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Abstract Number: 1922

Peripheral B Lymphocytes Secrete Both Interleukin 6 and Transforming Growth Factor-Beta and Potentiate Fibroblast Activation in Systemic Sclerosis

Nicolas Dumoitier1,2,3,4, Sebastien Lofek2,3,5, Alexis Regent6, Jonathan London7, Benjamin Chaigne1,2,3,8, Benjamin Terrier2,7,9,10, Nadine Varin-Blank4,11 and Luc Mouthon1,2,4,6, 1Infection, Immunité, Inflammation, Institut Cochin, INSERM U1016, Paris, France, 2CNRS, UMR8104, Paris, France, 3Université Paris Descartes, Paris, France, 4Labex Inflamex, Université Sorbonne Paris Cité, Paris, France, 5Infection,Immunité, Inflammation, Institut Cochin, INSERM U1016, Paris, France, 6Internal Medicine, National Referral Center for Rare Systemic Autoimmune Diseases, Hôpital Cochin, Paris, France, 7Internal Medecine, National Referral Center for Rare Systemic Autoimmune Diseases, Hôpital Cochin, Paris, France, 8Internal Medecine, National Referral Center for Rare Systemic Autoimmune Diseases, Hôpital Cochinl, Paris, France, 9Infection, Immunité, Inflammation, Institut Cochin INSERM U1016, Paris, France, 10Université Paris Descartes, paris, France, 11UFR SMBH, INSERM, UMR978, Bobigny, France

Meeting: 2015 ACR/ARHP Annual Meeting

Date of first publication: September 29, 2015

Keywords: B cells, Fibroblasts, interleukins (IL), systemic sclerosis and transforming growth factor

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Session Information

Date: Monday, November 9, 2015

Session Title: Systemic Sclerosis, Fibrosing Syndromes and Raynaud's - Pathogenesis, Animal Models and Genetics Poster I

Session Type: ACR Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose: Systemic sclerosis (SSc) is a rare connective tissue disease characterized by fibroblasts activation, increased extra-cellular matrix synthesis associated with autoimmunity. However, the potential role of B lymphocytes in the pathophysiology of SSc is still poorly defined.

Methods: Phenotyping of peripheral blood B cell subpopulations, activating and inhibitory receptor expressions, Interleukin 6 (IL-6) and transforming growth factor-beta (TGF-β) productions were characterized by flow cytometry and multiplex assay. Fibroblasts proliferation rate and collagen production upon incubation with supernatants of B cell isolated from patients or healthy controls were assessed using XTT, BrdU, Ki67 immunofluorescence staining and collagen assay, respectively.

Results:

Eighty patients (including 18 males) with SSc fulfilling the ACR/EULAR criteria, (22 with diffuse SSc, dSSc and 58 with limited cutaneous SSc, lSSc), and 21 healthy controls (HC) were studied. The proportion of IgD– CD27+ B cells was significantly lower for SSc patients (Mean±SD 19.33%±11.85) as for HC (25.91%±11.25) (p=0.041). Increased proportions of B cells expressing CD69 (8.29%±7.77 vs 2.36%±2.29 p=0.0017) and CD95 (47.02%±19.82 vs 28.93%±11.21 (p=0.0004)) were found in both lSSc and dSSc. Compared to HC and lSSc, B lymphocytes from dSSc patients also presented with higher proportions of cells positive for CD5 (24.12%±7.93 for dSSc vs 14.09%±6.58 for lSSc (p=0.0296) and 14.21%±5.34 for HC), CD86 (39.89%±22.11 for dSSc vs 17.72%±13.98 for lSSc (p=0.0007) and 11.68%±11.09 for HC), IL-6R (33.64%±23.12 for dSSc vs 17.91%±13.62 for lSSc (p<0.0001) and 12.08±8.68 for HC) and IL-21R (32.55%±20.19 for dSSc vs 5.76%±4.40 for lSSc (p<0.0001) and 5.93%±3.29 for HC). Intracellular flow cytometry identified a significantly increased proportion of IL-6 (24.53%±6.69 vs 13.23%±4.71 (p<0.0001)) and TGF-β (18.38%±10.22 vs 6.31%±3.62 (p<0.0001)) positive B lymphocytes in patients with SSc as compared to HC. Using multiplex assay, increased production of IL-6 (314.3 pg/ml±317.8 vs 6.10pg/ml±2.58 (p=0.0007)) and TGF-β (1020pg/ml±569 vs 163.8pg/ml±98.69 (p=0.0011)) altogether with reduced production of IL-10 (8.67pg/ml±6.22 vs 333.1pg/ml±123.7 (p=0.0008)) were detected upon stimulation of B lymphocytes isolated from SSc patients compared to HC. Fibroblast proliferation and collagen production were also significantly increased in the presence of B cell supernatant from SSc patients as compared to HC.

Conclusion: We characterized a differential activation of peripheral B cells of patients exhibiting dSSc with an upregulation of CD5, CD86, IL-6R and IL-21R compare to lSSc patients and HC. B lymphocytes of SSc patients also expressed more CD69 and CD95 compare to HC. Peripheral B lymphocytes secreted both interleukin 6 and transforming growth factor-beta, and activated fibroblasts in patients with SSc.


Disclosure: N. Dumoitier, None; S. Lofek, None; A. Regent, None; J. London, None; B. Chaigne, None; B. Terrier, Roche Pharmaceuticals, 5; N. Varin-Blank, None; L. Mouthon, Roche Pharmaceuticals, 5.

To cite this abstract in AMA style:

Dumoitier N, Lofek S, Regent A, London J, Chaigne B, Terrier B, Varin-Blank N, Mouthon L. Peripheral B Lymphocytes Secrete Both Interleukin 6 and Transforming Growth Factor-Beta and Potentiate Fibroblast Activation in Systemic Sclerosis [abstract]. Arthritis Rheumatol. 2015; 67 (suppl 10). https://acrabstracts.org/abstract/peripheral-b-lymphocytes-secrete-both-interleukin-6-and-transforming-growth-factor-beta-and-potentiate-fibroblast-activation-in-systemic-sclerosis/. Accessed March 3, 2021.
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