Article

Altered balance between self-reactive Th17 cells and Th10 cells and between full-length FOXP3 and FOXP3 splice variants in Hashimoto's thyroiditis

November 2014
Clinical & Experimental Immunology 180(1)

November 2014
180(1)

DOI:10.1111/cei.12557

Authors:

Birte Kristensen

University of Southern Denmark

Laszlo Hegedüs

University of Southern Denmark

Hans Ole Madsen

Copenhagen University Hospital

Terry J Smith

University of Michigan

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Th17 cells play a pathogenic role in autoimmune disease, while IL-10-producing T-helper (Th10) cells serve a protective role. The balance between the two subsets is regulated by the local cytokine milieu and by the relative expression of intact FOXP3 compared to FOXP3Δ2, missing exon 2. Th17- and Th10-cell differentiation has usually been studied using polyclonal stimuli, and little is known about the ability of physiologically relevant self-antigens to induce Th17- or Th10-cell differentiation in autoimmune thyroid disease. We subjected mononuclear cells from healthy donors and patients with Hashimoto's thyroiditis (HT) or Graves’ disease (GD) to polyclonal stimulation, or stimulation with human thyroglobulin (TG), human thyroid peroxidase (TPO), or E. coli lipopolysaccharide (LPS). TPO and LPS induced increased differentiation of naïve CD4+CD45RA+CD45R0- T cells from HT patients into Th17 cells. Th10-cell proportions were decreased in HT after polyclonal stimulation, but were comparable to those of healthy donors after antigen-specific stimulation. Taken together our data shows an increased Th17: Th10 ratio was found in HT patients after stimulation with thyroid specific self-antigens. We also observed an elevated baseline production of IL-6 and TGF-β1 and of mRNA encoding FOXP3Δ2 rather than intact FOXP3. This may contribute to the skewing towards Th17-cell responses in HT.

МОРФОГЕНЕЗ ЛІМФОЇДНОЇ ТКАНИНИ ЩИТОПОДІБНОЇ ЗАЛОЗИ У РАННЬОМУ ВІЦІ ЗА УМОВ ЕКСПЕРИМЕНТАЛЬНОГО ПРЕНАТАЛЬНОГО АНТИГЕННОГО НАВАНТАЖЕННЯ

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Jul 2021

У сучасній світовій літературі велика увага приділяється питанням загального впливу інфекційного навантаження на імунну систему організму, і, як наслідок, розвитку автоімунних станів щитоподібної залози. Т-лімфоцити є основними ефекторами адаптивної імунної системи і мають вирішальне значення для захисту організму від патогенних інфекційних збудників. Особливу увагу в цьому питанні привертає до себе вплив різноманітних чинників на організм вагітної, тобто пренатальна неспецифічна дія зовнішніх інфекційних антигенів на організм, які б мали віддалені прояви після народження. Мета – експериментально встановити особливості морфогенезу лімфоїдної тканини щитоподібної залози у ранньому постнатальному періоді онтогенезу за умов пренатальної дії стафілококового анатоксину. Матеріал і методи. Експериментально досліджено щитоподібні залози щурів лінії Вістар у віці від 1 до 14 доби постнатального розвитку (всього 90 тварин). Кожному плоду на 18 добу датованої вагітності під час лапаротомії, шляхом крізьматкової, крізьоболонкової підшкірної ін’єкції вводили стафілококовий анатоксин рідкий очищений адсорбований (10–14 одиниць зв’язування у 1 мл, розведений у 10 разів) в дозі 0,05 мл, тваринам контрольної групи аналогічно вводили 0,9 % розчин NaCl. Комплекс щитоподібної залози з ділянкою трахеї фіксували в 10 % розчині нейтрального забуференого формаліну протягом доби. Гістологічні зрізи товщиною 3–5 мкм забарвлювали гематоксиліном і еозином, імуногістохімічно із застосуванням моноклональних антитіл СD 3 (5В2), CD4 (5B4), CD8 (32-M4), CD20 (D-10) фірми Santa Cruz Biotechnology, Inc. із дофарбовуванням ядер гематоксиліном Майера. Фотодокументацію досліджуваних об’єктів виконано з використанням мікроскопа Carl Zeiss «Primo Star» з використанням камери AxioCam, комплекс морфометричних досліджень виконувався за допомогою програми Zeiss Zen (2017). Статистичний аналіз отриманих результатів проводили за допомогою персонального комп’ютера на базі операційної системи Windows XP за допомогою статистичного пакета «Statistica for Windows 6.0» (StatSoftInc.), програма Excel (Microsoft Office, USA). Результати. Внутрішньоплідне введення стафілококового анатоксину призвело до часткової лімфоцитарної інфільтрації строми щитоподібної залози і формування внутрішньоорганної дифузної лімфоїдної тканини здебільшого з СD 3+ та СD 4+ лімфоцитів (збільшення кількості Т-лімфоцитів за цей період у 75,1 раза), та кооперації її у вигляді лімфоїдного вузлика на 14 добу постнатального життя, клітинний склад якого був імуногістохімічно представлений СD 3+СD 4+ та СD 3+СD 8+ Т-лімфоцитами, що супроводжувалося розширенням і явищами стазу у венулах, активацією мастоцитів і макрофагів. Така морфологічна картина у щитоподібних залозах антигенпремійованих тварин наявна на периферії залози. Таким чином, інтенсивні зміни в імунологічному компартменті щитоподібної залози з формуванням дифузної лімфоїдної тканини у щурів раннього віку після пренатальної дії стафілококового анатоксину обумовлені генералізованим пренатальним антигенним навантаженням організму в цілому та є проявами реактивності у процесі формування антигеннезалежного диференціювання Т-лімфоцитів у тимусі.

The role behind the scenes of Tregs and Th17s in Hashimoto’s thyroiditis: Toward a pivotal role of FOXP3 and BACH2

Article

Full-text available

Dec 2022

In Hashimoto’s thyroiditis (HT), the genetic bases play a central role in determining development of the disease. In particular, the most frequent genes involved in the onset of HT are the Human Leukocyte Antigen (HLA). However, there are other genes and transcription factors in the autoimmune background of HT, both isolated and as part of autoimmune polyendocrine syndromes (APS). Recently more interest is being fueled toward BACH2 (BTB Domain and CNC Homolog 2), that promotes Tregs (T regulators lymphocytes) differentiation and enhances Treg-mediated immunity. The synergistic interaction between environmental agents and the aforementioned genes leads to the onset of autoimmunity and ultimately to damage of the thyroid gland. In this scenario, the role of Th17 (T helper-17 lymphocytes) and Treg cells is still less defined as compared to action of Th1 cells (T helper-1 lymphocytes) and cytotoxic lymphocytes (CD8 + T lymphocytes). Evidences show that an imbalance of Th17/Treg ratio represents a prognostic factor with respect to the gland damage. Moreover, the deficient ability of Treg to inhibit the proliferation of T cells against the self can break the immune balance. In light of these considerations, the use of genetic panels and the progress of immunotherapy could allow for better targeting treatment and preventive interventions in subjects with potential or early stage of HT.

Автоімунні захворювання щитоподібної залози та основні маркери їх патогенезу і діагностики

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Dec 2021
ENDOCRINOLOGY

Анотація Огляд присвячено оцінці основних гуморальних факторів у патогенезі автоімунних захворювань щитоподібної залози (АЗЩЗ) та можливості їх використання при діагностиці та складанні прогнозу. Показано, що клінічно різні тиреоїдит Хашимото (ТХ), хвороба Грейвса (ХГ) та офтальмопатія Грейвса (ОГ) тісно пов’язані патофізіологічно і мають подібні імуноопосередковані механізми — продукцію автоантитіл до тиреоїдних антигенів і лімфоїдну інфільтрацію тиреоїдної паренхіми. Втрата імунної толерантності до автоантигенів тиреоїдної пероксидази (thyroid peroxidase, TPO), тиреоглобуліну (thyroglobulin, Tg) та рецептору тиреотропного гормону (thyroid-stimulating hormone receptor, TSHR) є основою розвитку АЗЩЗ. Наголошується на ролі прозапальних та протизапальних цитокінів, які продукуються клітинами імунної системи та тиреоцитами. Цитокіни беруть участь в індукторній та ефекторній фазі імунної відповіді та запалення, відіграючи ключову роль у патогенезі АЗЩЗ. Значний вплив на розвиток і прогресування АЗЩЗ має дисбаланс між Th17- лімфоцитами, які підтримують автоімунну відповідь, та регуляторними Т-клітинами (regulatory T cells, Treg), які пригнічують автоімунний процес. Недостатність регуляторних В-клітин (regulatory B cells, Breg) та Тreg, які виробляють протизапальні цитокіни, порушує імунологічну толерантність і викликає аномальну продукцію прозапальних цитокінів, відіграє певну роль у патогенезі АЗЩЗ. Виявлення імунних клітин та антитиреоїдних антитіл у тканині щитоподібної залози (ЩЗ) та визначення рівнів прозапальних та протизапальних цитокінів у сироватці крові можуть дати інформацію про їх участь у патогенезі АЗЩЗ та можуть служити маркерами активності захворювання. Розглянуто діагностичне значення рівня цитокінів, тиреоїдних автоантитіл при ТХ, ХГ і ОГ та їх здатність відображати наявність та активність захворювання.

Peculiarities of innate and adaptive immunity in the pathogenesis of thyroid autoimmune diseases. Immunocorrection (part 1)

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Aug 2021

Abstract The review deals with the role of innate and adaptive, local and systemic immunity, cellular and humoral factors in the pathogenesis of autoimmune thyroid diseases (AITD). The importance of lymphoid infiltration of the thyroid gland, cytokines and autoantibodies, the role of thyroid hormones as modulators of the immune response, trace elements and dysregulation of apoptosis in the development of AITD is considered. Graves’ disease (GD) and Hashimoto’s thyroiditis (HT) have been shown to be closely pathophysiologically related and have similar immune-mediated mechanisms, such as the production of autoantibodies to thyroid antigens and lymphocytic infiltration of the thyroid gland. Loss of immune tolerance to thyroid peroxidase, thyroglobulin and thyroid-stimulating hormone autoantigens is the basis for the development of AITD. Emphasis is placed on the role of cytokines, which are produced by both immune system cells and thyroid follicular cells. Imbalance between Th17 lymphocytes and regulatory T cells (Treg) has a significant effect on the progression of AITD. An increase in Th17 lymphocytes may play a more important role in the pathogenesis of HT, whereas a decrease in Treg may be strongly involved in GD. Insufficiency of Treg that impairs immunological tolerance and causes abnormal cytokine production can lead to the initiation of apoptosis, which plays a role in the pathogenesis of GD and HT. Induction of apoptosis by HT leads to destruction of thyrocytes, while apoptosis in GD cause damage to thyroid infiltrating lymphocytes.

FOXP3 exon 2 controls T reg stability and autoimmunity

Article

Jun 2022

Differing from the mouse Foxp3 gene that encodes only one protein product, human FOXP3 encodes two major isoforms through alternative splicing—a longer isoform (FOXP3 FL) containing all the coding exons and a shorter isoform lacking the amino acids encoded by exon 2 (FOXP3 ΔE2). The two isoforms are naturally expressed in humans, yet their differences in controlling regulatory T cell phenotype and functionality remain unclear. In this study, we show that patients expressing only the shorter isoform fail to maintain self-tolerance and develop immunodeficiency, polyendocrinopathy, and enteropathy X-linked (IPEX) syndrome. Mice with Foxp3 exon 2 deletion have excessive follicular helper T (T FH ) and germinal center B (GC B) cell responses, and develop systemic autoimmune disease with anti-dsDNA and antinuclear autoantibody production, as well as immune complex glomerulonephritis. Despite having normal suppressive function in in vitro assays, regulatory T cells expressing FOXP3 ΔE2 are unstable and sufficient to induce autoimmunity when transferred into Tcrb -deficient mice. Mechanistically, the FOXP3 ΔE2 isoform allows increased expression of selected cytokines, but decreased expression of a set of positive regulators of Foxp3 without altered binding to these gene loci. These findings uncover indispensable functions of the FOXP3 exon 2 region, highlighting a role in regulating a transcriptional program that maintains T reg stability and immune homeostasis.

Автоімунні захворювання щитоподібної залози та основні маркери їх патогенезу і діагностики

Article

Full-text available

Dec 2021

Огляд присвячено оцінці основних гуморальних факторів у патогенезі автоімунних захворювань щитоподібної залози (АЗЩЗ) та можливості їх використання при діагностиці та складанні прогнозу. Показано, що клінічно різні тиреоїдит Хашимото (ТХ), хвороба Грейвса (ХГ) та офтальмопатія Грейвса (ОГ) тісно пов’язані патофізіологічно і мають подібні імуноопосередковані механізми — продукцію автоантитіл до тиреоїдних антигенів і лімфоїдну інфільтрацію тиреоїдної паренхіми. Втрата імунної толерантності до автоантигенів тиреоїдної пероксидази (thyroid peroxidase, TPO), тиреоглобуліну (thyroglobulin, Tg) та рецептору тиреотропного гормону (thyroid-stimulating hormone receptor, TSHR) є основою розвитку АЗЩЗ. Наголошується на ролі прозапальних та протизапальних цитокінів, які продукуються клітинами імунної системи та тиреоцитами. Цитокіни беруть участь в індукторній та ефекторній фазі імунної відповіді та запалення, відіграючи ключову роль у патогенезі АЗЩЗ. Значний вплив на розвиток і прогресування АЗЩЗ має дисбаланс між Th17- лімфоцитами, які підтримують автоімунну відповідь, та регуляторними Т-клітинами (regulatory T cells, Treg), які пригнічують автоімунний процес. Недостатність регуляторних В-клітин (regulatory B cells, Breg) та Тreg, які виробляють протизапальні цитокіни, порушує імунологічну толерантність і викликає аномальну продукцію прозапальних цитокінів, відіграє певну роль у патогенезі АЗЩЗ. Виявлення імунних клітин та антитиреоїдних антитіл у тканині щитоподібної залози (ЩЗ) та визначення рівнів прозапальних та протизапальних цитокінів у сироватці крові можуть дати інформацію про їх участь у патогенезі АЗЩЗ та можуть служити маркерами активності захворювання. Розглянуто діагностичне значення рівня цитокінів, тиреоїдних автоантитіл при ТХ, ХГ і ОГ та їх здатність відображати наявність та активність захворювання.

Correlation of hormonal and cytokines regulation in case of autoimmune thyroiditis

Article

Full-text available

Oct 2017

Victoria V. Zdor

Background. Studied immune aspects of the pathogenesis of autoimmune thyroiditis (AIT), which occupies the first place among human autoimmune pathologies. Treatment of the disease is based on thyroid hormones (TH) replacement therapy. TH are today considered to be super antigens in autoimmune inflammation of the thyroid gland. Aims. On the basis of complex assessment of hormonal and immunological markers (TSH, TH, Treg, the Th1-, Th2-, Th17-marker cytokines) with a research of possible interrelations of their indicators at patients with various clinical options of a current of AIT initially and against the background of replacement therapy of TH to define differences in functional activity of various types of immunocompetent cages depending on weight of inflammatory process for forecasting of a further clinical current of AIT, optimization of protocols of therapy and timely correction of strategy of treatment. Methods. In a prospective study, patients with AIT were evaluated for serum levels of cytokines and their receptors before initiating TH replacement therapy and on treatment by means of the ELISA modern methods with immuneсhemiluminescence and electroсhemiluminescence ways of detection. Results. Patients suffering from AIT showed an excess production of Th1-, Th2-, Th17- and Tregs marker cytokines with a deficiency of TGF-β1, closely connected with autoimmune hypothyroidism severity. Under pressure of TH therapy the indices of most cytokines decreased or improved, with the exception of IL-6, IL-8, IL-2, IFN-g, TNF-α. The greatest variations from the normal range were recorded in the complicated hypothyroidism. Conclusions. High serum TNF-α level in the onset of the disease is an important marker for the unfavourable AIT course and a predictor of hormone replacement therapy in case of its subclinical course. Safety indexes of functional thyroid epithelium are systemic levels of IL-8 and IL-22, their dynamic reduction in blood serum is an adverse factor, indicating a progressive loss of functionally active thyroid tissue and a possible increase of hypothyroidism in case of subclinical AIT course. If specific gravity and magnitudes of IL-1α, IL-6 and IFN-γ exceed manifold they can be considered to be predictors of AIT clinical course severity and autoimmune inflammation in the thyroid.

The Role of Iodine and Selenium in Autoimmune Thyroiditis

Article

Full-text available

Sep 2015
HORM METAB RES

Leonidas H Duntas

Iodine and selenium (Se) are both essential elements to thyroid hormone economy, while they represent key players in the development of autoimmune thyroiditis.Chronic high iodine intake has been associated in various studies with increased frequency of autoimmune thyroiditis. In susceptible individuals, iodine excess increases intra-thyroid infiltrating Th17 cells and inhibits T regulatory (TREG) cells development, while it triggers an abnormal expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in thyrocytes, thus inducing apoptosis and parenchymal destruction. As was shown in a mouse model, high iodine supply leads to changes in the immunogenicity of the thyroglobulin molecule, upregulation of vascular intercellular adhesion molecule-1 (ICAM-1), and reactive oxygen species (ROS) generation in the thyrocytes. Serum Se levels were found decreased in Hashimoto thyroiditis and especially in Graves' disease as well as in thyroid-associated ophthalmopathy patients, the levels being related to the pathogenesis and outcome. Selenium is strongly involved, via the variable selenoproteins, in antioxidant, redox, and anti-inflammatory processes. Selenium enhances CD4+/CD25 FOXP3 and T regulatory cells activity while suppressing cytokine secretion, thus preventing apoptosis of the follicular cells and providing protection from thyroiditis. Selenium supplementation may be useful in autoimmune thyroid diseases, though, while usually well-tolerated, it should not be universally recommended, and it is also likely to be helpful for those with low Se status and autoimmunity. Broadly speaking, the achievement and maintenance of "selenostasis" as well as adequate urinary iodine excretion are mandatory to control disease, while, putatively, they may additionally be critical to preventing disease. © Georg Thieme Verlag KG Stuttgart · New York.

The exploration of Hashimoto's Thyroiditis related miscarriage for better treatment modalities

Article

Full-text available

Aug 2020
Int J Med Sci

Hashimoto's thyroiditis (HT) is the most prevalent autoimmune thyroid disease (ATD) worldwide and is strongly associated with miscarriage and even recurrent miscarriage (RM). Moreover, with a deepening understanding, emerging evidence has shown that immune dysfunctions caused by HT conditions, including imbalanced subsets of CD4+ T-helper cells, B regulatory (Breg) cells, high expression levels of CD56dim natural killer (NK) cells, and cytokines, possibly play an important role in impairing maternal tolerance to the fetus. In recent years, unprecedented progress has been made in recognizing the specific changes in immune cells and molecules in patients with HT, which will be helpful in exploring the mechanism of HT-related miscarriage. Based on these findings, research investigating some potentially more effective treatments, such as selenium (Se), vitamin D3, and intravenous immunoglobulin (IVIG), has been well developed over the past few years. In this review, we highlight some of the latest advances in the possible immunological pathogenesis of HT-related miscarriage and focus on the efficacies of treatments that have been widely introduced to clinical trials or practice described in the most recent literature.

The value of central regulators of the immune response in the development of autoimmune thyroid diseases

Article

Full-text available

Jun 2020

The specific relationship between the endocrine and immune systems is represented by a numerous number of factors and mechanisms that form the structure and ensure the function of each of the two systems. For example, immunocompetent cells can produce immunologically active substances, as well as some hormones. On the other hand, immune cells are available to the effects of endogenous hormones. Currently, the so-called cross-regulation of endocrine and immune mechanisms in an equilibrium of pro-and anti-inflammatory responses has not been sufficiently studied. Among other autoimmune lesions, autoimmune thyreopathy occupies a significant place. The development of an autoimmune lesion of the thyroid gland is a complex process, which is the result of the interaction of infiltrating lymphocyte and thyrocyte tissue that can express a wide range of molecules involved in the immune response. Immunological and immunogenetic factors play a major role in the pathogenesis of autoimmune thyroid diseases, such as autoimmune thyroiditis and Graves disease. Despite the fact that more than 100 years have passed since the first description of autoimmune thyroiditis and Graves disease has been known for many centuries, the mechanisms of these pathologies are still not fully understood.

Apoptosis of blood lymphocytes in patients with autoimmune thyroiditis and its treatment

Article

Full-text available

Apr 2018

M.I. Sheremet

Мета — дослідити деякі ланки патогенезу розвитку вузлового зоба на тлі автоімунного тиреоїдиту (АІТ) шляхом вивчення активності процесів пероксидного окиснення ліпідів і білків, апоптозу лімфоцитів периферичної крові та рівня деяких цитокінів до та після оперативного втручання, розробити ефективні методи їх корекції. Матеріали та методи. Обстежено 80 жінок із вузловим зобом на тлі АІТ. Результати. Показано, що порушення програмованої загибелі лімфоцитів при АІТ проявляється збільшенням вмісту фактора некрозу пухлини α і CD95+ лімфоцитів на тлі зниження числа апоптичних клітин. Висновки. Після проведеного комплексного лікування відзначена позитивна динаміка порушених показників імунного статусу, насамперед нормалізація порушеного балансу субпопуляції апоптичних Т-лімфоцитів та антитіл до тиреоглобуліну та тиреоїдної пероксидази, зниження вмісту цитокінів і повернення в межі нормальних значень рівнів гормонів щитоподібної залози.

Crosstalk between alternative splicing and inflammatory bowel disease: Basic mechanisms, biotechnological progresses and future perspectives

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Full-text available

Nov 2023

Background Alternative splicing (AS) is an omnipresent regulatory mechanism of gene expression that enables the generation of diverse splice isoforms from a single gene. Recently, AS events have gained considerable momentum in the pathogenesis of inflammatory bowel disease (IBD). Methods Our review has summarized the complex process of RNA splicing, and firstly highlighted the potential involved molecules that target aberrant splicing events in IBD. The quantitative transcriptome analyses such as microarrays, next‐generation sequencing (NGS) for AS events in IBD have been also discussed. Results Available evidence suggests that some abnormal splicing RNAs can lead to multiple intestinal disorders during the onset of IBD as well as the progression to colitis‐associated cancer (CAC), including gut microbiota perturbations, intestinal barrier dysfunctions, innate/adaptive immune dysregulations, pro‐fibrosis activation and some other risk factors. Moreover, current data show that the advanced technologies, including microarrays and NGS, have been pioneeringly employed to screen the AS candidates and elucidate the potential regulatory mechanisms of IBD. Besides, other biotechnological progresses such as the applications of third‐generation sequencing (TGS), single‐cell RNA sequencing (scRNA‐seq) and spatial transcriptomics (ST), will be desired with great expectations. Conclusions To our knowledge, the current review is the first one to evaluate the potential regulatory mechanisms of AS events in IBD. The expanding list of aberrantly spliced genes in IBD along with the developed technologies provide us new clues to how IBD develops, and how these important AS events can be explored for future treatment.

Frequency of antithyroid antibodies in patients with primary biliary cholangitis

Article

Full-text available

Aug 2023
LABMEDICINE

Objective: Primary biliary cholangitis (PBC) is an autoimmune disease of liver that may be associated with other conditions, including autoimmune thyroid diseases. We aimed to investigate the frequency of anti-thyroperoxidase antibodies (TPO-Ab), antithyroglobulin antibodies (TG-Ab), and anti-thyrotropin receptor antibodies (TSHR-Ab) in Tunisian patients with PBC. Methods: Sera of 80 patients with PBC were collected over a 9-year period. A total of 189 healthy blood donors (HBD) were included in the control group. Measurements of TPO-Ab and TG-Ab were performed using indirect enzyme-linked immunosorbent assay (ELISA). Competitive ELISA was used to assess TSHR-Ab. Results: Antithyroid antibodies (ATA) were significantly more frequent in PBC patients than in the control group (13.7% vs 1.6%; P < 10-3). Out of 11 patients with ATA, 10 (90.9%) were female. Nine patients and 2 HBD had TPO-Ab (11.2% vs 1%; P < 10-3). TG-Ab were more frequent in patients than in healthy subjects but the difference was not statistically significant (6.2% vs 1.6%; P = .1). TPO-Ab and TG-Ab were present together in 3 patients (3.7%). TSHR-Ab were absent in patients and controls. Conclusion: This study shows that PBC is associated with a high frequency of ATA but not TG-Ab or TSHR-Ab.

Development of SARS-CoV-2 atypical thyroiditis in a patient with severe COVID-19

Article

Apr 2023

The novel coronavirus infection SARS-CoV-2 (COVID-19) has gone down in history as one of the deadliest pandemics in history. Since the first reports of SARS-CoV-2 infection, a lot of data has appeared regarding the features of the COVID-19 pathogenesis and the effect of the virus on various organs and tissues. It is known that SARS-CoV-2 can cause both pulmonary and systemic inflammation, causing multiple organ dysfunction. In addition, since the beginning of the pandemic, reports of the relationship between COVID-19 and thyroid dysfunction have continued to emerge. Numerous studies have demonstrated that the thyroid gland and the entire hypothalamus-pituitary-thyroid axis can be significant targets for the pathogenic effects of SARS-CoV-2.The classic thyroid dysfunctions in infectious diseases, also described in COVID-19, are subacute thyroiditis and secondary hypothyroidism. Of particular interest is the development of atypical thyroiditis-SARS-CoV-2 against the background of COVID-19, a condition first recorded during COVID-19, associated with formidable cardiovascular complications and high mortality in patients with COVID-19, not previously encountered in other viral infections.This article describes a clinical case of SARS-CoV-2 atypical thyroiditis in a patient with bilateral viral pneumonia in the acute period of COVID-19.

PROBIOTICS AS PROMISING THERAPEUTICS: A REVIEW OF LITERATURE

Article

Full-text available

Feb 2023

Scientific studies have been done in recent decades to investigate the potential impact of microorganisms on the central processes of life, including states of health and pathological disorders. They found that microbe colonization causes a variety of diseases. New safe probiotics made from non-viable microbes or bacterial-free extracts have recently been on the market. These products are known as postbiotics and paraprobiotics, respectively, which prevent the colonization of the intestines by pathogens and preserve the health of the resident microflora. It is now clearer than ever that probiotic strains can have a beneficial effect on human health. Various pieces of legislation addressing safety and efficacy issues have been drafted for substances that are often administered using food, dietary items, supplements, and medications. Moreover, regulatory bodies have addressed a variety of approaches to these items, including whether or not to allow claims regarding the diagnosis, prevention, or treatment of a condition. In order to evaluate the wide variety of bacterial and yeast strains, rigorous methods have been developed for evaluating potential adverse effects and conducting post-market surveillance. Probiotics have shown greater health benefits compared to synthetic drugs like antibiotics that interfere with the normal flora in the gut. Probiotics contribute to good food and nutrition as food is their major delivery vehicle. However, precise predictions of probiotic consumption rate call for additional consideration of various parameters. In light of the multifaceted nature of probiotics, this study presents a comprehensive review of these topics, including their uses in human medicine and their quality, safety, and regulation.

CD4+ T Cells Promote IgG Production in MHC-Independent and ICAM-1-Dependent Manners in Pristane-Induced Lupus Mice

Article

Full-text available

Jan 2022
MEDIAT INFLAMM

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production and chronic inflammation. The etiology and pathogenesis of SLE are complicated in which dysfunction of CD4+ T cells is largely engaged. In this study, we investigated the manners of CD4+ T cells in antibody production in a lupus-like mouse model through peritoneal injection of pristane reagent. With the increase in total IgG/IgM and autoantibody production after 6 months, CD4+ T cells exhibited activated phenotypes with the elevated CD44, ICOS, OX40, and PD-1 expression. Pristane injection induced the increase in IgM levels in both wild-type and T cell-deficient TCRα-/- mice whereas IgG, IgG1, and IgG2a production was impaired. When adoptively transferring CD4+ T cells into T cell-deficient mice or coculturing CD4+ T cells and B cells in vitro, it was found that CD4+ T cells derived from pristane-treated mice could help the production of total IgG as well as IgG1/IgG2a in a more efficient manner both in vivo and in vitro. While MHC was dispensable for IgG production, ICAM-1 likely functioned as an attenuating factor for IgG production. Our study thus reveals that CD4+ T cells in pristane-treated mice play important roles in IgG production, which implies the critical roles in the induction of pathological autoantibodies in MHC-independent and ICAM-1-dependent manners.

Morphofunctional transformations during the morphogenesis of the thyroid gland of the offspring of Wistar rats after intrauterine exposure to dexamethasone

Article

Full-text available

Dec 2021

Background. In recent years, the prevalence of thyroid pathologies of various origins among children in the world has reached a significantly high level. The use of glucocorticoids during pregnancy remains a debatable issue in obstetrics today, as they can both positively and negatively affect the processes of organ morphogenesis and be the cause of pathological conditions in the postnatal period. Objective: to establish the features of morphofunctional transformations during the morphogenesis of the thyroid gland of the offspring of rats at an early age in normal and after intrauterine action of dexamethasone. Methods. 108 thyroid glands of rats of 3 experimental groups were microscopically examined using histological and immunohistochemical methods, followed by statistical processing of the obtained results. Results. Against the background of high levels of total follicular thyrocytes per 1 day of life in animals that received prenatal dexamethasone, cytoplasmic expression of TgAb was expressed, which correlated with the indicators of nuclear and cytoplasmic Fox-1 expression. From the 7th to the 11th day, a decrease in the total number of thyrocytes per unit area was observed due to the accumulation of colloid in the follicles, an increase in Fox-1 cytoplasmic expression and a decrease in nuclear expression, against the background of increased proliferative activity. By day 21, Fox-1 cytoplasmic and nuclear expression were almost identical. There was a decrease in the intensity of TgAb expression in the cytoplasm of thyrocytes and its expression in the colloid, a decrease in the number of Ki-67 positive thyrocytes per conditional unit area compared with the previous observation period. Conclusion. It was found that prenatal exposure of dexamethasone causes the offspring accelerate the development of morphological structures of the thyroid gland, but functionally they are in a state of stress of both the synthesizing apparatus and the process of hormone excretion, which is expressed in the imbalance of immunohistochemical expression of Fox-1 and TgAb. Such thyrocytes with signs of disturbances in synthetic activity desquamate into the lumen of the follicles, while on the 11th day we compensatory increase in the proliferative activity of the thyroid epithelium.

SARS-CoV-2: Emerging Role in the Pathogenesis of Various Thyroid Diseases

Article

Full-text available

Nov 2021

Coronavirus disease-2019 (COVID-19) is asymptomatic in most cases, but it is impartible and fatal in fragile and elderly people. Heretofore, more than four million people succumbed to COVID-19, while it spreads to every part of the globe. Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) induces various dysfunctions in many vital organs including the thyroid by utilizing ACE2 as a receptor for cellular entry. Emerging reports clearly show the involvement of SARS-CoV-2 in diverse thyroid disorders. Thus, this review article aims to review comprehensively all the recent developments in SARS-CoV-2-induced pathogenesis of thyroid diseases. The review briefly summarizes the recent key findings on the mechanism of SARS-CoV-2 infection, the role of ACE2 receptor in viral entry, SARS-CoV-2-activated molecular signaling in host cells, ACE2 expression in the thyroid, cytokine storm, and its vital role in thyroid dysfunction and long-COVID in relation to thyroid and autoimmunity. Further, it extensively discusses rapidly evolving knowledge on the potential part of SARS-CoV-2 in emerging various thyroid dysfunctions during and post-COVID-19 conditions which include subacute thyroiditis, Graves’ diseases, Hashimoto’s thyroiditis, thyrotoxicosis, and other recent advances in further discerning the implications of this virus within thyroid dysfunction. Unraveling the pathophysiology of SARS-CoV-2-triggered thyroid dysfunctions may aid pertinent therapeutic options and management of these patients in both during and post-COVID-19 scenarios.

Article

May 2021
Expet Rev Clin Immunol

Introduction: During the COVID-19 pandemic thyroid gland alteration/dysfunction has been emerged as a possible endocrine complication. The present review is focused on inflammatory and autoimmune thyroid complications triggered by SARS-CoV-2 infection by searching through databases like MEDLINE and Scopus up to April 2021. Areas covered: Beside the occurrence of "non-thyroidal illness" in severe clinical conditions, alterations of thyroid function and structure may occur during COVID-19 as a consequence of either direct or indirect effects of SARS-CoV-2 infection. On the one hand, SARS-CoV-2 uses ACE2 as a receptor to infect the host cells and ACE2 is highly expressed by follicular thyroid cells. On the other hand, COVID-19 is associated with a systemic inflammatory and immune response, involving Th1/Th17/Th2 lymphocytes and proinflammatory cytokines, which resembles, at least in part, the immune activation that occurs in immune-mediated thyroid diseases. COVID-19-related thyroid disorders include destructive thyroiditis and onset or relapse of autoimmune thyroid disorders, leading to a broad spectrum of thyroid dysfunction ranging from thyrotoxicosis to hypothyroidism, that may worsen COVID-19 clinical course and affect prognosis. Expert opinion: Physicians should be aware of the possible occurrence of thyroid dysfunction during and after SARS-CoV-2 infection. Further longitudinal studies are warranted to evaluate potential long-term sequelae.

IL-8R2 Deficient Mice are More Resistant to Pulmonary Coccidioidomycosis

Article

Full-text available

Dec 2020
INFECT IMMUN

The pathology of human coccidioidomycosis is granulomatous inflammation with many neutrophils surrounding ruptured spherules, but the chemotactic pathways that draw neutrophils into the infected tissues are not known. We previously showed that formalin-killed spherules (FKS) stimulate mouse macrophages to secret MIP-2, which suggested that CXC ELR+ chemokines might be involved in neutrophil recruitment in vivo . To test that hypothesis, we intra-nasally infected IL-8R2 ( Cxcr2 ) deficient mice on a BALB/c background with C. immitis RS. IL-8R2 deficient mice had fewer neutrophils in infected lungs than controls, but unexpectedly the IL-8R2 deficient mice had fewer organisms in their lungs than the control mice. Infected IL-8R2 deficient mouse lungs had higher expression of genes associated with lymphocyte activation, including the Th1 and Th17-related cytokines Ifnγ and Il17a and the transcription factors Stat1 and Rorc . Additionally, bronchial alveolar lavage fluid from infected IL-8R2 deficient mice contained more IL-17A and IFNγ. We postulate that neutrophils in the lung directly or indirectly interfere with the development of a protective Th1/Th17 immune response to C. immitis at the site of infection.

Thyrotoxicosis in patients with COVID-19: the THYRCOV study

Article

Jul 2020

Objective. This study assessed thyroid function in patients affected by the coronavirus disease-19 (COVID-19), based on the hypothesis that the cytokine storm associated with COVID-19 may influence thyroid function and/or the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may directly act on thyroid cells, such as previously demonstrated for SARS-CoV-1 infection. Design and Methods. This single-center study was retrospective and consisted in evaluating thyroid function tests and serum interleukin-6 (IL-6) values in 287 consecutive patients (193 males, mean age 64.3±14.0 years, range: 27-92), hospitalized for COVID-19 in non-intensive care units. Results. Fifty-eight patients (20.2%) were found with thyrotoxicosis (overt in 31 cases), 15 (5.2%) with hypothyroidism (overt in only 2 cases), 214 (74.6%) with normal thyroid function. Serum thyrotropin (TSH) values were inversely correlated with age of patients (rho -0.27; p<0.001) and IL-6 (rho -0.41; p<0.001). In the multivariate logistic regression analysis, thyrotoxicosis resulted to be significantly associated with higher IL-6 (odds ratio 3.25, 95% confidence interval 1.97-5.36; p<0.001), whereas the association with age of patients was lost (p=0.09). Conclusions. This study provides a first evidence that COVID-19 may be associated with high risk of thyrotoxicosis in relationship with systemic immune activation induced by the SARS-CoV-2 infection.

Evaluation of Th1/Th2 Lymphocyte Balance in Peripheral Blood Mononuclear Cells of Patients with Hashimoto's Thyroiditis

Article

Full-text available

Mar 2020
IRAN J PUBLIC HEALTH

Background: The role of T cells in the pathogenesis of Hashimoto’s thyroiditis is well established, whereas the precise and likely the overlapping contributions of different T-cell subpopulations to thyroid injury are less understood. The purpose of this study was to assess the expression pattern of two lineage determining transcription factors, T-bet and GATA-3 that regulate differentiation of T cells into Th1 or Th2 cell fates, respectively. Moreover, the mRNA expression and plasma concentration of Th1(IFN-γ) and Th2(IL-4) cytokineswere analyzed. Methods: In this case-control study, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to determine the expression patterns of various transcripts in 20 patients (in Endocrinology Clinic, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran, Iran, in 2015) with Hashimoto’s thyroiditis (HT) and 22 healthy controls. Plasma IL-4 and IFN-γ concentrations were also measured using enzyme-linked immunosorbent assay. Results: T-bet gene expression was significantly lower in patients compared to healthy controls (P=0.014). The expression of IL-4 mRNAs was significantly increased in the peripheral blood mononuclear cells from patients as compared to normal controls (P=0.001). In addition, a marked increase in plasma IL-4 levels were observed in patient group compared to controls (P=0.043). Conclusion: Altered balance between Th1 and Th2 related transcription factors and cytokines may be implicated in the pathogenesis of Hashimoto’s thyroiditis.

The proportion of peripheral blood Tregs among the CD4+ T cells of autoimmune thyroid disease patients: a meta-analysis

Article

Dec 2019

Autoimmune thyroid disease (AITD) is characterized by a loss of self-tolerance to thyroid antigen. Tregs, whose proportions are controversial among CD4+ T cell from AITD patients (AITDs), are crucial in immune tolerance. Considering that drugs might affect Treg levels, we assumed that the differences originated from different treatment statuses. Thus, we performed a meta-analysis to explore proportions of Tregs in untreated and treated AITDs. PubMed, Embase and ISI Web of Knowledge were searched for relevant studies. Review Manager 5.3 and Stata 14.0 were used to conduct the meta-analysis. Subgroup analysis based on different diseases and cell surface markers was performed. Egger linear regression analysis was used to assess publication bias. Approximately 1,100 AITDs and healthy controls (HCs) from fourteen studies were included. Proportions of Tregs among CD4+ T cells of untreated AITDs were significantly lower than those in HCs (p = 0.002), but were not in treated patients (p = 0.40). Subgroup analysis revealed lower proportions of Tregs in untreated Graves’ disease patients (GDs) (p = 0.001) but did not show obvious differences in untreated Hashimoto’s thyroiditis patients (HTs) (p = 0.62). Furthermore, proportions of circulating FoxP3+ Tregs were reduced in untreated GDs (p < 0.00001) and HTs (p = 0.04). No publication bias was found. In this first meta-analysis exploring proportions of circulating Tregs among CD4+ T cells of AITDs with different treatment statuses, we found that Tregs potentially contribute to the pathogenesis of AITD but function differently in GD and HT. Remarkably, FoxP3+ Tregs, which were decreased in both diseases, might be promising targets for novel therapies.

Altered N-glycan profile of IgG-depleted serum proteins in Hashimoto's thyroiditis

Article

Full-text available

Oct 2019
BBA-GEN SUBJECTS

Background: Hashimoto's thyroiditis (HT) is an autoimmune disease characterized by chronic inflammation of thyroid gland. Although HT is the most common cause of hypothyroidism, the pathogenesis of this disease is not fully understood. Glycosylation of serum proteins was examined in HT only to a limited extent. The study was designed to determine the glycosylation pattern of IgG-depleted sera from HT patients. Methods: Serum N-glycans released by N-glycosidase F (PNGase F) digestion were analyzed by normal-phase high-performance liquid chromatography (NP-HPLC). N-glycan structures in each collected HPLC fraction were determined by liquid chromatography-mass spectrometry (LC-MS) and exoglycosidase digestion. Fucosylation and sialylation was also analyzed by lectin blotting. Results: The results showed an increase of monosialylated tri-antennary structure (A3G3S1) and disialylated diantennary N-glycan with antennary fucose (FA2G2S2). Subsequently, we analyzed the serum N-glycan profile by lectin blotting using lectins specific for fucose and sialic acid. We found a significant decrease of Lens culinaris agglutinin (LCA) staining in HT samples, which resulted from the reduction of α1,6-linked core fucose in HT serum. We also observed an increase of Maackia amurensis II lectin (MAL-II) reaction in HT due to the elevated level of α2,3-sialylation in HT sera. Conclusions: The detected alterations of serum protein sialylation might be caused by chronic inflammation in HT. The obtained results completed our previous IgG N-glycosylation analysis in autoimmune thyroid patients and showed that the altered N-glycosylation of serum proteins is characteristic for autoimmunity process in HT. General significance: Thyroid autoimmunity is accompanied by changes of serum protein sialylation.

Immunomodulatory effects of Calcitriol in acute spinal cord injury in rats

Article

Jul 2019
INT IMMUNOPHARMACOL

Pharmacological therapy options for spinal cord injury (SCI) in acute phase have so far been limited, thus we focused on Calcitriol, FDA-approved biologically active form of vitamin D whose neuroprotective effects are increasingly recognized, to ameliorating damage following acute SCI in rats. Calcitriol (1 μg/kg) treatment for 7 consecutive days after SCI was compared SCI control and Sham control rat groups. Calcitriol-treated group had significantly improved outcome in standard functional recovery evaluation test (BBB) 12 weeks after SCI compared to SCI control, which was confirmed by increased ventral horn motor neurons in Calcitriol-treated group. In addition, proliferation test performed on lymphocytes from spleen and lymph nodes one week after SCI showed that calcitriol injection has a significant regulatory effect on Division Index (DI) in response to MBP stimulation compared to control SCI groups, which was associated with significant reduction in IFN-γ and IL-17A secretion and leukocyte infiltration into injury site. Along with confirmation of immunoregulatory aspects of Calcitriol treatment against myelin antigens in SCI, this study has shown that reducing the extent of progressive tissue loss by Calcitriol therapy in acute phase, could result in better recovery after SCI.

Implications of immunotherapy with high-dose glatiramer acetate in acute phase of spinal cord injury in rats

Article

Apr 2019

Objective: Recently, many researches with different viewpoints have focused on application of immunotherapy agents in treatment of spinal cord injury (SCI) according to neuroprotective results in some neurodegenerative disease. Glatiramer acetate (GA) is the most commonly used drug for Multiple sclerosis (MS) patients that exerts an immunomodulatory effect against Myelin basic protein (MBP) antigen. Materials and methods: High-dose (2mg/kg) treatment of GA for 28 consecutive days after SCI was compared with its low-dose (0.5 mg/kg) treatment, SCI control and Sham control rat groups. Results: High-dose GA group had significantly worsened outcome in standard functional recovery evaluation test (BBB) 12 weeks after SCI compared to SCI control and low-dose GA groups, which was confirmed by augmented spinal cavity volume and reduced ventral horn motor neurons in high-dose GA group; however, there was no significant difference between low-dose GA and control SCI group. In addition, proliferation test performed on lymphocytes from spleen and lymph nodes one week after SCI showed that high-dose GA injection has more significant effect on Division Index (DI) in response to MBP stimulation compared to low-dose GA and control SCI groups, which was associated with significant increase in IFN-γ, IL-4, and IL-17A secretion. Conclusion: Along with confirmation of deleterious aspects of autoimmunity resulting from autoreactive lymphocytes against myelin antigens in SCI, this study has shown that high-dose immunotherapy using GA, especially in acute phase after SCI, overwhelms any neuroprotective effect of adoptive immune system.

Clinical, Immunological, and Molecular Heterogeneity of 173 Patients With the Phenotype of Immune Dysregulation, Polyendocrinopathy, Enteropathy, X-Linked (IPEX) Syndrome

Article

Full-text available

Nov 2018

Background: Immune Dysregulation, Polyendocrinopathy, Enteropathy, X-linked (IPEX) Syndrome is a rare recessive disorder caused by mutations in the FOXP3 gene. In addition, there has been an increasing number of patients with wild-type FOXP3 gene and, in some cases, mutations in other immune regulatory genes. Objective: To molecularly asses a cohort of 173 patients with the IPEX phenotype and to delineate the relationship between the clinical/immunologic phenotypes and the genotypes. Methods: We reviewed the clinical presentation and laboratory characteristics of each patient and compared clinical and laboratory data of FOXP3 mutation-positive (IPEX patients) with those from FOXP3 mutation-negative patients (IPEX-like). A total of 173 affected patients underwent direct sequence analysis of the FOXP3 gene while 85 IPEX-like patients with normal FOXP3 were investigated by a multiplex panel of “Primary Immune Deficiency (PID—related) genes.” Results: Forty-four distinct FOXP3 variants were identified in 88 IPEX patients, 9 of which were not previously reported. Among the 85 IPEX-like patients, 19 different disease-associated variants affecting 9 distinct genes were identified. Conclusions: We provide a comprehensive analysis of the clinical features and molecular bases of IPEX and IPEX-like patients. Although we were not able to identify major distinctive clinical features to differentiate IPEX from IPEX-like syndromes, we propose a simple flow-chart to effectively evaluate such patients and to focus on the most likely molecular diagnosis. Given the large number of potential candidate genes and overlapping phenotypes, selecting a panel of PID-related genes will facilitate a molecular diagnosis.

Alternative Splicing of FOXP3—Virtue and Vice

Article

Full-text available

Mar 2018

Reiner Mailer

FOXP3 is the lineage-defining transcription factor of CD4+ CD25+ regulatory T cells. While many aspects of its regulation, interaction, and function are conserved among species, alternatively spliced FOXP3 isoforms are expressed only in human cells. This review summarizes current knowledge about alternative splicing of FOXP3 and the specific functions of FOXP3 isoforms in health and disease. Future perspectives in research and the therapeutic potential of manipulating alternative splicing of FOXP3 are discussed.

Role of the T and B lymphocytes in pathogenesis of autoimmune thyroid diseases

Article

Full-text available

Feb 2018

Autoimmune thyroid disorders (AITD) broadly include Graves’ disease and Hashimoto’s thyroiditis which are the most common causes of thyroid gland dysfunctions. These disorders develop due to complex interactions between environmental and genetic factors and are characterized by reactivity to self-thyroid antigens due to autoreactive lymphocytes escaping tolerance. Both cell-mediated and humoral responses lead to tissue injury in autoimmune thyroid disease. The differentiation of CD4+ cells in the specific setting of immune mediators (for example cytokines, chemokines) results in differentiation of various T cell subsets. T cell identification has shown a mixed pattern of cytokine production indicating that both subtypes of T helper, Th1 and Th2, responses are involved in all types of AITD. Furthermore, recent studies described T cell subtypes Th17 and Treg which also play an essential role in pathogenesis of AITD. This review will focus on the role of the T regulatory (Treg) and T helper (Th) (especially Th17) lymphocytes, and also of B lymphocytes in AITD pathogenesis. However, we have much more to learn about cellular mechanisms and interactions in AITD before we can develop complete understanding of AITD pathophysiology.

Neuroprotective effect of (‑)‑epigallocatechin‑3‑gallate on autoimmune thyroiditis in a rat model by an anti‑inflammation effect, anti‑apoptosis and inhibition of TRAIL signaling pathway

Article

Nov 2017

Junfeng Li

(-)-Epigallocatechin-3-gallate (EGCG) is a polyphenol monomer compound extracted and separated from green tea, and is a key catechin in green tea. Recent research has identified that EGCG is equipped with important biological activities, including antitumor, antioxidant, anti-inflammation, blood fat reduction and radiation protection abilities. In the current study, it was investigated whether EGCG exerts a neuroprotective effect on AIT and examined the possible underlying mechanism. The present study sought to establish an experimental autoimmune thyroiditis (AIT) rat model and to investigate the neuroprotective effect of EGCG in this model. EGCG was demonstrated to inhibit urinary iodine values and thyroid pathological features in AIT model rats. Treatment with EGCG significantly reduced interleukin-1β, interferon-γ (INF-γ) and tumor necrosis factor-α (TNF-α) levels in the AIT rats through suppression of nuclear factor-κB (NF-κB) pathway. In addition, pretreatment with EGCG significantly increased B-cell lymphoma-2 protein expression, and suppressed caspase-3 activity and TNF-α-related apoptosis-inducing ligand (TRAIL) protein expression levels in the AIT model rats. In conclusion, these results suggested that the neuroprotective effect of EGCG protects against AIT through its anti-inflammatory ability, anti-apoptosis and TRAIL signaling pathway in model rats, and it may be used as a therapeutic agent against AIT caused by inflammation.

The Expression of T Cell FOXP3 and T-Bet Is Upregulated in Severe but Not Euthyroid Hashimoto’s Thyroiditis

Article

Full-text available

Jul 2016
MEDIAT INFLAMM

Hashimoto’s thyroiditis (HT) is an organ-specific autoimmune disorder characterized by progressive thyroid failure. Th1 and Treg subset of CD4 + cells have been implicated in the pathogenesis; however, less is known about their respective roles across the spectrum of HT clinical presentations. To shed more light on CD4 + subsets role in HT, we investigated the mRNA expression levels of several Th1/Treg-associated transcription factors (T-bet/ETS1, HIF1 α /BLIMP1/FOXP3) in peripheral blood T cells of 10 hypothyroid, untreated HT patients, 10 hypothyroid patients undergoing hormone replacement therapy, 12 euthyroid HT subjects, and 11 healthy controls by the qRT-PCR. Compared to euthyroid HT patients and controls, both hypothyroid (2.34-fold difference versus controls, P < 0.01 ) and thyroxine-supplemented patients (2.5-fold, P < 0.001 ) showed an increased FOXP3 mRNA expression in T cells. Similarly, mRNA expression levels of T-bet were upregulated in severely affected but not in euthyroid HT subjects (2.37-fold and 3.2-fold, hypothyroid and thyroxine-supplemented HT patients versus controls, resp., P < 0.01 ). By contrast, no differences in mRNA expression levels of ETS1, BLIMP1, and HIF1 α were observed across the study groups. In summary, severe but not euthyroid HT was associated with robust upregulation of T-bet and FOXP3 mRNA in peripheral T cells, independent of the thyroid hormone status but proportional to disease activity.

T regulatory (Treg) and T helper 17 (Th17) lymphocytes in thyroid autoimmunity

Article

Full-text available

Apr 2016
ENDOCRINE

Different immune cell subsets have a relevant role in the pathogenesis of and tissue damage seen in autoimmune thyroid diseases (AITD), including T regulatory (Treg) lymphocytes and T helper (Th) 17 cells. There are several types of CD4+ Treg cells (Foxp3+, CD69+, Tr1), which are able to prevent the appearance of autoimmune diseases, down regulating the immune response and the inflammatory phenomenon. However, despite their presence in peripheral blood and thyroid tissue from patients with AITD, these cells are apparently unable to put down the autoimmune process. Moreover, many reports indicate the involvement of Th17 cells in chronic inflammatory diseases, including AITD. Nevertheless, it is now evident that these lymphocytes show a remarkable plasticity, giving rise to anti-inflammatory (including Treg lymphocytes) and pro-inflammatory cell subtypes. Nowadays, both Treg and Th17 cells must be considered as key elements in the pathogenesis of AITD as well as plausible potential targets for the next generation of therapeutic options of this condition.

Immune stimulation improves endocrine and neural fetal outcomes in a model of maternofetal thyrotoxicosis

Article

Sep 2015

Characterization of Regulatory B Cells in Graves’ Disease and Hashimoto’s Thyroiditis

Article

Full-text available

May 2015
PLOS ONE

A hallmark of regulatory B cells is IL-10 production, hence their designation as IL-10+ B cells. Little is known about the ability of self-antigens to induce IL-10+ B cells in Graves' disease (GD), Hashimoto's thyroiditis (HT), or other autoimmune disease. Here we pulsed purified B cells from 12 HT patients, 12 GD patients, and 12 healthy donors with the thyroid self-antigen, thyroglobulin (TG) and added the B cells back to the remaining peripheral blood mononuclear cells (PBMCs). This procedure induced IL-10+ B-cell differentiation in GD. A similar tendency was observed in healthy donors, but not in cells from patients with HT. In GD, B cells primed with TG induced IL-10-producing CD4+ T cells. To assess the maximal frequency of inducible IL-10+ B cells in the three donor groups PBMCs were stimulated with PMA/ionomycin. The resulting IL-10+ B-cell frequency was similar in the three groups and correlated with free T3 levels in GD patients. IL-10+ B cells from both patient groups displayed CD25 or TIM-1 more frequently than did those from healthy donors. B-cell expression of two surface marker combinations previously associated with regulatory B-cell functions, CD24hiCD38hi and CD27+CD43+, did not differ between patients and healthy donors. In conclusion, our findings indicate that autoimmune thyroiditis is not associated with reduced frequency of IL-10+ B cells. These results do not rule out regulatory B-cell dysfunction, however. The observed phenotypic differences between IL-10+ B cells from patients and healthy donors are discussed.

Metabolic regulation of forkhead box P3 alternative splicing isoforms and their impact on health and disease

Article

Full-text available

Oct 2023

Forkhead Box P3 (FOXP3) is crucial for the development and suppressive function of human regulatory T cells (Tregs). There are two predominant FOXP3 splicing isoforms in healthy humans, the full-length isoform and the isoform lacking exon 2, with different functions and regulation mechanisms. FOXP3 splicing isoforms show distinct abilities in the cofactor interaction and the nuclear translocation, resulting in different effects on the differentiation, cytokine secretion, suppressive function, linage stability, and environmental adaptation of Tregs. The balance of FOXP3 splicing isoforms is related to autoimmune diseases, inflammatory diseases, and cancers. In response to environmental challenges, FOXP3 transcription and splicing can be finely regulated by T cell antigen receptor stimulation, glycolysis, fatty acid oxidation, and reactive oxygen species, with various signaling pathways involved. Strategies targeting energy metabolism and FOXP3 splicing isoforms in Tregs may provide potential new approaches for the treatment of autoimmune diseases, inflammatory diseases, and cancers. In this review, we summarize recent discoveries about the FOXP3 splicing isoforms and address the metabolic regulation and specific functions of FOXP3 splicing isoforms in Tregs.

Pathogenic role of Th17 cells in autoimmune thyroid disease and their underlying mechanisms

Article

Feb 2023

Autoimmune thyroid disease, encompassing Graves' disease and Hashimoto's thyroiditis, has a very complex etiology. Pathogenesis of the disease involves both genetic susceptibility and environmental triggers. Traditionally, imbalance of T helper cell 1 and 2 was thought to result in the immune disorders in Graves' disease and Hashimoto's thyroiditis. However, increasing evidence recently revealed the important role of T helper 17 cell and its relative cellular and secretory components in the pathogenesis and progression of autoimmune thyroid disease. This review is aimed to summarize the published studies on the involvement of T helper 17 cell in autoimmune thyroid disease and discuss the underlying regulatory mechanisms, which could possibly serve as the foundation of discovering new therapeutic targets.

IMMUNOHISTOCHEMICAL PARAMETERS OF TGAB AND FOX–1 EXPRESSION IN THE THYROID GLAND OF RATS AFTER PRENATAL ANTIGEN EXPOSURE

Article

Jan 2021

Hashimoto’s Thyroiditis

Chapter

Jan 2018

W M Wiersinga

Thyroid Autoimmunity

Chapter

Jan 2018

Autoimmune thyroid diseases and Th17/Treg lymphocytes

Article

Nov 2017
LIFE SCI

Years of researches have demonstrated that the imbalance of Th17 and Tregs contribute to the thyroid autoimmunity and the severity of autoimmune thyroid disease (AITD). The underlying mechanism comprises inherent genetic predisposition, abnormality of Th17 and Treg related biological molecules, and gut microbiota disorder. New therapeutic strategies have been developed to improve the Th17/Treg equilibrium, including regulation of intracellular signaling pathways, neutralization of Th17-related cytokines, as well as manipulation of Th17 and Treg specific transcription factors. Although a few of these agents are applied into AITD, the clinic prospect is promising.

Recent advances in understanding autoimmune thyroid disease: The tallest tree in the forest of polyautoimmunity

Article

Full-text available

Sep 2017

Autoimmune thyroid disease (AITD) is often observed together with other autoimmune diseases. The coexistence of two or more autoimmune diseases in the same patient is referred to as polyautoimmunity, and AITD is the autoimmune disease most frequently involved. The occurrence of polyautoimmunity has led to the hypothesis that the affected patients suffer from a generalized dysregulation of their immune system. The present review summarizes recent discoveries unravelling the immunological mechanisms involved in autoimmunity, ranging from natural autoimmunity to disease-specific autoimmunity. Furthermore, the clinical grounds for considering AITD in a setting of polyautoimmunity are explored. A better understanding of these may pave the way for designing new treatment modalities targeting the underlying immune dysregulation when AITD appears in the context of polyautoimmunity.

Hashimoto’s Thyroiditis

Chapter

Jan 2016

W M Wiersinga

Hashimoto’s thyroiditis is best defined as an organ-specific autoimmune disease, characterized by autoimmune-mediated destruction of the thyroid gland. Diagnostic criteria have changed dramatically since the first description in 1912; they now include the presence of antibodies against thyroid peroxidase (TPOAb) and thyroglobulin, hypoechogenicity on thyroid ultrasound, and often but not always hypothyroidism. Distinct pathologic phenotypes are recognized: goitrous and atrophic variants but also an IgG4-related variant, hashitoxicosis, juvenile thyroiditis, and silent or painless thyroiditis. With a prevalence of 10–12% in the general population, it is the most common autoimmune disease. The prevalence is higher in females than in males, increases with advancing age, and is highest in Whites and lowest in Blacks. The incidence of autoimmune hypothyroidism is about 350 cases/100,000/year for women and 60 cases/100,000/year for men in iodine-sufficient regions and 44 (females) and 12 (males) per 100,000 per year in iodine-deficient areas. Breakdown of self-tolerance against thyroid antigens may lead to thyroid autoimmunity. Loss of Treg inhibitory actions and gain of Th17 proinflammatory actions (reflected by a shift to higher values of Th17/Th10 ratio in peripheral blood) play a crucial role in the loss of tolerance against thyroid antigens. Cytotoxic CD8+ T cells directed against TPO and Tg mediate thyroid gland destruction, either by the granule exocytosis pathway or apoptosis (programmed cell death). TPOAb and TgAb may cause antibody-dependent cell-mediated cytotoxicity (ADCC) via complement-mediated lysis of thyrocytes. Hashimoto’s thyroiditis often runs in families as evident from a high sibling risk ratio of 28. Twin studies suggest genes contribute about 73% of the liability to the development of TPOAb and TgAb; environmental factors would thus contribute about 20–30%. Polymorphisms in TSHR, Tg, HLA, CTLA-4, IL2RA, and FOXP3 have all been associated with Hashimoto’s thyroiditis but account for only a small proportion of the heritability. Genome-wide association studies continue to detect novel genetic loci linked to TPOAb. Smoking and moderate alcohol consumption to a certain extent protect against Hashimoto’s thyroiditis. Low selenium or vitamin D intake are presumably related to a higher prevalence of TPOAb, but presently there is no convincing evidence that selenium or vitamin D supplementation may lower TPOAb concentration. Infections may provoke Hashimoto’s thyroiditis, but available epidemiological studies do not support a causative role.

The possible role of CD4+CD25highFoxp3+/CD4+IL-17A+ cell imbalance in the autoimmunity of patients with Hashimoto thyroiditis

Article

Mar 2015

Hashimoto thyroiditis (HT) is a prototypic organ-specific autoimmune thyroid disease, for which the exact etiology remains unclear. The aim of this study was to investigate dynamic changes in regulatory T cell (Treg) and T helper 17 cell (Th17) populations in patients with HT at different stages of thyroid dysfunction, as well as to analyze the possible correlation between the Treg/Th17 cell axis and autoimmune status in HT. We assessed thyroid function and autoantibody serology both in HT patients and in healthy controls (HCs) and divided HT patients into three subgroups according to thyroid function. We then determined the percentages of Treg and Th17 cells in peripheral blood mononuclear cells and analyzed mRNA expression of the Treg and Th17 cell-defining transcription factors Foxp3 and RORγt. In addition, serum levels of TGF-β and IL-17A were assessed. We found that the percentage of Treg cells, Foxp3 mRNA levels, and the ratio of Treg/Th17 cells were all significantly lower in HT patients, while Th17 cell percentages and RORγt mRNA levels were significantly higher. Interestingly, we also observed significant differences in these measurements between HT patient subgroups. Serum IL-17A levels were markedly increased in HT patients, while serum concentrations of TGF-β were lower, compared to HCs. The ratio of Treg/Th17 cells was negatively correlated with the levels of serum thyroperoxidase antibody, thyroglobulin antibody, and thyrotropin (TSH) in HT patients. Taken together, our data suggest that the balance between Treg and Th17 cells shifts in favor of Th17 cells during clinical progression of HT, which is negatively correlated with levels of thyroid-specific autoantibodies and TSH, implying that Treg/Th17 cell imbalance may contribute to thyroid damage in HT.

A High Frequency of Circulating Th22 and Th17 Cells in Patients with New Onset Graves’ Disease

Article

Full-text available

Jul 2013
PLOS ONE

T-helper (Th) 22 and Th17 cells are involved in the pathogenesis of autoimmune diseases. However, their roles in the pathogenesis of Graves'disease (GD) are unclear. This study is aimed at examining the frequency of peripheral blood Th22, Th17, and Th1 cells and the levels of plasma IL-22, IL-17, and IFN-γ in patients with GD. A total of 27 patients with new onset GD and 27 gender- and age-matched healthy controls (HC) were examined for the frequency of peripheral blood Th22, Th17, and IFN-γ cells by flow cytometry. The concentrations of plasma IL-22, IL-17, and IFN-γ were examined by enzyme-linked immunosorbent assay. The levels of serum TSHR antibodies (A-TSHR), free triiodothyronine (FT3), free thyroxine (FT4), and thyroid stimulating hormone (TSH) were examined by radioimmunoassay and chemiluminescent assay, respectively. The levels of serum TSAb were examined by enzyme-linked immunosorbent assay. In comparison with those in the HC, significantly elevated percentages of Th22 and Th17 cells, but not Th1 cells, and increased levels of plasma IL-22 and IL-17, but not IFN-γ, were detected in GD patients (P<0.0001, for both). The percentages of both Th22 and Th17 cells and the levels of plasma IL-22 and IL-17 were correlated positively with the levels of serum TSAb in GD patients (r = 0.7944, P<0.0001; r = 0.8110, P<0.0001; r = 0.7101, p<0.0001; r = 0.7407, p<0.0001, respectively). Th22 and Th17 cells may contribute to the pathogenesis of GD.

Human CD25 high Foxp3 pos regulatory T-cells differentiate into IL17 producing cells

Article

Full-text available

TGF- and IL-6 drive the production of IL-17 and IL-10 by T cells and restrain TH-17 cell–mediated pathology

Article

Full-text available

Nov 2007

Studies have shown that transforming growth factor- (TGF-) and interleukin 6 (IL-6) are required for the lineage commitment of pathogenic IL-17-producing T helper cells (TH-17 cells). Unexpectedly, here we found that stimulation of myelin-reactive T cells with TGF- plus IL-6 completely abrogated their pathogenic function despite upregulation of IL-17 production. Cells stimulated with TGF- plus IL-6 were present in the spleen as well as the central nervous system, but they failed to upregulate the proinflammatory chemokines crucial for central nervous system inflammation. In addition, these cells produced IL-10, which has potent anti-inflammatory activities. In contrast, stimulation with IL-23 promoted expression of IL-17 and proinflammatory chemokines but not IL-10. Hence, TGF- and IL-6 'drive' initial lineage commitment but also 'restrain' the pathogenic potential of TH-17 cells. Our findings suggest that full acquisition of pathogenic function by effector TH-17 cells is mediated by IL-23 rather than by TGF- and IL-6.

Pathogen-induced human TH17 cells produce IFN-?? or IL-10 and are regulated by IL-1??

Article

Full-text available

Apr 2012
NATURE

IL-17-producing CD4+ T helper cells (TH17) have been extensively investigated in mouse models of autoimmunity. However, the requirements for differentiation and the properties of pathogen-induced human TH17 cells remain poorly defined. Using an approach that combines the in vitro priming of naive T cells with the ex vivo analysis of memory T cells, we describe here two types of human TH17 cells with distinct effector function and differentiation requirements. Candida albicans-specific TH17 cells produced IL-17 and IFN-γ, but no IL-10, whereas Staphylococcus aureus-specific TH17 cells produced IL-17 and could produce IL-10 upon restimulation. IL-6, IL-23 and IL-1β contributed to TH17 differentiation induced by both pathogens, but IL-1β was essential in C. albicans-induced TH17 differentiation to counteract the inhibitory activity of IL-12 and to prime IL-17/IFN-γ double-producing cells. In addition, IL-1β inhibited IL-10 production in differentiating and in memory TH17 cells, whereas blockade of IL-1β in vivo led to increased IL-10 production by memory TH17 cells. We also show that, after restimulation, TH17 cells transiently downregulated IL-17 production through a mechanism that involved IL-2-induced activation of STAT5 and decreased expression of ROR-γt. Taken together these findings demonstrate that by eliciting different cytokines C. albicans and S. aureus prime TH17 cells that produce either IFN-γ or IL-10, and identify IL-1β and IL-2 as pro- and anti-inflammatory regulators of TH17 cells both at priming and in the effector phase.

Increased Circulating Pro-Inflammatory Cytokines and Th17 Lymphocytes in Hashimoto's Thyroiditis

Article

Full-text available

Dec 2009
J CLIN ENDOCR METAB

Context: Th17 lymphocytes play an important role in different chronic inflammatory and autoimmune conditions. Aim: The aim of the study was to explore the status of Th17 cells in patients with autoimmune thyroid diseases (AITD). Design: We assessed the serum levels and in vitro synthesis of IL-17 and IL-22 and of different cytokines (IL-6, IL-15, and IL-23) involved in the differentiation of Th17 cells in the peripheral blood and thyroid glands of 26 patients with AITD, eight with Graves' disease, and 18 with Hashimoto's thyroiditis (HT) as well as 10 healthy controls. Results: We found enhanced levels of T cells synthesizing IL-17 and IL-22 in the peripheral blood from AITD patients, mainly in those with HT. In addition, a stronger expression of IL-17 and IL-22 and an enhanced number of IL-23R(+) cells was detected in thyroid glands from HT patients compared with Graves' disease or controls. Furthermore, increased concentrations of IL-6 and IL-15 were detected in sera from HT patients, whereas serum levels of IL-23 tended to be higher in these patients. Finally, an enhanced in vitro differentiation of T lymphocytes into Th17 cells induced by IL-23/IL-6 was observed in AITD patients. Accordingly, a strong induction of RORC2 gene was detected in lymphocytes from HT patients when stimulated with IL-23. Conclusion: Our results indicate that there is an increased differentiation of Th17 lymphocytes and an enhanced synthesis of Th17 cytokines in AITD, mainly in HT. These phenomena may have an important role in the pathogenesis of thyroid autoimmunity.

Human memory FOXP3+ Tregs secrete IL17 ex vivo and constitutively express the TH17 lineage-specific transcription factor RORgammat

Article

Full-text available

Jun 2009
P NATL ACAD SCI USA

Recent studies have suggested a close relationship between CD4(+)FOXP3(+) regulatory T cells (Tregs) and proinflammatory IL-17-producing T helper cells (T(H)17) expressing the lineage-specific transcription factor RORgamma t. We report here the unexpected finding that human memory Tregs secrete IL-17 ex vivo and constitutively express RORgamma t. IL-17-secreting Tregs share some phenotypic and functional features with conventional T(H)17 cells, expressing high levels of CCR4 and CCR6 and low levels of CXCR3. However, unlike conventional T(H)17 cells, they express low levels of CD161 and mostly fail to cosecrete IL-22 and TNF-alpha ex vivo. Ex vivo secretion of IL-17 and constitutive expression of RORgamma t by human memory Tregs suggest that, in addition to their well-known suppressive functions, these cells likely play additional, as yet undescribed, proinflammatory functions.

Regulatory T Cell Suppression Is Potentiated by Target T Cells in a Cell Contact, IL-35- and IL-10-Dependent Manner

Article

Full-text available

Jun 2009
J IMMUNOL

Regulatory T cells (T(reg)) are believed to suppress conventional T cell (T(conv)) proliferation in vitro in a contact-dependent, cytokine-independent manner, based in part on experiments in which T(reg) and T(conv) are separated by a permeable membrane. We show that the production of IL-35, a novel inhibitory cytokine expressed by natural T(reg), increases substantially following contact with T(conv). Surprisingly, T(reg) were able to mediate potent suppression of T(conv) across a permeable membrane when placed in direct contact with T(conv) in the upper chamber of a Transwell plate. Suppression was IL-35 and IL-10 dependent, and T(conv) activation was required for maximal potentiation of T(reg) suppression. These data suggest that it is the induction of suppression, rather than the function of T(reg) that is obligatorily contact dependent.

Systematic Method for Determining an Ideal Housekeeping Gene for Real-Time PCR Analysis

Article

Full-text available

Jan 2009
J Biomol Tech

To standardize the amount of biological material between samples (e.g., number of cells or amount of tissue) for quantitative real-time reverse transcriptase PCR (qRT-PCR), the cycle of the target gene at which expression is detected (the cycle threshold, or Ct) is divided by the Ct of a gene either thought to be unaffected by experimental conditions or similarly expressed among donors. Genes that maintain cellular structure or homeostasis, referred to as housekeeping genes, or 18S ribosomal RNA are often used for this purpose. Although unstable or inconsistent housekeeping gene expression will misrepresent experimental effects on target gene expression, housekeeping genes are often chosen arbitrarily rather than systematically. We designed a simple and systematic approach towards selection of housekeeping genes based on Ct variance (as reflected by the standard deviation) and normality of distribution. We validated this approach by comparing stability and consistency of expression of 11 housekeeping genes across different types of cells, experimental treatments, and human donors. Finally, we demonstrated the consequences of inconsistent housekeeping gene expression on the calculation of target gene expression, and conclude that validation of stability of housekeeping gene expression by considering both distribution normality and standard deviation is straightforward and critical for proper experimental design.

Malignant Tregs express low molecular splice forms of FOXP3 in Sezary syndrome

Article

Full-text available

Oct 2008
LEUKEMIA

Sézary syndrome (SS) is an aggressive variant of cutaneous T-cell lymphoma. During disease progression, immunodeficiency develops; however, the underlying molecular and cellular mechanisms are not fully understood. Here, we study the regulatory T cell (Treg) function and the expression of FOXP3 in SS. We demonstrate that malignant T cells in 8 of 15 patients stain positive with an anti-FOXP3 antibody. Western blotting analysis shows expression of two low molecular splice forms of FOXP3, but not of wild-type (wt) FOXP3. The malignant T cells produce interleukin-10 and TGF-beta and suppress the growth of non-malignant T cells. The Treg phenotype and the production of suppressive cytokines are driven by aberrant activation of Jak3 independent of the FOXP3 splice forms. In contrast to wt FOXP3, the low molecular splice forms of FOXP3 have no inhibitory effect on nuclear factor-kappaB (NF-kappaB) activity in reporter assays which is in keeping with a constitutive NF-kappaB activity in the malignant T cells. In conclusion, we show that the malignant T cells express low molecular splice forms of FOXP3 and function as Tregs. Furthermore, we provide evidence that FOXP3 splice forms are functionally different from wt FOXP3 and not involved in the execution of the suppressive function. Thus, this is the first description of FOXP3 splice forms in human disease.

Immunoregulatory Role of IL-10 during Superantigen-Induced Hyporesponsiveness in Vivo

Article

Full-text available

Feb 1997

Injection of the superantigen staphylococcal enterotoxin A (SEA) to mice rapidly elicits production of the Th1-related pro-inflammatory cytokines IL-2, TNF, and IFN-gamma, while repeated SEA challenges transduce a hyporesponsive state characterized by T cell deletions and anergy in the remaining SEA-reactive T cells. In the present study we show that exposure to SEA promotes the development of immunoregulatory IL-10-producing Th cells. Serum IL-10 was undetectable in mice given a priming injection of SEA, but rose to considerable levels when a SEA challenge was administered 4 days later. This coincided with maintained IL-2 production and superinduction of TNF, IFN-gamma, and IL-4. Interestingly, administration of a second SEA challenge resulted in high serum levels of IL-10, but the production of all other studied cytokines (IL-2, TNF, IFN-gamma, and IL-4) was impaired. The IL-10 production was sustained after a third and a fourth SEA challenge in the complete absence of IL-2, IFN-gamma, and IL-4. Pretreatment of mice with neutralizing anti-lL-10 mAb before the SEA challenges substantially enhanced IFN-gamma and TNF serum levels, but failed to rescue IL-2 production. Depletion of T cells with anti-CD4 or anti-CD8 mAb indicated that CD4+ T cells were crucial for SEA-induced IL-10 production. This finding was confirmed using CD4- and CD8- knockout mice. We conclude that repeated SEA challenges promote an IL-10-producing Th cell subset ("Th10" profile), which exerts an immunoregulatory effect by suppressing IFN-gamma and TNF production.

Ex Vivo Isolation and Characterization of CD4+CD25+ T cells with Regulatory Properties from Human Blood

Article

Full-text available

Jun 2001
J EXP MED

It has been known for years that rodents harbor a unique population of CD4(+)CD25(+) "professional" regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4(+)CD25(+)CD45RO(+) T cells (mean 6% of CD4(+) T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4(+)CD25(+) T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte-associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4(+)CD25(+) T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4(+)CD25(+) T cells released IL-10, and in coculture experiments suppressed the activation and proliferation of CD4(+) and CD8(+) T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4(+)CD25(+) T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer.

Novel mutations of FOXP3 in two Japanese patients with immune dysregulation, polyendocrinopathy, enteropathy, X linked syndrome (IPEX) [8]

Article

Full-text available

Jan 2002
J MED GENET

Editor—Immune dysregulation, polyendocrinopathy, enteropathy, X linked syndrome (IPEX), also known as X linked autoimmunity-allergic dysregulation syndrome (XLAAD), is characterised by enteropathy and involvement of the endocrine system, such as insulin dependent diabetes mellitus (IDDM) and thyroiditis, which develop in association with autoantibodies in early infancy (MIM 304930, 304790).1 2 IPEX has been mapped to chromosome Xp11.23-Xq13.3.3 4 Recent studies have indicated that FOXP3 , a member of forkhead/winged-helix proteins, is a causative gene for both IPEX and an equivalent mouse, scurfy .5-8 Human FOXP3 consists of 11 exons and encodes 431 amino acids containing a zinc finger (Zn) domain, a leucine zipper (Zip) motif, and a forkhead domain.6 8 We have previously reported two unrelated Japanese patients with X linked autoimmune enteropathy associated with tubulonephropathy and endocrinopathy.2 9 10 We report here novel mutations in the FOXP3 gene of these patients. Clinical and laboratory findings of our patients have been previously reported.2 9 10 Briefly, patient 1, a boy, now 11 years old, was diagnosed as having autoimmune …

Hori S, Nomura T, Sakaguchi SControl of regulatory T cell development by the transcription factor FOXP3+. Science 299:1057-1061

Article

Full-text available

Mar 2003
SCIENCE

Regulatory T cells engage in the maintenance of immunological self-tolerance by actively suppressing self-reactive lymphocytes. Little is known, however, about the molecular mechanism of their development. Here we show that Foxp3, which encodes a transcription factor that is genetically defective in an autoimmune and inflammatory syndrome in humans and mice, is specifically expressed in naturally arising CD4+ regulatory T cells. Furthermore, retroviral gene transfer of Foxp3 converts naïve T cells toward a regulatory T cell phenotype similar to that of naturally occurring CD4+ regulatory T cells. Thus, Foxp3 is a key regulatory gene for the development of regulatory T cells.

CD4+CD25+ T regulatory cells control anti-islet CD8+ T cells through TGF- -TGF- receptor interactions in type 1 diabetes

Article

Full-text available

Oct 2003

Pancreatic lymph node-derived CD4+CD25+ T regulatory (Treg) cells inhibit in situ differentiation of islet-reactive CD8+ T cells into cytotoxic T lymphocytes, thereby preventing diabetes progression. The mechanism by which these Treg cells suppress anti-islet CD8+ T cells is unknown. Here, we show by using a CD8+ T cell-mediated model of type 1 diabetes that transforming growth factor (TGF)-beta-TGF-beta receptor signals are critical for CD4+CD25+ Treg cell regulation of autoreactive islet-specific cytotoxic T lymphocytes. Transgenic expression of tumor necrosis factor alpha from birth to 25 days of age in the islets of B6 mice that constitutively express CD80 on their beta cells results in accumulation of CD4+CD25+TGF-beta+ cells exclusively in the islets and pancreatic lymph nodes, which delays diabetes progression. In contrast, expression of tumor necrosis factor alpha until 28 days of age prevents islet accumulation of CD4+CD25+TGF-beta+ Treg cells, resulting in acceleration to diabetes. Furthermore, adoptive transfer experiments demonstrated that CD4+CD25+ Treg cells could not control naïve or activated islet-reactive CD8+ T cells bearing a dominant negative TGF-beta receptor type II. Our data demonstrate that, in vivo, TGF-beta signaling in CD8+ T cells is critical for CD4+CD25+ Treg cell suppression of islet-reactive CD8+ T cells in type 1 diabetes.

CD25- T Cells Generate CD25+Foxp3+ Regulatory T Cells by Peripheral Expansion

Article

Full-text available

Jan 2005

Naturally occurring CD4(+) regulatory T cells are generally identified through their expression of CD25. However, in several experimental systems considerable T(reg) activity has been observed in the CD4(+)CD25(-) fraction. Upon adoptive transfer, the expression of CD25 in donor-derived cells is not stable, with CD4(+)CD25(+) cells appearing in CD4(+)CD25(-) T cell-injected animals and vice versa. We show in this study that CD25(+) cells arising from donor CD25(-) cells upon homeostatic proliferation in recipient mice express markers of freshly isolated T(reg) cells, display an anergic state, and suppress the proliferation of other cells in vitro. The maintenance of CD25 expression by CD4(+)CD25(+) cells depends on IL-2 secreted by cotransferred CD4(+)CD25(-) or by Ag-stimulated T cells in peripheral lymphoid organs.

Foxp3 interacts with nuclear factor of activated T cells and NF-kappa B to repress cytokine gene expression and effector functions of T helper cells

Article

Full-text available

May 2005

Scurfy mice, which are deficient in a functional Foxp3, exhibit a severe lymphoproliferative disorder and display generalized over-production of cytokines. Here, we show that, among the Foxp transcriptional factor family, which includes Foxp1, Foxp2, and Foxp3, only Foxp3 has the ability to inhibit IL-2, IL-4, and IFN-γ production by primary T helper cells. We found that Foxp3 physically associates with the Rel family transcription factors, nuclear factor of activated T cells (NFAT) and NF-κB, and blocks their ability to induce the endogenous expression of their target genes, including key cytokine genes. More importantly, T cells derived from scurfy mice have a dramatic increase in nuclear factor of activated T cells (NFAT) and NF-κB transcriptional activity compared with the T cells derived from WT mice. Furthermore, complementation of Foxp3 in scurfy-derived T cells lowers the NFAT and NF-κB transcriptional activity to the physiological level. Finally, we show that myelin proteolipid protein-specific autoreactive T cells transduced with Foxp3 cannot mediate experimental autoimmune encephalomyelitis, providing further support that Foxp3 suppresses the effector function of autoreactive T cells. Foxp3 has already been associated with the generation of CD4⁺CD25⁺ regulatory T cells; our data additionally demonstrate that Foxp3 suppresses the effector functions of T helper cells by directly inhibiting the activity of two key transcription factors, NFAT and NF-κB, which are essential for cytokine gene expression and T cell functions. • experimental autoimmune encephalomyelitis • transcription factors • CD4+ T cells • interleukins

Pillars Article: Foxp3 Programs the Development and Function of CD4+CD25+ Regulatory T Cells. Nat. Immunol. 2003. 4: 330-336

Article

Feb 2017
J IMMUNOL

Pillars Article: Control of Regulatory T Cell Development by the Transcription Factor Foxp3. Science 2003. 299: 1057-1061

Article

Feb 2017
J IMMUNOL

T helper cell type 1 (Th1), Th2 and Th17 responses to myelin basic protein and disease activity in multiple sclerosis (vol 125, pg 161, 2008)

Article

Nov 2008

Foxp3 programs the development and function of CD4+CD25+ regulatory T cells

Article

Jan 2003

Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells

Article

Jan 2006
NATURE

Immunologic self-tolerance Maintained by activated T cells expressing 11-2 receptor α-chains (CD25++)

Article

Apr 2011

Approximately 10% of peripheral CD4+ cells and less than 1% of CD8+ cells in normal unimmunized adult mice express the IL-2 receptor a-chain (CD25++) molecules. When CD4+ cell suspensions prepared from BALB/c nu/+ mice lymph nodes and spleens were depleted of CD25++ cells by specific mAb and C, and then inoculated into BALB/c athymic nude (nu/nu) mice, all recipients spontaneously developed histologically and serologically evi dent autoimmune diseases (such as thyroiditis, gastritis, insulitis, sialoadenitis, adrenalitis, oophoritis, glomerulo nephritis, and polyarthritis); some mice also developed graft-vs-host-like wasting disease. Reconstitution of CD4+ cells within a limited period after transfer of CD4+ cells prevented these autoimmune de velopments in a dose-dependent fashion, whereas the reconstitution several days later, or inoculation of an equivalent dose of CD8+ cells, was far less efficient for the prevention. When nu/nu mice were transplanted with allogeneic skins or immunized with xenogeneic proteins at the time of CD25++ cell inoculation, they showed significantly heightened immune responses to the skins or proteins, and reconstitution of CD4+ cells normalized the responses. Taken together, these results indicate that CD4+ cells contribute to maintaining self-tolerance by down-regulating immune response to self and non-self Ags in an Ag-nonspecific manner, pre sumably at the T cell activation stage; elimination/reduction of CD4+ cells relieves this general suppres sion, thereby not only enhancing immune responses to non-self Ags, but also eliciting autoimmune responses to certain self-Ags. Abnormality of this T cell-mediated mechanism of peripheral tolerance can be a possible cause of various autoimmune diseases.

Th1 And Th2 Cells: Different Patterns Of Lymphokine Secretion Lead To Different Functional Properties

Article

Jan 1989

T R Mosmann

Impairment of Regulatory T-Cell Function in Autoimmune Thyroid Disease

Article

Feb 2013
THYROID

Background: Autoimmune thyroid disease (AITD) pathogenesis may result from a loss of immune tolerance to thyroid antigens. Regulatory T cells (Tregs) control immune responses, prevent excessive inflammation, and may be dysfunctional in AITD. We investigated the role of Tregs in Hashimoto's thyroiditis (HT) and Graves' disease (GD), complicated by Down syndrome (DS). Our goal was to identify differences in CD4(+)CD25(high) Treg function or number in patients with GD and HT, compared to healthy controls (HC). Methods: Treg number was assessed by flow cytometric analysis in samples from 20 AITD patients (seven GD, 13 HT), nine HC, and seven individuals with DS, a genetic disorder associated with multiple autoimmune disorders including AITD. Treg function was assessed by the inhibition of proliferation (radioactive thymidine incorporation into DNA) of blood-derived T effector (Teff) cells by Tregs in a coculture. Various methods of stimulation were contrasted. Cytokine levels were determined in conditioned media from the co-cultures. Results: No differences were found in the frequency of Tregs as a percentage of CD4(+) cells between AITD and HC. AITD Tregs were less capable of inhibiting the proliferation of Teff cells when compared to HC; however, the impairment was dependent on the type of stimulation used. DS patients without AITD exhibited normal Treg function. We observed few differences in cytokine production between HC and AITD patients. Conclusions: Tregs from AITD patients are partly dysfunctional, possibly explaining their autoimmunity. Future work will elucidate the diagnostic potential and pathophysiology of Tregs in AITD.

B-cell exposure to self-antigen induces IL-10 producing B cells as well as IL-6- and TNF-α-producing B-cell subsets in healthy humans

Article

Jul 2012
CLIN IMMUNOL

Human B cells are able to secrete IL-10 after stimulation with mitogens, but their ability to produce IL-10 and regulate T-cell responses after stimulation with self-antigens is unclear. We co-cultured thyroglobulin-pulsed B cells from healthy donors with autologous T cells and observed production of IL-10 and TGF-β, in addition to TNF-α and IL-6. Pulsing with foreign antigen, tetanus toxoid (TT), induced a Th1-response with minimal IL-10 production. After thyroglobulin-pulsing, 1.10±0.50% of B cells and 1.00±0.20% of CD4(+) T cells produced IL-10, compared to 0.29±0.19% of B cells (P=0.01) and 0.13±0.15% of CD4(+) T cells (P=0.006) following TT-pulsing. Thyroglobulin-stimulated, IL-10-secreting B cells were enriched within CD5(+) and CD24(high) cells. While thyroglobulin-pulsed B cells induced only modest proliferation of CD4(+) T cells, B cells pulsed with TT induced vigorous proliferation. Thus, B cells mediate self-antigen-specific IL-10, TNF-α and IL-6 production in co-cultures with T cells and contribute actively to these cytokine secretions.

Type 1 T regulatory cells

Article

Mar 2002
IMMUNOL REV

Suppression by T regulatory (Tr) cells is essential for induction of tolerance. Many types of Tr cells have been described in a number of systems, and their biology has been the subject of intensive investigation. Although many aspects of the mechanisms by which these cells exert their effects remain to be elucidated, it is well established that Tr cells suppress immune responses via cell-to-cell interactions and/or the production of interleukin (IL)-10 and transforming growth factor (TGF)-β. Type-1 T regulatory (Tr1) cells are defined by their ability to produce high levels of IL-10 and TGF-β. Tr1 cells specific for a variety of antigens arise in vivo, but may also differentiate from naive CD4+ T cells in the presence of IL-10 in vitro. Tr1 cells have a low proliferative capacity, which can be overcome by IL-15. Tr1 cells suppress naive and memory T helper type 1 or 2 responses via production of IL-10 and TGF-β. Further characterisation of Tr1 cells at the molecular level will define their mechanisms of action and clarify their relationship with other subsets of Tr cells. The use of Tr1 cells to identify novel targets for the development of new therapeutic agents, and as a cellular therapy to modulate peripheral tolerance, can be foreseen.

FoxP3 mRNA splice forms in synovial CD4+ T cells in rheumatoid arthritis and psoriatic arthritis

Article

May 2012
APMIS

Ryder LR, Bartels EM, Woetmann A, Madsen HO, Ødum N, Bliddal H, Danneskiold-Samsøe B, Ribel-Madsen S, Ryder LP. FoxP3 mRNA splice forms in synovial CD4+ T cells in rheumatoid arthritis and psoriatic arthritis. APMIS 2012; 120: 387–96. Our aim was to elucidate the relative amount of the different splice forms of FoxP3 mRNA in CD4+ T cells in peripheral blood (PB) compared to synovial fluid (SF) in RA and PsA patients. FoxP3 mRNA was measured using a quantitative real-time PCR method. CD4+ T cells were isolated from 17 paired samples of PB and SF from RA and PsA patients, and PB from 10 controls. FoxP3fl and FoxP3Δ2 mRNA was significantly increased (6.7 and 2.1-fold, respectively) in PB CD4+ T cells from RA patients compared to controls. FoxP3fl and Δ2 mRNA in SF CD4+ T cells was increased compared to controls in sero-negative RA and PsA, but not in sero-positive RA patients, who had a high FoxP3 expression in both PB and SF. The FoxP3Δ2Δ7 mRNA was barely detectable in patient samples, and not at all in healthy individuals. We provide evidence of an increased expression of FoxP3 splice forms in synovial CD4+ T cells from RA patients. A skewed, high expression profile of FoxP3, but not CTLA-4, in sero-negative RA and PsA, indicates that synovial CD4+ T cells may represent unique subsets of T cells which have been induced locally or selectively recruited to the joint.

Inhibition of Immunoglobulin-Secreting Cells by Antithyroid Drugs*

Article

Dec 1981

Previous studies have indicated that titers of thyroid-specific autoantibodies decrease in patients with autoimmune thyroid disease during antithyroid drug therapy. In keeping with these observations has been the accumulating evidence that such drugs may have an immunosuppressive action both in vivo and in vitro. To further analyze the mechanism of such activity, we have used an indirect hemolytic plaque assay based on protein A-coated sheep red blood cells and a specific plaque-forming cell (PFC) assay based on human thyroglobulin-linked sheep red blood cells. When freshly prepared peripheral mononuclear cells were exposed to increasing concentrations of methimazole or propylthiouracil (PTU), there was no significant effect on spontaneous plaque-forming cell responses (PFC). However, after incubation for more than 6 days with methimazole or PTU, there was a marked dose-related inhibition of pokeweed mitogen stimulated PFC, the degree of inhibition varying from one individual to another. Overall, there were 31.5 +/- 9.6% and 32.5 +/- 5.2% reductions in plaque formation with 10(-4) M methimazole or PTU, respectively, a level which probably approximates intrathyroidal thionamide concentrations in patients. A similar inhibitory response was seen in the human thyroglobulin-PFC assay using peripheral mononuclear cells from patients with autoimmune thyroiditis, confirming the potential inhibition of thyroid-specific autoantibody production by these compounds. Furthermore, at 10(-4) M there was no effect of PTU or methimazole on cell survival, as assessed by trypan blue exclusion. These data suggested that the thionamide drugs interfered with immunoglobulin production, rather than secretion, at concentrations which were likely to be present intrathyroidally in many treated patients.

Differentiation Imbalance of Th1/ Th17 in Peripheral Blood Mononuclear Cells Might Contribute to Pathogenesis of Hashimoto’s Thyroiditis

Article

Sep 2010
SCAND J IMMUNOL

T helper 17(Th17) cell is a new subset of CD4(+) T cells that produce a proinflammatory cytokine interleukin-17 (IL-17). Th17 cells have recently been shown to play a critical role in many autoimmune diseases that had previously been thought to be Th1 dominant. Although Hashimoto's thyroiditis (HT) was thought to be a Th1-type disease, the contributions of Th17 cells to the pathogenesis remain unclear. In this study, we investigated the expression levels of Th1/Th17 cell-associated factors in peripheral blood mononuclear cells (PBMC) and plasma from patients with HT by quantitative real-time polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). Our results showed that the expression levels of Th1 cells-related T-bet and interferon-gamma (IFN-gamma) mRNA in PBMC from HT significantly decreased. However, the mRNA of Th17 coherent retinoic acid-related orphan nuclear receptor gamma t (RORgammat) and IL-17 in patients with HT increased. In addition, a negative correlation between T-bet and RORgammat mRNA expression was found in patients with HT, and the similar phenomena also appeared on the levels of mRNA and plasma concentration between IFN-gamma and IL-17. It suggested that Th17 cells rather than Th1 cells predominated among patients suffering from HT, and Th17 cells might be involved in the pathogenesis of HT.

Expression of full-length and splice forms of FoxP3 in rheumatoid arthritis

Article

Aug 2010
SCAND J RHEUMATOL

The aim of our study was to compare the presence of full-length and alternative splice forms of FoxP3 mRNA in CD4 cells from rheumatoid arthritis (RA) patients and healthy controls. A quantitative real-time polymerase chain reaction (QRT-PCR) method was used to measure the amount of FoxP3 mRNA full-length and splice forms. CD4-positive T cells were isolated from peripheral blood from 50 RA patients by immunomagnetic separation, and the FoxP3 mRNA expression was compared with the results from 10 healthy controls. We observed an increased expression of full-length FoxP3 mRNA in RA patients when compared to healthy controls, as well as an increase in CD25 mRNA expression, but no corresponding increase in CTLA-4 mRNA expression. The presence of an alternative splice form of FoxP3 lacking exon 2 was confirmed in both RA patients and healthy controls, but with no significant difference in expression between the two groups. There was a positive correlation between the amount of FoxP3 mRNA and the clinical inflammation parameters C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), and a negative correlation between FoxP3 mRNA and the dose of methotrexate (MTX) given to the patients. RA patients express more full-length FoxP3 than healthy controls in peripheral blood CD4-positive cells, suggesting an increased number of regulatory T cells (Tregs). However, no concomitant increase in CTLA-4 expression was seen. We therefore propose that the Tregs are left unable to suppress the ongoing inflammation due to a deficiency in CTLA-4 needed for cell contact-dependent suppression.

Potent intestinal Th17 priming through peripheral lipopolysaccharide-based immunization

Article

Jul 2010
J LEUKOCYTE BIOL

The data provide impetus for using TLR-based peripheral vaccination to augment specific Th17 immunity in the gut mucosa. Lipopolysaccharide (LPS) is a potent natural adjuvant, commonly used to amplify Th1 responses. Here, we report that systemic immunization using LPS generates large numbers of specific Th17 cells in murine small intestinal lamina propria. The priming of these Th17 cells required IL-23p19 production by bone marrow-derived cells. In contrast, IL-23 had no impact on Th1 differentiation or overall numbers of Ag-specific regulatory T cells. Experiments using T-cell adoptive transfers revealed a previously unappreciated mechanism for how Th17 responses are amplified in vivo: stimulation through LPS expanded precommitted Th17 cells rather than causing Th17 differentiation. Second, LPS drove Th17 cell expansion independently of IL-23, demonstrating that this cytokine is not necessary for expansion and possibly functions at an earlier stage in Th17 priming. Our data provide an impetus for using LPS-based peripheral vaccination to augment specific T-cell-mediated immunity in the gut mucosa.

The self-antigen, thyroglobulin, induces antigen-experienced CD4 + T cells from healthy donors to proliferate and promote production of the regulatory cytokine, interleukin-10, by monocytes

Article

Oct 2009
IMMUNOLOGY

Thyroglobulin (TG), as autoantigen, induces in vitro proliferation of T and B cells from normal individuals, but the cytokine production differs from that in patients with autoimmune thyroid disease. Here, we investigate whether normal T cells responding to TG are naive, or have previously encountered TG in vivo, using their responses to classic primary and secondary antigens, keyhole limpet haemocyanin (KLH) and tetanus toxoid (TT), respectively, for comparison. While TG elicited T-cell proliferation kinetics typical of a secondary response, the cytokine profile was distinct from that for TT. Whereas TT induced pro-inflammatory cytokines [interleukin-2 (IL-2)/interferon-gamma (IFN-gamma)/IL-4/IL-5], TG evoked persistent release of the regulatory IL-10. Some donors, however, also responded with late IFN-gamma production, suggesting that the regulation by IL-10 could be overridden. Although monocytes were prime producers of IL-10 in the early TG response, a few IL-10-secreting CD4(+) T cells, primarily with CD45RO(+) memory phenotype, were also detected. Furthermore, T-cell depletion from the mononuclear cell preparation abrogated monocyte IL-10 production. Our findings indicate active peripheral tolerance towards TG in the normal population, with aberrant balance between pro- and anti-inflammatory cytokine responses for some donors. This observation has implications for autoantigen recognition in general, and provides a basis for investigating the dichotomy between physiological and pathological modes of auto-recognition.

Increases of the Th1/Th2 Cell Ratio in Severe Hashimoto's Disease and in the Proportion of Th17 Cells in Intractable Graves' Disease

Article

May 2009
THYROID

T helper type 1 (Th1), Th2, and Th17 cells produce interferon (IFN)-gamma, interleukin (IL)-4, and IL-17A, respectively. We reported that IFN-gamma and IL-4 gene polymorphisms, which are related to higher IFN-gamma and lower IL-4 production, respectively, are more frequent in patients with severe Hashimoto's disease (HD) than in those mild HD. We now aim to investigate the proportion of peripheral Th1, Th2, and Th17 cells in patients with autoimmune thyroid disease (AITD). We studied 17 patients with HD who developed hypothyroidism and were treated with l-thyroxine, referred to as severe HD; 17 untreated patients with HD who were euthyroid, referred to as mild HD; 18 euthyroid patients with Graves' disease (GD) who remained positive for anti-thyrotropin receptor antibody (TRAb) despite being treated with anti-thyroid drugs for more than 5 years, referred to as intractable GD; and 17 patients with GD who were euthyroid and negative for TRAb for more than 2 years after cessation of anti-thyroid drugs, referred to as GD in remission; and 10 control subjects without AITD. By the definitions in this study Th1 cells were CD4(+)IFN-gamma(+)IL-4(-)IL-17A(-) cells, Th2 cells were CD4(+)IFN-gamma(-)IL-4(+)IL-17A(-) cells, and CD4(+)IFN-gamma(-)IL-4(-)IL-17A(+) cells were Th17 cells. The proportion of peripheral Th1 cells was higher in patients with severe HD than in patients with mild HD (p < 0.05), and the proportion of peripheral Th2 cells was lower in patients with severe HD than in patients with mild HD (p < 0.001). Therefore the Th1/Th2 ratio was higher in severe than in mild HD patients (p < 0.001). The proportion of peripheral Th17 cells in patients with AITD was higher than in control subjects and the proportion of these cells in patients with intractable GD was higher than in patients with GD in remission (p < 0.05). The peripheral Th1/Th2 cell ratio is related to the severity of HD, and the proportion of Th17 cells is related to the intractability of GD. We hypothesize that these patterns of peripheral Th cell subsets may be expressed within the thyroid.

Naturally-existing CD4(+)CD25(+)Foxp3(+) regulatory T cells are required for tolerance to experimental autoimmune thyroiditis induced by either exogenous or endogenous autoantigen

Article

May 2009
J AUTOIMMUN

Murine experimental autoimmune thyroiditis (EAT) is a model for Hashimoto's thyroiditis, an organ-specific autoimmune disease characterized by mononuclear cell infiltration and destruction of the thyroid gland. Susceptibility to EAT is MHC-linked, and influenced by CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs). Treg depletion enables thyroiditis induction with mouse thyroglobulin (mTg) in traditionally-resistant mice and mTg-induced, Treg-mediated tolerance protects against EAT induction in genetically-susceptible mice. Here, we demonstrate the existence of naturally-existing CD4(+)CD25(+)Foxp3(+) Tregs (nTregs) influencing thyroiditis development in naive susceptible mice and that induction of thyroiditis in these mice involves overcoming peripheral homeostatic immune suppression by nTregs. Additionally we demonstrate that nTregs are required for induction of antigen-specific tolerance, indicating that induced EAT tolerance is a result of activation of naturally-existing nTregs rather than de novo generation of induced Tregs (iTregs). Examination of several potential costimulatory molecules previously described as involved in peripheral activation of Tregs demonstrates a critical role indeed for CTLA-4 in the activation of nTregs leading to development of EAT tolerance and providing a mechanism for mTg-induced Treg activation during tolerance induction. Together, these data reinforce the important role of Tregs in mediating self-tolerance, and illuminate a potential mechanism for their therapeutic expansion in induced tolerance.

Regulatory T cells in Graves' disease

Article

Mar 2009
CLIN ENDOCRINOL

Graves' disease (GD) involves auto-immunity against thyroid cell antigens, but the reasons for induction of auto-immunity are uncertain. We wished to determine whether there was a deficiency of regulatory T cells in patients with active GD. Venous blood samples were obtained from patients with GD before and after treatment, and controls, and peripheral blood mononuclear cells were prepared. Regulatory T cells were enumerated by Fluorescent Activated Cell sorting (FACS) in nineteen patients with untreated GD, 9 patients 6-8 weeks post RAI therapy, and 30 control subjects. Twenty-one patients with active GD prior to control of hyperthyroidism, 23 euthyroid controls without known autoimmune thyroid disease, and 10 patients who were euthyroid 6-12 months after RAI treatment were studied for expression of genes found in regulatory T cells by real-time Polymerase Chain reaction (PCR). Percent distribution of CD4+, CD4+CD25+ and CD4+ CD25+(int-hi) CD127+(lo) regulatory T cells was similar in active GD patients and control subjects. The number of CD25+ and CD4+ CD25+(int-hi) CD127+(lo) cells was similar in GD patients and control subjects, but was lower in recently treated patients. Messenger RNA was prepared from PBMC, and reverse transcribed. Copy DNA abundance was evaluated by Real Time PCR using appropriate primers, for GAPDH (glyceraldehyde phosphate dehydrogenase) as a control housekeeping gene, and 5 genes related to function of regulatory T cells. Message RNA for Gadd45 alpha, Gadd45beta (growth arrest and damage inducible proteins), GITR (glucocorticoid inducible TNF receptor) and CD25 (IL-2R subunit) was more abundant in patients with active GD than in normal controls, and FoxP3 mRNA level was equal to that in controls. Message RNA levels in patients treated and euthyroid for 6 months were also greater than or equal to values in controls. This study provides evidence that there is no deficit in T regulatory cells during active GD, or during the months post therapy.

IL-17-producing human peripheral regulatory T cells retain suppressive function

Article

Feb 2009
BLOOD

Although implicated in antagonistic functions, both regulatory T cells (Tregs) and Th17 effector cells play an important role in controlling autoimmune pathogenesis. Paradoxically, recent studies indicate that Tregs have the capacity to produce interleukin-17 (IL-17), although the ability of these cells to retain their suppressive function remains unknown. Here we report that human Tregs within the CD4(+)CD45RA(-)CD25(high)CCR6(+)HLA-DR(-)FoxP3(+) population produce IL-17 when activated in the presence of the proinflammatory cytokines IL-1beta and IL-6, whereas IL-17 secretion was inhibited by TGFbeta. To assess the ability of a single Treg to secrete IL-17 and to suppress in vitro immune function, we isolated clones from this population. We found that IL-17(+)/FoxP3(+) Treg clones retain suppressive function and exhibit the plasticity to secrete IL-17 or suppress depending on the nature of the stimulus provided. IL-17 production by these Treg clones was accompanied by sustained FoxP3 expression and concomitant, but reversible, loss of suppressive activity. Our data demonstrate that at the single cell level a subset of in vitro suppressive FoxP3(+) cells can be driven to secrete IL-17 under inflammatory conditions. These findings suggest a new mechanism by which inflammation can drive Tregs to secrete IL-17, thereby dampening suppression and promoting an inflammatory milieu.

Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins

Article

May 1986

A panel of antigen-specific mouse helper T cell clones was characterized according to patterns of lymphokine activity production, and two types of T cell were distinguished. Type 1 T helper cells (TH1) produced IL 2, interferon-gamma, GM-CSF, and IL 3 in response to antigen + presenting cells or to Con A, whereas type 2 helper T cells (TH2) produced IL 3, BSF1, and two other activities unique to the TH2 subset, a mast cell growth factor distinct from IL 3 and a T cell growth factor distinct from IL 2. Clones representing each type of T cell were characterized, and the pattern of lymphokine activities was consistent within each set. The secreted proteins induced by Con A were analyzed by biosynthetic labeling and SDS gel electrophoresis, and significant differences were seen between the two groups of T cell line. Both types of T cell grew in response to alternating cycles of antigen stimulation, followed by growth in IL 2-containing medium. Examples of both types of T cell were also specific for or restricted by the I region of the MHC, and the surface marker phenotype of the majority of both types was Ly-1+, Lyt-2-, L3T4+, Both types of helper T cell could provide help for B cells, but the nature of the help differed. TH1 cells were found among examples of T cell clones specific for chicken RBC and mouse alloantigens. TH2 cells were found among clones specific for mouse alloantigens, fowl gamma-globulin, and KLH. The relationship between these two types of T cells and previously described subsets of T helper cells is discussed.

TH1 and TH2 Cells: Different Patterns of Lymphokine Secretion Lead to Different Functional Properties

Article

Feb 1989

Enhanced production of IFNγ by thyroid-derived T cell clones from patients with Hashimoto's thyroiditis

Article

Sep 1987

T lymphocytes from thyroid infiltrate and peripheral blood (PB) of four patients with Hashimoto's thyroiditis (HT) were analysed at clonal level for their ability to secrete interleukin 2 (IL-2) and gamma-interferon (gamma-IFN). As controls, T cell clones from PB of four normal donors and from spleen of two trauma victims were used. While no abnormality was found in the capacity to produce IL-2, the proportion of gamma-IFN-producing (IFN-P) T cell clones derived from HT infiltrates was significantly higher (P less than 0.0005) than that of IFN-P clones derived from normal or patient PB. Most of CD4+ and CD8+ IFN-P clones from thyroid infiltrates, as well as a proportion of CD4+ PB-derived clones of patients with HT, released higher amounts of gamma-IFN than control clones. A relationship could be demonstrated between high gamma-IFN production and natural killer (NK) activity in T cell clones from thyroid and PB of HT patients. In fact, the percentage of IFN-P clones with NK potential (NK+) was remarkably higher (P less than 0.0005) in thyroid infiltrates than in normal spleen or PB. The proportion of IFN-P NK+ clones from patient PB was also significantly increased (P less than 0.02) but, unlike thyroid-derived clones in which the majority of IFN-P NK+ clones were CD8+, most PB-derived IFN-P NK+ clones from the same patients expressed the CD4+ phenotype. Almost all thyroid NK+ clones could be triggered to produce more gamma-IFN, while gamma-IFN synthesis by NK-negative thyroid clones was comparable to that of control clones. In view of the multiple effects ascribed to gamma-IFN in the cascade of events leading to immune responses, the abnormal potential to gamma-IFN secretion shown by intrathyroidal T lymphocytes may be of importance in the pathogenesis of autoimmune thyroiditis.

Carbimazole and the Autoimmune Response in Graves' Disease

Article

Sep 1980

Microsomal antibodies and antibodies directed toward the receptor for thyroid-stimulating hormone (TSH) decreased in parallel while patients with Graves' disease were taking carbimazole, whereas no significant changes were observed during treatment with placebo or propranolol. The changes in autoantibody levels during carbimazole treatment were independent of changes in serum thyroxine and could have been due to a direct effect of the drug on autoantibody synthesis. Evidence for this suggestion was provided when low doses of methimazole (the active metabolite of carbimazole) were found to inhibit thyroid-autoantibody production in cultured lymphocytes. Since thyroid lymphocytes are probably a major site of thyroid-antibody synthesis in Graves' disease and methimazole is concentrated in the thyroid during treatment, a local action of the drug on antibody production seems likely. This possibility could be important in the use of carbimazole to control hyperthyroidism.

Immunologic Self-Tolerance Maintained by Activated T Cells Expressing IL-2 Receptor alpha-Chains (CD25) Breakdown of a Single Mechanism of Self-Tolerance Causes Various Autoimmune Diseases

Article

Sep 1995

Approximately 10% of peripheral CD4+ cells and less than 1% of CD8+ cells in normal unimmunized adult mice express the IL-2 receptor alpha-chain (CD25) molecules. When CD4+ cell suspensions prepared from BALB/c nu/+ mice lymph nodes and spleens were depleted of CD25+ cells by specific mAb and C, and then inoculated into BALB/c athymic nude (nu/nu) mice, all recipients spontaneously developed histologically and serologically evident autoimmune diseases (such as thyroiditis, gastritis, insulitis, sialoadenitis, adrenalitis, oophoritis, glomerulonephritis, and polyarthritis); some mice also developed graft-vs-host-like wasting disease. Reconstitution of CD4+CD25+ cells within a limited period after transfer of CD4+CD25- cells prevented these autoimmune developments in a dose-dependent fashion, whereas the reconstitution several days later, or inoculation of an equivalent dose of CD8+ cells, was far less efficient for the prevention. When nu/nu mice were transplanted with allogeneic skins or immunized with xenogeneic proteins at the time of CD25- cell inoculation, they showed significantly heightened immune responses to the skins or proteins, and reconstitution of CD4+CD25+ cells normalized the responses. Taken together, these results indicate that CD4+CD25+ cells contribute to maintaining self-tolerance by down-regulating immune response to self and non-self Ags in an Ag-nonspecific manner, presumably at the T cell activation stage; elimination/reduction of CD4+CD25+ cells relieves this general suppression, thereby not only enhancing immune responses to non-self Ags, but also eliciting autoimmune responses to certain self-Ags. Abnormality of this T cell-mediated mechanism of peripheral tolerance can be a possible cause of various autoimmune diseases.

Intrathyroidal cytokine gene expression in Hashimoto's thyroiditis

Article

Oct 1996

Hashimoto's thyroiditis (HT) is an organ-specific autoimmune disease in which cytokines are likely to have a role in the initiation and perpetuation of the disease. Using the reverse transcription-polymerase chain reaction (RT-PCR) we analysed the cytokine profile in four HT tissue samples. Furthermore, cell fractionation was carried out on two tissue samples and cytokine profile was studied in CD4+ and CD8+ T cells, in addition to the residual cellular infiltrate composed of CD4- and CD8- cells. Our results showed IL-1 beta, IL-4 and IL-6 mRNA expression in three out of four tissue samples, whereas IL-1 alpha, IL-2, IL-8, IL-10, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) were expressed in all tissue samples studied. Expression of IL-1 alpha and IL-1 beta was absent in both CD4+ and CD8+ subsets. However, IL-2, IL-4, IL-6, IL-8, IFN-gamma and TNF-alpha mRNA were detected in both CD4+ and CD8+ subsets. IL-10 was expressed in the CD4+ subset in one sample, whereas it was negative in both CD8+ fractions. All the cytokines studied were expressed in the residual infiltrate. These results suggest a mixed Th1 and Th2 response in HT, both of which may have an important role in the pathophysiology of the thyroid destructive process through cell-mediated cytotoxicity, and/or humoral autoimmunity manifested by autoantibody production.

The Expanding Universe of T-Cell Subsets: Th1, Th2 and More

Article

Apr 1996
Immunol Today

Since their discovery nearly ten years ago, T helper 1 (Th1) and Th2 subsets have been implicated in the regulation of many immune responses. In this article, Tim Mosmann and Subash Sad discuss the increasing number of T-cell subsets defined by cytokine patterns; the differentiation pathways of CD4+ and CD8+ T cells; the contribution of other cell types to these patterns; and the cytokine interactions during infection and pregnancy.

Removal of Endotoxin from Recombinant Protein Preparations

Article

Sep 1997
CLIN BIOCHEM

To develop an effective method to remove endotoxin from large scale E. coli recombinant protein purifications. Triton X-114 phase separation, affinity chromatography utilizing immobilized polymyxin B or immobilized histidine, were used to remove endotoxin from purified preparations of recombinant CK-BB, CK-MB, CK-MM, myoglobin, and cardiac troponin I. Endotoxin levels were measured by a Limulus Amebocyte Lysate gel-clot assay. The immunoactivity of these protein preparations was determined by BIAcore analysis using a panel of in-house generated monoclonal antibodies and by a Stratus Fluorometric Analyzer. In the case of troponin I, the BIAcore was also utilized to measure troponin C interactions. Phase separation with Triton X-114 was the most effective method in reducing the amount of endotoxin present in the protein preparations compared to either polymyxin B or histidine affinity chromatography. With Triton X-114, the reduction in endotoxin levels was greater than 99% and recovery of the proteins after endotoxin removal was greater than 90%. All three procedures for removing endotoxin had no deleterious effects on the immunoactivity of majority proteins when tested with a panel of monoclonal antibodies. Troponin I also retained its ability to bind to troponin C in the presence of Ca2+. Recombinant CK-BB and CK-MM which were expressed in the soluble fraction of E. coli cell lysates, contained significantly higher endotoxin levels than recombinant CK-MB, myoglobin and cardiac troponin I which were expressed in the form of inclusion bodies. Of the three methods tested, Triton X-114 phase separation was the most effective way of removing endotoxin from recombinant proteins.

Balance of Th1/Th2 Cytokines in Thyroid Autoantibody Synthesis in vitro

Article

Feb 1999

Thyroid-infiltrating lymphocytes spontaneously synthesize IgG class thyroid autoantibodies while blood lymphocytes require activation to produce the same autoantibodies. Surprisingly, the stimulus commonly used to induce autoantibodies by blood lymphocytes, Pokeweed mitogen (PWM), inhibits autoantibody synthesis by thyroid lymphocytes. To address this dichotomy, we investigated the Th1: Th2 cytokine balance in relation to thyroid peroxidase (TPO) autoantibody production in cultures of thyroid, lymph node and blood lymphocytes. The characteristic PWM effect on TPO autoantibody production by thyroid and blood lymphocytes (10 day cultures) was confirmed. Cytokine measurements in these cultures revealed that PWM increased IFN-gamma production by thyroid, lymph node and blood lymphocytes. However, PWM enhanced IL-4 levels in lymphocytes from blood and lymph node but not in thyroid lymphocytes. Moreover, the IL-4: IFN-gamma ratios in short- and long-term cultures were higher for PBMC and lymph node lymphocytes than for thyroid lymphocytes. In summary, PWM shifts the cytokine balance towards Th2 for blood lymphocytes and towards Th1 for thyroid lymphocytes. The shift towards Th1 in the target organ is associated with reduced autoantibody synthesis. Our observations suggest that "immune deviation" towards Th2 as a form of therapy in Graves' disease could project the patient from the frying pan into the fire.

The Immunomodulatory Effects of Anti-thyroid Drugs are Mediated via Actions on Thyroid Cells, Affecting Thyrocyte-immunocyte Signalling A Review

Article

May 2001

Robert Volpé

The mechanism of action of the immunosuppressive effects of antithyroid drugs has remained a matter of controversy, despite our earlier contention that such effects in vivo were indirect; ie., it was our view that the drugs were acting on the thyroid cells, reducing their thyroid hormone production and other activities, with a consequent reduction in thyrocyte-immunocyte signalling. The reduction in the activation of CD4+ cells,the increased number and activation of CD8+ (and CD8+CDllb+) cells, and the reduction of soluble interleukin-2 receptors, thought once to be direct effects of the medication, are now shown to be due to amelioration of the hyperthyroidism. Thus the reduction in thyroid hormone production induced by the drugs is central to these actions. In addition, the iodination of thyroglobulin is inhibited by these agents, which may affect antigen presentation by the thyrocyte. Furthermore, there is now evidence that the thionamides interfere with thyrocyte expression of such molecules as Class I antigen, interleukin-1, interleukin-6, prostaglandin E2, and heat shock protein. The expression of thyrocyte Class II antigen is probably not inhibited by these drugs, although one group has shown that lectin-stimulated thyrocyte Class II expression is diminished by this treatment; this group postulated that this effect might be mediated by reduced interferon gamma production by T lymphocytes, but in vitro experiments do not corroborate this proposal. In any event, the actions as described of the effects of antithyroid drugs on the thyroid cells (particularly normalization of thyroid function) would certainly suffice to explain the diminution of thyroid antibodies (including thyroid stimulating antibody), the reduced immunological response, and the increased remission rate in Graves disease as a consequence of antithyroid drug therapy, without the need to invoke a direct immunosuppressive effect.

Determinants of autoimmune thyroid disease

Article

Oct 2001

Anthony P Weetman

Graves' disease and other autoimmune syndromes affecting the thyroid are the archetypes of organ-specific autoimmunity. Despite intensive research, the relative contribution of genetic and environmental factors to disease pathogenesis is not clear. Here, the latest developments in understanding the determinants of these diseases are discussed.

Fontenot JD, Gavin MA, Rudensky AYFoxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol 4:330-336

Article

May 2003

CD4+CD25+ regulatory T cells are essential for the active suppression of autoimmunity. Here we report that the forkhead transcription factor Foxp3 is specifically expressed in CD4+CD25+ regulatory T cells and is required for their development. The lethal autoimmune syndrome observed in Foxp3-mutant scurfy mice and Foxp3-null mice results from a CD4+CD25+ regulatory T cell deficiency and not from a cell-intrinsic defect of CD4+CD25- T cells. CD4+CD25+ regulatory T cells rescue disease development and preferentially expand when transferred into neonatal Foxp3-deficient mice. Furthermore, ectopic expression of Foxp3 confers suppressor function on peripheral CD4+CD25- T cells. Thus, Foxp3 is a critical regulator of CD4+CD25+ regulatory T cell development and function.

Functional lymphocyte subset assessment of the Th1/Th2 profile in patients with autoimmune thyroiditis by flowcytometric analysis of peripheral lymphocytes

Article

Jan 2004
J BIOL REG HOMEOS AG

Hashimoto's thyroiditis (HT) results from a parenchymal infiltration by Th1 T cell clones that ultimately may cause tissue destruction. We analysed here whether the quantitative assessment of the cytokine profile in peripheral lymphocytes could help for the evaluation of patients with HT. We added to a flow cytometric evaluation of lymphocyte subpopulations, an assay to identify specific Th1 and Th2 functional subsets. Whole blood diluted in RPMI was activated with PMA and ionomycin for 4h at 37 degrees C in the presence of brefeldin. Immunophenotyping of samples was performed with PerCP-Cy5.5-conjugated CD3, and APC-conjugated CD4 antibodies. After staining for surface antigens, red cells were lysed and white cells were permeabilized. Intracellular cytokines were detected with FITC-conjugated INFgamma (Th1) and PE-conjugated IL-4 (Th2) monoclonal antibodies (mabs). Twenty-three consecutive patients were selected based on the detection of anti TPO ab concentration equal or higher than 600 UI/L. They were compared to 17 healthy control subjects (with undetectable TPO abs). The lymphocyte count and the proportion of the different lymphocyte subsets (T cells, B cells, NK cells, T-CD4+, T-CD8+, Th1 CD3, Th2 CD3, Th1 CD4, Th2 CD4 cells) were alike when both groups were compared. A significant difference appeared when the Th1/Th2 ratio measured on CD3 T cells was considered. This ratio was significantly increased in HT patients when compared to controls (24.91+/-2.9 vs. 15.5+/-1.4 (mean +/- SEM); p <0.05). When the same analysis was performed on CD4 T cells, the Th1/Th2 ratio was again higher in HT patients, although without reaching significance (13.5+/-1.9 vs. 8.8+/-0.6; p >0.05). Our data indicate that the Th1 context analysed in peripheral lymphocytes is dominant in HT patients. Flowcytometry could be used as a diagnostic tool to better understand the pathogeny and the outcome of destructive autoimmune thyroiditis.

Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins

Article

Aug 2005

A panel of antigen-specific mouse helper T cell clones was characterized according to patterns of lymphokine activity production, and two types of T cell were distinguished. Type 1 T helper cells (T(H)1) produced IL 2, interferon-gamma, GM-CSF, and IL 3 in response to antigen + presenting cells or to Con A, whereas type 2 helper T cells (T(H)2) produced IL 3. BSF1, and two other activities unique to the T(H)2 subset, a mast cell growth factor distinct from IL 3 and a T cell growth factor distinct from IL 2. Clones representing each type of T cell were characterized, and the pattern of lymphokine activities was consistent within each set. The secreted proteins induced by Con A were analyzed by biosynthetic labeling and SDS gel electrophoresis, and significant differences were seen between the two groups of T cell line. Both types of T cell grew in response to alternating cycles of antigen stimulation, followed by growth in IL 2-containing medium. Examples of both types of T cell were also specific for or restricted by the I region of the MHC, and the surface marker phenotype of the majority of both types was Ly-1(+), Lyt-2(-), L3T4(+). Both types of helper T cell could provide help for B cells, but the nature of the help differed. T(H)1 cells were found among examples of T cell clones specific for chicken RBC and mouse alloantigens. T(H)2 cells were found among clones specific for mouse alloantigens, fowl gamma-globulin, and KLH. The relationship between these two types of T cells and previously described subsets of T helper cells is discussed.

Altered balance between self-reactive Th17 cells and Th10 cells and between full-length FOXP3 and FOXP3 splice variants in Hashimoto's thyroiditis

Abstract

No full-text available

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Thyroperoxidase, thyroglobulin, Na+/I- symporter, pendrin in thyroid autoimmunity

Immune mechanisms involved in the coexistence of oral lichen planus and autoimmune thyroid diseases