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Arthritis & Rheumatology DOI 10.1002/art.39811

Altered Innate Lymphoid Cells subsets in human lymph node biopsies during the at risk and earliest phase of rheumatoid arthritis Javier Rodríguez-Carrio1,2,3*, MSc, PhD, Janine S Hähnlein1,3*, MSc, Tamara H Ramwadhdoebe1,3, MSc, Johanna F Semmelink1,3, MSc, Ivy Y. Choi1, MD, Krijn P van Lienden4, MD, PhD, Mario Maas4, Prof., MD, PhD, Danielle M Gerlag1,5, MD, PhD, Paul P Tak1,6, Prof., MD, PhD, Teunis B H Geijtenbeek3, Prof. PhD, Lisa G M van Baarsen1,3, PhD.

1Amsterdam

Rheumatology & immunology Center (ARC), Department of Clinical Immunology and Rheumatology, Academic

Medical Center, University of Amsterdam, Amsterdam, Netherlands 2Area

of Immunology, Department of Functional Biology, University of Oviedo, Asturias, Spain

3Department

of Experimental Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands

4Department

of Radiology, Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands

5Current

affiliation: Clinical Unit Cambridge, GlaxoSmithKline, Cambridge, U.K.

6Current

affiliations: Ghent University, Ghent, Belgium and University of Cambridge, Cambridge, U.K. and GlaxoSmithKline,

Stevenage, U.K. *authors contributed equally

Corresponding author:

Lisa G. M. van Baarsen, PhD Amsterdam Rheumatology & immunology Center (ARC) Department of Clinical Immunology and Rheumatology and the Department of Experimental Immunology Academic Medical Center / University of Amsterdam Meibergdreef 9, Amsterdam, 1105 AZ The Netherlands E-mail: [email protected]

Running title: ILCs subsets in LN biopsies obtained during the earliest phases of rheumatoid arthritis Conflict of interest: none of the authors has any potential financial conflict of interest related to this manuscript.

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1002/art.39811 © 2016 American College of Rheumatology Received: Mar 31, 2016; Revised: Jun 01, 2016; Accepted: Jul 07, 2016 This article is protected by copyright. All rights reserved.

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Arthritis & Rheumatology

ABSTRACT

Objectives: Innate lymphoid cells (ILCs) are emerging mediators of immunity and accumulation of inflammatory ILC populations can occur in inflammatory-mediated conditions. Since early lymph node (LN) activation has been shown in rheumatoid arthritis (RA), we aimed to investigate the frequency and distribution of ILCs in LN biopsies obtained during the earliest phases of RA. Methods: Twelve early RA patients, 12 individuals with IgM-rheumatoid factor and/or anti-citrullinated protein antibodies without arthritis (RA-risk group) and 7 healthy controls underwent ultrasound-guided inguinal LN biopsy. ILCs subsets and the expression of VCAM and ICAM by LN endothelial cells and fibroblasts were analyzed by flow cytometry. Results: Although no differences in total ILCs (Lin-CD45+/lowCD127+) frequency were found, the distribution of the ILC populations changed among groups. RA patients showed lower LTi (c-Kit+NKp44-ILCs) and increased both ILC1 (c-Kit-NKp44-ILCs) and ILC3 (c-Kit+NKp44+ILCs) counts compared with controls (p