Characterization of Breg Cells

Study Overview
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Brief Summary
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Recently, it has been shown that B cells could also have regulatory functions through the secretion of interleukin 10 (IL-10). They are called the B regulatory cells (Breg). In the mouse model the most commonly used of rheumatoid arthritis, collagen-induced arthritis (CIA), the transfer Breg helps prevent the development of CIA and cure established arthritis. The investigators have recently shown that Breg were decreased in patients with RA compared to controls and that the rate of Breg was inversely correlated with disease activity and autoantibody. These results thus suggest that the lack of IL-10 secretion by B cells plays an important role in the pathophysiology of RA. Nevertheless, in humans, the Breg remain poorly understood. The main objective of this project is to better characterize the B capable of producing IL-10 both in subjects with RA and controls. Understanding which induces the secretion of IL-10 by B could allow to consider new therapeutic approaches in autoimmune diseases, including in RA. The investigators therefore aim to identify nutrient transporters, chemokine receptors, genes and surface proteins differentially expressed between Breg and other B cells in patients with RA and in controls.

Detailed Description
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Rational: Rheumatoid arthritis (RA), the most common inflammatory joint disease, is often associated with irreversible joint destruction and can involve the prognosis of patients. If treatments to stabilize the disease are now available, research continues to try to permanently cure the disease. It is well established that the B cells have a pathogenic role in RA. More recently, it has been shown that B cells could also have regulatory functions through the secretion of interleukin 10 (IL-10). They are called the B regulatory cells (Breg). In the mouse model the most commonly used of rheumatoid arthritis, collagen-induced arthritis (CIA), the transfer Breg helps prevent the development of CIA and cure established arthritis. The investigators have recently shown that Breg were decreased in patients with RA compared to controls and that the rate of Breg was inversely correlated with disease activity and autoantibody levels. These results thus suggest that the lack of IL-10 secretion by B cells plays an important role in the pathophysiology of RA. Nevertheless, in humans, the Breg remain poorly understood. The project's main objective is to better characterize the B capable of producing IL-10 both in subjects with RA and controls. Understanding which induces the secretion of IL-10 by B could allow to consider new therapeutic approaches in autoimmune diseases, including in RA. Objectives: Principal: To identify nutrient transporters and chemokine receptors differentially expressed between Breg and other B cells in patients with RA. Secondary: To identify genes and surface proteins differentially expressed between Breg and other B Lymphocytes (BL) in patients with RA. To identify nutrient transporters, chemokine receptors, genes and surface proteins differentially expressed between Breg and other BL in healthy subjects. To compare the expression of nutrient transporters, chemokine receptors, genes and surface proteins between Breg Breg controls and subjects with RA. Methods: Design: Cross-sectional study involving bicentric rheumatology services in Montpellier and Nîmes to recruitment; our research team at the Translational IGMM Nîmes and immunology laboratory for biological analyzes. Population: RA: patient meets the criteria ACR (American College of Rheumatology) -EULAR (European League Against Rheumatism) 2010, naïve and biotherapy with corticosteroids less than 10 mg / day, stable for at least a week. Controls: matched for age and sex to RA patients, with no systemic disease. Endpoints Main: the average flow cytometry fluorescence intensity nutrient carriers and the percentage of BL expressing the different chemokine receptors Secondary: transcriptome analysis and proteomics surface with cytometric confirmation of protein expression of RNA and proteins identified. Number of subjects: 50 controls and 50 RA patients (10 each for each of the 3 methods of comparison Breg / B IL10- and 10 each for each of the two stages of validation). Each patient will have a visit. The expected study duration is 2 years. Statistical analysis: Comparing ratios B + IL-10 / IL-10-B between RA patients and controls by Student or Mann-Whitney tests. Expected Results and Prospects: This project will allow us to better define and understand the Breg in patients with RA and in controls. If the investigators can find specific extracellular markers for Breg, this will simplify the further study of these cells. Understanding allowing BL becoming regulator and this explains the lack of IL-10 by the BL in RA could open new therapeutic perspectives.

Official Title
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Characterization of B Regulatory (Breg) Cells in Healthy Subjects and in Rheumatoid Arthritis (RA)

Conditions
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Healthy Volunteers, Arthritis, Rheumatoid

Intervention / Treatment
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* Other: blood sampling


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: RA responding to ACR/EULAR 2010 criteria Exclusion Criteria: steroid> 10 mg/d previous use of biological disease-modifying antirheumatic drug (DMARD) age<18 years

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Other: Nutrient transporters study<br>Blood sampling from 10 patients vith RA and 10 control patient for biological analysis and measure of nutrient transporters expression | Other: blood sampling<br>* Blood sample retrieval for biological and genetic analysis and comparison<br>|
| Other: Chemokine receptors study<br>Blood sampling from 10 patients vith RA and 10 control patient for biological analysis and measure of chemokine receptors expression | Other: blood sampling<br>* Blood sample retrieval for biological and genetic analysis and comparison<br>|
| Other: Genes study<br>Blood sampling from 10 patients vith RA and 10 control patient for biological analysis and measure of genes expression | Other: blood sampling<br>* Blood sample retrieval for biological and genetic analysis and comparison<br>|
| Other: Protein surface study<br>Blood sampling from 10 patients vith RA and 10 control patient for biological analysis and measure of protein surface expression | Other: blood sampling<br>* Blood sample retrieval for biological and genetic analysis and comparison<br>|
| Other: Protein surface & genes data validation<br>Blood sampling from 10 patients vith RA and 10 control patient for biological analysis and measure for validation of the data from protein surface study and genes study arms analysis. | Other: blood sampling<br>* Blood sample retrieval for biological and genetic analysis and comparison<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Identification of nutrient transporters' and of chemokine receptors' differentially expressed between Breg and other B cells in patients with RA | Comparison between Breg (B IL-10+) and IL-10 non secreting B lymphocytes (B IL-10 -) in RA patient of : Medium fluorescence intensity of B lymphocytes for ASCT2, Glut1, PiT1, PiT2, RFT1&3 Percentage of B lymphocytes expressing chemokine receptors CCR4, CCR7, CCR9, CXCR3, CXCR4 and CXCR5 | after analysis of the blood sample from the subject selected for the primary outcome mesure. Estimated at half a year after subject recruitement started | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Identification of genes and surface receptors expressed differentially between Breg and others B cells in patients with RA |  | Estimated at 6 month after end of subject recruitement | 
| Identification of nutrient transporters and chemokine receptors expressed differentially between Breg and others B cells in control patients |  | Estimated at 6 month after end of subject recruitement | 
| : Identification of genes and surface receptors expressed differentially between Breg and others B cells in control patients |  | Estimated at 6 month after end of subject recruitement | 
| Comparison of nutrient transporters, chemokine receptors, gene and protein surface expression expressed between Breg in patients with RA and Breg in control patients |  | Estimated at 6 month after end of subject recruitement | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Regulatory B cells (Breg), Genes, Cell surface proteins, Nutrient transporters, Chemokine receptors