For our understanding of the pathogenesis of rheumatoid arthritis (RA), it is important to elucid... more For our understanding of the pathogenesis of rheumatoid arthritis (RA), it is important to elucidate the mechanisms underlying early stages of synovitis. Here, synovial cytokine production was investigated in patients with very early arthritis. Synovial biopsies were obtained from patients with at least one clinically swollen joint within 12 weeks of symptom onset. At an 18-month follow-up visit, patients who went on to develop RA, or whose arthritis spontaneously resolved, were identified. Biopsies were also obtained from patients with RA with longer symptom duration (>12 weeks) and individuals with no clinically apparent inflammation. Synovial mRNA expression of 117 cytokines was quantified using PCR techniques and analysed using standard and novel methods of data analysis. Synovial tissue sections were stained for CXCL4, CXCL7, CD41, CD68 and von Willebrand factor. A machine learning approach identified expression of mRNA for CXCL4 and CXCL7 as potentially important in the cla...
Immunotherapy using short immunogenic peptides of disease-related autoantigens restores immune to... more Immunotherapy using short immunogenic peptides of disease-related autoantigens restores immune tolerance in preclinical disease models. We studied safety and mechanistic effects of injecting human leukocyte antigen-DR4(DRB1*0401)-restricted immunodominant proinsulin peptide intradermally every 2 or 4 weeks for 6 months in newly diagnosed type 1 diabetes patients. Treatment was well tolerated with no systemic or local hypersensitivity. Placebo subjects showed a significant decline in stimulated C-peptide (measuring insulin reserve) at 3, 6, 9, and 12 months versus baseline, whereas no significant change was seen in the 4-weekly peptide group at these time points or the 2-weekly group at 3, 6, and 9 months. The placebo group's daily insulin use increased by 50% over 12 months but remained unchanged in the intervention groups. C-peptide retention in treated subjects was associated with proinsulin-stimulated interleukin-10 production, increased FoxP3 expression by regulatory T cells...
This manuscript is a companion paper to Amara et al. [1]. Data shown here include detailed clinic... more This manuscript is a companion paper to Amara et al. [1]. Data shown here include detailed clinical characteristics from anonymized patients, the Ig subclass data generated from B cells sorted from four individual patients, tables detailing variable gene region sequences from sorted cells linked to the patient information and the sequence yields from individual patients. Furthermore a URL link to the RNAseq datasets submitted to GEO is included.
The clinical efficacy of B cell targeting therapies highlights the pathogenic potential of B cell... more The clinical efficacy of B cell targeting therapies highlights the pathogenic potential of B cells in inflammatory diseases. Expression of Fc Receptor like 4 (FcRL4) identifies a memory B cell subset, which is enriched in the joints of patients with rheumatoid arthritis (RA) and in mucosa-associated lymphoid tissue. The high level of RANKL production by this B cell subset indicates a unique pathogenic role. In addition, recent work has identified a role for FcRL4 as an IgA receptor, suggesting a potential function in mucosal immunity. Here, the contribution of FcRL4+ B cells to the specific autoimmune response in the joints of patients with RA was investigated. Single FcRL4+ and FcRL4- B cells were sorted from synovial fluid and tissue from RA patients and their immunoglobulin genes characterized. Levels of hypermutation in the variable regions in both populations were largely consistent with memory B cells selected by an antigen- and T cell-dependent process. Recombinant antibodies...
Synovial fibroblasts play a key role in joint destruction and regulation of the inflammatory infi... more Synovial fibroblasts play a key role in joint destruction and regulation of the inflammatory infiltrate in established rheumatoid arthritis (RA). The mechanisms by which this occurs in the earliest stages of RA are largely unknown. We investigated the role of Dickkopf-related protein 1 (DKK1) produced by synovial fibroblasts of patients with very early rheumatoid arthritis (VeRA). Fibroblasts were isolated from the disease-modifying anti-rheumatic drug-naive Birmingham early arthritis cohort of patients with new onset of clinically apparent arthritis and inflammatory symptoms of ≤12 weeks' duration, who at follow-up had either resolving arthritis or RA. Endothelial fibroblast co-cultures were formed using porous filters, and lymphocyte adhesion to co-cultures was assessed using phase-contrast microscopy. DKK1 gene expression and secretion were quantified by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Synovial fibroblasts from patie...
Background The importance of the B cell lineage in RA pathogenesis is highlighted by the clinical... more Background The importance of the B cell lineage in RA pathogenesis is highlighted by the clinical effectiveness of rituximab. Potential mechanisms by which B cells drive disease processes in RA include their ability to produce autoantibodies, act as antigen-presenting cells, or secrete cytokines. Our recent findings suggest a pro-inflammatory and destructive role for this cell type via production of cytokines including RANKL, TNF-α and IL-6 [1]. A memory B cell subset normally resident in epithelial niches which is characterised by expression of the inhibitory receptor FcRL4 has previously been described to express RANKL [2]. Objectives As we previously made the observation that B cells in the RA synovium produce RANKL, we sought to determine whether RANKL-expressing cells belonged to the FcRL4+ B cell subset. Methods Synovial fluid, peripheral blood and synovial tissue samples were obtained from patients fulfilling 1987 ACR criteria for RA. Mononuclear cells were isolated from synovial fluid and peripheral blood by density gradient centrifugation, stained with antibodies against CD19, CD20, CD27, IgD, CD11c, RANKL, FcRL4, CD95 and CD21, and analysed by flow cytometry. Mononuclear synovial fluid cells were stained with antibodies against CD19 and FcRL4 and sorted using a Mo-Flo™ cell sorter. Taqman™ microfluidic cards were used to detect mRNA expression of 48 cytokines by real-time PCR in these samples. Immunofluorescence staining was performed on 5 µm frozen tissue sections with antibodies against CD20, RANKL and FcRL4. Results FcRL4-expressing B cells, comprising 4-22% of the B cell population, were detected in the synovial fluid at a significantly higher frequency compared to matched peripheral blood (n=12, p=0.0004). RANKL was significantly enriched in the FcRL4+ B cell population compared to the FcRL4- population (p=0.006). Both FcRL4+ B cells and RANKL+ B cells were predominantly switched IgD-CD27+/- memory B cells, expressing significantly higher levels of CD11c and CD95, and lower CD21 levels than the FcRL4- or the RANKL- population. FcRL4+ B cells expressing RANKL were also present in synovial tissue (n=4). At the mRNA level, sorted FcRL4+ B cells were found to express considerably higher levels of both RANKL and TNF compared to the FcRL4- fraction (n=5). Conclusions We have identified a novel subset of B cells in the RA synovium which is characterised by expression of FcRL4 and has not previously been described in RA or any other chronic inflammatory joint condition. FcRL4+ B cells produce RANKL, and express high levels of TNF mRNA, indicating a destructive, pro-inflammatory role for this B cell subpopulation in RA pathogenesis. References Yeo L, Toellner KM, Salmon M, Filer A, Buckley CD, Raza K, Scheel-Toellner D: Cytokine mRNA profiling identifies B cells as a major source of RANKL in rheumatoid arthritis. Ann Rheum Dis 2011, 70(11):2022-2028. Ehrhardt GR, Hsu JT, Gartland L, Leu CM, Zhang S, Davis RS, Cooper MD: Expression of the immunoregulatory molecule FcRH4 defines a distinctive tissue-based population of memory B cells. J Exp Med 2005, 202(6):783-791. Acknowledgements The University of Birmingham has filed a patent on the subject of this work. Disclosure of Interest None Declared
Background In the earliest clinically apparent phases of arthritis, it is difficult to distinguis... more Background In the earliest clinically apparent phases of arthritis, it is difficult to distinguish patients who will develop RA from those whose disease will resolve. We hypothesised that synovial tissue cytokine expression would differ between early patients with these distinct outcomes. As control groups we included patients with newly presenting established RA and individuals with no evidence of inflammatory arthritis. Methods Synovial tissue biopsies were obtained from 18 early synovitis patients who eventually developed RA (according to 1987 ACR criteria within 18 month follow-up) and 13 early synovitis patients with self-limiting disease; all samples were collected within 12 weeks of the onset of any musculoskeletal symptom attributed to inflammatory arthritis. In addition, 12 patients with established RA (according to 1987 ACR criteria with a symptom duration of >3 months at sample collection), and 12 control subjects undergoing arthroscopy with no macroscopic or radiological evidence of inflammatory pathology were studied. No individuals were taking DMARD therapy at the time of sample collection. mRNA expression of 135 cytokines and related molecules (including interleukins, TNF family members, chemokines, growth factors, interferons and adipokines) was determined by real-time PCR using microfluidic cards. The Kruskal-Wallis test and Dunn’s post-test were used for comparison of 4 groups, and the Mann-Whitney test was used for paired analysis. Results Of the 135 cytokines analysed, 20 genes were differentially expressed between the 4 groups (early resolving synovitis, early RA, established RA and controls), 16 genes were not detected, and 99 genes did not show statistically significant differences between groups. Significant differences included higher levels of IFN-b in established RA compared to early RA. Levels of IL-1a, IL-2 and IL-21 were significantly higher in established RA than early resolving synovitis patients. Conversely, XCL1 and VEGFa were higher in early resolving synovitis patients than established RA. The levels of many cytokines and chemokines were elevated in the synovium of early and established arthritis patients compared to control subjects. 11 genes were significantly different between ACPA-positive and ACPA-negative RA patients. Conclusions The profile of cytokine mRNA expression in patients with synovitis is consistently different to that of uninflamed synovial tissue. Although significant differences exist between cytokine expression levels in different outcome groups and time points in RA disease, for the majority of cytokines studied, expression is similar between groups and may reflect the extent of inflammation rather than showing specificity for its outcome in early arthritis. Disclosure of Interest None Declared
Interleukin-6 (IL-6) is a proinflammatory cytokine with regulatory effects on the survival and di... more Interleukin-6 (IL-6) is a proinflammatory cytokine with regulatory effects on the survival and differentiation of T cells. It exerts its biologic function in 2 ways: by directly binding to the IL-6 receptor (IL-6R; CD126) or via trans-signaling, in which soluble IL-6R/IL-6 complexes bind to the signaling component CD130. This study was undertaken to assess the expression and regulation of CD126 and CD130 and determine how these affect the response of CD4+ T cells to IL-6 in the joints of patients with rheumatoid arthritis (RA). Flow cytometry and immunofluorescence microscopy were used to determine the expression, function, and regulation of CD126 and CD130 in CD4+ T cells from the peripheral blood (PB), synovial fluid (SF), and synovial tissue of RA patients. Compared to the findings in RA PB, CD4+ T cells in the SF and synovial tissue expressed low levels of CD126. In contrast, whereas CD4+ T cell expression of CD130 was minimal in the SF, its level in the synovial tissue was high. Consistent with this phenotype, synovial tissue T cells responded to trans-signaling by soluble IL-6R/IL-6 complexes, whereas no response was evident in CD4+ T cells from the SF. Down-regulation of both receptor components in SF T cells could be explained by exposure to high levels of IL-6. Increased levels of CD130 messenger RNA and protein in synovial tissue CD4+ T cells suggested that CD130 is up-regulated locally. Among a range of cytokines tested, only IL-10 induced CD130 expression in T cells. The inflamed microenvironment in the synovial tissue maintains responsiveness to IL-6 trans-signaling through the up-regulation of CD130 expression in CD4+ T cells, and this process may be driven by IL-10.
ObjectivesTo examine how rituximab may result in the inhibition of joint destruction in rheumatoi... more ObjectivesTo examine how rituximab may result in the inhibition of joint destruction in rheumatoid arthritis (RA) patients.MethodsTwenty-eight patients with active RA were treated with rituximab. Radiographs of hands and feet before and 1 year after therapy were assessed using the Sharp–van der Heijde score (SHS). Expression of bone destruction markers was evaluated by immunohistochemistry and immunofluorescence of synovial biopsies obtained before and 16 weeks after the initiation of treatment. Serum levels of osteoprotegerin, receptor activator of nuclear factor κB ligand (RANKL), osteocalcin and cross-linked N-telopeptides of type I collagen (NTx) were measured by ELISA before and 16 weeks post-treatment.ResultsAfter 1 year, the mean (SD) change in total SHS was 1.4 (10.0). Sixteen weeks after treatment there was a decrease of 99% in receptor activator of nuclear factor κB-positive osteoclast precursors (p=0.02) and a decrease of 37% (p=0.016) in RANKL expression in the synovium ...
Summary In type 1 diabetes (T1D), autoreactive cytotoxic CD8+ T cells are implicated in the destr... more Summary In type 1 diabetes (T1D), autoreactive cytotoxic CD8+ T cells are implicated in the destruction of insulin-producing β cells. The HLA-B*3906 and HLA-A*2402 class I genes confer increased risk and promote early disease onset, suggesting that CD8+ T cells that recognize peptides presented by these class I molecules on pancreatic β cells play a pivotal role in the autoimmune response. We examined the frequency and phenotype of circulating preproinsulin (PPI)-specific and insulin B (InsB)-specific CD8+ T cells in HLA-B*3906+ children newly diagnosed with T1D and in high-risk HLA-A*2402+ children before the appearance of disease-specific autoantibodies and before diagnosis of T1D. Antigen-specific CD8+ T cells were detected using human leucocyte antigen (HLA) class I tetramers and flow cytometry was used to assess memory status. In HLA-B*3906+ children with T1D, we observed an increase in PPI5–12-specific transitional memory CD8+ T cells compared to non-diabetic, age- and HLA-mat...
For our understanding of the pathogenesis of rheumatoid arthritis (RA), it is important to elucid... more For our understanding of the pathogenesis of rheumatoid arthritis (RA), it is important to elucidate the mechanisms underlying early stages of synovitis. Here, synovial cytokine production was investigated in patients with very early arthritis. Synovial biopsies were obtained from patients with at least one clinically swollen joint within 12 weeks of symptom onset. At an 18-month follow-up visit, patients who went on to develop RA, or whose arthritis spontaneously resolved, were identified. Biopsies were also obtained from patients with RA with longer symptom duration (>12 weeks) and individuals with no clinically apparent inflammation. Synovial mRNA expression of 117 cytokines was quantified using PCR techniques and analysed using standard and novel methods of data analysis. Synovial tissue sections were stained for CXCL4, CXCL7, CD41, CD68 and von Willebrand factor. A machine learning approach identified expression of mRNA for CXCL4 and CXCL7 as potentially important in the cla...
Immunotherapy using short immunogenic peptides of disease-related autoantigens restores immune to... more Immunotherapy using short immunogenic peptides of disease-related autoantigens restores immune tolerance in preclinical disease models. We studied safety and mechanistic effects of injecting human leukocyte antigen-DR4(DRB1*0401)-restricted immunodominant proinsulin peptide intradermally every 2 or 4 weeks for 6 months in newly diagnosed type 1 diabetes patients. Treatment was well tolerated with no systemic or local hypersensitivity. Placebo subjects showed a significant decline in stimulated C-peptide (measuring insulin reserve) at 3, 6, 9, and 12 months versus baseline, whereas no significant change was seen in the 4-weekly peptide group at these time points or the 2-weekly group at 3, 6, and 9 months. The placebo group's daily insulin use increased by 50% over 12 months but remained unchanged in the intervention groups. C-peptide retention in treated subjects was associated with proinsulin-stimulated interleukin-10 production, increased FoxP3 expression by regulatory T cells...
This manuscript is a companion paper to Amara et al. [1]. Data shown here include detailed clinic... more This manuscript is a companion paper to Amara et al. [1]. Data shown here include detailed clinical characteristics from anonymized patients, the Ig subclass data generated from B cells sorted from four individual patients, tables detailing variable gene region sequences from sorted cells linked to the patient information and the sequence yields from individual patients. Furthermore a URL link to the RNAseq datasets submitted to GEO is included.
The clinical efficacy of B cell targeting therapies highlights the pathogenic potential of B cell... more The clinical efficacy of B cell targeting therapies highlights the pathogenic potential of B cells in inflammatory diseases. Expression of Fc Receptor like 4 (FcRL4) identifies a memory B cell subset, which is enriched in the joints of patients with rheumatoid arthritis (RA) and in mucosa-associated lymphoid tissue. The high level of RANKL production by this B cell subset indicates a unique pathogenic role. In addition, recent work has identified a role for FcRL4 as an IgA receptor, suggesting a potential function in mucosal immunity. Here, the contribution of FcRL4+ B cells to the specific autoimmune response in the joints of patients with RA was investigated. Single FcRL4+ and FcRL4- B cells were sorted from synovial fluid and tissue from RA patients and their immunoglobulin genes characterized. Levels of hypermutation in the variable regions in both populations were largely consistent with memory B cells selected by an antigen- and T cell-dependent process. Recombinant antibodies...
Synovial fibroblasts play a key role in joint destruction and regulation of the inflammatory infi... more Synovial fibroblasts play a key role in joint destruction and regulation of the inflammatory infiltrate in established rheumatoid arthritis (RA). The mechanisms by which this occurs in the earliest stages of RA are largely unknown. We investigated the role of Dickkopf-related protein 1 (DKK1) produced by synovial fibroblasts of patients with very early rheumatoid arthritis (VeRA). Fibroblasts were isolated from the disease-modifying anti-rheumatic drug-naive Birmingham early arthritis cohort of patients with new onset of clinically apparent arthritis and inflammatory symptoms of ≤12 weeks' duration, who at follow-up had either resolving arthritis or RA. Endothelial fibroblast co-cultures were formed using porous filters, and lymphocyte adhesion to co-cultures was assessed using phase-contrast microscopy. DKK1 gene expression and secretion were quantified by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Synovial fibroblasts from patie...
Background The importance of the B cell lineage in RA pathogenesis is highlighted by the clinical... more Background The importance of the B cell lineage in RA pathogenesis is highlighted by the clinical effectiveness of rituximab. Potential mechanisms by which B cells drive disease processes in RA include their ability to produce autoantibodies, act as antigen-presenting cells, or secrete cytokines. Our recent findings suggest a pro-inflammatory and destructive role for this cell type via production of cytokines including RANKL, TNF-α and IL-6 [1]. A memory B cell subset normally resident in epithelial niches which is characterised by expression of the inhibitory receptor FcRL4 has previously been described to express RANKL [2]. Objectives As we previously made the observation that B cells in the RA synovium produce RANKL, we sought to determine whether RANKL-expressing cells belonged to the FcRL4+ B cell subset. Methods Synovial fluid, peripheral blood and synovial tissue samples were obtained from patients fulfilling 1987 ACR criteria for RA. Mononuclear cells were isolated from synovial fluid and peripheral blood by density gradient centrifugation, stained with antibodies against CD19, CD20, CD27, IgD, CD11c, RANKL, FcRL4, CD95 and CD21, and analysed by flow cytometry. Mononuclear synovial fluid cells were stained with antibodies against CD19 and FcRL4 and sorted using a Mo-Flo™ cell sorter. Taqman™ microfluidic cards were used to detect mRNA expression of 48 cytokines by real-time PCR in these samples. Immunofluorescence staining was performed on 5 µm frozen tissue sections with antibodies against CD20, RANKL and FcRL4. Results FcRL4-expressing B cells, comprising 4-22% of the B cell population, were detected in the synovial fluid at a significantly higher frequency compared to matched peripheral blood (n=12, p=0.0004). RANKL was significantly enriched in the FcRL4+ B cell population compared to the FcRL4- population (p=0.006). Both FcRL4+ B cells and RANKL+ B cells were predominantly switched IgD-CD27+/- memory B cells, expressing significantly higher levels of CD11c and CD95, and lower CD21 levels than the FcRL4- or the RANKL- population. FcRL4+ B cells expressing RANKL were also present in synovial tissue (n=4). At the mRNA level, sorted FcRL4+ B cells were found to express considerably higher levels of both RANKL and TNF compared to the FcRL4- fraction (n=5). Conclusions We have identified a novel subset of B cells in the RA synovium which is characterised by expression of FcRL4 and has not previously been described in RA or any other chronic inflammatory joint condition. FcRL4+ B cells produce RANKL, and express high levels of TNF mRNA, indicating a destructive, pro-inflammatory role for this B cell subpopulation in RA pathogenesis. References Yeo L, Toellner KM, Salmon M, Filer A, Buckley CD, Raza K, Scheel-Toellner D: Cytokine mRNA profiling identifies B cells as a major source of RANKL in rheumatoid arthritis. Ann Rheum Dis 2011, 70(11):2022-2028. Ehrhardt GR, Hsu JT, Gartland L, Leu CM, Zhang S, Davis RS, Cooper MD: Expression of the immunoregulatory molecule FcRH4 defines a distinctive tissue-based population of memory B cells. J Exp Med 2005, 202(6):783-791. Acknowledgements The University of Birmingham has filed a patent on the subject of this work. Disclosure of Interest None Declared
Background In the earliest clinically apparent phases of arthritis, it is difficult to distinguis... more Background In the earliest clinically apparent phases of arthritis, it is difficult to distinguish patients who will develop RA from those whose disease will resolve. We hypothesised that synovial tissue cytokine expression would differ between early patients with these distinct outcomes. As control groups we included patients with newly presenting established RA and individuals with no evidence of inflammatory arthritis. Methods Synovial tissue biopsies were obtained from 18 early synovitis patients who eventually developed RA (according to 1987 ACR criteria within 18 month follow-up) and 13 early synovitis patients with self-limiting disease; all samples were collected within 12 weeks of the onset of any musculoskeletal symptom attributed to inflammatory arthritis. In addition, 12 patients with established RA (according to 1987 ACR criteria with a symptom duration of >3 months at sample collection), and 12 control subjects undergoing arthroscopy with no macroscopic or radiological evidence of inflammatory pathology were studied. No individuals were taking DMARD therapy at the time of sample collection. mRNA expression of 135 cytokines and related molecules (including interleukins, TNF family members, chemokines, growth factors, interferons and adipokines) was determined by real-time PCR using microfluidic cards. The Kruskal-Wallis test and Dunn’s post-test were used for comparison of 4 groups, and the Mann-Whitney test was used for paired analysis. Results Of the 135 cytokines analysed, 20 genes were differentially expressed between the 4 groups (early resolving synovitis, early RA, established RA and controls), 16 genes were not detected, and 99 genes did not show statistically significant differences between groups. Significant differences included higher levels of IFN-b in established RA compared to early RA. Levels of IL-1a, IL-2 and IL-21 were significantly higher in established RA than early resolving synovitis patients. Conversely, XCL1 and VEGFa were higher in early resolving synovitis patients than established RA. The levels of many cytokines and chemokines were elevated in the synovium of early and established arthritis patients compared to control subjects. 11 genes were significantly different between ACPA-positive and ACPA-negative RA patients. Conclusions The profile of cytokine mRNA expression in patients with synovitis is consistently different to that of uninflamed synovial tissue. Although significant differences exist between cytokine expression levels in different outcome groups and time points in RA disease, for the majority of cytokines studied, expression is similar between groups and may reflect the extent of inflammation rather than showing specificity for its outcome in early arthritis. Disclosure of Interest None Declared
Interleukin-6 (IL-6) is a proinflammatory cytokine with regulatory effects on the survival and di... more Interleukin-6 (IL-6) is a proinflammatory cytokine with regulatory effects on the survival and differentiation of T cells. It exerts its biologic function in 2 ways: by directly binding to the IL-6 receptor (IL-6R; CD126) or via trans-signaling, in which soluble IL-6R/IL-6 complexes bind to the signaling component CD130. This study was undertaken to assess the expression and regulation of CD126 and CD130 and determine how these affect the response of CD4+ T cells to IL-6 in the joints of patients with rheumatoid arthritis (RA). Flow cytometry and immunofluorescence microscopy were used to determine the expression, function, and regulation of CD126 and CD130 in CD4+ T cells from the peripheral blood (PB), synovial fluid (SF), and synovial tissue of RA patients. Compared to the findings in RA PB, CD4+ T cells in the SF and synovial tissue expressed low levels of CD126. In contrast, whereas CD4+ T cell expression of CD130 was minimal in the SF, its level in the synovial tissue was high. Consistent with this phenotype, synovial tissue T cells responded to trans-signaling by soluble IL-6R/IL-6 complexes, whereas no response was evident in CD4+ T cells from the SF. Down-regulation of both receptor components in SF T cells could be explained by exposure to high levels of IL-6. Increased levels of CD130 messenger RNA and protein in synovial tissue CD4+ T cells suggested that CD130 is up-regulated locally. Among a range of cytokines tested, only IL-10 induced CD130 expression in T cells. The inflamed microenvironment in the synovial tissue maintains responsiveness to IL-6 trans-signaling through the up-regulation of CD130 expression in CD4+ T cells, and this process may be driven by IL-10.
ObjectivesTo examine how rituximab may result in the inhibition of joint destruction in rheumatoi... more ObjectivesTo examine how rituximab may result in the inhibition of joint destruction in rheumatoid arthritis (RA) patients.MethodsTwenty-eight patients with active RA were treated with rituximab. Radiographs of hands and feet before and 1 year after therapy were assessed using the Sharp–van der Heijde score (SHS). Expression of bone destruction markers was evaluated by immunohistochemistry and immunofluorescence of synovial biopsies obtained before and 16 weeks after the initiation of treatment. Serum levels of osteoprotegerin, receptor activator of nuclear factor κB ligand (RANKL), osteocalcin and cross-linked N-telopeptides of type I collagen (NTx) were measured by ELISA before and 16 weeks post-treatment.ResultsAfter 1 year, the mean (SD) change in total SHS was 1.4 (10.0). Sixteen weeks after treatment there was a decrease of 99% in receptor activator of nuclear factor κB-positive osteoclast precursors (p=0.02) and a decrease of 37% (p=0.016) in RANKL expression in the synovium ...
Summary In type 1 diabetes (T1D), autoreactive cytotoxic CD8+ T cells are implicated in the destr... more Summary In type 1 diabetes (T1D), autoreactive cytotoxic CD8+ T cells are implicated in the destruction of insulin-producing β cells. The HLA-B*3906 and HLA-A*2402 class I genes confer increased risk and promote early disease onset, suggesting that CD8+ T cells that recognize peptides presented by these class I molecules on pancreatic β cells play a pivotal role in the autoimmune response. We examined the frequency and phenotype of circulating preproinsulin (PPI)-specific and insulin B (InsB)-specific CD8+ T cells in HLA-B*3906+ children newly diagnosed with T1D and in high-risk HLA-A*2402+ children before the appearance of disease-specific autoantibodies and before diagnosis of T1D. Antigen-specific CD8+ T cells were detected using human leucocyte antigen (HLA) class I tetramers and flow cytometry was used to assess memory status. In HLA-B*3906+ children with T1D, we observed an increase in PPI5–12-specific transitional memory CD8+ T cells compared to non-diabetic, age- and HLA-mat...
Uploads