Synovial fluid neutrophils in oligoarticular juvenile idiopathic arthritis have an altered phenotype and impaired effector functions

Abstract Background Neutrophils are the most prevalent immune cells in the synovial fluid in inflamed joints of children with oligoarticular juvenile idiopathic arthritis (JIA). Despite this, little is known about neutrophil function at the site of inflammation in JIA and how local neutrophils contribute to disease pathogenesis. This study aimed to characterize the phenotype and function of synovial fluid neutrophils in oligoarticular JIA. Methods Neutrophils obtained from paired blood and synovial fluid from patients with active oligoarticular JIA were investigated phenotypically (n = 17) and functionally (phagocytosis and oxidative burst, n = 13) by flow cytometry. In a subset of patients (n = 6), blood samples were also obtained during inactive disease at a follow-up visit. The presence of CD206-expressing neutrophils was investigated in synovial biopsies from four patients by immunofluorescence. Results Neutrophils in synovial fluid had an activated phenotype, characterized by increased CD66b and CD11b levels, and most neutrophils had a CD16hi CD62Llowaged phenotype. A large proportion of the synovial fluid neutrophils expressed CD206, a mannose receptor not commonly expressed by neutrophils but by monocytes, macrophages, and dendritic cells. CD206-expressing neutrophils were also found in synovial tissue biopsies. The synovial fluid neutrophil phenotype was not dependent on transmigration alone. Functionally, synovial fluid neutrophils had reduced phagocytic capacity and a trend towards impaired oxidative burst compared to blood neutrophils. In addition, the effector functions of the synovial fluid neutrophils correlated negatively with the proportion of CD206+ neutrophils. Conclusions Neutrophils in the inflamed joint in oligoarticular JIA were altered, both regarding phenotype and function. Neutrophils in the synovial fluid were activated, had an aged phenotype, had gained monocyte-like features, and had impaired phagocytic capacity. The impairment in phagocytosis and oxidative burst was associated with the phenotype shift. We speculate that these neutrophil alterations might play a role in the sustained joint inflammation seen in JIA.

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Synovial Fluid Neutrophils in Oligoarticular Juvenile Idiopathic Arthritis Have an Altered Phenotype and Impaired Effector Functions

10.21203/rs.3.rs-200749/v1 ◽

2021 ◽

Author(s):

Sabine Arve-Butler ◽

Tobias Schmidt ◽

Anki Mossberg ◽

Elisabet Berthold ◽

Birgitta Gullstrand ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Oxidative Burst ◽

Joint Inflammation ◽

Mannose Receptor ◽

Inactive Disease ◽

Phagocytic Capacity ◽

Effector Functions ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

And Function

Abstract BackgroundNeutrophils are the most prevalent immune cells in synovial fluid in inflamed joints of children with oligoarticular juvenile idiopathic arthritis (JIA). Despite this, little is known about neutrophil function at the site of inflammation in JIA and how local neutrophils contribute to disease pathogenesis. This study aimed to characterize phenotype and function of synovial fluid neutrophils in oligoarticular JIA.Methods Neutrophils obtained from paired blood and synovial fluid from patients with active oligoarticular JIA were investigated phenotypically (n=17) and functionally (phagocytosis and oxidative burst, n=13) by flow cytometry. In a subset of patients (n=6), blood samples were also obtained during inactive disease at a follow-up visit. Presence of CD206-expressing neutrophils was investigated in synovial biopsies from four patients by immunofluorescence. ResultsNeutrophils in synovial fluid had an activated phenotype, characterized by increased CD66b and CD11b levels, and most neutrophils had a CD16hi CD62Llow aged phenotype. A large proportion of the synovial fluid neutrophils expressed CD206, a mannose receptor not commonly expressed by neutrophils but by monocytes, macrophages and dendritic cells. CD206 expressing neutrophils were also found in synovial tissue biopsies. The synovial fluid neutrophil phenotype was not dependent on transmigration alone. Functionally, synovial fluid neutrophils had reduced phagocytic capacity and a trend towards impaired oxidative burst compared to blood neutrophils. In addition, the effector functions of the synovial fluid neutrophils correlated negatively with the proportion of CD206+ neutrophils.ConclusionsNeutrophils in the inflamed joint in oligoarticular JIA were altered, both regarding phenotype and function. Neutrophils in the synovial fluid were activated, had an aged phenotype, had gained monocyte-like features and had impaired phagocytic capacity. The impairment in phagocytosis and oxidative burst was associated to the phenotype shift. We speculate that these neutrophil alterations might play a role in the sustained joint inflammation seen in JIA.

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Children with oligoarticular Juvenile Idiopathic Arthritis have skewed synovial monocyte polarization pattern with functional impairment – a distinct inflammatory pattern for oligoarticular juvenile arthritis

10.21203/rs.3.rs-29879/v2 ◽

2020 ◽

Author(s):

Tobias Schmidt ◽

Elisabet Berthold ◽

Sabine Arve-Butler ◽

Birgitta Gullstrand ◽

Anki Mossberg ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Mrna Expression ◽

Mrna Level ◽

Western World ◽

Polarization Pattern ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

Monocytes And Macrophages ◽

Anti Inflammatory

Abstract Background Juvenile idiopathic arthritis (JIA) is an umbrella term of inflammatory joint diseases in children. Oligoarthritis is the most common form in the Western world, representing roughly 60% of all patients. Monocytes and macrophages play an important role in adult arthritides, but their role in oligoarticular JIA is less studied. Polarization highly influences monocytes’ and macrophages’ effector functions, broadly separated into pro-inflammatory M1 or anti-inflammatory M2 phenotypes. Here, we set out to investigate the polarization pattern and functional aspects of synovial monocytes in oligoarticular juvenile idiopathic arthritis (JIA). Methods Paired synovial fluid, blood samples (n=13) and synovial biopsies (n=3) were collected from patients with untreated oligoarticular JIA. Monocytes were analyzed for polarization markers by flow cytometry and qPCR. Effector function was analyzed by a phagocytosis assay. Polarization of healthy monocytes was investigated by stimulation with synovial fluid in vitro . Monocyte/macrophage distribution, polarization and mRNA expression were investigated in biopsies by immunohistochemistry, immunofluorescence and in situ hybridization. Results Children with oligoarticular JIA have polarized synovial fluid monocytes of a specific M1(IFNγ)/M2(IL-4)-like pattern. This was evidenced by increased surface expression of CD40 (p<0.001), CD86 (p<0.001) and CD206 (p<0.001), but not CD163, as compared to paired circulating monocytes. Additionally, polarization was extensively explored at the mRNA level and synovial fluid monocytes differentially expressed classical markers of M1(IFNγ)/M2(IL-4) polarization compared to circulating monocytes. Synovial fluid monocytes were functionally affected, as assessed by reduced capacity to phagocytose (p<0.01). Synovial fluid induced M2 markers (CD16 and CD206), but not M1 (CD40) or CD86 in healthy monocytes and did not induce cytokine production. Single and co-expression of surface CD40 and CD206, as well as mRNA expression of IL-10 and TNF, was observed in monocytes/macrophages in synovial biopsies. Conclusion Children with untreated oligoarticular JIA have similar and distinct synovial fluid monocyte polarization pattern of mixed pro- and anti-inflammatory features. This pattern was not exclusively a result of the synovial fluid milieu as monocytes/macrophages in the synovial membrane show similar patterns. Our study highlights a distinct polarization pattern in oligoarticular JIA, which could be utilized for future treatment strategies.

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Concentration Of Survivin In Children With Oligo- And Poly-Articular Juvenile Idiopathic Arthritis (JIA): Diagnostic And Prognostic Value – Single Center Study.

10.21203/rs.3.rs-26777/v3 ◽

2021 ◽

Author(s):

Joanna Lipinska ◽

Marcin Kaszkowiak ◽

Beata Malachowska ◽

Joanna Swidrowska-Jaros ◽

Elzbieta Smolewska

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Disease Activity ◽

Joint Inflammation ◽

Elisa Test ◽

Control Group ◽

Healthy Children ◽

Tnf Α ◽

Prognostic Utility ◽

Radiological Damage

Abstract AIM: The goal of the study was to assess the diagnostic and prognostic utility of survivin in patients with Juvenile Idiopathic Arthritis (JIA).METHODS: Seventy children with JIA – 59 newly diagnosed and 11 biologically treated (46 girls and 17 boys) aged 1.5-18 years and 29 healthy children as a control group, appropriately matched in terms of sex and age, were included into the study. The disease activity was established on the basis of JADAS-27 criteria. Concentration of survivin was assessed by an ELISA test in serum and also 18 matched synovial fluid samples collected from patients with JIA.RESULTS: Children with JIA were divided according to the subtype of the JIA. In 65.7% of patients oligoarthritis was diagnosed. The largest group comprised children of low disease activity (62.9%) according to JADAS-27. The serum concentration of survivin was significantly higher in children with JIA compared to the controls (p<0.001). Concentration of survivin was higher among patients positive for anti-cyclic citrullinated peptide autoantibodies (ACPA) (p=0.001). In all synovial fluid samples the concentration of survivin was higher than in matched serum (p=0.003). Serum survivin concentration was not significantly associated with radiological damage status or active synovitis assessed by joint ultrasonography. Survivin level was not significantly associated with disease duration time or treatment with TNF-α inhibitors in DMARD’s non-responders. CONCLUSIONS: Survivin should be considered as a biomarker of joint inflammation helpful in the diagnosis of oligo- and polyatricular JIA and probably not dependent of treatment with TNF-α inhibitors.

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Pathogenesis of juvenile idiopathic arthritis

Oxford Textbook of Rheumatology ◽

10.1093/med/9780199642489.003.0060_update_002 ◽

2013 ◽

pp. 437-443

Author(s):

David Bending ◽

Kiran Nistala ◽

Lucy R. Wedderburn ◽

Elizabeth C. Rosser

Keyword(s):

Juvenile Idiopathic Arthritis ◽

T Regulatory Cells ◽

Joint Inflammation ◽

Disease Pathogenesis ◽

Activated T Cells ◽

Effector Cells ◽

T Effector Cells ◽

Key Factor ◽

Autoimmune Conditions ◽

And Function

Although the term juvenile idiopathic arthritis (JIA) encompasses a heterogeneous group of diseases, they all share a common pathological hallmark: inflammation of the synovium. Highly activated T cells, monocytes, and neutrophils are attracted to the joint and secrete mediators that not only perpetuate inflammation, but also may attenuate immune regulation. In the oligoarticular and polyarticular forms of JIA, which are thought to be autoimmune conditions, dysregulated adaptive immunity is a likely factor in disease pathogenesis; the nature of the interactions between T-effector cells (Teffs) and T-regulatory cells (Tregs) is probably a key factor in controlling disease progression. Factors that affect the frequency and function of Tregs and/or the sensitivity of Teffs to mechanisms of immune suppression will therefore impact the disease course. In the systemic form of JIA, however, dysregulation of innate immune pathways appears more central to disease pathogenesis, resulting in augmented levels of interleukins IL-1β‎‎, IL-6, and IL-18. In the end, a final common pathological pathway in JIA is the activation of monocytes and neutrophils which are the principal mediators of joint inflammation and damage. This is supported by the fact that the therapies that have targeted innate cytokine pathways have shown greater success in the treatment of JIA.

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Children with oligoarticular Juvenile Idiopathic Arthritis have skewed synovial monocyte polarization pattern with functional impairment – a distinct inflammatory pattern for oligoarticular juvenile arthritis

10.21203/rs.3.rs-29879/v1 ◽

2020 ◽

Author(s):

Tobias Schmidt ◽

Elisabet Berthold ◽

Sabine Arve-Butler ◽

Birgitta Gullstrand ◽

Anki Mossberg ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Mrna Expression ◽

Mrna Level ◽

Western World ◽

Polarization Pattern ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

Monocytes And Macrophages ◽

Anti Inflammatory

Abstract Background Juvenile idiopathic arthritis (JIA) is an umbrella term of inflammatory joint diseases in children. Oligoarthritis is the most common form in the Western world, representing roughly 60% of all patients. Monocytes and macrophages play an important role in adult arthritides, but their role in oligoarticular JIA is less studied. Polarization highly influences monocytes’ and macrophages’ effector functions, broadly separated into pro-inflammatory M1 or anti-inflammatory M2 phenotypes. Here, we set out to investigate the polarization pattern and functional aspects of synovial monocytes in oligoarticular juvenile idiopathic arthritis (JIA). Methods Paired synovial fluid, blood samples (n = 13) and synovial biopsies (n = 3) were collected from patients with untreated oligoarticular JIA. Monocytes were analyzed for polarization markers by flow cytometry and qPCR. Effector function was analyzed by a phagocytosis assay. Polarization of healthy monocytes was investigated by stimulation with synovial fluid in vitro. Monocyte/macrophage distribution, polarization and mRNA expression were investigated in biopsies by immunohistochemistry, immunofluorescence and in situ hybridization. Results Children with oligoarticular JIA have polarized synovial fluid monocytes of a specific M1(IFNg)/M2(IL-4)-like pattern. This was evidenced by increased surface expression of CD40 (p < 0.001), CD86 (p < 0.001) and CD206 (p < 0.001), but not CD163, as compared to paired circulating monocytes, and extensively explored at the mRNA level were synovial fluid monocytes differentially expressed classical markers of M1(IFNg)/M2(IL-4) polarization. Synovial fluid monocytes were functionally affected, as assessed by reduced capacity to phagocytose (p < 0.01). Synovial fluid induced M2 (CD16 and CD206), but not M1 (CD40 and CD86) markers in healthy monocytes and did not induce cytokine production. Single and co-expression of surface CD40 and CD206, as well as mRNA expression of IL-10 and TNF, was observed in monocytes/macrophages in synovial biopsies. Conclusion Children with untreated oligoarticular JIA have similar and distinct synovial fluid monocyte polarization pattern of mixed pro- and anti-inflammatory features. This pattern was not exclusively a result of the synovial fluid milieu as monocytes/macrophages in the synovial membrane show similar patterns. Our study highlights a distinct polarization pattern in oligoarticular JIA, which could be utilized for future treatment strategies.

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Pathogenesis of juvenile idiopathic arthritis

10.1093/med/9780199642489.003.0060 ◽

2013 ◽

Author(s):

David Bending ◽

Kiran Nistala ◽

Lucy R. Wedderburn

Keyword(s):

Juvenile Idiopathic Arthritis ◽

T Regulatory Cells ◽

Joint Inflammation ◽

Innate Immune ◽

Disease Pathogenesis ◽

Activated T Cells ◽

Key Factor ◽

Autoimmune Conditions ◽

And Function ◽

Systemic Form

Although the term juvenile idiopathic arthritis (JIA) encompasses a heterogeneous group of diseases, they all share a common pathological hallmark: inflammation of the synovium. Highly activated T cells, monocytes, and neutrophils are attracted to the joint and secrete mediators that not only perpetuate inflammation but also may attenuate immune regulation. In the oligoarticular and polyarticular forms of JIA, which are thought to be autoimmune conditions, dysregulated adaptive immunity is a likely factor in disease pathogenesis; the nature of the interactions between T effector (Teff) cells and T regulatory cells (Treg) is probably a key factor in controlling disease progression. Factors that affect the frequency and function of Tregs and/or the sensitivity of Teffs to mechanisms of immune suppression will therefore impact on the disease course. In the systemic form of JIA, however, dysregulation of innate immune pathways appears more central to disease pathogenesis resulting in augmented levels of interleukins IL-1β‎, IL-6, and IL-18. In the end, a final, common pathological pathway in JIA is the activation of monocytes and neutrophils, which are the principal mediators of joint inflammation and damage. This is supported by the fact that the therapies that have targeted innate cytokine pathways have shown greater success in the treatment of JIA.

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Biologic predictors of extension of oligoarticular juvenile idiopathic arthritis as determined from synovial fluid cellular composition and gene expression

Arthritis & Rheumatism ◽

10.1002/art.27284 ◽

2010 ◽

Vol 62 (3) ◽

pp. 896-907 ◽

Cited By ~ 44

Author(s):

Patricia J. Hunter ◽

Kiran Nistala ◽

Nipurna Jina ◽

Ayad Eddaoudi ◽

Wendy Thomson ◽

...

Keyword(s):

Gene Expression ◽

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Cellular Composition ◽

Oligoarticular Juvenile Idiopathic Arthritis

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Concentration of survivin in children with oligo- and polyarticular juvenile idiopathic arthritis (JIA): diagnostic and prognostic value—a single-center study

Arthritis Research & Therapy ◽

10.1186/s13075-021-02424-y ◽

2021 ◽

Vol 23 (1) ◽

Author(s):

Joanna Lipinska ◽

Marcin Kaszkowiak ◽

Beata Malachowska ◽

Joanna Swidrowska-Jaros ◽

Elzbieta Smolewska

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Synovial Fluid ◽

Disease Activity ◽

Joint Inflammation ◽

Elisa Test ◽

Control Group ◽

Healthy Children ◽

Tnf Α ◽

Prognostic Utility ◽

Radiological Damage

Abstract Aim The goal of the study was to assess the diagnostic and prognostic utility of survivin in patients with juvenile idiopathic arthritis (JIA). Methods Seventy children with JIA—59 newly diagnosed and 11 biologically treated (46 girls and 17 boys) aged 1.5–18 years and 29 healthy children as a control group, appropriately matched in terms of sex and age, were included in the study. The disease activity was established on the basis of the JADAS-27 criteria. The concentration of survivin was assessed by an ELISA test in serum and also 18 matched synovial fluid samples collected from patients with JIA. Results Children with JIA were divided according to the subtype of the JIA. In 65.7% of patients, oligoarthritis was diagnosed. The largest group comprised children of low disease activity (62.9%) according to JADAS-27. The serum concentration of survivin was significantly higher in children with JIA compared to the controls (p < 0.001). The concentration of survivin was higher among patients positive for anti-cyclic citrullinated peptide autoantibodies (ACPA) (p = 0.001). In all synovial fluid samples, the concentration of survivin was higher than in matched serum (p = 0.003). Serum survivin concentration was not significantly associated with radiological damage status or active synovitis assessed by joint ultrasonography. Survivin level was not significantly associated with disease duration time or treatment with TNF-α inhibitors in DMARD’s non-responders. Conclusions Survivin should be considered as a biomarker of joint inflammation helpful in the diagnosis of oligo- and polyarticular JIA and probably not dependent on treatment with TNF-α inhibitors.

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