FOXP3 expression in peripheral blood and synovial cells of patients with juvenile idiopathic arthritis: relationship with IL-17 at cytokine and molecular level

Abstract Regulatory T (Treg) cells are often found in human tumors; however, their functional characteristics have been difficult to evaluate due to low cell numbers and the inability to adequately distinguish between activated and Treg cell populations. Using a novel approach, we examined the intracellular cytokine production capacity of tumor-infiltrating T cells in the single-cell suspensions of enzymatically digested tumors to differentiate Treg cells from effector T cells. Similar to Treg cells in the peripheral blood of healthy individuals, tumor-infiltrating FOXP3+CD4 T cells, unlike FOXP3− T cells, were unable to produce IL-2 and IFN-γ upon ex vivo stimulation, indicating that FOXP3 expression is a valid biological marker for human Treg cells even in the tumor microenvironment. Accordingly, we enumerated FOXP3+CD4 Treg cells in intratumoral and peritumoral sections of metastatic melanoma tumors and found a significant increase in proportion of FOXP3+CD4 Treg cells in the intratumoral compared with peritumoral areas. Moreover, their frequencies were 3- to 5-fold higher in tumors than in peripheral blood from the same patients or healthy donors, respectively. These findings demonstrate that the tumor-infiltrating CD4 Treg cell population is accurately depicted by FOXP3 expression, they selectively accumulate in tumors, and their frequency in peripheral blood does not properly reflect tumor microenvironment.

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Th1 and Th17 Predominance in the Enthesitis-related Arthritis Form of Juvenile Idiopathic Arthritis

The Journal of Rheumatology ◽

10.3899/jrheum.081179 ◽

2009 ◽

Vol 36 (8) ◽

pp. 1730-1736 ◽

Cited By ~ 16

Author(s):

ANKIT MAHENDRA ◽

RAMNATH MISRA ◽

AMITA AGGARWAL

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Peripheral Blood ◽

Th17 Cells ◽

Transforming Growth Factor ◽

Health Assessment ◽

Treg Cells ◽

Th2 Cells ◽

Median Frequency ◽

Enthesitis Related Arthritis

Objective.A Th1 biased immune response in synovial fluid has been reported in children with polyarticular and extended oligoarticular-type juvenile idiopathic arthritis (JIA). We investigated T cell phenotypes including Th1, Th2, Th17, and Treg with emphasis on Th17 and Treg, in order to differentiate cytokines in the enthesitis-related arthritis (ERA) form of JIA.Methods.The frequencies of Th1, Th2, Th17, and Treg cells were determined by flow cytometry in peripheral blood (PB) and synovial fluid from patients with ERA and healthy subjects. Levels of interleukin 1ß (IL-1ß), IL-6, IL-21, IL-23, and transforming growth factor ß (TGF-ß), cytokines that influence Th17 lineage cells, were measured in paired plasma and synovial fluid (SF) samples by ELISA. Frequencies are expressed as percentages and cytokine levels as pg/ml.Results.There were no differences in blood samples in the frequency of Th1, Th2, Th17, and Treg cells between patients and controls. In paired samples, the median frequency of CD4+IFN-γ+ (20.49 vs 4.03; p < 0.005) and CD4+IL-17+ (2.27 vs 0.57; p < 0.01) cells was significantly higher in SF compared to PB, respectively; whereas the frequency of CD4+IL-4+ (1.79 vs 2.29; p < 0.04) cells was significantly reduced in the SF compared to PB. There was no difference in the frequency of regulatory T cells. Patients receiving methotrexate had fewer Th2 cells, whereas the Childhood Health Assessment Questionnaire score had a negative association with the frequency of Treg. Median levels of IL-1ß (p < 0.008), IL-6 (p < 0.0001), and IL-17 (p < 0.0001) were higher in SF than in plasma and levels of TGF-ß were lower (p < 0.001). Levels of IL-21 were similar in SF and plasma, whereas IL-23 was undetectable.Conclusion.In patients with ERA, peripheral blood Th1, Th2, Th17, and Treg cells were unchanged, but Th1 and Th17 cells were increased and Th2 cells were reduced in the SF compared to blood. Elevated IL-1ß and IL-6 in SF may be responsible for increased Th17 cells.

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The immunoregulatory function of peripheral blood CD71+ erythroid cells in systemic-onset juvenile idiopathic arthritis

Scientific Reports ◽

10.1038/s41598-021-93831-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hikaru Kanemasa ◽

Masataka Ishimura ◽

Katsuhide Eguchi ◽

Tamami Tanaka ◽

Etsuro Nanishi ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Inflammatory Diseases ◽

Active Phase ◽

Serum Levels ◽

Cell Interaction ◽

Erythroid Cells ◽

Peripheral Blood Mononuclear ◽

Systemic Onset

AbstractCD71+ erythroid cells (CECs) are recognized to have an immunoregulatory function via direct cell–cell interaction and soluble mediators. Circulating CECs appear in newborns or patients with hemolytic and cardiopulmonary disorders. To assess the biological role of CECs in systemic inflammation, we studied the gene expression and function in systemic-onset juvenile idiopathic arthritis (SoJIA). Peripheral blood mononuclear cells of SoJIA patients expressed upregulated erythropoiesis-related genes. It represented the largest expansion of CECs during active phase SoJIA among other inflammatory diseases. Despite the opposing roles of erythropoietin and hepcidin in erythropoiesis, both serum levels were in concert with the amounts of SoJIA-driven CECs. Circulating CECs counts in inflammatory diseases were positively correlated with the levels of C-reactive protein, IL-6, IL-18, or soluble TNF receptors. Co-culture with active SoJIA-driven CECs suppressed secretions of IL-1β, IL-6, and IL-8 from healthy donor monocytes. The top upregulated gene in SoJIA-driven CECs was ARG2 compared with CECs from cord blood controls, although cytokine production from monocytes was suppressed by co-culture, even with an arginase inhibitor. CECs are driven to the periphery during the acute phase of SoJIA at higher levels than other inflammatory diseases. Circulating CECs may control excessive inflammation via the immunoregulatory pathways, partly involving arginase-2.

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Identification of potential peripheral blood diagnostic biomarkers for patients with juvenile idiopathic arthritis by bioinformatics analysis

Rheumatology International ◽

10.1007/s00296-016-3607-z ◽

2016 ◽

Vol 37 (3) ◽

pp. 423-434 ◽

Cited By ~ 1

Author(s):

Zhi-qiang Tu ◽

Hai-yan Xue ◽

Wei Chen ◽

Lan-fang Cao ◽

Wei-qi Zhang

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Peripheral Blood ◽

Bioinformatics Analysis ◽

Diagnostic Biomarkers

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Long-term response of juvenile idiopathic arthritis after conditioning with 8 Gy total body irradiation followed by autologous peripheral blood stem cells: Case report

Pediatric Transplantation ◽

10.1111/j.1399-3046.2009.01129.x ◽

2010 ◽

Vol 14 (6) ◽

pp. E65-E69 ◽

Cited By ~ 1

Author(s):

Ann Woolfrey ◽

Jan Storek ◽

Suzanne Bowyer ◽

Robert Nelson ◽

Michael Robertson ◽

...

Keyword(s):

Stem Cells ◽

Juvenile Idiopathic Arthritis ◽

Case Report ◽

Total Body Irradiation ◽

Peripheral Blood ◽

Peripheral Blood Stem Cells ◽

Blood Stem Cells ◽

Total Body ◽

Term Response

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Calcineurin Free Protocol Is Associated With Increase in FOXP3 Expression in the Peripheral Blood of Recipients of Extended Criteria Donor Kidneys.

Transplantation Journal ◽

10.1097/00007890-201407151-00935 ◽

2014 ◽

Vol 98 ◽

pp. 297-298

Author(s):

M. Abbud-Filho ◽

C. Dias ◽

H. Caldas ◽

R. Nunes ◽

C. Mazeti ◽

...

Keyword(s):

Peripheral Blood ◽

Foxp3 Expression ◽

Extended Criteria Donor

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IL-2 Therapy Promotes the Expansion of Human CD4+CD25+ Regulatory T Cells and Selectively Upregulates the Expression of FOXP3 in T Cells In Vivo.

Blood ◽

10.1182/blood.v106.11.1257.1257 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1257-1257

Author(s):

Emmanuel Zorn ◽

Erik A. Nelson ◽

Mehrdad Mohseni ◽

Despina Litsa ◽

Haesook Kim ◽

...

Keyword(s):

Gene Expression ◽

T Cells ◽

Regulatory T Cells ◽

Nk Cells ◽

Quantitative Pcr ◽

Peripheral Blood ◽

Fold Increase ◽

Foxp3 Expression ◽

Prolonged Administration

Abstract Recombinant IL-2 has been used extensively in clinical trials to enhance a wide range of immune responses. Overall this strategy has had limited efficacy. Recent evidence suggests that IL-2 plays a key role in the generation and maintenance of CD4+CD25+ regulatory T cells (Treg) in vivo. In our study, we investigated the effect of prolonged administration of recombinant IL-2 on Treg in vivo. In a retrospective analysis, we first examined CD4+CD25+ Treg in blood samples collected from 21 cancer patients before and after they started continuous treatment with IL-2 at a dose of 2 X 105 U/m2/day for 3 months. Nine patients received IL-2 beginning 3 months after CD6 T cell depleted allogeneic bone marrow transplantation (BMT) for CML. The remaining 12 patients received IL-2 as treatment for advanced solid tumors. Overall toxicity was minimal and none of the transplant patients developed GVHD following IL-2 administration. Previous reports demonstrated that this prolonged treatment with low-dose IL-2 resulted in the expansion of CD56+CD3− NK cells in peripheral blood. Further analysis showed that 15 patients exhibited an expansion of Treg in peripheral blood 26 to 77 days after beginning IL-2 as demonstrated by an increase in the CD4+CD25+/CD3+ ratio (median fold increase 2.68; range 1.3 to 59). Three patients had no significant change and 3 patients demonstrated a decreased Treg/CD3 ratio. Using RNA from the same samples we also measured the expression of the Treg specific transcription factor FOXP3 by quantitative PCR. Nineteen of 21 patients showed a marked increase in FOXP3 expression following IL-2 treatment (8.38 median fold increase; range 1.4 to 41.5). Only 2 of 21 patients had lower FOXP3 expression after IL-2 administration. Since IL-2 treatment resulted in the expansion of NK cells as well as Treg, we purified CD56+CD3− NK cells and CD4+ T cells from patient samples collected post-IL-2 treatment, and measured FOXP3 gene expression in both subsets. In 4 analyzed cases, FOXP3 was selectively expressed in CD4+ T cells. Further analysis of purified Treg and NK cells incubated with IL-2 in vitro confirmed that FOXP3 expression was selectively induced in Treg, and also suggested that the in vivo increase in FOXP3 expression resulted from both Treg expansion and up-regulation of gene expression at the single cell level. To study the duration of the IL-2 effect, we analyzed additional samples collected 2 to 8 months after IL-2 treatment was completed. Nine of 10 patient samples tested showed a decrease in the CD4+CD25+/CD3+ ratio (1.39 median fold decrease; range 1.13 to 15.02). Using quantitative PCR, expression of FOXP3 decreased for 6 of 8 patients tested (10.76 median fold decrease; range 1.22 to 88.31). These results indicate that prolonged administration of IL-2 promotes the expansion of CD4+CD25+ Treg in vivo and also has a direct effect on FOXP3 expression. Although administration of IL-2 has previously been used to enhance T and NK cell responses, this study demonstrates that IL-2 therapy predominantly reinforces the regulatory component of the immune response, and may provide a means for controlling immune reactions in vivo.

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