bluuebunny/crossref_metadata_2025_split huggingface dataset:
Achievement of a synergistic adjuvant effect on arthritis induction by activation of innate immunity and forcing the immune response toward the Th1 phenotype
Authors: Hanyecz Anita , Berlo Suzanne E. , Szántó Sándor , Broeren Chris P. M. , Mikecz Katalin , Glant Tibor T. 
Published: May 2004

AbstractObjectiveTo apply and analyze the mechanisms of action of dimethyldioctadecylammonium bromide (DDA), a powerful adjuvant that does not have the side effects of the conventionally used Freund's adjuvants, in proteoglycan‐induced arthritis (PGIA) and collagen‐induced arthritis (CIA).MethodsPGIA and CIA were generated using standard immunization protocols with cartilage proteoglycan aggrecan (PG) or human type II collagen (CII) emulsified with Freund's complete adjuvant (CFA), and compared with PGIA and CIA generated using immunization protocols in which the same antigens were used in combination with the adjuvant DDA. Immune responses to immunizing and self PGs and CII, and the incidence, severity, and onset of arthritis were monitored throughout the experiments. In addition, a new, inexpensive, and powerful method of inducing arthritis using crude cartilage extracts is described.ResultsA significantly reduced onset period and a more severe arthritis were achieved in BALB/c mice immunized with cartilage PGs in DDA. PGs from bovine, ovine, and porcine cartilage, which otherwise have no effect or have only a subarthritogenic effect, and crude extracts of human osteoarthritic cartilage induced a 100% incidence with a very high arthritis score in BALB/c mice. The overall immune responses to either CII or PG were similar in antigen/CFA‐immunized and antigen/DDA‐immunized animals, but the Th1/Th2 balance shifted significantly toward a Th1 bias in DDA‐injected animals with either PGIA or CIA.ConclusionDDA, which was first used in autoimmune models, is a potent nonirritant adjuvant, which eliminates all undesired side effects of the Freund's adjuvants. DDA exerts a strong stimulatory effect via the activation of nonspecific (innate) immunity and forces the immune regulation toward Th1 dominance. These lines of evidence also suggest the possibility that seemingly innocuous compounds may exert an adjuvant effect in humans and may create the pathophysiologic basis of autoimmunity in susceptible individuals via the activation/stimulation of innate immunity.
Antiangiogenic effects of anti–tumor necrosis factor α therapy with infliximab in psoriatic arthritis
Authors: Cañete Juan D. , Pablos José L. , Sanmartí Raimon , Mallofré Carmen , Marsal Sara , Maymó´ Joan , Gratacós Jordi , Mezquita Jovita , Mezquita Cristobal , Cid Maria C. 
Published: May 2004

AbstractObjectiveNeovascularization, with an increased number of synovial vessels with a characteristic morphology, seems to contribute to the progression of psoriatic arthritis (PsA). Accordingly, angiogenesis may be an important therapeutic target in PsA. The aim of this study was to analyze the effects of infliximab on angiogenesis in the synovial membrane of patients with PsA who responded to this therapy.MethodsThe study group comprised 9 patients with PsA who were selected for the presence of active polyarthritis (including knee synovitis) despite methotrexate therapy. Clinical and biologic evaluations were performed at each visit. Arthroscopy and synovial biopsies were performed at week 0, before infliximab therapy was initiated, and at week 8, after administration of 3 intravenous infusions of infliximab (5 mg/kg). We used immunohistochemistry to identify changes in infiltrating cells and in the angiogenesis modulators αvβ3 integrin, vascular endothelial growth factor (VEGF), angiopoietin 2 (Ang‐2), flt‐1 (VEGF receptor 1 [VEGFR‐1]), kinase insert domain receptor [KDR]/flk‐1 (VEGFR‐2), and stromal cell–derived factor 1 (SDF‐1). Neovascularization was assessed by automated histomorphometry of CD31+ vessels and by measuring αvβ3 expression.ResultsRapid and significant clinical and biological improvement were observed after treatment in all patients. In the synovium, infliximab therapy induced a significant reduction in macrophages, the CD31+ vascular area, αvβ3+ neovessels/Ulex europaeus agglutinin+ vessels, VEGF and its receptor KDR/flk‐1 (VEGFR‐2), and SDF‐1+ vessels. Expression of flt‐1 (VEGFR‐1), and SDF‐1 in lining cells showed a nonsignificant reduction, whereas expression of Ang‐2 increased. In 3 patients, reverse transcription–polymerase chain reaction confirmed the changes in some of these markers at the messenger RNA level.ConclusionThese results show consistent changes in several factors involved in angiogenesis regulation, in parallel with the clinical response to infliximab in patients with PsA. The pattern of reduced VEGF with increased Ang‐2 suggests vascular regression as a potential mechanism underlying the antiangiogenic effect of infliximab.
Ribozymes that inhibit the production of matrix metalloproteinase 1 reduce the invasiveness of rheumatoid arthritis synovial fibroblasts
Authors: Rutkauskaite Edita , Zacharias Wolfgang , Schedel Jörg , Müller‐Ladner Ulf , Mawrin Christian , Seemayer Christian A. , Alexander Dorothea , Gay Renate E. , Aicher Wilhelm K. , Michel Beat A. , Gay Steffen , Pap Thomas 
Published: May 2004

AbstractObjectiveTo investigate whether retroviral gene transfer of ribozymes targeting matrix metalloproteinase 1 (MMP‐1) inhibits the production of MMP‐1 in rheumatoid arthritis synovial fibroblasts (RASFs) and reduces the invasiveness of these cells in vivo.MethodsMMP‐1–specific ribozymes (RzMMP‐1) were designed and cloned into the pLNSX retroviral vector. Cleavage of MMP‐1 was determined in vitro, and the most effective ribozyme was selected for further investigation. RASFs were transduced with replication‐deficient viruses carrying RzMMP‐1 or with empty viruses (mock). Quantitative polymerase chain reaction with cleavage site–spanning fluorescent probes was used to measure the levels of MMP‐1, MMP‐9, and MMP‐13 messenger RNA. In addition, protein levels of MMP‐1 in cell culture supernatants were determined by enzyme linked immunosorbent assay. The effects of stimulation with lipopolysaccharide (LPS) and tumor necrosis factor α (TNFα) on the production of MMP‐1 were assessed accordingly. The invasiveness of RzMMP‐1–transduced, mock‐transduced, and untransduced RASFs was analyzed in the SCID mouse in vivo model of RA.ResultsTransduction of RASFs with RzMMP‐1 significantly decreased the production of MMP‐1 in RASFs without affecting other MMPs, such as MMP‐9 and MMP‐13. RzMMP‐1 not only reduced the spontaneous production of MMP‐1, but also prevented the LPS‐ and TNFα‐induced increase in MMP‐1 production. Inhibition of MMP‐1 was maintained for at least 2 months and was accompanied by a significant reduction of the invasiveness of RASFs in the SCID mouse model of RA.ConclusionIntracellular expression of ribozymes constitutes a feasible tool for inhibiting the production of matrix‐degrading enzymes. Inhibition of MMP‐1 alone results in a significant reduction of cartilage invasion by RASFs.
Requirement of dying cells and environmental adjuvants for the induction of autoimmunity
Authors: Bondanza Attilio , Zimmermann Valérie S. , Dell'Antonio Giacomo , Cin Elena Dal , Balestrieri Genesio , Tincani Angela , Amoura Zahir , Piette Jean‐Charles , Sabbadini Maria Grazia , Rovere‐Querini Patrizia , Manfredi Angelo A. 
Published: May 2004

AbstractObjectiveCells commonly die without eliciting autoimmunity. However, dying cells are a potential initiating stimulus for systemic lupus erythematosus (SLE). Our goal was to verify whether immune adjuvants influence the autoimmunity induction that ensues following in vivo injection of dying cells.MethodsMice were immunized with apoptotic thymocytes in the presence of artificial moieties, such as Freund's incomplete adjuvant (IFA), or natural adjuvants, such as dendritic cells (DCs). Renal involvement and the development of autoantibodies were monitored.ResultsApoptotic cells failed to induce clinical disease or to sustain production of autoantibodies in (NZB × NZW)F1 mice. In contrast, autoimmunity developed in the presence of IFA or DCs. The characteristics of the adjuvant influenced the array of autoantibodies, the kinetics of their development, and the severity of the disease. DCs were required for induction of anti–β2‐glycoprotein I IgG. Adjuvants alone did not elicit disease.ConclusionA “two‐hit” signal composed of autoantigens and adjuvants initiates systemic autoimmunity. Moreover, environmental signals at the site of clearance of dead cells shape the features and the severity of the autoimmune disease. Strategies aimed at preventing the accumulation of dying cells and at modulating endogenous adjuvants may be beneficial for the treatment of SLE.
Proteomic surveillance of autoimmunity in osteoarthritis: Identification of triosephosphate isomerase as an autoantigen in patients with osteoarthritis
Authors: Xiang Yang , Sekine Taichi , Nakamura Hiroshi , Imajoh‐Ohmi Shinobu , Fukuda Hiroyuki , Nishioka Kusuki , Kato Tomohiro 
Published: May 2004

AbstractObjectiveAutoimmunity to proteins, such as type II collagen and cartilage intermediate layer protein, that are produced by chondrocytes has been reported in patients with osteoarthritis (OA) as well as in patients with rheumatoid arthritis (RA). However, it remains to be determined whether the overall specificities of the autoimmunity differ between OA and RA patients. This study sought to clarify the differences by applying proteomic surveillance for the detection of autoantigens comprehensively.MethodsSerum samples were obtained from 20 patients with OA, 20 patients with RA, and 20 healthy volunteers. Human chondrocyte proteins were separated from the sera by 2‐dimensional electrophoresis, and antigenic protein spots were detected by Western blotting. The antigenic proteins were then identified by mass fingerprinting. The antigenicity of the identified proteins was confirmed and the prevalence of the autoantibodies in the OA, RA, and other disease groups was determined with the use of recombinant proteins. In addition, autoepitopes were mapped on the antigens.ResultsNineteen protein spots were recognized only by the OA sera, but not by the RA sera. One of these proteins was identified as triosephosphate isomerase (TPI). IgG‐type anti‐TPI autoantibodies were detected in 24.7% of the serum samples and 24.1% of the synovial fluid samples from the patients with OA, whereas <6% of the RA and systemic lupus erythematosus samples were positive for anti‐TPI. In addition, multiple autoepitopes were identified on TPI.ConclusionThe overall profile of autoimmunity in OA differs from that in RA, which may reflect the OA‐specific pathologic role of autoimmunity. The autoantibody to TPI, detected predominantly in the OA samples and produced by the antigen‐driven mechanism, has the potential to be used as a diagnostic marker for OA.
Defective costimulatory function is a striking feature of antigen‐presenting cells in an HLA–B27–transgenic rat model of spondylarthropathy
Authors: Hacquard‐Bouder Cécile , Falgarone Géraldine , Bosquet Antoine , Smaoui Faïza , Monnet Dominique , Ittah Marc , Breban Maxime 
Published: May 2004

AbstractObjectiveA disease resembling the human spondylarthropathies develops in HLA–B27–transgenic rats. This disease in rats is mediated by CD4+ T cells, but antigen‐presenting cells (APCs) may also play a role. Dendritic cells (DCs) have been reported to be defective in allogeneic mixed lymphocyte culture in this model. Here, we further investigated the functional defect of APCs.MethodsDCs and B cells from nontransgenic, HLA–B27 (33–3)–transgenic, and HLA–B7 (120–4)–transgenic rats were used to stimulate T cells. Surface expression of HLA–B transgene and rat molecules on APCs and the formation of conjugates between DCs and T cells were monitored by flow cytometry.ResultsWe observed a strikingly defective stimulation of allogeneic and syngeneic T lymphocytes by APCs from HLA–B27 but not HLA–B7 rats, even if stimulation was driven in the presence of anti–T cell receptor (TCR) antibody. We found no evidence that HLA–B27 DCs were immature, lacked production of some diffusible factor, or produced an inhibitory factor for T cells. When comparing the levels of expression of class II major histocompatibility complex, CD2, intercellular adhesion molecule 1, lymphocyte function–associated antigen 1, B7, and CD40 molecules at the surface of DCs from 33‐3, 120‐4, and nontransgenic rats, we found little difference. However, HLA–B27–transgenic DCs formed fewer conjugates with T cells than did nontransgenic DCs. Furthermore, the proportion of conjugates formed between DCs and T cells, as well as the difference between nontransgenic and HLA–B27–transgenic DCs, were in large part reduced by blocking CD86 on DCs.ConclusionWe confirmed defective stimulation of T cells by APCs in HLA–B27 rats, the mechanism of which appears to implicate APC/T cell contact, independent of TCR engagement. In addition, decreased use of the CD86 costimulatory molecule by B27 DCs was observed. Impaired costimulatory function could result in a loss of tolerance toward microbial flora in this model.
Increased tyrosine phosphorylation mediates the cooling‐induced contraction and increased vascular reactivity of Raynaud's disease
Authors: Furspan Philip B. , Chatterjee Soumya , Freedman Robert R. 
Published: May 2004

AbstractObjectiveIncreased levels of protein tyrosine kinase (PTK) are mechanistically associated with increased contractile responsiveness to cooling. This study tests the hypothesis that increased PTK activity mediates the increased vascular reactivity to agonists and cooling associated with primary Raynaud's disease (RD).MethodsThe response of dermal arterioles isolated from control (n = 29) and RD (n = 29) subjects to contractile and dilatory agents at 37°C and 31°C was characterized using the microvessel perfusion technique. Fluorescence immunohistochemistry was used to measure tyrosine phosphorylation.ResultsAt 37°C, arteries from RD patients exhibited similar sensitivity to the specific α2‐adrenergic agonist UK 14,304, to serotonin, and to angiotensin II. At 31°C, however, the response to all 3 agonists was greater in the arterioles from the RD patients than in those from the control subjects. Agonist‐induced contraction at both temperatures was reversed by cumulative addition of the PTK inhibitors genistein (1–30 μM) and tyrphostin 47 (0.1–10 μM). All arterioles from control subjects relaxed slightly in response to cooling, whereas more than half of those from RD patients contracted. This cooling‐induced contraction was reversed by the cumulative addition of genistein. The 3 agonists elicited large increases in tyrosine phosphorylation only in arterial segments from RD patients at 31°C. Cooling from 37°C to 31°C elicited a large increase in tyrosine phosphorylation in arterioles from RD patients, but not those from control subjects. All increases in tyrosine phosphorylation could be prevented by genistein.ConclusionIncreased tyrosine phosphorylation mediates cooling‐induced contraction and the increased vascular reactivity of skin arterioles from individuals with RD.
Expression of toll‐like receptor 2 on CD16+ blood monocytes and synovial tissue macrophages in rheumatoid arthritis
Authors: Iwahashi Mitsuhiro , Yamamura Masahiro , Aita Tetsushi , Okamoto Akira , Ueno Akiko , Ogawa Norio , Akashi Sachiko , Miyake Kensuke , Godowski Paul J. , Makino Hirofumi 
Published: May 2004

AbstractObjectiveCD16 (IgG Fcγ receptor type IIIA [FcγRIIIA])–expressing CD14+ monocytes express high levels of Toll‐like receptor 2 (TLR‐2) and are able to efficiently produce proinflammatory cytokines such as tumor necrosis factor α (TNFα). To understand the role of CD16 and TLR‐2 in monocyte and macrophage activation in rheumatoid arthritis (RA), we investigated the expression of TLR‐2 on CD16+ blood monocytes and synovial tissue macrophages and the effect of CD16 and TLR‐2 activation on cytokine production.MethodsThe expression of CD14, CD16, TLR‐2, and TLR‐4 on blood monocytes was measured by flow cytometric analysis. CD16 and TLR‐2 expression in RA synovial tissue was detected by 2‐color immunofluorescence labeling. CD16+ mature monocytes were prepared by incubating blood monocytes in plastic plates for 24 hours. These adhered monocytes were stimulated with lipoteichoic acid (LTA), anti‐FcγRIII antibody, and Hsp60 for 5 hours, and culture supernatants were measured for various cytokines by immunoassay. The activation of NF‐κB was detected by electrophoretic mobility shift assay.ResultsThe frequency of CD16+ cells in all blood monocytes was significantly increased in patients with RA compared with healthy controls. TLR‐2 was expressed at higher levels on CD16+ monocytes than on CD16− monocytes, while TLR‐4 was expressed similarly on both monocytes. In RA synovial tissue, CD16+/TLR‐2+ cells were distributed mainly in the lining layer. TLR‐2 expression on monocytes was enhanced by macrophage colony‐stimulating factor (M‐CSF) and interleukin‐10 (IL‐10), but was reduced by transforming growth factor β1, while CD16 expression was inducible by these cytokines. Adhered monocytes (∼50% CD16+) produced TNFα, IL‐1β, IL‐6, IL‐8, IL‐12 p40, IL‐1 receptor antagonist, and IL‐10 after LTA stimulation. This cytokine response was inhibited significantly by anti–TLR‐2 antibody and partly by anti–TLR‐4 antibody. Anti‐FcγRIII antibody stimulation markedly enhanced the LTA‐induced TNFα response. Hsp60 could stimulate TNFα production by adhered monocytes, which was inhibited similarly by anti–TLR‐2 antibody and anti–TLR‐4 antibody. NF‐κB activation in adhered monocytes was induced by LTA, but this NF‐κB activity was not augmented by anti‐FcγRIII antibody stimulation.ConclusionThese results suggest that CD16+ monocytes and synovial tissue macrophages with high TLR‐2 expression may be induced by M‐CSF and IL‐10, and their production of TNFα could be simulated by endogenous TLR ligands such as Hsp60 and FcγRIIIA ligation by small immune complexes in RA joints.
Down‐regulation of the aberrant expression of the inflammation mediator high mobility group box chromosomal protein 1 in muscle tissue of patients with polymyositis and dermatomyositis treated with corticosteroids
Authors: Ulfgren Ann‐Kristin , Grundtman Cecilia , Borg Kristian , Alexanderson Helene , Andersson Ulf , Harris Helena Erlandsson , Lundberg Ingrid E. 
Published: May 2004

AbstractObjectiveHigh mobility group box chromosomal protein 1 (HMGB‐1) is an endogenous nuclear protein that can be translocated to the cytoplasm and then released extracellularly. It can induce tumor necrosis factor and interleukin‐1 production in myeloid cells. Increased expression of these 2 cytokines has been observed in muscle tissue of patients with polymyositis (PM) and dermatomyositis (DM). The present study was therefore undertaken to investigate how HMGB‐1 is expressed in muscle tissue of patients with myositis and, if so, whether such expression is modulated by prednisolone therapy.MethodsMuscle biopsy specimens from 5 patients with PM and 4 patients with DM, obtained before and 3–6 months after initiation of prednisolone therapy, were assessed by conventional microscopic evaluation and computerized image analysis, and HMGB‐1 expression was investigated by immunohistochemical staining. Muscle biopsy specimens from 7 healthy controls were also studied.ResultsCytoplasmic HMGB‐1 expression was detected in infiltrating rounded mononuclear cells, vascular endothelial cells, and muscle fibers of PM and DM patients. Extracellular staining surrounding the inflammatory cells was also observed. After treatment with high‐dose prednisolone, cytoplasmic and extracellular HMGB‐1 expression was significantly reduced, coinciding mainly with a decreased number of infiltrating inflammatory cells. Cytoplasmic HMGB‐1 expression was still evident in endothelial cells and muscle fibers. No HMGB‐1 expression was observed in muscle tissue from healthy controls.ConclusionThe cytoplasmic and extracellular distribution of HMGB‐1 in muscle tissue may indicate an important role of this proinflammatory molecule in the pathogenesis of PM and DM. Furthermore, our findings indicate that systemically administered high‐dose corticosteroids selectively down‐regulate aberrant expression of HMGB‐1 in mononuclear inflammatory cells in vivo.
An increased transforming growth factor β receptor type I:Type II ratio contributes to elevated collagen protein synthesis that is resistant to inhibition via a kinase‐deficient transforming growth factor β receptor type II in scleroderma
Authors: Pannu Jaspreet , Gore‐Hyer Elizabeth , Yamanaka Masayoshi , Smith Edwin A. , Rubinchik Semyon , Dong Jian‐Yun , Jablonska Stefania , Blaszczyk Maria , Trojanowska Maria 
Published: May 2004

AbstractObjectiveAberrant transforming growth factor β (TGFβ) signaling has been implicated in the pathogenesis of scleroderma (systemic sclerosis [SSc]), but the contribution of specific components in this pathway to SSc fibroblast phenotype remains unclear. This study was undertaken to delineate the role of TGFβ receptor type I (TGFβRI) and TGFβRII in collagen overexpression by SSc fibroblasts.MethodsPrimary dermal fibroblasts from SSc patients and healthy adults were studied (n = 10 matched pairs). Adenoviral vectors were generated for TGFβRI (AdTGFβRI), TGFβRII (AdTGFβRII), and kinase‐deficient TGFβRII (AdΔkRII). TGFβRI basal protein levels were analyzed by 35S‐methionine labeling/immunoprecipitation and immunohistochemistry. Type I collagen and TGFβRII basal protein levels were analyzed by Western blot and newly secreted collagen by 3H‐proline incorporation assay.ResultsAnalysis of endogenous TGFβRI and TGFβRII protein levels revealed that SSc TGFβRI levels were increased 1.7‐fold (P = 0.008; n = 7) compared with levels in healthy controls, while TGFβRII levels were decreased by 30% (P = 0.03; n = 7). This increased TGFβRI:TGFβRII ratio correlated with SSc collagen overexpression. To determine the consequences of altered TGFβRI:TGFβRII ratio on collagen expression, healthy fibroblasts were transduced with AdTGFβRI or AdTGFβRII. Forced expression of TGFβRI in the range corresponding to elevated SSc TGFβRI levels increased basal collagen expression in a dose‐dependent manner, while similar TGFβRII overexpression had no effect, although transduction of fibroblasts at higher multiplicities of infection led to a marked reduction of basal collagen levels. Blockade of TGFβ signaling via AdΔkRII resulted in ∼50% inhibition of basal collagen levels in healthy fibroblasts and in 5 of 9 SSc cell lines. A subset of SSc fibroblasts (4 of 9 cell lines) was resistant to this treatment. SSc fibroblasts with the highest levels of TGFβRI were the least responsive to collagen inhibition via ΔkRII.ConclusionThis study indicates that an increased TGFβRI:TGFβRII ratio may underlie aberrant TGFβ signaling in SSc and contribute to elevated basal collagen production, which is insensitive to TGFβ signaling blockade via ΔkRII.