CD4 CD25high regulatory T cells are not impaired in patients with primary Sjögren's syndrome

doi:10.1016/j.jaut.2005.01.015

Journal of Autoimmunity

Volume 24, Issue 3, May 2005, Pages 235-242

https://doi.org/10.1016/j.jaut.2005.01.015 Get rights and content

Abstract

In animal models of autoimmunity, CD4 CD25^high T cells play a key role in the control of the autoimmune process. Few studies have investigated the role of these cells in human autoimmune diseases. We aimed to investigate CD4 CD25^high T cells in the peripheral blood of patients with primary Sjögren's syndrome (pSS). The proportion of blood CD4 CD25^high T cells was determined by flow cytometry in 21 patients with pSS as determined by the American–European consensus group criteria and two groups of controls (18 patients with lumbar back pain of mechanical origin and 15 healthy blood donors). The suppressive function of CD4 CD25 cells was assessed using co-culture assays. The Vbeta repertoire of CD4 CD25 T cells was examined by flow cytometry. The proportion of CD4 CD25 T cells depended on age in patients and controls. In an age-matched comparison, no significant difference was observed in the proportion of total CD4 CD25^low T cells between patients with pSS and controls (P=0.36). In contrast, the pool of CD4 CD25^high was significantly increased in patients with pSS (8.5% vs 4.1% in controls, P=0.04). There was a slight but not significant higher proportion of CD4 CD25^high cells in patients with a more active disease. CD4 CD25 T cells in patients with pSS effectively suppressed the proliferation of CD4 CD25⁻ autologous responder T cells. The Vbeta repertoire of regulatory T cells from patients with pSS was polyclonal and was not significantly restricted as compared with that in controls. Functional CD4 CD25^high regulatory cells are increased in patients with established pSS, through a reactive feedback, despite ongoing autoimmunity. These results suggest that pSS does not occur as a result of reduced level of CD4 CD25^high regulatory T cells, nor as a defect of inhibition of proliferation of responder cells.

Introduction

Primary Sjögren's syndrome (pSS) is an autoimmune disorder characterized by lymphocytic infiltration of the salivary and lacrimal glands leading to xerostomia and xerophthalmia. This syndrome, whose prevalence is estimated to be 0.2% to 0.5% in the general population, could be nearly as common as rheumatoid arthritis and is often complicated by extra-glandular manifestations, the most serious being non-Hodgkin's lymphoma [1], [2]. The pathogenesis of pSS is dominated by target-organ T cell infiltrates; destruction of the glandular epithelial cells, which are subjected to signals inducing their apoptosis; and polyclonal activation of B cells [3], [4].

A constitutive and high level of expression of the IL-2 receptor alpha chain (CD 25) defines a subset of naturally arising CD4 regulatory cells, the CD4 CD25^high T cells. The CD4 CD25^high T cells were first characterized in animals, as a population of suppressor T cells that maintain peripheral immune tolerance by inhibiting the activation and expansion of self-reactive T cells. In vitro suppression requires T cell receptor (TCR)-mediated activation of CD4 CD25^high T cells and is mediated through a cell-contact mechanism, independent of cytokines [5], [6], [7]. Recently, Foxp3, a transcription factor, has been reported to be specifically expressed by this subset of T cells [8]. The expansion of Foxp3-expressing CD4 CD25^high cells could be dependent on TGF-beta [9], [10]. Unfortunately, no cell-surface markers can absolutely discriminate between regulatory T cells and activated T cells: a consistent feature of proposed surface markers of these regulatory T cells, including CD25, cytotoxic T-lymphocyte antigen 4 (CTLA-4) and glucocorticoid-induced tumor necrosis factor receptor family-related gene (GITR), is that they can all be acquired in CD4 CD25⁻ responder T cells with cellular activation [11].

Very few studies have investigated the potential role of these cells in human autoimmune diseases. To date, to individualize regulatory CD4 CD25 T cells in the absence of specific phenotypic markers, one must consider only CD4 CD25 T cells with a high expression level of CD25 and ensure that these CD4 CD25^high T cells exhibit a functional activity of suppression. With these restrictions, some studies tried to characterize human blood CD4 CD25^high cells, which could represent 1–10% of CD4⁺ T cells in human peripheral blood [12], [13], [14], [15], [16]. The aim of the present study was to determine the number, function and repertoire of CD4 CD25^high circulating cells in patients with pSS.

Section snippets

Patients

Twenty-one patients with pSS as determined by the American–European consensus group criteria (four criteria, including positive results for anti-SSA/SSB antibodies or focus score ≥1; mean age 53.3±15.3 years; mean disease duration 11.5±6.8 years) were studied. Nine patients had extraglandular involvement of disease (presence or confirmed records of synovitis, myositis, vasculitis, or skin, neurological or renal involvement) and eight were treated with prednisone and/or methotrexate. Eight

Increased proportion of CD4 CD25^high cells in pSS

Unaware of the possibility of an age bias, we first compared the population of CD4 CD25^high cells in patients with pSS, in patients with lumbar back pain of mechanical origin and in healthy blood donors, who were markedly younger. The CD4 CD25^high cell population was significantly increased in patients with pSS (8.5±11.5%) compared with blood donors (3.3±1.4%, P=0.007) (Table 1). Since healthy blood donors were younger, we investigated whether an age bias could explain these results. Indeed,

Discussion

This study demonstrated that the CD4 CD25^high T cell population was unexpectedly increased in patients with pSS. The experimental data definitely show that we dealt with suppressive cells, since functional assays demonstrated that proliferation of CD4 CD25⁻ responder cells were suppressed by 54–87%, when they were co-cultured with CD4 CD25 cells at a ratio of 1 to 1. The higher numbers of CD4 CD25^high T cells in patients with pSS must be interpreted with caution, since CD4 CD25^high regulatory T

Acknowledgements

We acknowledge Laurence Meyer and Beatrice Ducot (INSERM U292, Department of Biostatistics and Public Health, Bicêtre Hospital), for helpful statistical discussions.

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Regulatory T (Treg) cells with immunosuppressive properties have a pivotal role in the maintenance of immune homeostasis. Alterations in Treg cell functionality appear to be of great importance in the development of immune senescence and contribute to increased susceptibility to immune-mediated diseases with age.
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Sjögren's syndrome (SS) is a chronic autoimmune disease that primarily affects the exocrine glands, resulting in their functional impairment. In SS, lymphocytic infiltration of salivary and lacrimal glands, and deposition of several types of autoantibodies, mainly anti-SS-A (anti-Ro) and anti-SS-B (anti-La), lead to chronic inflammation, with xerostomia and keratoconjunctivitis sicca. In its primary form (pSS), SS does not involve additional connective tissue diseases, whereas in its secondary and more common form (sSS), SS presents in association with other rheumatic autoimmune diseases, mainly rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and systemic sclerosis (SSc). As in most autoimmune diseases, environmental, hormonal and genetic factors are implicated in SS pathogenesis. In SS T cells predominate in mild lesions, whereas B cells predominate in advanced lesions. Th1, Th2, Th17, follicular helper T (Tfh) cells and regulatory cells (Tregs/Bregs), with their characteristic cytokine profiles, have been implicated in the pathogenesis of SS. It has been suggested that Th1 and Th17 cells initiate SS and, as the disease progresses, Th2 and Tfh cells predominate. It is assumed that, as in all autoimmune and inflammatory conditions, tolerance defects contribute to SS pathogenesis. It is intriguing that in SS it remains unclear which types of regulatory cells are functional and whether they ameliorate or worsen the disease. In this review we present a comprehensive update on SS with emphasis on immune system involvement, and suggest new insights into SS immunopathogenesis.
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¹: JEG was supported by The Institut National Pour la Santé et la Recherche Médicale (INSERM).

²: These authors participated equally in the work.

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CD4 CD25high regulatory T cells are not impaired in patients with primary Sjögren's syndrome

Abstract

Introduction

Section snippets

Patients

Increased proportion of CD4 CD25high cells in pSS

Discussion

Acknowledgements

Blood

Semin Arthritis Rheum

Blood

J Autoimmun

Blood

Blood

Blood

Immunity

Immunity

Blood

Increased risk of lymphoma in sicca syndrome

Ann Intern Med

Pathogenesis and treatment of Sjogren's syndrome

Curr Opin Rheumatol

A paragon of self-tolerance: CD25+CD4+ regulatory T cells and the control of immune responses

Arthritis Res Ther

CD4+CD25+ suppressor T cells: more questions than answers

Nat Rev Immunol

Regulatory T cells under scrutiny

Nat Rev Immunol

Control of regulatory T cell development by the transcription factor Foxp3

Science

TGF-{beta} regulates in vivo expansion of Foxp3-expressing CD4+CD25+ regulatory T cells responsible for protection against diabetes

Proc Natl Acad Sci U S A

Conversion of peripheral CD4+CD25− naive T cells to CD4+CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3

J Exp Med

CD4 CD25^high regulatory T cells are not impaired in patients with primary Sjögren's syndrome

Increased proportion of CD4 CD25^high cells in pSS