Rheumatoid Synovitis Formation of Germinal Centers in T Cells in the +
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The Role of CD8+ CD40L+ T Cells in the Formation of Germinal Centers in Rheumatoid Synovitis This information is current as Ulf G. Wagner, Paul J. Kurtin, Andrea Wahner, Marc of October 1, 2021. Brackertz, Daniel J. Berry, Jörg J. Goronzy and Cornelia M. Weyand J Immunol 1998; 161:6390-6397; ; http://www.jimmunol.org/content/161/11/6390 Downloaded from References This article cites 32 articles, 18 of which you can access for free at: http://www.jimmunol.org/content/161/11/6390.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on October 1, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1998 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Role of CD81 CD40L1 T Cells in the Formation of Germinal Centers in Rheumatoid Synovitis1 Ulf G. Wagner,*§ Paul J. Kurtin,‡ Andrea Wahner,* Marc Brackertz,* Daniel J. Berry,† Jo¨rg J. Goronzy,* and Cornelia M. Weyand2* In rheumatoid synovitis, lymphocytes can be arranged in follicular structures resembling secondary lymphoid follicles. To un- derstand the organizing principles of this ectopic lymphoid tissue, the cellular components contributing to synovial follicles were examined. In 9 of 24 synovial tissue biopsies, lymphoid aggregates were found consisting of CD41 T cells and CD201 B cells. In four of the nine patients, the follicular centers were occupied by CD231 CD211 cellular networks representing follicular dendritic cells involved in germinal center reactions. In five patients, CD231 cells were absent from the centers of the aggregates, suggesting that fully developed germinal centers are generated in only a subset of patients. To identify factors involved in the regulation of the synovial microarchitecture, cell populations contributing to the follicles were quantified by digital image analysis of immu- Downloaded from nostained tissue and by flow cytometry of tissue-derived lymphocytes. Proportions of CD41, CD201, and CD681 cell subsets were surprisingly invariant, irrespective of the presence or absence of CD231 follicular dendritic cells. Instead, tissue biopsies with CD231 germinal center-like regions could be distinguished from those with CD232 T cell-B cell aggregates by a fourfold increase in the frequency of tissue-infiltrating CD81 T cells, a fraction of which expressed CD40 ligand (CD40L). The data suggest a previously unsuspected role of CD81 lymphocytes in modulating germinal center formation and raise the possibility that CD81 1 CD40L T cells are involved in aggravating pathologic immune responses in rheumatoid synovitis. The Journal of Immunology, http://www.jimmunol.org/ 1998, 161: 6390–6397. n rheumatoid arthritis (RA3), the synovial membrane of af- The follicular formations adopted by T cells and B cells in rheu- fected joints undergoes a series of changes that result in the matoid synovium have microscopic characteristics reminiscent of I formation of a proliferative tissue exhibiting invasive and germinal centers (GCs) (4, 5). GCs are specialized microanatom- destructive features. Although the factors that initiate rheumatoid ical structures that are required for the generation of high-affinity synovitis have not been identified, recruitment of T cells, B cells, Abs and the selection of memory B cells in response to protein Ags and macrophages into the membrane, hypertrophy, and hyperpla- (6–8). Immunization with Ag induces activation of T cells and B sia of synovial lining cells and a shift in the phenotype and func- cells in the T cell-rich areas of secondary lymphoid tissues with by guest on October 1, 2021 tion of synovial fibroblasts are accepted as principal events (1, 2). cognate- and costimulation-dependent expansion of Ag-specific In general, lymphocytes are infrequent in the hyperplastic lining lymphocytes (9). Activated T cells and B cells from these popu- layer, which is composed of a mixture of highly activated macro- lations migrate into adjacent B cell zones to form GCs (10). GCs phage-like cells and proliferating resident fibroblast-like cells. arise when B cells accumulate among the processes of follicular Lymphocytes migrate into the sublining tissue, where they accu- dendritic cells (FDCs) and undergo intense proliferation, apopto- mulate around blood vessels or infiltrate into the stroma. Evidence sis, and V(D)J gene hypermutation. The current paradigm holds suggests that the process of tissue infiltration is organized, because that somatically mutated cell surface Igs are probed on Ags bound the lymphocytes acquire a defined structural organization. T cells to FDCs and that B cells producing high-affinity Abs are selected and B cells are often organized into follicular structures that can to survive, while B cells that fail to recognize Ag die by apoptotic resemble lymphoid follicles. Besides their pseudofollicular orga- programmed cell death (6, 11). Only 10% of GC cells are CD31 nization, tissue-infiltrating lymphocytes may also be involved in T cells, virtually all of which express the CD4 marker (12). CD81 forming typical granulomas containing giant cells (3). T cells account for ,3% of the T cells. Although they represent a minority of GC cells, CD41 T cells are requisite for GC formation. In murine studies, CD41 T cells migrating into the GC bear ab Departments of *Medicine, Division of Rheumatology, †Orthopedic Surgery, and ‡Laboratory Medicine and Pathology, Mayo Clinic and Foundation, Rochester, MN TCRs reactive to the Ag driving the GC reaction (13, 14). 55905; and §Department of Medicine IV, University of Leipzig, Leipzig, Germany Molecular mechanisms involved in GC formation are beginning Received for publication April 13, 1998. Accepted for publication July 21, 1998. to be understood. Patients with X-linked immunodeficiency caus- The costs of publication of this article were defrayed in part by the payment of page ing hyper-IgM production do not generate secondary Ab re- charges. This article must therefore be hereby marked advertisement in accordance sponses, lack memory B cells, and possess lymph nodes with pri- with 18 U.S.C. Section 1734 solely to indicate this fact. mary follicles lacking GC reactions (15). The molecular defect 1 Supported in part by grants from the National Institutes of Health (RO1 AR41974 and RO1 AR42527), by a National Arthritis Foundation Biomedical Science grant underlying this syndrome has been localized to the CD40 ligand (AF #16), and by the Mayo Foundation. U.G.W. was the recipient of fellowships from (CD40L) gene (16, 17). According to the current model, CD40- the Deutscher Akademischer Austauschdienst (DAAD) and the Stiftung Familie Klee. CD40L interactions are critical for the recruitment of GC precur- 2 Address correspondence and reprint requests to Dr. C. M. Weyand, Mayo Clinic, sors, with CD40L1 T cells and interdigitating dendritic cells pro- 200 First Street SW, Rochester, MN 55905; E-mail address: weyand.cornelia@ mayo.edu viding signals to B cell precursors in extrafollicular areas (18–20). 3 B cell memory generation appears to be regulated by the interac- Abbreviations used in this paper: RA, rheumatoid arthritis; CD40L, CD40 ligand; 1 1 FDC, follicular dendritic cell; GC, germinal center. tion of CD40 centrocytes with CD40L intrafollicular T cells Copyright © 1998 by The American Association of Immunologists 0022-1767/98/$02.00 The Journal of Immunology 6391 Table I. Demographics of the study population ME); and anti-IgD (A093, 1:1000, Dako). Anti-CD40L (anti-gp39, Ancell, Bayport, MN) was used at a dilution of 1:500, and slides were incubated overnight at 4°C. Secondary species-specific Abs were applied for 30 min Disease Age at Disease Rheumatoid Duration at room temperature followed by detection with the streptavidin-biotin Patient Sex Onset (yr) Factor (yr) Biopsy Site complex immunoperoxidase or alkaline phosphatase technique. For light microscopy, 3-amino-9-ethylcarbazole (AEC; Sigma, St. Louis, MO) or 1 Female 59 1 11 Elbow 3,39-diaminobenzidine tetrahydrochloride (DAB; Sigma) substrate solu- 2 Female 59 2 5 Knee tions were used. For digital fluorescence imaging, the Vectastain ABC kit 3 Female 26 1 12 Hip and alkaline phosphatase substrate kit I (Vector Red, Vector Laboratories, 4 Male 75 2 4 Knee Burlinghame, CA) were used for detection. For two-color immunohisto- 5 Female 37 1 38 Knee chemistry, slides were washed for 10 min in 1% Triton X-100 in PBS and 6 Female 54 1 2 Wrist blocked with 5% normal goat serum before adding the second Ab. Sec- 7 Male 64 1 10 Knee tions were counterstained with hematoxylin and permanently mounted 8 Female 17 1 1 Shoulder in Cytoseal-280 (Stephens Scientific, Riverdale, NJ). 9 Male 48 1 10 Elbow Digital imaging Fluorescence-stained tissue sections from serial sections were scanned us- ing a confocal microscope (LSM310, Carl Zeiss, Oberkochen, Germany) (21, 22). Other molecules implicated in GC reactions include the with an argon-krypton laser, excitation 568 nm. Fluorescence signals for integrin, VLA-4, on lymphocytes interacting with VCAM-1 on each Ab were translated into pseudocolors. For analysis of spatial relation- FDCs, possibly facilitating the adherence of B cells to the FDC ships, images from the immunohistochemical analysis for CD68, CD4, CD8, CD20, and CD23 expression were overlaid. Correct positioning of reticulum (23). Additionally, mice deficient in either lymphotoxin Downloaded from a a the images was ensured by placing the digitized scans on a light micro- (24), TNF- (25), or the type-I TNFR (26) fail to develop sec- scope background that showed tissue landmarks and cellular infiltrates.