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

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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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/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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AB0166 Sex-Specific Gene Expression Differences in Oligoarticular Juvenile Idiopathic Arthritis

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2016-eular.3049 ◽

2016 ◽

Vol 75 (Suppl 2) ◽

pp. 953.1-953

Author(s):

R.C. Chiaroni-Clarke ◽

R.A. Chavez ◽

J.E. Munro ◽

R.C. Allen ◽

J.D. Akikusa ◽

...

Keyword(s):

Gene Expression ◽

Juvenile Idiopathic Arthritis ◽

Specific Gene ◽

Specific Gene Expression ◽

Oligoarticular Juvenile Idiopathic Arthritis

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Th1 polarization defines the synovial fluid T cell compartment in oligoarticular juvenile idiopathic arthritis

JCI Insight ◽

10.1172/jci.insight.149185 ◽

2021 ◽

Author(s):

Amélie M. Julé ◽

Kacie J. Hoyt ◽

Kevin Wei ◽

Maria Gutierrez-Arcelus ◽

Maria L. Taylor ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

T Cell ◽

Synovial Fluid ◽

Cell Compartment ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

Th1 Polarization

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

Arthritis Research & Therapy ◽

10.1186/s13075-021-02483-1 ◽

2021 ◽

Vol 23 (1) ◽

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 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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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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Outcome of children with oligoarticular juvenile idiopathic arthritis compared to polyarthritis on methotrexate- data of the German BIKER registry

Pediatric Rheumatology ◽

10.1186/s12969-021-00522-4 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

A. Raab ◽

T. Kallinich ◽

D. Huscher ◽

I. Foeldvari ◽

F. Weller-Heinemann ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Disease Activity ◽

Chronic Arthritis ◽

Primary Objective ◽

Minimal Disease Activity ◽

Serious Adverse Events ◽

Disease Modifying Drugs ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

Minimal Disease ◽

Evidence Based Guidelines

Abstract Background Oligoarticular juvenile idiopathic arthritis (oligoJIA) is the most commonly diagnosed category of chronic arthritis in children. Nevertheless, there are no evidence- based guidelines for its treatment, in particular for the use of methotrexate (MTX). The primary objective of this analysis is to evaluate the outcomes in patients with persistent oligoJIA compared to those with extended oligoJIA and rheumatoid factor (RF) negative polyarthritis treated with methotrexate. Methods Patients with persistent or extended oligoJIA or RF negative PA recorded in the Biologics in Pediatric Rheumatology Registry (BiKeR), receiving methotrexate for the first time were included in the analyses. Efficacy was determined using the Juvenile Arthritis Disease Activity Score 10 (JADAS 10). Safety assessment included the documentation of adverse and serious adverse events. Results From 2005 through 2011, 1056 patients were included: 370 patients with persistent oligoJIA, 221 patients with extended oligoJIA and 467 patients with RF negative PA. Therapeutic efficacy was observed following the start of methotrexate. Over a period of 24 months JADAS-minimal disease activity (JADAS ≤2) was reached in 44% of patients with persistent oligoJIA, 38% with extended oligoJIA, 46% with RF negative PA, JADAS-remission defined as JADAS ≤1 was reached in 33% of patients with persistent oligoJIA, 29% with extended oligoJIA and 35% (RF negative PA). Patients with extended oligoJIA achieved JADAS remission significantly later and received additional biologic disease-modifying drugs significantly more often than patients with persistent oligoJIA or RF negative PA (p < 0.001). Tolerability was comparable. New onset uveitis occurred in 0.3 to 2.2 per 100 patient years. Conclusions Patients with persistent oligoJIA taking methotrexate are at least as likely to enter remission as patients with extended oligo JIA or polyarticular JIA. Patients with extended oligoJIA achieved JADAS remission significantly later. Within 2 years, almost half of the patients with persistent oligoJIA achieved JADAS-minimal disease activity.

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POS0578 CORRELATION BETWEEN EXPRESSION LEVELS OF MIR-155 AND MIR-223 IN SYNOVIAL FLUID AND ULTRASOUND SCORES FOR ACTIVE SYNOVITIS IN RHEUMATOID ARTHRITIS PATIENTS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.3828 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 522.3-523

Author(s):

R. Shumnalieva ◽

D. Kachakova ◽

R. Kaneva ◽

Z. Kolarov ◽

S. Monov

Keyword(s):

Rheumatoid Arthritis ◽

Gene Expression ◽

Clinical Practice ◽

Synovial Fluid ◽

Disease Activity ◽

Power Doppler ◽

Joint Inflammation ◽

Expression Levels ◽

Ultrasound Features ◽

Local Expression

Background:MicroRNAs (miRNAs) are a class of small, non-coding RNAs that negatively regulate gene expression at posttranscriptional level. In rheumatoid arthritis studies have shown that miRNA are differentially expressed systemically as well as locally in the inflamed joints [1,2]. The correlation between their systemic or local expression levels and scores for disease activity and progression in RA make them possible candidate for biomarkers in the clinical practice.Objectives:To analyze the expression levels of miR-155 and miR-223 in synovial fluid (SF) from RA patients in regard to the ultrasound scores for disease activity.Methods:A total number of 48 RA patients according to the 1987 ACR criteria were included in the study. Expression levels of miR-155 and miR-223 SF were determined by qPCR (SybrGreen technology) and compared to healthy controls (HCs). Relative changes of gene expression levels of the studied miRNAs were calculated by 2-ΔΔCt method. Musculoskeletal ultrasound (MSUS) examination was performed by two independent examiners on ESAOTE, MyLab60 using both grey scale and power Doppler technic. A semi-quantitative assessment of the peripheral joints was performed for detecting joint inflammation and determining the grade of synovial thickening and the degree of vascularization. Ultrasound features for active disease were correlated to the local expression of the studied miRNAs. SPSS was used for statistical analysis.Results:RA SF showed overexpression of miR-155 (in 79.17%, p=1.63x10-4) and of miR-223 (in 79.17%, p=1.64x10-3) when compared to HCs and both miRNAs could be used to differentiate RA patients from HCs (р=8.0х10-5 and р=2.8х10-4, respectively). When we analyzed the correlation between the diagnosis, the expression of miRNAs and the changes on the musculoskeletal ultrasound examination we found a statistically significant correlation between the presence of synovitis and the degree of the power Doppler signal on MSUS and the local expression of miR-223 (p=6.19 x 10-4 and p=0.003, respectively). SF levels of miR-223 correlated also with the degree of synovial hypertrophy on MSUS (p=0.013). The results for miRNA-155 were not statistically significant.Conclusion:The correlation between the local expression of miR-223 and the ultrasound features of active joint inflammation shows that this miRNA might be a better candidate for local disease biomarker than miR-155. Further analysis with larger sets is needed to confirm if altered local miRNA expression could be used in the clinical practice as biomarker for disease activity especially in cases with subclinical synovitis.References:[1]Filková M, Aradi B, Šenolt L, et al. Association of circulating miR-223 and miR-16 with disease activity in patients with early rheumatoid arthritis. ARD, 2014; 73: 1898-1904.[2]Kriegsmann, M., Randau, T.M., Gravius, S. et al. Expression of miR-146a, miR-155, and miR-223 in formalin-fixed paraffin-embedded synovial tissues of patients with rheumatoid arthritis and osteoarthritis. Virchows Arch, 2016; 469, 93–100.Acknowledgements:The study was supported by Grant 14-D/2012 and Grant 60/2013 funded by Medical University-Sofia.Disclosure of Interests:Russka Shumnalieva: None declared, Darina Kachakova: None declared, Radka Kaneva: None declared, Zlatimir Kolarov Speakers bureau: Amgen, Pfizer, Novartis, Abbvie, Roche, Astra-Zeneka, Simeon Monov Speakers bureau: Amgen, Pfizer, Novartis, Abbvie, Roche, Astra-Zeneka

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P080 Acroosteolysis: a rare complication or a co-occurrence with Juvenile Idiopathic Arthritis?

Rheumatology ◽

10.1093/rheumatology/keab722.072 ◽

2021 ◽

Vol 60 (Supplement_5) ◽

Author(s):

Makhlouf Yasmine ◽

Miladi Saoussen ◽

Fazaa Alia ◽

Sellami Mariem ◽

Souabni Leila ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Psoriatic Arthritis ◽

Rare Complication ◽

Plain Radiograph ◽

Limited Range ◽

Acute Phase Reactants ◽

Limited Range Of Motion ◽

Macular Rash ◽

Oligoarticular Juvenile Idiopathic Arthritis ◽

Dysmorphic Syndrome

Abstract Background Acroosteolysis refers to a destructive process involving the distal phalanges of the fingers and toes that may extend to metacarpals or metatarsals. Rarely idiopathic, the diagnosis of primary acroosteolysis requires ruling out other causes. Juvenile idiopathic arthritis is an exceptional aetiology of acroosteolysis occurring mainly in psoriatic arthritis. Here by a case of juvenile idiopathic arthritis associated with acroosteolysis of the toes. Methods A 13-year-old girl with no past medical history, presented to our department of rheumatology with oligoarthritis affecting both wrists and knees. She had no familiar history of psoriasis nor rheumatic diseases. She described a dull ache and recurring swelling of knees evolving for >6 years associated with a macular rash of the chest without fever. On examination, the knees were swollen with a limited range of motion of < 90°. Examination of the spine and sacroiliac joints was unremarkable. There was no deformity, no dysmorphic syndrome nor ligamentous hyper laxity. The mucocutaneous examination was normal. Similarly, there was no hepatosplenomegaly or swollen lymph nodes. Laboratory investigations showed high acute phase reactants and normal blood count. Rheumatoid factor, anti-cyclic citrullinated peptide antibodies and antinuclear antibodies were also negative. Besides, she was negative for HLAB-27. Ophthalmic examination did not show any sequelae of uveitis. Results Plain radiograph of the feet revealed bone resorption of the second and fifth distal phalanges without signs of reconstruction. Other secondary causes of acroosteolysis were ruled out. The diagnosis of oligoarticular juvenile idiopathic arthritis was made. In view of the involvement of the distal phalanges, the phenotype of psoriatic arthritis was probable. The patient was initially treated with non-steroidal anti-inflammatory drugs as well as intraarticular injections of corticosteroids in knees. As the flares persisted, she was put on Methotrexate at a dosage of 15 mg per week with marked clinical improvement. Conclusion Our case illustrates a possible occurrence of acroosteolysis of the feet in the field of an active juvenile idiopathic arthritis. It is important to rule out other causes and make a rapid diagnosis in order to ensure appropriate management decisions.

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