Lymphoid Chemokine B Cell-Attracting Chemokine-1 (CXCL13) Is Expressed in Germinal Center of Ectopic Lymphoid Follicles Within the Synovium of Chronic This information is current as Patients of September 26, 2021. Kenrin Shi, Kenji Hayashida, Motoharu Kaneko, Jun Hashimoto, Tetsuya Tomita, Peter E. Lipsky, Hideki Yoshikawa and Takahiro Ochi

J Immunol 2001; 166:650-655; ; Downloaded from doi: 10.4049/jimmunol.166.1.650 http://www.jimmunol.org/content/166/1/650 http://www.jimmunol.org/ References This article cites 37 articles, 13 of which you can access for free at: http://www.jimmunol.org/content/166/1/650.full#ref-list-1

Why The JI? Submit online.

• Rapid Reviews! 30 days* from submission to initial decision

by guest on September 26, 2021 • No Triage! Every submission reviewed by practicing scientists

• Fast Publication! 4 weeks from acceptance to publication

*average

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 © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Lymphoid Chemokine B Cell-Attracting Chemokine-1 (CXCL13) Is Expressed in Germinal Center of Ectopic Lymphoid Follicles Within the Synovium of Chronic Arthritis Patients1

Kenrin Shi,* Kenji Hayashida,2* Motoharu Kaneko,* Jun Hashimoto,* Tetsuya Tomita,* Peter E. Lipsky,† Hideki Yoshikawa,* and Takahiro Ochi*

A unique feature in inflammatory tissue of (RA) is the formation of ectopic lymphoid aggregates with germinal center (GC)-like structures that can be considered to contribute to the pathogenesis of RA, because local production of the autoantibody, rheumatoid factor, is thought to be a causative factor in tissue damage. However, the factors governing the formation of GC in RA are presently unknown. To begin to address this, the expression of B cell attracting chemokine (BCA-1)

(CXCL13), a potent chemoattractant of B cells, was examined in the synovium of patients with RA or with (OA). Downloaded from Expression of BCA-1 mRNA was detected in all RA samples, but in only one of five OA samples. Lymphoid follicles were observed in four of seven RA samples and in two of eight OA samples, and in most of them BCA-1 protein was detected in GC. BCA-1 was not detected in tissues lacking lymphoid follicles. Notably, BCA-1 was detected predominantly in follicular dendritic cells in GC. CD20-positive B cells were aggregated in regions of BCA-1 expression, but not T cells or . These data suggest that BCA-1 produced by follicular dendritic cells may attract B cells and contribute to the formation of GC-like structures in chronic arthritis. The Journal of Immunology, 2001, 166: 650–655. http://www.jimmunol.org/

cell attracting chemokine-1 (BCA-1)3 (CXCL13; Ref. disease by leading to the local production of the autoantibody, 1), a ligand of CXCR-5, is classified as a lymphoid che- rheumatoid factor. B mokine and is a potent B cell chemoattractant, because The mechanism underlying the formation of lymphocyte aggre- CXCR-5 is highly expressed by naive B cells (2, 3). BCA-1 is gates and GC-like structures in RA synovium has not been eluci- thought to be expressed predominantly by follicular dendritic cells dated. In the present study, we explored the hypothesis that local (FDC) in secondary lymphoid tissues and organs such as lymph expression of BCA-1 might play a role. BCA-1 expression in sy- nodes and spleen and to play an important role in attracting naive novia of RA and osteoarthritis (OA) was assessed, and its distri- B cells to form germinal centers (GC) in such organs (4, 5). bution and relationship to the location of T cells, B cells, macro- by guest on September 26, 2021 Rheumatoid arthritis (RA) is a chronic inflammatory autoim- phages, and FDC were examined. BCA-1 was expressed by FDC mune disease. Within the inflamed synovium, there is an accumu- in GC of RA and OA synovia and was found in regions of B cell lation of lymphoid and myeloid cells, including T cells, B cells, aggregation. Therefore, BCA-1 may play a central role in attract- and monocytes. The local activation of these cells is thought es- ing B cells to form GC in the synovium of patients with chronic sential in perpetuating chronic inflammation and accelerating joint arthritis. damage (6, 7). T cells and B cells in RA synovium frequently accumulate underneath the synovial surface and organize into lym- phocyte aggregates with certain features of GC (8, 9), including Materials and Methods the local differentiation of FDC and plasma cells (PC) and the Tissue samples production of large amounts of Ab (10, 11). The development of Tissue samples were obtained from seven patients with RA (mean age, these structures appears to contribute to the pathogenesis of the 54.4 years; range, 29–71 years) and eight patients with OA (mean age, 74.1 years; range, 65–86 years) during surgeries. RA was diagnosed according to the American College of (ACR) criteria for the classification of RA (12), and OA was diagnosed according to ACR criteria for the classification of OA of the knee (13). Tissue samples of normal *Department of Orthopedic Surgery, Osaka University Medical School, Suita, Osaka, Japan; and †National Institute of Arthritis and Musculoskeletal and Skin Diseases, inguinal lymph nodes were obtained from nonarthritic patients during other Bethesda, MD 20892 surgeries. Received for publication July 10, 2000. Accepted for publication October 9, 2000. The costs of publication of this article were defrayed in part by the payment of page Primary Abs charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Goat anti-human BCA-1 (R&D Systems, Minneapolis, MN) was used to 1 This work was supported by a grant from the Organization for Pharmaceutical detect BCA-1. For detection of FDC, Abs for CD21 (clone 1F8; Dako, Safety and Research, Tokyo, Japan. Glostrup, Denmark) and for dendritic reticulum cells (DRC) (clone R4/23; 2 Address correspondence and reprint requests to Dr. Kenji Hayashida, Department of Dako) were used. mAbs directed against CD3-positive T cells (clone PS1; Orthopedic Surgery, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka Medac, Hamburg, Germany), CD20-positive B cells (clone L26; Dako), 565-0871, Japan. E-mail address: [email protected] and CD68-positive macrophages (clone KP1; Dako) were also used. 3 Abbreviations used in this paper: BCA-1, B cell attracting chemokine-1; ACR, Primary Abs against another lymphoid chemokine, secondary lymphoid American College of Rheumatology; DRC, dendritic reticulum cell; FDC, follicular tissue chemokine (SLC) (CCL21; R&D Systems), were also used for im- dendritic cell; GC, germinal center; OA, osteoarthritis; PC, plasma cell; RA, rheu- munohistochemistry in RA synovium as well as in normal inguinal lymph matoid arthritis; SLC, secondary lymphoid tissue chemokine. nodes as a control.

Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 The Journal of Immunology 651

RNA isolation and RT-PCR RA and OA tissues obtained at surgery were minced and weighed. One milligram of the tissue was dissolved with 1 ml of Isogen (Nippongene, Tokyo, Japan) and sonicated. RNA was extracted in accordance with the company’s directions. Two micrograms of the extracted RNA was treated with DNase I (Life Technologies, Rockville, MD) to eliminate DNA and reverse-transcribed by SuperScript II reverse-transcriptase (Life Technol- ogies) at 42°C for 70 min using oligo (dT) as a primer (Amersham Phar- macia Biotech, Piscataway, NJ). PCR was conducted with Taq DNA poly- merase (AmpliTaq Gold; Perkin-Elmer Applied Biosystems, Foster City, ␮ CA) using 1–3 l of cDNA (1.5 mM of MgCl2) on a PCR thermal cycler (PC-700; Astec, Fukuoka, Japan). The primer pair for BCA-1 was CAG AAT CCT CTG GAA CTT GAG G (5Ј) and CTT CCA GAC ATT CGG AGA CC (3Ј), and that of ␤-actin was GTC CTC TCC CAA GTC CAC ACA (5Ј) and CTG GTC TCA AGT CAG TGT ACA GGT AA (3Ј). Annealing temperatures used for the amplifications were 56°C for ␤-actin and 58°C for BCA-1. PCR products were resolved by electrophoresis on 1.5% agarose gels and identified with ethidium bromide staining. To adjust the amount of cDNA of each sample precisely, ␤-actin expression was examined first using 26–30 cycles of RT-PCR to amplify 0.1, 0.2, and 0.3 ␮l of cDNA. After resolving the PCR products on agarose gels and iden-

tifying the relevant bands with ethidium bromide, the optimal amounts of Downloaded from cDNA for analysis were determined. Chemokine expression in this amount of cDNA was examined using 35, 38, and 40 cycles of PCR amplification, and the results in the linear part of the amplification curve are indicated in Fig. 1. Immunohistochemistry

Tissues from RA and OA patients were embedded in Tissue Tek (Sakura, http://www.jimmunol.org/ Tokyo, Japan), and stored at Ϫ80°C until use. The tissue was cut into 4- to 7-␮m sections with a cryostat, dried at room temperature for 1 h, and fixed in 2% paraformaldehyde at 4°C for 8 min. Normal inguinal lymph nodes were treated in the same manner. For detection of CD3 and CD20, tissue sections were boiled in 10 mM FIGURE 2. BCA-1 is focally expressed in lymphoid follicles of RA sodium citrate buffer (pH 6.0) in a 90°C water bath for 90 min to unmask the Ags. synovium. Sections of follicular (A–D) and diffuse (E and F) lymphoid After blocking endogenous peroxidase and nonspecific Ags, primary aggregation infiltrates in RA synovium, as well as in OA synovium (G and Abs were applied onto tissue sections. Tissues were incubated for2hat H), are shown. Focal expression of BCA-1 is observed in lymphoid folli- room temperature for detection of CD3, CD20, CD21, DRC, and CD68 and cles in RA synovium (B and D), whereas almost no expression is recog- for 24 h at 4°C for detection of BCA-1 and SLC. Isotype-matched Abs nized in the area of diffuse lymphocytic aggregation in RA (F)orOA by guest on September 26, 2021 were used for control staining. synovium (H). A, E, and G, Hematoxylin and eosin staining. B, D, F, and Thereafter, detection using the streptavidin biotin-peroxidase complex H, Immunohistochemistry for BCA-1. Higher magnification of the area technique (Histofine SAB-PO Kit; Nichirei, Tokyo, Japan) was conducted. outlined in B is shown in D. C, Immunohistochemistry with control goat Finally, the sections were developed in 3,3Ј-diaminobenzidine tetrahydro- IgG. Original magnification: A–C and E–H, ϫ200; D, ϫ600. chloride (Dojindo Laboratories, Kumamoto, Japan) and counterstained with hematoxylin. FITC-conjugated rabbit anti-goat IgG (Wako, Osaka, Japan) was used Two-color immunofluorescence staining as the secondary Ab for BCA-1, and rhodamine-conjugated rabbit anti- To determine the expression of BCA-1 by FDC, two-color immunofluo- mouse Igs (Dako) for CD21 or DRC. Secondary Abs were diluted together rescence staining was performed with CD21 and with DRC, respectively. and applied onto each tissue section for 30 min at room temperature. Fi- Two primary Abs raised in different species (BCA-1, goat; CD21 and nally, slides were coverslipped with PBS-glycerol with antifading agents DRC, mouse) were diluted together and applied onto tissue sections for ( p-phenylenediamine dihydrochloride; Sigma, St. Louis, MO) and ob- 24 h at 4°C. Isotype-matched Abs were used for negative control. served under a fluorescence microscope (E800; Nikon, Tokyo, Japan) equipped with a standard mercury lamp power supply. Results BCA-1 mRNA is expressed in RA and OA tissues Expression of BCA-1 mRNA was assessed by RT-PCR using mRNA directly extracted from RA and OA tissues (Fig. 1). All samples from RA patients expressed BCA-1, whereas only one of five samples from OA patients expressed BCA-1.

BCA-1 is expressed in lymphoid follicles of RA and OA synovium The location of BCA-1 protein expression was determined by im- FIGURE 1. BCA-1 mRNA is uniformly expressed in RA synovium. All munohistochemistry (Fig. 2). BCA-1 was mostly expressed in five RA and one of five OA samples expressed BCA-1 mRNA. Total RNA lymphoid follicles in RA synovium, but no expression of BCA-1 extracted from freshly isolated synovial tissue of patients with RA or OA was reverse-transcribed and subjected to PCR using specific primers. The was recognized in areas of diffuse lymphoid cell infiltration. No- optimal amount of cDNA, which was determined for each sample by ␤-ac- tably, BCA-1 was also detected in a few OA synovia with follic- tin expression, was subjected to PCR for detecting BCA-1. PCR products ular lymphocytic infiltrates, but not so obviously as was observed were electrophoresed on agarose gel and identified with ethidium bromide in RA. No expression of BCA-1 was detected in OA synovia with- staining. out lymphoid follicle formation. 652 BCA-1 IS EXPRESSED IN CHRONIC INFLAMMATORY ARTHRITIS

FIGURE 3. BCA-1-expressing cells reside within B cell aggregates. Immunohistochemistry of sections derived from RA synovium either of follicular (A–E) or of diffuse (F–J) lymphoid infiltrates was conducted to detect BCA-1 (A and F), CD20 (B and G), CD3 (C and H), and CD68 (D and I). BCA-1 expression is focally observed in the central area of lymphoid follicles (A), where CD20-positive B cells are densely accumulated (B). CD3-positive T cells Downloaded from are aggregated at the boundary of the follicle (C), and CD68-positive macrophages do not present remarkable expression within the follicle (D). BCA-1 expression was not recognized in the area of diffuse lymphocytic aggregation in RA synovium (F), where CD3-positive T cells predominantly infiltrate (H), as compared with CD20-positive B cells (G). E and J, Hematoxylin and eosin staining. Original magnification: ϫ200.

Histological characteristics of the synovium and BCA-1 most all lymphoid follicles. CD3-positive T cells were aggregated

expression at the boundary of lymphoid follicles, and CD68-positive macro- http://www.jimmunol.org/ Histological characteristics of each sample were assessed to determine phages did not present remarkable expression within lymphoid fol- whether it contained a follicular and/or diffuse lymphocyte infiltrate licles (Fig. 3). BCA-1 expression was almost compatible with the (Table I). Of seven RA samples, four contained follicular and five accumulation of B cells, and the positivity, which was expressed as diffuse infiltrates. Three of the samples contained both, and one con- the ratio of the number of BCA-1-positive follicles to the number tained no lymphocyte infiltrates. Of eight OA samples, two contained of follicles with B cell accumulation, was 100% in RA and 60% in follicular, one diffuse, and five no lymphocytic infiltration. OA (Table I). Notably, aggregations of CD20-positive cells were BCA-1 was detected in each of the synovia with follicles, but not found in regions that did not express BCA-1 in RA. not in those without. The expression of BCA-1 was examined in by guest on September 26, 2021 each lymphoid follicle, and BCA-1 positivity was calculated as the BCA-1 is expressed in FDC in GC ratio of the number of BCA-1-positive follicles to the total number of follicles. The ratio was 92% in RA and 60% in OA. BCA-1-positive cells were usually found in the central area of lymphoid follicles, which could be regarded as GC due to the BCA-1 is expressed in the area of B cell accumulation in accumulation of B cells. CD21-positive cells and DRC-positive lymphoid follicles cells were also enriched in those regions (Fig. 4). Furthermore, Immunohistohemistry for CD3, CD20, or CD68 demonstrated an two-color immunofluorescence staining demonstrated that BCA- accumulation of CD20-positive B cells in the central area of al- 1-positive cells were observed in the center of lymphoid follicles

Table I. Histological characteristics of patients synovial tissue

Age (yr)/ Total No. of No. of BCA-1- No. of Follicles with B Case Sex Folliculara Diffusea Follicles (A)b Positive Follicles (B)b Cell Accumulation (C)c B/A (%)b B/C (%)c

RA 1 57/F ϩϩ 8 7 7 88 100 2 71/F ϩϩ ϩ 20 19 19 95 100 3 59/F ϪϪ 4 63/F Ϫϩϩ 5 53/F Ϫϩ 6 29/F ϩϩ 6 5 5 83 100 7 49/F ϩϪ 3 3 3 100 100 RA total 37 34 34 92 100 OA 1 78/F ϪϪ 2 86/M ϪϪ 3 66/F ϪϪ 4 74/F ϪϪ 5 73/F ϩϪ 32 36767 6 79/F ϪϪ 7 65/F Ϫϩ 8 72/F ϩϪ 21 25050 OA total 5 3 5 60 60

a The synovium of each patient was histologically characterized as having a follicular or diffuse infiltrate by hemotoxylin and eosin staining. b Total number of follicles (A) and BCA-1-positive follicles (B) were counted, and BCA-1 positivity was given as the ratio B/A (%). c Number of follicles with B cell accumulation (C) was counted after staining for CD20, and BCA-1 positivity was given as the ratio B/C (%). The Journal of Immunology 653

FIGURE 4. BCA-1 is expressed in FDCs in the GC. Immunohistochemistry of serial sections derived from RA synovium with follicular lymphocytic aggregates was conducted for the detection of BCA-1 (B and C), CD21 (D), and DRC (E). A subset of CD21-positive and DRC-positive cells express BCA-1 in lymphoid follicles in RA synovium. Higher magnification of the area out- lined in B is shown in C. A, Hematoxylin and eosin staining. Original magnification: A, B, D, and E, ϫ200; C, ϫ400.

and colocalized with the expression of CD21 and DRC, respec- phoid follicle in chronic inflammation, it could contribute to Ag tively (Fig. 5). These findings demonstrated that BCA-1 is ex- presentation in situ, to clonal expansion of Ag-specific B and T Downloaded from pressed by FDC in GC, the B cell-rich area of lymphoid follicles. cells, and to rendering an acute inflammation into a chronic one (10, 11, 14). SLC is not detected in synovium of RA RA is a representative chronic inflammatory arthritis, the patho- Immunohistochemistry for another lymphoid chemokine, SLC, logical mechanism of which could be thought of as being an acute showed no notable expression in RA synovium. SLC was not de- inflammation at the onset, followed by progression of the chronic tected even in the area that was rich in CD3-positive T cells in RA process (6). Although the mechanism underlying the change of the http://www.jimmunol.org/ synovium, although it was observed to express in T cell-rich areas inflammatory course from acute to chronic has not been elucidated, of normal inguinal lymph nodes that we examined as a positive an immune response to Ag proteins derived from joint structures control (Fig. 6). might be one of the causes of prolonged inflammation. From this point of view, the formation of lymphoid follicles, especially those Discussion with GC, which is a typical histological feature of RA synovium, In the present study, we demonstrated that BCA-1 was expressed, seems an important process in the development of chronic arthritis. most likely by FDC, in GC in the synovium of RA as well as OA The literature illustrates that lymphoid follicles with GC consist and could contribute to lymphoid follicle formation. This paper is of naive B cells in the central zone surrounded by mature B cells, the first demonstration of BCA-1 expression in ectopic lymphoid PCs, and CD4-positive T cells in RA synovium, and there, naive by guest on September 26, 2021 follicles of chronic inflammatory sites. B cells are specifically selected, activated, and clonally expanded, Lymphoid follicles are usually observed in secondary lymphoid exhibiting isotype switching and somatic hypermutation (8–11). tissues and organs, such as spleen, tonsils, lymph nodes, and mu- PCs in RA synovium have been reported to synthesize high cosa-associated lymphoid tissues, and are thought to be essential amounts of Igs, including rheumatoid factor, which is thought to structures for the normal immune response in which Ag-specific B participate in the pathogenesis of tissue damage (6, 15–17). More- and T cells are expanded against Ags. Lymphoid follicle formation over, rheumatoid synoviocytes are reported to support the terminal is also observed ectopically in chronic inflammatory sites. Al- differentiation of B cells into mature PCs in vitro (18). Based on though there is no knowledge of the true role of an ectopic lym- this knowledge, B cell development in GC might be one of the essential courses in the pathogenesis of RA. In the formation of GC as well as in the events necessary for B cell development, FDCs are supposed to be involved (19). It is also reported that FDC contribute to the apoptosis of B cells in GC via Fas-Fas ligand (20). As shown in Fig. 3, B cells are localized around BCA-1 expression in the central area of lymphoid follicles in RA synovium. Moreover, FDC characterized by CD21 and DRCs has been observed to express BCA-1 in GC of the follicles (Figs. 4 and 5). These results indicate that FDC might play crucial roles in constructing GC in RA synovium and that BCA-1 pro- duced by FDC might act as a potent chemoattractant of naive B cells into the synovium. In this study, BCA-1 expression was recognized in all RA sam- ples, although the formation of lymphoid follicles was observed histologically in four of seven RA patients. The most explanatory reason for this discrepancy could be the sampling issue; the part FIGURE 5. Cells expressing BCA-1 are identical with FDCs in the GC. without lymphoid follicles might be used for histological analysis, Colocalization of BCA-1 with CD21 and DRCs in the central area of the whereas another part of such tissues might include lymphoid fol- lymphoid follicle is shown. Two-color immunofluorescence staining of BCA-1 with CD21 (A and B) and with DRC (C and D) was performed, re- licles. A divergence in transcription and translation might also be spectively. BCA-1 expression (A and C; FITC, green staining) was observed present. in FDC characterized by specific staining with CD21 and DRC (B and D, Recently, several lymphoid chemokines, which are mainly ex- respectively; rhodamine, red staining). Original magnification: ϫ400. pressed in secondary lymphoid tissues and organs, have been 654 BCA-1 IS EXPRESSED IN CHRONIC INFLAMMATORY ARTHRITIS

synovium of OA in the early stage as well as the restricted TCR usage of V␤-chain among those T cells. These findings suggest that OA can be regarded as one of the chronic inflammatory dis- eases with a certain immune response resembling autoimmune dis- eases like RA, although the clinical feature of inflammation is not so severe. In this study, formation of a few lymphoid follicles with BCA-1 expression was recognized in two of eight OA synovia. The ag- gregation of CD20-positive B cells was also observed in those lymphoid follicles. The synovium of these patients apparently showed inflammatory changes including villous formation and looked like RA synovium. Although we diagnosed these patients as OA simply by their clinical features and by ACR criteria for the classification of OA, they might have possessed inflammatory FIGURE 6. Another lymphoid chemokine, SLC, is not expressed in RA properties similar to those in RA in their pathological process. synovium. Tissue sections from normal inguinal lymph nodes and RA sy- Further follow-up will be necessary to diagnose these patients novia were examined for detection of SLC by immunohistochemistry. Ex- precisely. pression of SLC was observed in lymph nodes where CD3-expressing T In this study, we examined the expression of lymphoid chemo- cells were observed (A and B). However, SLC was not detected in RA kines other than BCA-1 in RA synovium. Because the expression Downloaded from synovia even in the area that was rich in CD3-positive T cells (C and D). of SLC was detected by immunohistochemistry in T cell-rich areas A and C, Immunohistochemistry for SLC. B and D, Immunohistochemistry of normal lymph nodes but not in RA synovium, even in the area ϫ for CD3. Original magnification: 200. with abundant T cells, SLC might not contribute to T cell accu- mulation in RA synovium. Many chemokines have been reported and they are classified

found, and their role in the organization of secondary lymphoid into two groups as pro-inflammatory chemokines and lymphoid http://www.jimmunol.org/ tissue has been elucidated (4, 5). Their potent and con- chemokines. BCA-1 has been classified as a lymphoid chemokine stitutive expression in lymphoid organs suggest that lymphoid che- so far, but the results of this study showed its marked expression mokines may play important roles in the generation and mainte- in chronic inflammatory sites. BCA-1 may play an important role nance of the architecture of the . in the induction of immune response in chronic inflammatory dis- BCA-1 was first reported in 1998 with its specific expression in eases, especially in autoimmune diseases such as RA. Further in- lymphoid organs, especially in GC in secondary lymphoid tissues vestigation of lymphoid chemokines in inflammatory conditions and organs (2, 4). BCA-1 is a potent chemoattractant of B cells and will be necessary to better understand the immunological patho- is produced by FDC in lymphoid organs. Because CXCR5, the genesis in such diseases. specific receptor of BCA-1, is mostly expressed by B cells, FDC by guest on September 26, 2021 may produce BCA-1 to attract B cells and initiate the formation of References GC in lymphoid organs. Recently, BCA-1 expression was recog- 1. Zlotnik, A., and O. Yoshie. 2000. Chemokines: a new classification system and nized in mucosa-associated lymphoid tissues with Helicobacter their role in immunity. Immunity 12:121. pylori infection (21). In addition, BCA-1 was detected in chronic 2. Legler, D. F., M. Loetscher, R. S. Roos, I. Clark-Lewis, M. Baggiolini, and ␣ B. Moser. 1998. B cell-attracting chemokine 1, a human CXC chemokine ex- inflammation induced by lymphotoxin- in a transgenic mouse pressed in lymphoid tissues, selectively attracts B lymphocytes via BLR1/ model (22). These reports suggest that BCA-1 may contribute to CXCR5. J. Exp. Med. 187:655. the development of chronic inflammation. 3. Gunn, M. D., V. N. Ngo, K. M. Ansel, E. H. Ekland, J. G. Cyster, and L. T. Williams. 1998. A B-cell-homing chemokine made in lymphoid follicles The origin of FDC is controversial. Kapasi et al. (23) reported activates Burkitt’s lymphoma receptor-1. Nature 391:799. that precursors of FDC exist in bone marrow, move into target 4. Cyster, J. G. 1999. Chemokines and cell migration in secondary lymphoid organs. organs, and differentiate into FDC, while Humphery et al. (24) and Science 286:2098. 5. Melchers, F., A. G. Rolink, and C. Schaniel. 1999. The role of chemokines in Yamakawa et al. (25) did not support the idea that FDC originate regulating cell migration during humoral immune responses. Cell 99:351. from bone marrow. According to previous reports using knockout 6. Harris, E. D., Jr. 1990. Rheumatoid arthritis: pathophysiology and implications mice, lymphotoxin-␣ and TNFRI play a crucial role in establishing for therapy. N. Engl. J. Med. 322:1277. FDC in vivo (26–30). 7. Krane, S. M. 1993. Mechanisms in tissue destruction in rheumatoid arthritis. In Arthritis and Allied Conditions, 12th Ed. D. J. McCarty and W. J. Koopman, eds. RA fibroblast-like synoviocytes are known to produce pro-in- Lea & Febiger, London, U.K., p. 763. flammatory chemokines such as IL-8, monocyte chemoattractant 8. Klimiuk, P. A., J. J. Goronzy, J. Bjornsson, R. D. Beckenbaugh, and C. M. Weyand. 1997. Tissue patterns distinguish variants of rheumatoid protein-1, epithelial neutrophil activating peptide-78, synovitis. Am. J. Pathol. 151:1311. inflammatory protein-1␤, and RANTES (31–35). Furthermore, 9. Wagner, U. G., P. J. Kurtin, A. Wahner, M. Brackertz, D. J. Berry, J. J. Goronzy, Lindhout et al. (36) reported that RA fibroblast-like synoviocytes and C. M. Weyand. 1998. The role of CD8ϩCD40Lϩ T cells in the formation of stimulated with TNF-␣ and IL-1␤ had intrinsic properties of FDC. germinal centers in rheumatoid synovitis. J. Immunol. 161:6390. 10. Schroder, A. E., A. Greiner, C. Seyfert, and C. Berek. 1996. Differentiation of B However, in this study, BCA-1 was not expressed in fibroblast-like cells in the nonlymphoid tissue of the synovial membrane of patients with rheu- cells in synovium. Moreover, RA fibroblast-like synoviocytes matoid arthritis. Proc. Natl. Acad. Sci. USA 93:221. ␤ 11. Kim, H. J., V. Krenn, G. Steinhauser, and C. Berek. 1999. Plasma cell develop- stimulated with pro-inflammatory including IL-1 , ment in synovial germinal centers in patients with rheumatoid and reactive ar- TNF-␣, and lymphotoxin-␣ did not show BCA-1 expression (data thritis. J. Immunol. 162:3053. not shown). Further investigation is necessary to address the prob- 12. Arnett, F. C., S. M. Edworthy, D. A. Bloch, D. J. McShane, J. F. Fries, N. S. Cooper, L. A. Healey, S. R. Kaplan, M. H. Liang, H. S. Luthra, et al. 1988. lem of which cell in RA synovium is the precursor of FDC. The American Rheumatism Association 1987 revised criteria for the classifica- Recently, several investigators have reported inflammatory fea- tion of rheumatoid arthritis. Arthritis Rheum. 31:315. 13. Altman, R., E. Asch, D. Bloch, G. Bole, D. Borenstein, K. Brandt, W. Christy, tures of OA. Krenn et al. (37) demonstrated highly mutated VH T. D. Cooke, R. Greenwald, M. Hochberg, et al. 1986. Development of criteria genes in B cells, with a characteristic arrangement of B cells and for the classification and reporting of osteoarthritis: classification of osteoarthritis PCs. Nakamura et al. (38) reported the infiltration of T cells in the of the knee. Arthritis. Rheum. 29:1039. The Journal of Immunology 655

14. Stott, D. I., F. Hiepe, M. Hummel, G. Steinhauser, and C. Berek. 1998. Antigen- photoxin ␣/␤ and are required for stromal cell expression driven clonal proliferation of B cells within the target tissue of an autoimmune of homing chemokines in B and T cell areas of the spleen. J. Exp. Med. 189:403. disease. J. Clin. Invest. 102:938. 28. Tkachuk, M., S. Bolliger, B. Ryffel, G. Pluschke, T. A. Banks, S. Herren, 15. Panush, R. S., N. E. Bianco, and P. H. Schur. 1971. Serum and synovial fluid IgG, R. H. Gisler, and M. H. Kosco-Vilbois. 1998. Crucial role of tumor necrosis IgA and IgM antigammaglobulins in rheumatoid arthritis. Arthritis Rheum. 14: factor receptor 1 expression on nonhematopoietic cells for B cell localization 737. within the splenic white pulp. J. Exp. Med. 187:469. 16. Bankhurst, A. D., G. Husby, and R. C. Williams, Jr. 1976. Predominance of T 29. Gonzalez, M., F. Mackay, J. L. Browning, M. H. Kosco-Vilbois, and R. J. Noelle. cells in the lymphocytic infiltrates of synovial tissues in rheumatoid arthritis. 1998. The sequential role of lymphotoxin and B cells in the development of Arthritis. Rheum. 19:555. splenic follicles. J. Exp. Med. 187:997. 17. Vaughan, J. H. 1993. Pathogenetic concepts and origins of rheumatoid factor in 30. Fu, Y. X., G. Huang, Y. Wang, and D. D. Chaplin. 1998. B lymphocytes induce rheumatoid arthritis. Arthritis Rheum. 36:1. the formation of follicular dendritic cell clusters in a lymphotoxin ␣-dependent 18. Dechanet, J., P. Merville, I. Durand, J. Banchereau, and P. Miossec. 1995. The fashion. J. Exp. Med. 187:1009. ability of synoviocytes to support terminal differentiation of activated B cells may explain plasma cell accumulation in rheumatoid synovium. J. Clin. Invest. 102: 31. Koch, A. E., S. L. Kunkel, J. C. Burrows, H. L. Evanoff, G. K. Haines, 938. R. M. Pope, and R. M. Strieter. 1991. Synovial tissue macrophage as a source of 19. Lindhout, E., G. Koopman, S. T. Pals, and C. de Groot. 1997. Triple check for the chemotactic cytokine IL-8. J. Immunol. 147:2187. antigen specificity of B cells during germinal centre reactions. Immunol. Today 32. Koch, A. E., S. L. Kunkel, L. A. Harlow, B. Johnson, H. L. Evanoff, 18:573. G. K. Haines, M. D. Buridick, R. M. Pope, and R. M. Strieter. 1992. Enhanced 20. Hur, D. Y., D. J. Kim, S. Kim, Y. I. Kim, D. Cho, D. S. Lee, Y. Hwang, production of monocyte chemoattractant protein-1 in rheumatoid arthritis. J. Clin. K. W. Bae, K. Y. Chang, and W. J. Lee. 2000. Role of follicular dendritic cells Invest. 90:772. in the apoptosis of germinal center B cells. Immunol. Lett. 72:107. 33. Koch, A. E., S. L. Kunkel, L. A. Harlow, D. D. Mazarakis, G. K. Haines, 21. Mazzucchelli, L., A. Blaser, A. Kappeler, P. Scharli, J. A. Laissue, M. Baggiolini, M. D. Burdick, R. M. Pope, R. M. Strieter. 1994. Epithelial neutrophil activating and M. Uguccioni. 1999. BCA-1 is highly expressed in Helicobacter pylori- peptide-78: a novel chemotactic cytokine for neutrophils in arthritis. J. Clin. induced mucosa-associated lymphoid tissue and gastric lymphoma. J. Clin. In- Invest. 94:1012. vest. 104:49. 34. Koch, A. E., S. L. Kunkel, M. R. Shah, R. Fu, D. D. Mazarakis, G. K. Haines, 22. Hjelmstrom, P., J. Fjell, T. Nakagawa, R. Sacca, C. A. Cuff, and N. H. Ruddle. M. D. Burdick, R. M. Pope, and R. M. Strieter. 1995. Macrophage inflammatory Downloaded from 2000. Lymphoid tissue homing chemokines are expressed in chronic inflamma- protein-1␤: a C-C chemokine in osteoarthritis. Clin. Immunol. Immunopathol. tion. Am. J. Pathol. 156:1133. 77:307. 23. Kapasi, Z. F., D. Qin, W. G. Kerr, M. H. Kosco-Vilbois, L. D. Shultz, J. G. Tew, 35. Robinson, E., E. C. Keystone, T. J. Schall, N. Gillett, and E. N. Fish. 1995. and A. K. Szakal. 1998. Follicular dendritic cell (FDC) precursors in primary Chemokine expression in rheumatoid arthritis (RA): evidence of RANTES and lymphoid tissues. J. Immunol. 160:1078. macrophage inflammatory protein (MIP)-1␤ production by synovial T cells. Clin. 24. Humphrey, J. H., D. Grennan, and V. Sundaram. 1999. The origin of follicular Exp. Immunol. 101:398. dendritic cells in the mouse and the mechanism of trapping of immune complexes 36. Lindhout, E., M. van Eijk, M. van Pel, J. Lindeman, H. J. Dinant, and

on them. Histol. Histopathol. 14:135. http://www.jimmunol.org/ 25. Yamakawa, M., Y. Imai, M. Dobashi, and T. Kasajima. 2000. Development of C. de Groot. 1999. Fibroblast-like synoviocytes from rheumatoid arthritis patients follicular dendritic cells: a study using short-term bone marrow cell grafting in have intrinsic properties of follicular dendritic cells. J. Immunol. 162:5949. SCID mice. Eur. J. Immunol. 14:859. 37. Krenn, V., F. Hensel, H. J. Kim, M. M. Souto Carneiro, P. Starostik, G. Ristow, 26. Matsumoto, M., Y. X. Fu, H. Molina, G. Huang, J. Kim, D. A. Thomas, A. Konig, H. P. Vollmers, and H. K. Muller-Hermelink. 1999. Molecular IgV(H) M. H. Nahm, and D. D. Chaplin. 1997. Distinct roles of lymphotoxin ␣ and the analysis demonstrates highly somatic mutated B cells in synovialitis of osteoar- type I tumor necrosis factor (TNF) receptor in the establishment of follicular thritis: a degenerative disease is associated with a specific, not locally generated dendritic cells from non-bone marrow-derived cells. J. Exp. Med. 186:1997. immune response. Lab. Invest. 79:1377. 27. Ngo, V. N., H. Korner, M. D. Gunn, K. N. Schmidt, D. S. Riminton, 38. Nakamura, H., S. Yoshino, T. Kato, J. Tsuruha, and K. Nishioka. 1999. T-cell M. D. Cooper, J. L. Browning, J. D. Sedgwick, and J. G. Cyster. 1999. Lym- mediated inflammatory pathway in osteoarthritis. Osteoarthritis 7:401. by guest on September 26, 2021