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CD8 T cells are required for the formation of ectopic germinal centers in rheumatoid synovitis.

Kang YM, Zhang X, Wagner UG, Yang H, Beckenbaugh RD, Kurtin PJ, Goronzy JJ, Weyand CM - J. Exp. Med. (2002)

Bottom Line: In the absence of CD8 T cells, follicular dendritic cells disappeared, production of lymphotoxin-alpha1beta2 markedly decreased, and immunoglobulin (Ig) secretion ceased.Perifollicular IFN-gamma+ CD8 T cells were rare in secondary lymphoid tissues but accounted for the majority of IFN-gamma+ cells in synovial infiltrates.We propose that CD8+ T cells regulate the structural integrity and functional activity of GCs in ectopic lymphoid follicles.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
The assembly of inflammatory lesions in rheumatoid arthritis is highly regulated and typically leads to the formation of lymphoid follicles with germinal center (GC) reactions. We used microdissection of such extranodal follicles to analyze the colonizing T cells. Although the repertoire of follicular T cells was diverse, a subset of T cell receptor (TCR) sequences was detected in multiple independent follicles and not in interfollicular zones, suggesting recognition of a common antigen. Unexpectedly, the majority of shared TCR sequences were from CD8 T cells that were highly enriched in the synovium and present in low numbers in the periphery. To examine their role in extranodal GC reactions, CD8 T cells were depleted in human synovium-SCID mouse chimeras. Depletion of synovial CD8 T cells caused disintegration of the GC-containing follicles. In the absence of CD8 T cells, follicular dendritic cells disappeared, production of lymphotoxin-alpha1beta2 markedly decreased, and immunoglobulin (Ig) secretion ceased. Immunohistochemical studies demonstrated that these CD8 T cells accumulated at the edge of the mantle zone. Besides their unique localization, they were characterized by the production of interferon (IFN)-gamma, lack of the pore-forming enzyme perforin, and expression of CD40 ligand. Perifollicular IFN-gamma+ CD8 T cells were rare in secondary lymphoid tissues but accounted for the majority of IFN-gamma+ cells in synovial infiltrates. We propose that CD8+ T cells regulate the structural integrity and functional activity of GCs in ectopic lymphoid follicles.

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Follicular TCR β-chain sequences preferentially derive from CD8 T cells. Synovial tissue was selected from five patients with RA who had multiple ectopic GCs in the synovial membrane. (A) The distribution of CD4 T cells (left), CD20 B cells (middle), and IgD+ cells (right) is shown in a representative example. The mantle zone is indicated by an asterisk (right). Follicles were microdissected and TCR β-chain sequences were obtained (Tables II and III). In parallel, synovial tissue and peripheral CD4 and CD8 cells were purified by FACS®. cDNA was tested for the presence of these TCR β-chain sequences by PCR-ELISA using N-D-N–specific oligonucleotides as probes. (B) In four of the five patients, the majority of the TCR β-chain sequences were detected in the CD8, and not the CD4, population. (C) To define the localization of the CD8 cells in synovial follicles, tissue sections were stained with anti-CD8 (brown) and anti-CD23 (red, expressed on follicular dendritic cells) mAb. CD8 T cells were found in the perifollicular zone, sometimes within the mantle zone, and only occasionally in the GC. Original magnifications: (left) × 100 and (right) × 400.
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fig1: Follicular TCR β-chain sequences preferentially derive from CD8 T cells. Synovial tissue was selected from five patients with RA who had multiple ectopic GCs in the synovial membrane. (A) The distribution of CD4 T cells (left), CD20 B cells (middle), and IgD+ cells (right) is shown in a representative example. The mantle zone is indicated by an asterisk (right). Follicles were microdissected and TCR β-chain sequences were obtained (Tables II and III). In parallel, synovial tissue and peripheral CD4 and CD8 cells were purified by FACS®. cDNA was tested for the presence of these TCR β-chain sequences by PCR-ELISA using N-D-N–specific oligonucleotides as probes. (B) In four of the five patients, the majority of the TCR β-chain sequences were detected in the CD8, and not the CD4, population. (C) To define the localization of the CD8 cells in synovial follicles, tissue sections were stained with anti-CD8 (brown) and anti-CD23 (red, expressed on follicular dendritic cells) mAb. CD8 T cells were found in the perifollicular zone, sometimes within the mantle zone, and only occasionally in the GC. Original magnifications: (left) × 100 and (right) × 400.

Mentions: Synovial tissue samples were collected from a series of 81 patients undergoing synovectomy or arthroplasty. In a subset of 21 patients (Table I), lymphoid aggregates with follicular dendritic cells and characteristic cellular compartmentalization were identified as ectopic GCs (Fig. 1 A); five of these patients were selected for microdissection analysis. All of these patients had multiple GCs in the synovial tissue, while T cell–B cell aggregates without follicular dendritic cells were not detected. In each of these tissues, T cell–B cell clusters were isolated and cDNA was amplified with TCR-BV– and -BC–specific primers. Amplified products from three such clusters from each patient were cloned and sequenced. In all patients and in all dissected follicles, the TCR repertoire in the follicle was diverse. A typical example of the diversity of TCR sequences isolated from three distinct follicles of patient 624 is given in Table II. For each BV family in each of the follicles, 9–15 TCR β-chains were sequenced. The vast majority of the sequences were unique; only few of the TCR sequences were seen repeatedly.


CD8 T cells are required for the formation of ectopic germinal centers in rheumatoid synovitis.

Kang YM, Zhang X, Wagner UG, Yang H, Beckenbaugh RD, Kurtin PJ, Goronzy JJ, Weyand CM - J. Exp. Med. (2002)

Follicular TCR β-chain sequences preferentially derive from CD8 T cells. Synovial tissue was selected from five patients with RA who had multiple ectopic GCs in the synovial membrane. (A) The distribution of CD4 T cells (left), CD20 B cells (middle), and IgD+ cells (right) is shown in a representative example. The mantle zone is indicated by an asterisk (right). Follicles were microdissected and TCR β-chain sequences were obtained (Tables II and III). In parallel, synovial tissue and peripheral CD4 and CD8 cells were purified by FACS®. cDNA was tested for the presence of these TCR β-chain sequences by PCR-ELISA using N-D-N–specific oligonucleotides as probes. (B) In four of the five patients, the majority of the TCR β-chain sequences were detected in the CD8, and not the CD4, population. (C) To define the localization of the CD8 cells in synovial follicles, tissue sections were stained with anti-CD8 (brown) and anti-CD23 (red, expressed on follicular dendritic cells) mAb. CD8 T cells were found in the perifollicular zone, sometimes within the mantle zone, and only occasionally in the GC. Original magnifications: (left) × 100 and (right) × 400.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2193749&req=5

fig1: Follicular TCR β-chain sequences preferentially derive from CD8 T cells. Synovial tissue was selected from five patients with RA who had multiple ectopic GCs in the synovial membrane. (A) The distribution of CD4 T cells (left), CD20 B cells (middle), and IgD+ cells (right) is shown in a representative example. The mantle zone is indicated by an asterisk (right). Follicles were microdissected and TCR β-chain sequences were obtained (Tables II and III). In parallel, synovial tissue and peripheral CD4 and CD8 cells were purified by FACS®. cDNA was tested for the presence of these TCR β-chain sequences by PCR-ELISA using N-D-N–specific oligonucleotides as probes. (B) In four of the five patients, the majority of the TCR β-chain sequences were detected in the CD8, and not the CD4, population. (C) To define the localization of the CD8 cells in synovial follicles, tissue sections were stained with anti-CD8 (brown) and anti-CD23 (red, expressed on follicular dendritic cells) mAb. CD8 T cells were found in the perifollicular zone, sometimes within the mantle zone, and only occasionally in the GC. Original magnifications: (left) × 100 and (right) × 400.
Mentions: Synovial tissue samples were collected from a series of 81 patients undergoing synovectomy or arthroplasty. In a subset of 21 patients (Table I), lymphoid aggregates with follicular dendritic cells and characteristic cellular compartmentalization were identified as ectopic GCs (Fig. 1 A); five of these patients were selected for microdissection analysis. All of these patients had multiple GCs in the synovial tissue, while T cell–B cell aggregates without follicular dendritic cells were not detected. In each of these tissues, T cell–B cell clusters were isolated and cDNA was amplified with TCR-BV– and -BC–specific primers. Amplified products from three such clusters from each patient were cloned and sequenced. In all patients and in all dissected follicles, the TCR repertoire in the follicle was diverse. A typical example of the diversity of TCR sequences isolated from three distinct follicles of patient 624 is given in Table II. For each BV family in each of the follicles, 9–15 TCR β-chains were sequenced. The vast majority of the sequences were unique; only few of the TCR sequences were seen repeatedly.

Bottom Line: In the absence of CD8 T cells, follicular dendritic cells disappeared, production of lymphotoxin-alpha1beta2 markedly decreased, and immunoglobulin (Ig) secretion ceased.Perifollicular IFN-gamma+ CD8 T cells were rare in secondary lymphoid tissues but accounted for the majority of IFN-gamma+ cells in synovial infiltrates.We propose that CD8+ T cells regulate the structural integrity and functional activity of GCs in ectopic lymphoid follicles.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
The assembly of inflammatory lesions in rheumatoid arthritis is highly regulated and typically leads to the formation of lymphoid follicles with germinal center (GC) reactions. We used microdissection of such extranodal follicles to analyze the colonizing T cells. Although the repertoire of follicular T cells was diverse, a subset of T cell receptor (TCR) sequences was detected in multiple independent follicles and not in interfollicular zones, suggesting recognition of a common antigen. Unexpectedly, the majority of shared TCR sequences were from CD8 T cells that were highly enriched in the synovium and present in low numbers in the periphery. To examine their role in extranodal GC reactions, CD8 T cells were depleted in human synovium-SCID mouse chimeras. Depletion of synovial CD8 T cells caused disintegration of the GC-containing follicles. In the absence of CD8 T cells, follicular dendritic cells disappeared, production of lymphotoxin-alpha1beta2 markedly decreased, and immunoglobulin (Ig) secretion ceased. Immunohistochemical studies demonstrated that these CD8 T cells accumulated at the edge of the mantle zone. Besides their unique localization, they were characterized by the production of interferon (IFN)-gamma, lack of the pore-forming enzyme perforin, and expression of CD40 ligand. Perifollicular IFN-gamma+ CD8 T cells were rare in secondary lymphoid tissues but accounted for the majority of IFN-gamma+ cells in synovial infiltrates. We propose that CD8+ T cells regulate the structural integrity and functional activity of GCs in ectopic lymphoid follicles.

Show MeSH
Related in: MedlinePlus