Peripheral regulatory T cells and anti-inflammatory cytokines in children with juvenile idiopathic arthritis

Background Juvenile idiopathic arthritis (JIA) is a chronic, heterogenous inflammatory disease of unclear pathogenesis. JIA is hypothesized to be linked to a defective immune regulation. Anti-inflammatory cytokines belong to the best known regulatory factors. T-regulatory cells are a crucial cellular component of immune tolerance. One of their functions is synthesis of interleukin 10 (IL-10) and transforming growth factor beta1 (TGF-ß1).The aim of this study was to determine the proportion of T-regulatory cells (CD4+CD25highFOXP3+) in peripheral blood, and serum levels of TGF-ß1 and IL-10 in patients with JIA.Methods: The study included 25 patients with newly diagnosed JIA: oligoarthritis (n=17) and polyarthritis (n=8). Control group was comprised of 17 healthy children. CD4+CD25highFOXP3+ T cells in peripheral blood were quantified by means of three-color flow cytometry. Serum concentrations of TGF-ß1 and IL-10 were estimated with ELISA.Results: The proportion of peripheral CD4+CD25highFOXP3+cells in patients with JIA was significantly higher than in the controls (p=0.04). The two groups did not differ significantly in terms of their TGF-ß1 and IL-10 concentrations.Conclusions: At the time of the diagnosis, children with JIA present with elevated proportion of T-regulatory cells (CD4+CD25highFOXP3+) in peripheral blood. Anti-inflammatory cytokines, IL-10 and TGF-ß1, are not upregulated in the serum of patients with JIA, and therefore should not be considered as biomarkers of this condition.

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JAK Inhibition Reduces CD25 high CD27+ FOXp3+ T Regulatory Cells and Causes a Silencing Of T Effector Cells In Patients With Myeloproliferative Neoplasms Whilst Promoting a TH17 Phenotype

Blood ◽

10.1182/blood.v122.21.4092.4092 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4092-4092 ◽

Cited By ~ 6

Author(s):

Clodagh Keohane ◽

Shahram Y Kordasti ◽

Thomas Seidl ◽

Pilar Perez Abellan ◽

Nicholas Shaun B. Thomas ◽

...

Keyword(s):

T Cells ◽

Inflammatory Cytokines ◽

Peripheral Blood ◽

T Regulatory Cells ◽

Myeloproliferative Neoplasms ◽

Healthy Controls ◽

Regulatory Cells ◽

Effector Cells ◽

T Effector Cells

Abstract Background The myeloproliferative neoplasms (MPN), in particular myelofibrosis, are associated with elevated levels of inflammatory cytokines and constitutional symptoms. Treatment with JAK inhibitors (JAKi) have lead to marked improvement in symptoms and splenomegaly. Signaling through the JAK pathway is critical for T cell development and differentiation. However the baseline immune signature remains largely undescribed in MPN as does the effect of JAK inhibition on the immune subsets in this disease. Materials and Methods The % and absolute number of CD4+ T cell subsets (TH1, TH2 and TH17 and Foxp3+ T regulatory cells) in peripheral blood (PB) were investigated by flow cytometry. T cells were stimulated and stained intracellularly for IFNg, IL-4, IL-17 & TNFα. Tregs were defined as CD4+ CD25highCD27+FOXp3+. The serum level of 30 cytokines was also measured by Luminex. Patients received either ruxolitinib (n=21) or SAR302503 (n=13) as JAKi. Results We analysed 50 MPN patients (30 Myelofibrosis, 15 Polycythemia Vera, 5 Essential Thrombocythemia) and 14 healthy donors (HD). 34 patients were treated with JAKi and sequential PB samples were obtained at 1, 3, 6 and 12 month intervals (median follow up 6 months). Tregs are significantly lower in MPN patients compared to HD and drop further following treatment (p<0.0001 and p=0.0049 respectively). There was no difference at baseline in the T effector subsets between the groups including TH1, TH2 and TH17 secreting cells but there was a significant increase in TH17 following JAKi therapy (fig 1a). JAKi resulted in a significant decrease (p=0.03) in CD4 T cells secreting pro-inflammatory cytokines at 3 months follow up although this was less evident at 6 months follow up and occurred irrespective of disease response to treatment. This silencing was confirmed by both intracellular staining and luminex assay of supernatants including a significant decrease in Interleukin-2 receptor (IL-2r) p=0.0007, Interferon gamma induced protein (IP-10) p=0.0006, monokine induced by gamma interferon (MIG) p=0.0008 and hepatocyte growth factor (HGF) p=0.0009. This finding was reproduced in-vitro in healthy peripheral blood mononuclear cells (PBMCs). PBMCs were treated with the JAKi ruxolitinib (100-300NM) in the presence or absence of plate bound anti-CD3/28 stimulation and cultured for 5 days. Tregs were reduced in number and there was a considerable increase in the percentage of “cytokine negative” or “silent” T effector cells by FACS analysis compared to untreated or vehicle treated cells (median of 42 % of CD4 to 91% of CD4) (fig 1b). This finding was reproduced by Luminex cytokine assay of supernatants. Western blot demonstrated a reduction in pSTAT3 in ruxolitinib treated cells. To assess the effect of JAKi on Treg function, healthy isolated Tregs were treated with ruxolitinib and co cultured with CFSE labeled autologous T effector cells. Short term JAKi treated Tregs were unable to suppress the proliferation of T effector cells compared to Tregs treated with vehicle alone. Similarly, proliferation rate and function of Tregs was reduced following 4 weeks expansion in the presence of ruxolitinib compared to expanded Tregs in the presence of ATRA and rapamycin as a control. Conclusions Tregs are significantly lower in MPN patients compared to healthy controls in keeping with the inflammatory environment of MPN and decrease further with JAKi. Surprisingly, T effector numbers were not significantly different to healthy controls at baseline and TH1 and TH2 subsets did not change with therapy. However, secretion of proinflammatory cytokines from these cells was blocked with JAKi both invivo and invitro resulting in a functional silencing of T effectors. Interestingly, TH17 subsets increase with treatment possibly representing a polarization from a Treg phenotype to a TH17 phenotype, suggesting the re-establishment of immune-surveillance against the malignant clone. Further investigation is required to confirm the hypothesis that these expanded TH17 cells originate from Tregs or previously “silenced” CD4 T cells. Disclosures: Harrison: Novartis: Honoraria; Sanofi : Honoraria.

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СONCENTRATION OF ANTI-INFLAMATORY CYTOKINES IN CELL CULTURE SUPERNATANTS IN CHILDREN WITH JUVENILE IDIOPATHIC ARTHRITIS

Medical Immunology (Russia) ◽

10.15789/1563-0625-2019-4-725-736 ◽

2019 ◽

Vol 21 (4) ◽

pp. 725-736

Author(s):

I. M. Krivolapova ◽

I. A. Pashnina ◽

V. A. Chereshnev

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Inflammatory Cytokines ◽

Whole Blood ◽

Peripheral Blood ◽

Blood Cells ◽

Blood Cultures ◽

Juvenile Arthritis ◽

Peripheral Blood Cells ◽

Healthy Children ◽

Anti Inflammatory

Juvenile idiopathic arthritis is a chronic inflammatory disease of the joints in children, mainly of autoimmune or auto-inflammatory nature. It is a heterogeneous group, which includes different subtypes of the disease. Different mechanisms may play role in the pathogenesis of distinct subtypes of juvenile arthritis. However, a long-term imbalance of pro- and anti-inflammatory cytokines is important for all subtypes of disease. The aim of the present study was to determine spontaneous and stimulated anti-inflammatory cytokines production by peripheral blood cells from the children with juvenile idiopathic arthritis. Patients of 2 to 17 years old with different subtypes of juvenile idiopathic arthritis (n = 99) and healthy children without signs of autoimmune diseases (control, n = 31) were examined. Spontaneous and phytohemagglutinin-stimulated concentrations of IL-1ra, IL-4, IL-10, TGF-β in supernatants of whole-blood cultures were determined by ELISA. Differences in the spontaneous and mitogen-stimulated secretion of the cytokines in patients with different subtypes of juvenile arthritis have not been revealed. The spontaneous IL-1ra, IL-4 and IL-10 production by blood cells in the common group of patients with juvenile idiopathic arthritis was similar to the controls. The median value of spontaneous TGF-β concentration in the patients was below the detection level, whereas blood cells of healthy children had a higher potential of spontaneous TGF-β production. IL-4 and IL-10 production after incubation of peripheral blood cells with phytohemagglutinin in patients and in the control group did not differ from the controls, while IL-1ra and TGF-β synthesis was significantly lower than in healthy children.The spontaneous and/or stimulated production of IL-1ra, TGF-β by blood cells in children with juvenile idiopathic arthritis reflects the pathogenic significance of these cytokines in disease. Stimulation of cells can reveal a latent deficiency in the synthesis of cytokines, which is not evident when determining its concentration in serum or supernatants of spontaneous whole-blood cultures.

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T regulatory cells in childhood arthritis – novel insights

Expert Reviews in Molecular Medicine ◽

10.1017/erm.2013.14 ◽

2013 ◽

Vol 15 ◽

Cited By ~ 18

Author(s):

Anne M. Pesenacker ◽

Lucy R. Wedderburn

Keyword(s):

T Cells ◽

Juvenile Idiopathic Arthritis ◽

Regulatory T Cells ◽

T Regulatory Cells ◽

Current Knowledge ◽

Autoimmune Arthritis ◽

Regulatory Cells ◽

New Developments ◽

Childhood Arthritis ◽

Recent Developments

In recent years, there have been many new developments in the field of regulatory T cells (Treg), challenging the consensus on their behaviour, classification and role(s) in disease. The role Treg might play in autoimmune disease appears to be more complex than previously thought. Here, we discuss the current knowledge of regulatory T cells through animal and human research and illustrate the recent developments in childhood autoimmune arthritis (juvenile idiopathic arthritis (JIA)). Furthermore, this review summarises our understanding of the fields and assesses current and future implications for Treg in the treatment of JIA.

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Allergen-specific IL-10-secreting type I T regulatory cells, but not CD4+CD25+Foxp3+ T cells, are decreased in peripheral blood of patients with persistent allergic rhinitis

Clinical Immunology ◽

10.1016/j.clim.2010.03.006 ◽

2010 ◽

Vol 136 (2) ◽

pp. 292-301 ◽

Cited By ~ 37

Author(s):

Demin Han ◽

Chengshuo Wang ◽

Wei Lou ◽

Yili Gu ◽

Yang Wang ◽

...

Keyword(s):

T Cells ◽

Allergic Rhinitis ◽

Peripheral Blood ◽

T Regulatory Cells ◽

Type I ◽

Regulatory Cells ◽

Persistent Allergic Rhinitis

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Imbalance of Th17 and T-regulatory cells in peripheral blood and synovial fluid in treatment naïve children with juvenile idiopathic arthritis

Central European Journal of Immunology ◽

10.5114/ceji.2014.42128 ◽

2014 ◽

Vol 1 ◽

pp. 71-76 ◽

Cited By ~ 6

Author(s):

Joanna Szymańska-Kałuża ◽

Barbara Cebula-Obrzut ◽

Piotr Smolewski ◽

Jerzy Stanczyk ◽

Elżbieta Smolewska

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Peripheral Blood ◽

T Regulatory Cells ◽

Regulatory Cells ◽

Treatment Naïve

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Cytokines in the blood of patients with type 2 diabetes mellitus depending on the level of overweight/obesity (literature review and own data)

INTERNATIONAL JOURNAL OF ENDOCRINOLOGY ◽

10.22141/2224-0721.17.7.2021.244969 ◽

2021 ◽

Vol 17 (7) ◽

pp. 534-551

Author(s):

K.P. Zak ◽

V.V. Popova ◽

V.L. Orlenko ◽

O.V. Furmanova ◽

N.D. Tronko

Keyword(s):

Type 2 Diabetes ◽

Inflammatory Cytokines ◽

Proinflammatory Cytokines ◽

Peripheral Blood ◽

Control Group ◽

Obese Women ◽

Cytokine Network ◽

Tnf Α ◽

Anti Inflammatory

The paper analyzes the current literature data and the results of our own researches concerning the state of the cytokine network: pro- and anti-inflammatory cytokines (interleukin (IL) 1α, IL-1β, IL-4, IL-6, IL-10, IL-17 and tumor necrosis factor (TNF) α), α- and β-chemokines, including IL-8 and IL-16, as well as adipokines (leptin and adiponectin) in the peripheral blood of patients with type 2 diabetes (T2D) with normal and increased body weight/obesity. It has been shown that patients with T2D are characterized by an increased content of proinflammatory cytokines (IL-1, IL-6, IL-17, TNFα), α- and β-chemokines in the peripheral blood, including IL-8 and IL-16, as well as leptin with a decrease in adiponectin content. In lean patients (with body mass index (BMI) < 25.5 kg/m2) compared to lean normoglycemic individuals from the control group (BMI < 25.5 kg/m2), there is a small but significant increase in IL-1β, IL-6, IL-17, TNFα and leptin, which, as BMI increases, significantly increases in severe obesity (BMI > 30.0 kg/m2), especially in obese women (BMI > 35.0 kg/m2). Similarly, an increase in proinflammatory cytokines is observed in normoglycemic people, but not as significant as in T2D. Less clear data were obtained when during determination of the anti-inflammatory cytokines IL-4 and IL-10, which is explained by a significant polymorphism of their genes, and both protective and compensatory effects on pro-inflammatory cytokine rise. In T2D patients, especially those with obesity, there is an increase in the leptin level and a decrease in the adiponectin content. The severity of the course and the percentage of mortality are closely associated with the BMI of patients. The effectiveness of the fight against an increase in the incidence of T2D should be primarily aimed at preventing obesity, and in case of already developed T2D — at reducing concomitant obesity. The analysis of the data presented also suggests that a sharp increase in the content of pro-inflammatory cytokines (so called cytokine storm) observed in patients with T2D and obesity infected with COVID-19, is a consequence of the summation and potentiation of already existing inflammatory process.

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Changes In Tumor Infiltrating Lymphocytes Of Peripheral Blood And Tissue During Chemotherapy In Patients With Gastric Cancer

The American Journal of Medical Sciences and Pharmaceutical Research ◽

10.37547/tajmspr/volume03issue03-03 ◽

2021 ◽

Vol 03 (03) ◽

pp. 20-31

Author(s):

Gulnoz Golibovna Khakimova ◽

◽

Golib Abdulloevich Khakimov ◽

Shakhnoz Golibovna Khakimova ◽

Aziz Timurovich Khakimov ◽

...

Keyword(s):

Gastric Cancer ◽

T Cells ◽

Gastric Adenocarcinoma ◽

Peripheral Blood ◽

Tumor Tissue ◽

T Regulatory Cells ◽

Regulatory Cells ◽

Cytotoxic Potential ◽

Local Immunity ◽

Systemic Immunity

To study the state of systemic immunity and local immunity before and during chemotherapy in patients with gastric adenocarcinoma. From 2017 to 2018 at the Tashkent city branch of the Republican specialized scientific and practical medical center of oncology and radiology 20 primary metastatic patients with gastric adenocarcinoma received chemotherapy. The sampling of biological material (peripheral blood, tumor tissue) was carried out twice (before treatment and during the first control examination, after 3 courses). The percentage of the degree of infiltration of tumor tissue by lymphocytes (CD45+CD14-TILs) was estimated by flow cytometry; T cells (CD3+CD19-TILs); B cells (CD3-CD19+TILs); NK cells (CD3-CD16+CD56+TILs); CD16 and CD8 effector cells and their cytotoxic potential (CTP) (CD16+Perforin+TILs; CD16CTPTILs), (CD8+Perforin+TILs; CD8CTPTILs); regulatory T cells - NKT cells (CD3+CD16+CD56+TILs), CD4 (CD4+CD25+CD127-TILs) and CD8 (CD8+CD11b-CD28-TILs) regulatory cells and these parameters of systemic results. The factor of a favorable prognosis for PFS in patients with metastatic gastric cancer in the peripheral blood was an increase in the number of CD8 + T-regulatory cells (5.1% - 12.1%, p = 0.019), and in tumor tissue - an increase in the perforin potential of effector CD16 cells (0.5% - 4.9%, p = 0.030) and their cytotoxic potential (13.2% - 55.7%, p = 0.011). When assessing the changes in the indices of local immunity during chemotherapy, it was noted a negative effect of an increase of T cells (22.0% - -9.7%, p = 0.012), NKT - cells (207.9% - -13.8%, p = 0.002) and CD4 + T-regulatory cells (190.7% - -25.2%, p = 0.002). In contrast, an increase in the level of effector CD16 cells during chemotherapy increases the likelihood of surviving PFS - 9 months (-69.5% - 9.1%, p = 0.013). Indicators of local and systemic immunity serve as additional prognostic factors for gastric cancer.

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