    Detection of GAD65 antibodies in diabetes and other autoimmune diseases using a simple radioligand assay

    Diabetes

    (1994)
  • Cited by (0)

    View full text