ovalbumin and Arthritis--Rheumatoid

To observe the effect of heat-tonifying needling on Keap1-Nrf2/ARE/HO-1 signal transduction pathway in knee synovium in rabbits with cold syndrome type rheumatoid arthritis (RA), so as to explore its mechanisms underl-ying improvement of RA.. New Zealand rabbits were randomly divided into normal control, RA model, uniform reinforcing-reducing acupuncture, twisting reinforcing acupuncture and heat-tonifying acupuncture groups, with 6 rabbits in each group. The cold syndrome type RA model was established by subcutaneous injection of mixture fluid of ovalbumin and Freund's complete adjuvant at the shoulder-back as well as injection of mixture of ovalbumin and normal saline into knee-joint cavity combined with ice-compress freezing. Acupuncture stimulation (uniform reinforcing-reducing, or twisting reinforcing or heat-tonifying) was applied to bilateral "Zusanli"(ST36) for 1 min with the needle retained for 30 min, once a day for 7 consecutive days. The general conditions of rabbits in each group were recorded, the thermal pain threshold (TPT) and perimeter of knee joints was measured. Conditions of the synovium in the knee cavity, hydrops, blood flow signal, articular surface, and related muscles were observed by using a color Doppler ultrasonic diagnostic apparatus, and the blood flow signals inside the synovium (image scores) were divided into 0 (no signals), I (1 or 2 dot-like signal), II (less than half) ad III (more than half). After H.E. staining, the pathological changes (0-3 points) were assessed according to the state of inflammatory cell infiltration, and hyperplasia of synovial matrix and coating cells. The expression levels of Keap1, Nrf2, HO-1 and GSH-PX1 mRNAs in the knee synovium were detected by quantitative real-time PCR, and the expression of knee synovial HO-1 protein was measured by Western blot.. In comparison with the normal control group, the model group had a significant increase in the perimeter, pathological score, expression of Nrf2, HO-1 mRNAs and HO-1 protein (. Heat-tonifying, uniform reinforcing-reducing and twirling reinforcing needling manipulations may relieve pain and improve pathological state in RA rabbits, which may be associated with their functions in raising the ability of anti-oxidative stress by regulating Keap1-Nrf2/ARE/ HO-1 signaling pathway, the therapeutic effect of heat-tonifying needling is superior to that of uniform reinforcing-reducing and twirling reinforcing needling.

Topics: Acupuncture Therapy; Animals; Arthritis, Rheumatoid; Hot Temperature; Kelch-Like ECH-Associated Protein 1; NF-E2-Related Factor 2; Ovalbumin; Pain Threshold; Rabbits; RNA, Messenger; Signal Transduction; Syndrome

Autoimmune diseases affect over 4% of the world's population. Treatments are generally palliative or use broad spectrum immunosuppressants to reduce symptoms and disease progression. In some diseases, antibodies generated to a single autoantigen are the major cause of pathogenic inflammation, suggesting that treatments to induce tolerance to the autoantigen could be therapeutic. Here we report the development of hybrid nanoparticles (NPs) that induce tolerance in both T cells and B cells. The NPs comprise a lipid monolayer encapsulating a PLGA core loaded with rapamycin that promotes development of regulatory T cells (Tregs). The lipid monolayer displays the protein antigen and a ligand of the B cell inhibitory co-receptor CD22 (CD22L) that act together to suppress activation of B cells recognizing the antigen. We demonstrate that the hybrid NPs decorated with ovalbumin (OVA) elicit tolerance to OVA in naı̈ve mice, as judged by low OVA-specific antibody titers after the challenge. In the K/BxN mouse model of rheumatoid arthritis caused by B and T cell-dependent responses to the self-antigen glucose-6-phosphate-isomerase (GPI), we show that GPI hybrid NPs delay development of disease, with some treated mice remaining arthritis-free for 300 days. We provide evidence that the mechanism of rheumatoid arthritis suppression involves induction of B cell tolerance, as measured by low anti-GPI antibodies and decreased plasma cell populations, and T cell tolerance, as measured by increased Tregs. The results show the potential of this versatile NP platform for inducing immune tolerance to a self-antigen and suppressing autoimmune disease.

Topics: Animals; Arthritis, Rheumatoid; Autoantigens; Autoimmune Diseases; Immune Tolerance; Lipids; Mice; Nanoparticles; Ovalbumin; Polylactic Acid-Polyglycolic Acid Copolymer

Topics: Animals; Arthritis, Rheumatoid; B-Lymphocytes; Immune Tolerance; Immunosuppressive Agents; Liposomes; Lymphocyte Activation; Mice, Inbred C57BL; Nanoparticles; Ovalbumin; Polylactic Acid-Polyglycolic Acid Copolymer; Sialic Acid Binding Ig-like Lectin 2; Sirolimus; T-Lymphocytes, Regulatory

Autoimmune diseases resulting from MHC class II-restricted autoantigen-specific T cell immunity include the systemic inflammatory autoimmune conditions rheumatoid arthritis and vasculitis. While currently treated with broad-acting immunosuppressive drugs, a preferable strategy is to regulate antigen-specific effector T cells (Teffs) to restore tolerance by exploiting DC antigen presentation. We targeted draining lymph node (dLN) phagocytic DCs using liposomes encapsulating 1α,25-dihydroxyvitamin D3 (calcitriol) and antigenic peptide to elucidate mechanisms of tolerance used by DCs and responding T cells under resting and immunized conditions. PD-L1 expression was upregulated in dLNs of immunized relative to naive mice. Subcutaneous administration of liposomes encapsulating OVA323-339 and calcitriol targeted dLN PD-L1hi DCs of immunized mice and reduced their MHC class II expression. OVA323-339/calcitriol liposomes suppressed expansion, differentiation, and function of Teffs and induced Foxp3+ and IL-10+ peripheral Tregs in an antigen-specific manner, which was dependent on PD-L1. Peptide/calcitriol liposomes modulated CD40 expression by human DCs and promoted Treg induction in vitro. Liposomes encapsulating calcitriol and disease-associated peptides suppressed the severity of rheumatoid arthritis and Goodpasture's vasculitis models with suppression of antigen-specific memory T cell differentiation and function. Accordingly, peptide/calcitriol liposomes leverage DC PD-L1 for antigen-specific T cell regulation and induce antigen-specific tolerance in inflammatory autoimmune diseases.

Topics: Adoptive Transfer; Animals; Anti-Glomerular Basement Membrane Disease; Antigen Presentation; Arthritis, Rheumatoid; B7-H1 Antigen; Calcitriol; Cell Differentiation; CHO Cells; Cricetulus; Dendritic Cells; Disease Models, Animal; Female; HLA-DR Antigens; Humans; Immune Tolerance; Immunodominant Epitopes; Immunologic Memory; Injections, Subcutaneous; Liposomes; Lymph Nodes; Mice; Mice, Transgenic; Ovalbumin; Peptide Fragments; Phagocytosis; Severity of Illness Index; T-Lymphocytes

Rheumatoid arthritis (RA) preferentially affects women, with the peak incidence coinciding with estrogen decrease in menopause. Estrogen (E2) may therefore have intrinsic immune-regulatory properties that vanish with menopause. Fc sialylation is a crucial factor determining the inflammatory effector function of antibodies. We therefore analyzed whether E2 affects immunoglobulin G (IgG) sialylation.. Postmenopausal (ovariectomized) mice were immunized with ovalbumin and treated with E2 or vehicle. Total and ovalbumin-specific IgG concentrations, sialylation, and Fcγ receptor expression were analyzed. Postmenopausal women with RA receiving hormone replacement therapy, including E2, or no treatment were analyzed for IgG sialylation. Furthermore, effects of E2 on the expression of the sialylation enzyme β-galactoside α2,6-sialyltransferase 1 (St6Gal1) were studied in mouse and human antibody-producing cells.. E2 treatment significantly increased Fc sialylation of total and ovalbumin-specific IgG in postmenopausal mice. Furthermore, E2 led to increased expression of inhibitory Fcγ receptor IIb on bone marrow leukocytes. Treatment with E2 also increased St6Gal1 expression in mouse and human antibody-producing cells, providing a mechanistic explanation for the increase in IgG-Fc sialylation. In postmenopausal women with RA, treatment with E2 significantly increased the Fc sialylation of IgG.. E2 induces anti-inflammatory effector functions in IgG by inducing St6Gal1 expression in antibody-producing cells and by increasing Fc sialylation. These observations provide a mechanistic explanation for the increased risk of RA in conditions with low estrogen levels such as menopause.

Topics: Animals; Arthritis, Rheumatoid; B-Lymphocytes; beta-D-Galactoside alpha 2-6-Sialyltransferase; Estrogen Replacement Therapy; Estrogens; Female; Gene Expression Regulation, Enzymologic; Humans; Immunoglobulin G; Mice, Inbred C57BL; Middle Aged; Ovalbumin; Ovariectomy; Postmenopause; Receptors, IgG; Sialic Acids; Sialyltransferases

Effector memory T lymphocytes (T

Topics: Adult; Allergens; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Line; Cells, Cultured; Female; Humans; Hypersensitivity, Delayed; Immunomodulation; Kv1.3 Potassium Channel; Leukocytes, Mononuclear; Mice; Middle Aged; Ovalbumin; Peptides; Polyethylene Glycols; Potassium Channel Blockers; Rats; Rats, Inbred Lew; Scorpion Venoms; Spleen; T-Lymphocytes; Terpenes; Young Adult

Rheumatoid arthritis (RA) is an autoimmune connective tissue disease, associated with the presence of anti-citrullinated protein antibodies (ACPA). These antibodies have been found in approximately 70% of patients suffering from RA and they are currently used for diagnosis of RA. Although they exhibit an absolute need for citrulline for antibody reactivity, no precise cognate antigen for these antibodies has been determined. In this study, we analyzed the reactivity of ACPA to various citrullinated peptides by modified enzyme-linked immunosorbent assays, in order to determine the dependency of specific amino acids for antibody reactivity. A non-human protein (ovalbumin) and antigens directly related to RA were used as templates for synthesis of non-modified and citrullinated peptides, becoming potential target epitopes. Mainly peptides containing a Cit-Gly motif were recognized by ACPAs, while no particular amino acids N-terminal of citrulline were found to be essential for antibody reactivity. Moreover, ACPA reactivity was not restricted to antigens known to be associated with ACPA-positive RA alone, but also to proteins without relation to RA, primarily illustrating that any protein in theory can be turned into an RA autoantigen, by introducing Cit-Gly motifs. Knowledge about the interaction between ACPAs and their citrullinated targets is important for understanding autoimmune ACPA responses in RA, which are known to contribute to the pathophysiology.

Topics: Amino Acid Motifs; Arthritis, Rheumatoid; Autoantibodies; Autoantigens; Autoimmune Diseases; Citrulline; Epitopes; Humans; Ovalbumin; Peptides

Rheumatoid arthritis is a chronic autoimmune pathology characterized by the proliferation and inflammation of the synovium. Boron neutron capture synovectomy (BNCS), a binary treatment modality that combines the preferential incorporation of boron carriers to target tissue and neutron irradiation, was proposed to treat the pathological synovium in arthritis. In a previous biodistribution study, we showed the incorporation of therapeutically useful boron concentrations to the pathological synovium in a model of antigen-induced arthritis (AIA) in rabbits, employing two boron compounds approved for their use in humans, i.e., decahydrodecaborate (GB-10) and boronophenylalanine (BPA). The aim of the present study was to perform low-dose BNCS studies at the RA-1 Nuclear Reactor in the same model. Neutron irradiation was performed post intra-articular administration of BPA or GB-10 to deliver 2.4 or 3.9 Gy, respectively, to synovium (BNCS-AIA). AIA and healthy animals (no AIA) were used as controls. The animals were followed clinically for 2 months. At that time, biochemical, magnetic resonance imaging (MRI) and histological studies were performed. BNCS-AIA animals did not show any toxic effects, swelling or pain on palpation. In BNCS-AIA, the post-treatment levels of TNF-α decreased in four of six rabbits and IFN-γ levels decreased in five of six rabbits. In all cases, MRI images of the knee joint in BNCS-AIA resembled those of no AIA, with no necrosis or periarticular effusion. Synovial membranes of BNCS-AIA were histologically similar to no AIA. BPA-BNCS and GB-10-BNCS, even at low doses, would be therapeutically useful for the local treatment of rheumatoid arthritis.

Topics: Animals; Arthritis, Rheumatoid; Boron Neutron Capture Therapy; Disease Models, Animal; Female; Ovalbumin; Rabbits; Radiobiology; Radiotherapy Dosage; Safety; Synovectomy; Synovial Membrane

Neutrophil extracellular traps (NETs) are web-like structures released by activated neutrophils. Recent studies suggest that NETs play an active role in driving autoimmunity and tissue injury in diseases including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The purpose of this study was to investigate if celastrol, a triterpenoid compound, can inhibit NET formation induced by inflammatory stimuli associated with RA and SLE. We found that celastrol can completely inhibit neutrophil oxidative burst and NET formation induced by tumor necrosis factor alpha (TNFα) with an IC50 of 0.34 µM and by ovalbumin:anti-ovalbumin immune complexes (Ova IC) with an IC50 of 1.53 µM. Celastrol also completely inhibited neutrophil oxidative burst and NET formation induced by immunoglobulin G (IgG) purified from RA and SLE patient sera. Further investigating into the mechanisms, we found that celastrol treatment downregulated the activation of spleen tyrosine kinase (SYK) and the concomitant phosphorylation of mitogen-activated protein kinase kinase (MAPKK/MEK), extracellular-signal-regulated kinase (ERK), and NFκB inhibitor alpha (IκBα), as well as citrullination of histones. Our data reveals that celastrol potently inhibits neutrophil oxidative burst and NET formation induced by different inflammatory stimuli, possibly through downregulating the SYK-MEK-ERK-NFκB signaling cascade. These results suggest that celastrol may have therapeutic potentials for the treatment of inflammatory and autoimmune diseases involving neutrophils and NETs.

Topics: Arthritis, Rheumatoid; Cells, Cultured; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Extracellular Traps; Humans; I-kappa B Proteins; Immunoglobulin G; Inflammation; Intracellular Signaling Peptides and Proteins; Lupus Erythematosus, Systemic; MAP Kinase Kinase Kinases; Neutrophils; NF-KappaB Inhibitor alpha; Ovalbumin; Pentacyclic Triterpenes; Phosphorylation; Protein-Tyrosine Kinases; Respiratory Burst; Syk Kinase; Tripterygium; Triterpenes; Tumor Necrosis Factor-alpha

Regulatory T (Treg) cells play a crucial role in preventing autoimmune diseases and are an ideal target for the development of therapies designed to suppress inflammation in an antigen-specific manner. Type 1 regulatory T (Tr1) cells are defined by their capacity to produce high levels of interleukin 10 (IL-10), which contributes to their ability to suppress pathological immune responses in several settings. The aim of this study was to evaluate the therapeutic potential of collagen type II-specific Tr1 (Col-Treg) cells in two models of rheumatoid arthritis (RA) in mice.. Col-Treg clones were isolated and expanded from collagen-specific TCR transgenic mice. Their cytokine secretion profile and phenotype characterization were studied. The therapeutic potential of Col-Treg cells was evaluated after adoptive transfer in collagen-antibody- and collagen-induced arthritis models. The in vivo suppressive mechanism of Col-Treg clones on effector T-cell proliferation was also investigated.. Col-Treg clones are characterized by their specific cytokine profile (IL-10(high)IL-4(neg)IFN-γ(int)) and mediate contact-independent immune suppression. They also share with natural Tregs high expression of GITR, CD39 and granzyme B. A single infusion of Col-Treg cells reduced the incidence and clinical symptoms of arthritis in both preventive and curative settings, with a significant impact on collagen type II antibodies. Importantly, injection of antigen-specific Tr1 cells decreased the proliferation of antigen-specific effector T cells in vivo significantly.. Our results demonstrate the therapeutic potential of Col-Treg cells in two models of RA, providing evidence that Col-Treg could be an efficient cell-based therapy for RA patients whose disease is refractory to current treatments.

Topics: Adoptive Transfer; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Collagen Type II; Flow Cytometry; Humans; Immunophenotyping; Interferon-gamma; Interleukin-10; Interleukin-4; Mice, Congenic; Mice, Inbred BALB C; Mice, Inbred DBA; Mice, Transgenic; Ovalbumin; Peptide Fragments; T-Lymphocytes, Regulatory; Treatment Outcome

Rheumatoid arthritis (RA) is a polyarticular inflammatory, angiogenic disease. Synovial angiogenesis contributes to inflammation in RA. In this study we have developed an arthritic model in rats using a novel angiogenic protein (NAP), isolated from human synovial fluid of RA patients. We produced anti-NAP monoclonal antibodies (mAbs) and investigated the therapeutic efficacy of the same in adjuvant-induced or NAP-induced arthritis as a model of human RA. The treatment of arthritic rats with anti-NAP mAbs resulted in effective amelioration of paw oedema, radiological arthritic characteristics, serum levels of vascular endothelial growth factor (VEGF) and NAP, compared to that of untreated arthritic animals. Further, profiling of angiogenic markers such as synovial microvessel density, angiogenesis, CD31, VEGF and fms-like tyrosine kinase (Flt1) by immunohistochemistry both in arthritic and anti-NAP mAb-treated animals revealed the efficacy of mAb as an anti-angiogenic functional antibody. Therefore, NAP may be an attractive target to design anti-angiogenic and anti-arthritic therapies to control the pathogenesis of arthritis.

Topics: Adult; Aged; Angiogenesis Inducing Agents; Animals; Antibodies, Monoclonal; Arthritis, Experimental; Arthritis, Rheumatoid; Calcium-Binding Proteins; Cells, Cultured; Disease Models, Animal; Disease Progression; Female; Humans; Immunotherapy; Male; Membrane Proteins; Middle Aged; Ovalbumin; Rats; Rats, Wistar; Synovial Membrane

Urokinase-type plasminogen activator (u-PA) has been implicated in fibrinolysis, cell migration, latent cytokine activation, cell activation, T-cell activation, and tissue remodeling, all of which are involved in the development of rheumatoid arthritis. Previously, u-PA has been reported to play a protective role in monoarticular arthritis models involving mBSA as the antigen, but a deleterious role in the systemic polyarticular collagen-induced arthritis (CIA) model. The aim of the current study is to determine how u-PA might be acting in systemic arthritis models.. The CIA model and bone marrow chimeras were used to determine the cellular source of u-PA required for the arthritis development. Gene expression of inflammatory and destructive mediators was measured in joint tissue by quantitiative PCR and protein levels by ELISA. The requirement for u-PA in the type II collagen mAb-induced arthritis (CAIA) and K/BxN serum transfer arthritis models was determined using u-PA(-/-) mice. Neutrophilia was induced in the peritoneal cavity using either ovalbumin/anti-ovalbumin or the complement component C5a.. u-PA from a bone marrow-derived cell was required for the full development of CIA. The disease in u-PA(-/-) mice reconstituted with bone marrow from C57BL/6 mice was indistinguishable from that in C57BL/6 mice, in terms of clinical score, histologic features, and protein and gene expression of key mediators. u-PA(-/-) mice were resistant to both CAIA and K/BxN serum transfer arthritis development. u-PA(-/-) mice developed a reduced neutrophilia and chemokine production in the peritoneal cavity following ovalbumin/anti-ovalbumin injection; in contrast, the peritoneal neutrophilia in response to C5a was u-PA independent.. u-PA is required for the full development of systemic arthritis models involving immune complex formation and deposition. The cellular source of u-PA required for CIA is bone marrow derived and likely to be of myeloid origin. For immune complex-mediated peritonitis, and perhaps some other inflammatory responses, it is suggested that the u-PA involvement may be upstream of C5a signaling.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Bone Marrow Cells; Collagen; Cytokines; Female; Gene Expression; Hindlimb; Immune Complex Diseases; Immunohistochemistry; Joints; Male; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Ovalbumin; Peritonitis; Urokinase-Type Plasminogen Activator

Topics: Adolescent; Adult; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Female; Humans; Intestinal Absorption; Intestinal Mucosa; Intestine, Small; Male; Middle Aged; Ovalbumin; Rheumatic Diseases; Young Adult

We have investigated the mechanisms underlying IL-15-induced neutrophil migration into inflamed tissues. IL-15 induced neutrophil migration to the peritoneal cavity in mice in a time- and dose-dependent manner. The cell migration was not induced in IL-18-/-, MIP-1alpha (CCL3)-/-, TNFR1-/- or 5-LOX-/- mice but was normal in IFN-gamma-/- mice. IL-15-induced neutrophil migration was inhibited by anti-MIP-2 (CXCL2) antibody or MK886 (leukotriene synthesis inhibitor). IL-18-induced neutrophil migration was also dependent on TNFR1, MIP-1alpha, MIP-2 and leukotriene. Consistent with this observation, IL-15 induced IL-18 production, and IL-15 or IL-18 injection induced the production of MIP-2, MIP-1alpha, TNF-alpha and LTB4. In an antigen-specific inflammation model, ovalbumin (OVA)-induced neutrophil migration was completely inhibited by soluble IL-15Ralpha (sIL-15Ralpha) or anti-MIP-2 antibody. Furthermore, cell migration was absent in IL-18-/-, MIP-1alpha-/-, TNFR1-/-, or 5-LOX-/- mice. OVA challenge induced the release of MIP-2, MIP-1alpha, TNF-alpha and LTB4 in the peritoneal cavity in an IL-15- and IL-18-dependent manner. We also found that neutrophils from the peripheral blood and synovial fluid of patients with rheumatoid arthritis produced substantial amounts of IL-18 and LTB4 following activation by IL-15. Together, these results demonstrate that IL-15 plays an important role in antigen-induced neutrophil migration during inflammation, triggering a sequential OVA, IL-15, IL-18, MIP-2, MIP-1alpha, TNF-alpha, LTB4 and neutrophil migration signaling cascade.

Topics: Animals; Arachidonate 5-Lipoxygenase; Arthritis, Rheumatoid; Chemokine CCL3; Chemokine CXCL2; Chemotaxis, Leukocyte; Humans; Injections, Intraperitoneal; Interleukin-15; Interleukin-18; Leukotriene Antagonists; Leukotrienes; Mice; Mice, Knockout; Neutrophils; Ovalbumin; Receptors, Tumor Necrosis Factor, Type I; Recombinant Proteins; Signal Transduction; Synovial Fluid; Tumor Necrosis Factor-alpha

We examined whether oral administration of lipopolysaccharide (LPS) from Escherichia coli reactivated antigen-induced arthritis (AIA) in mice that is one of models of human rheumatoid arthritis. To induce AIA, mice were immunized by subcutaneous injection of ovalbumin (OVA) emulsified with complete Freund's adjuvant into the base of the tail (day 0) followed by intraarticular injection of OVA on day 21. To investigate the ability of LPS to reactivate AIA, varying doses of LPS were p.o. administered 48 h after the challenge injection. The results showed that administration of LPS was followed by reactivation of AIA in a dose-related fashion. The reactivation of AIA by LPS was associated with increases in interferon-gamma, interleukin-1beta and tumour necrosis factor-alpha. Polymyxin B sulfate given immediately before administration of LPS suppressed the reactivation of AIA. These findings suggest that LPS from intestinal bacteria may play a role in the reactivation of joint inflammation in which immune responses to pathogenic antigens are involved.

Topics: Animals; Anti-Bacterial Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Disease Models, Animal; Hindlimb; Immunoglobulin G; Interferon-gamma; Interleukin-4; Joints; Lipopolysaccharides; Male; Mice; Mice, Inbred DBA; Ovalbumin; Polymyxin B; Spleen

Fibrin deposits adhered to the synovial surface are typical of rheumatoid joints. Since fibrin appears to have a role in arthritis perpetuation our aim was to investigate how these deposits are formed and the consequences of their adhesion to the tissue.. The appearance of fibrin aggregates either free in the synovial fluid or attached to the membrane was studied in rabbits with antigen-induced arthritis by histological techniques at different time points from challenge. In the fixed synovial membranes areas of fibrin-bound synovium were evaluated by qualitative variables to obtain a sequential profile of morphological changes.. Fibrin aggregates appeared from the initial stages of the disease in the synovial effusion. Later on, they were localized on the synovial surface and progressive changes were noted at the fibrin-tissue interface, ending with the invasion of the aggregates by synovial cells and their incorporation into the tissue.. Fibrin aggregates generated inside the joint cavity may constitute a source of activation and acquisition of invasiveness of the synovial fibroblasts, a process to explore within the perpetuating mechanisms of rheumatoid arthritis.

Topics: Animals; Arthritis, Rheumatoid; Fibrin; Fibrinogen; Fibroblasts; Fibronectins; Hindlimb; Immunohistochemistry; Models, Animal; Ovalbumin; Rabbits; Synovial Fluid; Synovial Membrane; Synovitis; Time Factors; Tissue Adhesions

Much evidence implicates IL-8 as a major mediator of inflammation and joint destruction in rheumatoid arthritis. The effects of IL-8 and its related ligands are mediated via two receptors, CXCR1 and CXCR2. In the present study, we demonstrate that a potent and selective nonpeptide antagonist of human CXCR2 potently inhibits (125)I-labeled human IL-8 binding to, and human IL-8-induced calcium mobilization mediated by, rabbit CXCR2 (IC(50) = 40.5 and 7.7 nM, respectively), but not rabbit CXCR1 (IC(50) = >1000 and 2200 nM, respectively). These data suggest that the rabbit is an appropriate species in which to examine the anti-inflammatory effects of a human CXCR2-selective antagonist. In two acute models of arthritis in the rabbit induced by knee joint injection of human IL-8 or LPS, and a chronic Ag (OVA)-induced arthritis model, administration of the antagonist at 25 mg/kg by mouth twice a day significantly reduced synovial fluid neutrophils, monocytes, and lymphocytes. In addition, in the more robust LPS- and OVA-induced arthritis models, which were characterized by increased levels of proinflammatory mediators in the synovial fluid, TNF-alpha, IL-8, PGE(2), leukotriene B(4), and leukotriene C(4) levels were significantly reduced, as was erythrocyte sedimentation rate, possibly as a result of the observed decreases in serum TNF-alpha and IL-8 levels. In vitro, the antagonist potently inhibited human IL-8-induced chemotaxis of rabbit neutrophils (IC(50) = 0.75 nM), suggesting that inhibition of leukocyte migration into the knee joint is a likely mechanism by which the CXCR2 antagonist modulates disease.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Arthritis, Rheumatoid; Chemotaxis, Leukocyte; Chronic Disease; Female; Humans; In Vitro Techniques; Interleukin-8; Lipopolysaccharides; Neutrophils; Ovalbumin; Rabbits; Receptors, Interleukin-8B; Recombinant Proteins; Urea

The purpose of this study was to demonstrate a technique, in a pilot study, for measuring abnormal capillary permeability in synovial tissue of rabbit arthritic knees using dynamic MRI with a gadolinium-based blood pool agent. Arthritis, simulating rheumatoid arthritis, was induced in knees of 8 rabbits by intra-articular injection of carrageenan (n = 4) or ovalbumin (n = 4). Sequential fat presaturated T1-weighted Spoiled Grass images were obtained before and up to 30 min after intravenous administration of albumin-(Gd-DTPA)30. Estimates of synovial tissue plasma-volume (PV), fractional-leak-rate (FLR), and permeability-surface-area-product (PS) were computed. Histologic correlation was obtained in the corresponding regions. Dynamic MRI showed extravasation of albumin-(Gd-DTPA)30 into hypertrophic synovium in six of the eight arthritic knees. Histologic examination of these six knees showed markedly inflamed synovium. The two knees that did not show abnormal vascular permeability contained non-hypertrophic synovium. None of the rabbits showed abnormal permeability in muscle. MRI derived microvascular characteristics (PV, FLR and PS) correlated positively (r2 = 0.51, 0.97 and 0.86) with the histology. Factors involving the structural and functional microvascular characteristics of synovial tissue can be estimated non-invasively using albumin-(Gd-DTPA)30. This technique may be useful for monitoring disease progression and treatment response in rheumatoid arthritis.

Topics: Albumins; Animals; Arthritis, Rheumatoid; Capillary Permeability; Carrageenan; Contrast Media; Gadolinium DTPA; Image Processing, Computer-Assisted; Knee Joint; Magnetic Resonance Imaging; Male; Ovalbumin; Pilot Projects; Rabbits; Synovial Membrane

Minimally invasive synovectomy techniques have been unsuccessful due to lack of selectivity. The purpose of this study was to evaluate the potential of photodynamic therapy to destroy diseased synovium in an antigen-induced arthritis model.. Three sets of experiments evaluated the biodistribution and treatment effects of Photofrin (PF) in rabbits with bilateral knee antigen-induced arthritis. The first set of experiments evaluated the biodistribution of PF in articular tissues of 30 rabbits from 6-72 hours after systemic injection of 2 mg/kg. In the second series of experiments, light was delivered to the knee joint via cleaved optical fibers, whereas for the third, light was delivered via a 600 microm diffusion tip fiber. Tissues were harvested at 2 and 4 weeks posttreatment.. The biodistribution experiments demonstrated maximal uptake in inflamed synovium at 48 hours and a lack of uptake in normal tissues. With bare cleaved fibers, necrosis was observed in one specimen at 2 weeks and was absent in all specimens at 4 weeks. In the third experiment, synovial necrosis was observed in 3 of 7 specimens at 2 weeks and 3 of 8 at 4 weeks. No damage to articular cartilage or periarticular tissues was seen with either mode of light delivery.. These studies indicate that selective destruction of synovium can be achieved with PF and suggest that optimization of light delivery techniques will play an important role in development of this new technique.

Topics: Analysis of Variance; Animals; Arthritis, Rheumatoid; Cartilage, Articular; Dihematoporphyrin Ether; Disease Models, Animal; Hyperplasia; Knee Joint; Microscopy, Fluorescence; Necrosis; Ovalbumin; Photochemotherapy; Rabbits; Synovial Membrane; Synovitis

To compare the alterations in proteoglycan metabolism in antigen induced arthritis and polycation induced arthritis and to determine the involvement of interleukin-1 (IL-1) in the cartilage degradation that occurs in these models of rheumatoid arthritis (RA).. The time course for loss of proteoglycan into the synovial fluid (SF) and inhibition of proteoglycan synthesis, as well as depletion of articular cartilage proteoglycan content, was compared in rabbit antigen arthritis and polycation arthritis. The ability of recombinant IL-1 receptor antagonist IL-1ra to block the acute cartilage loss at 24 h in these models was investigated, compared to its ability to block the cartilage breakdown induced by direct administration of IL-1 in rabbits.. Initial loss of cartilage proteoglycan was accompanied by release of high levels of glycosaminoglycan (GAG) into the SF and decrease in proteoglycan synthetic rates in both antigen and polycation induced arthritis SF GAG rapidly returned to control levels, while proteoglycan synthesis and cartilage proteoglycan content remained depressed, suggesting that the inhibition in proteoglycan synthesis prevented recovery to normal levels. GAG loss from the cartilage into the SF in response to IL-1 injection, as well as other effects of IL-1 challenge, was blocked in a dose dependent manner by IL-1ra administered either intraarticularly (ED50 = 160 ng) or intravenously (iv) (ED50 = 0.09mg/kg). In the antigen induced arthritis model, IL-1ra (20 mg/kg, iv -2h) inhibited GAG release by 40%, whereas in polycation induced arthritis no inhibition was observed even with repeated administration of high doses of inhibitor.. These studies suggest that sustained depression of proteoglycan synthesis may be responsible for the chronic depletion of articular cartilage proteoglycan in the antigen and the polycation model of RA. However, while IL-1 may play a role in the initial breakdown of articular cartilage in antigen induced arthritis, it does not appear to be involved in polycation induced arthritis in the rabbit.

Topics: Animals; Arthritis, Rheumatoid; Cartilage, Articular; Disease Models, Animal; Glycosaminoglycans; Interleukin 1 Receptor Antagonist Protein; Male; Ovalbumin; Polyamines; Polyelectrolytes; Proteoglycans; Rabbits; Receptors, Interleukin-1; Sialoglycoproteins; Synovial Fluid

Lymphocytic beta 1,4-galactosyltransferase (beta 1,4-GalTase, EC 2.4.1.38) activity was measured in B cells using a neoglyco-protein, N-acetylglucosamine-phenylisothiocyanate-bovine serum albumin (GlcNAc-pITC-BSA), as an acceptor substrate in a novel enzyme-linked immunosorbent assay (ELISA)-based method. This assay proved to be much simpler to use than the lengthy and expensive radiochemical assays commonly used, and has the additional advantage that it specifically detects the enzyme mediating transfer via the Gal beta 1,4GlcNAc linkage. A F(ab')2 antibody against GalTase was able to specifically inhibit the reaction. Greater sensitivity for beta 1,4-GalTase activity was obtained using GlcNAc-pITC-BSA as an acceptor substrate rather than ovalbumin. Low levels of beta-galactosidase activity were detectable in lymphocyte cell lysates at acidic pH, although such activity was not detectable at the neutral pH used in the beta 1,4-GalTase activity assay. Using this assay with the GlcNAc-pITC-BSA acceptor, similar beta 1,4-GalTase activities were observed in CD19+ B cells from patients with rheumatoid arthritis (RA) to those seen in normal control individuals.

Topics: Adult; Aged; Antigens, CD19; Arthritis, Rheumatoid; B-Lymphocytes; Carbohydrate Conformation; Enzyme-Linked Immunosorbent Assay; Female; Humans; Isothiocyanates; Male; Middle Aged; N-Acetyllactosamine Synthase; Ovalbumin; Serum Albumin; Substrate Specificity; Thiocyanates

We used synthetic peptides to analyze the human natural antibody response to V beta determinants. A major determinant recognized by IgM and IgG autoantibodies of clinically healthy individuals as well as those suffering from rheumatoid arthritis (RA) was a peptide corresponding to the first complementarity-determining region (CDR1). The natural IgM response of RA patients to this synthetic autoepitope was significantly elevated relative to that shown by healthy individuals. The levels of IgM reactivity to determinants corresponding to this region decreased with increasing age. By contrast, IgG autoantibodies to certain V beta CDR1 peptides increased markedly with age. In order to determine whether the CDR1 V beta determinant might be involved in immunization, we immunized rabbits with a human peptide that is greater than 80% identical to the homologous sequence derived from a rabbit V beta gene. As a control, the rabbits were immunized with a peptide of equal length derived from the N-terminus of the human V beta chain. Like humans, rabbits tended to have high levels of autoantibodies to the CDR1 peptide but not to the N-terminal segment. Following immunization, the rabbits produced strong IgG responses to the N-terminal peptide. By contrast, immunization with the CDR1 peptide resulted in levels of IgG antibody less than or equal to the natural activity in unimmunized rabbits. These studies indicate that the CDR1 region of Tcr V beta is a widely recognized autoantigenic portion of the Tcr that most probably functions as a regulatory epitope in man and other species.

Topics: Adult; Aged; Aged, 80 and over; Aging; Amino Acid Sequence; Arthritis, Rheumatoid; Autoantibodies; Autoimmune Diseases; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin G; Immunoglobulin M; Immunoglobulin Variable Region; Immunoglobulins, Intravenous; Lupus Erythematosus, Systemic; Male; Middle Aged; Molecular Sequence Data; Ovalbumin; Receptors, Antigen, T-Cell, alpha-beta; Sequence Alignment; Sequence Homology, Amino Acid

Microbiochemical assays of the proteoglycan and collagen content of articular cartilage and synovial lining have been performed on tissue sections taken from rabbits with antigen-induced arthritis. This experimental arthritis is a close analogue of the natural disease-rheumatoid arthritis. Animals were killed at intervals during the first 21 days following induction of arthritis to assess changes in the composition of the extracellular matrices of the synovial lining and articular cartilage during the early development of this experimental lesion. In confirmation of earlier studies these microbiochemical assays revealed a rapid and significant loss of proteoglycan from the articular cartilage. This loss was, however, not uniform but was restricted to the intermediate zone of the cartilage. Over the period studied, there was only a slight loss of proteoglycan from the superficial zone of the cartilage facing the joint cavity. These findings demonstrate that, at least in this model, cartilage proteoglycan loss is not due to the action of proteases present in the synovial fluid. Moreover it suggests that the chondrocytes in the mid-zone of the cartilage are responsive to those signals stimulating proteoglycan breakdown. There was no significant loss of collagen from the cartilage over the time period of this study. The synovial lining from arthritic joints, in contrast, showed a progressive increase in both proteoglycan and collagen.

Topics: Animals; Arthritis, Rheumatoid; Azo Compounds; Cartilage, Articular; Collagen; Disease Models, Animal; Femur; Male; Ovalbumin; Phenazines; Picrates; Proteoglycans; Rabbits; Staining and Labeling

We have investigated a role for T cells in chronic antigen-induced arthritis in rats employing a monoclonal antibody (R73 mAb) against the T cell receptor alpha beta. Treatment with R73 mAb from the time of intra-articular antigenic challenge blocked completely the induction of chronic, but not acute ovalbumin-induced arthritis in sensitized rats. Histologically, treatment-controlled arthritic rats exhibited marked hyperplasia of synovial membrane with pronounced infiltration of inflammatory cells including alpha beta + T cells in the chronic phase of arthritis. In contrast, R73 mAb-treated rats had almost normal joint histology. Treatment with R73 mAb after onset of arthritis was also effective in suppressing the progression of chronic antigen-induced joint inflammation. The preventive and suppressive effects of the mAb on chronic antigen-induced arthritis were associated with marked depletion of alpha beta + T cells in peripheral blood. The DTH but not the humoral response to ovalbumin in sensitized rats was suppressed significantly by R73 mAb. Thus, alpha beta + T cells appear to have a central role in both induction and progression of chronic antigen-induced arthritis.

Topics: Animals; Antibodies; Antibodies, Monoclonal; Arthritis, Rheumatoid; Chronic Disease; Disease Models, Animal; Female; Ovalbumin; Rats; Rats, Inbred Strains; Receptors, Antigen, T-Cell, alpha-beta; Receptors, Antigen, T-Cell, gamma-delta

A method is described for the characterization of immune complex components by dot blot analysis. After isolation by chromatographic techniques and precipitation with polyethylene glycol, immune complexes were dissociated in 0.1 M phosphate (pH 2) and bound to a nitrocellulose membrane in a dot blot unit. Biotinylated probes were then used to identify the following immune complex components: specific antigens, biologically active antibodies, antibody isotypes, antibody subclasses, antibody idiotypes, and rheumatoid factors. This nonradioactive procedure takes less than 2 h to perform and has been used to analyze immune complexes isolated from sera (rabbit and human) and synovial fluid (human).

Topics: Acids; Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antigen-Antibody Complex; Arthritis, Rheumatoid; Buffers; Chemical Precipitation; Collodion; Complement C1q; Electrophoresis; Humans; Hydrogen-Ion Concentration; Immunoblotting; Immunoglobulin G; Immunoglobulin Isotypes; Ovalbumin; Rabbits; Synovial Fluid

Platelet activating factor (PAF-acether) is a potent pro-inflammatory mediator. The possible involvement of this molecule in the pathogenesis of chronic erosive arthritis has been investigated using an animal model, antigen-induced arthritis in the rabbit, which closely resembles rheumatoid arthritis. The arthritic joint fluids from rabbits with antigen-induced arthritis contained low levels of PAF-acether in the acute stages of the disease. However, PAF-acether was not detectable in the chronic stages of the lesion. The biologically inactive precursor/metabolite of PAF-acether, lyso-PAF-acether, was detectable in both control and arthritic joint washes. However, the levels of lyso-PAF-acether in the arthritic joint fluids were significantly elevated above those of control in the acute stages of the disease, but not in the chronic stages. Intra-articular injection of PAF-acether at doses up to 100 times the levels detected in the acute stages of this model did not induce joint swelling or leucocyte accumulation in normal rabbits. This study suggest that PAF-acether may contribute to the acute phase of antigen-induced arthritis but is less likely to be involved in the chronic processes.

Topics: Animals; Arthritis; Arthritis, Rheumatoid; Chronic Disease; Disease Models, Animal; Humans; Inflammation; Injections, Intra-Articular; Male; Ovalbumin; Platelet Activating Factor; Rabbits; Synovial Fluid

Injection of ovalbumin into subcutaneous air pouches prepared on the backs of rats previously sensitised to the antigen resulted in the induction of a small and transient accumulation of inflammatory fluid with a predominantly polymorph cell infiltrate. Challenge of pouches of appropriately sensitised rats with Bordetella pertussis vaccine, on the other hand, resulted in a larger and more prolonged accumulation of fluid and cells with a predominantly mononuclear presence. When intact homologous femoral head cartilage was implanted in these inflamed pouches proteoglycan loss was found to be not different from similar implants in non-inflamed pouches. Coating the cartilage with human heat-aggregated immunoglobulin G prior to implantation in air pouches was also found to be without effect on subsequent proteoglycan loss.

Topics: Animals; Arthritis, Rheumatoid; Arthus Reaction; Cartilage, Articular; Female; Male; Neutrophils; Ovalbumin; Pertussis Vaccine; Rats; Rats, Inbred Strains

The arthritogenic effect of dietary cow's milk, egg albumin and soya milk were compared in Old English rabbits. The 12-week cow's milk feeding regimen produced the highest incidence of significant joint lesions. Lesions were evident but mild at 5 weeks and the synovitis had resolved by 32 weeks. It is suggested that the transient nature of the synovitis may be due to the development of specific secretory IgA antibodies which were detectable in faecal pellet extracts. Sandy Lop rabbits were less susceptible to the arthritogenic effect than were Old English rabbits. Dietary ovalbumin was less arthritogenic than cow's milk despite high titres of serum and synovial fluid antibodies and immune complexes. The rabbits were 'tolerant' to dietary soya due probably to pre-existing levels of soya protein in their diet. Lewis and Wistar strain rats, CBA, Balb/c and C57/BL6 mice fed on cow's milk for 3 months did not develop serum antibodies or synovial lesions. It is suggested that this allergic synovitis is not a model for early rheumatoid joint disease because of the transience of the lesions and lack of stimulation of rheumatoid factor. It may well, however, be a model for the arthralgia seen in patients with certain food allergies.

Topics: Animals; Antibodies, Anti-Idiotypic; Antigen-Antibody Complex; Arthritis, Rheumatoid; Autoantibodies; Cattle; Collagen; Complement Activating Enzymes; Complement C1q; Diet; Female; Glycine max; Immunoglobulin G; Leukocyte Count; Mice; Mice, Inbred Strains; Milk; Ovalbumin; Rabbits; Rats; Rats, Inbred Lew; Rats, Inbred Strains; Synovial Fluid; Synovial Membrane; T-Lymphocytes; Time Factors

The much favoured ovalbumin antigen induced model of arthritis in rabbits is widely used in rheumatoid arthritis (RA) research. When examined histologically, it was found to have important deficiencies as parallels to the human disease. After sensitization to ovalbumin, 2 intraarticular challenge doses of a magnitude at each end of the spectrum used by investigators were used in 152 rabbits. The effects of the high and low dose challenges were examined histologically with particular attention to the articular cartilage. With high doses, the gross and histological changes in the knee joint were remarkably akin to acute cartilage necrosis rather than RA1. In the low dose, a milder smoldering arthritis was produced. These observations suggest that, depending on the challenge dose used, there is a tremendous variability in the kind of arthritis produced by the antigen induced arthritis model. Furthermore, it is suggested that previous conclusions about the pathophysiology and immunology of RA drawn from the models that produce a rapid and severe arthritis should be reexamined.

Topics: Animals; Antigens; Arthritis, Rheumatoid; Cartilage; Disease Models, Animal; Evaluation Studies as Topic; Female; Immunization; Inflammation; Injections, Intra-Articular; Lipid Metabolism; Male; Ovalbumin; Rabbits; Research Design; Synovitis

We report six patients with coeliac disease in whom arthritis was prominent at diagnosis and who improved with dietary therapy. Joint pain preceded diagnosis by up to three years in five patients and 15 years in one patient. Joints most commonly involved were lumbar spine, hips, and knees (four cases). In three cases there were no bowel symptoms. All were seronegative. X-rays were abnormal in two cases. HLA-type A1, B8, DR3 was present in five and B27 in two patients. Circulating immune complexes showed no consistent pattern before or after treatment. Coeliac disease was diagnosed in all patients by jejunal biopsy, and joint symptoms in all responded to a gluten-free diet. Gluten challenge (for up to three weeks) failed to provoke arthritis in three patients tested. In a separate study of 160 treated coeliac patients attending regular follow up no arthritis attributable to coeliac disease and no ankylosing spondylitis was identified, though in a control group of 100 patients with Crohn's disease the expected incidence of seronegative polyarthritis (23%) and ankylosing spondylitis (5%) was found (p less than 0.01). Arthritis appears to be a rare manifestation of coeliac disease. This relationship may provide important clues to the role of gastrointestinal antigens in rheumatic diseases.

Topics: Adolescent; Adult; Antibodies, Anti-Idiotypic; Antigen-Antibody Complex; Arthritis, Rheumatoid; Celiac Disease; Female; Gliadin; Hand; Humans; Male; Middle Aged; Ovalbumin; Radiography; Reticulin

Antigen-specific suppressor cell activity of peripheral blood mononuclear cells was investigated in 20 patients with rheumatoid arthritis (RA) and 16 age- and sex-matched healthy controls. Suppressor cell activity was generated by priming peripheral blood mononuclear cells with high dose antigen (ovalbumin) and adding the washed primed or control (unprimed) cells to autologous, optimally stimulated, target plaque forming cell (PFC) cultures. The ability of the primed cells to interfere with an optimal ovalbumin specific PFC response in the target culture was used as a measure of antigen-specific suppressor cell activity. The results demonstrated that the mean (+/- SE) PFC response of the rheumatoid patients (669 +/- 76 PFC/10(6) cells) was not statistically different from that of the normal controls (722 +/- 83 PFC/10(6) cells), P = 0.1. However, reduced suppressor cell activity was observed in the rheumatoid patients relative to controls (46.4 +/- 4.2% versus 64.6 +/- 2.7% suppression, respectively; P < 0.001). No correlation was demonstrated between suppressor cell activity in rheumatoid patients and disease activity or therapy.

Topics: Adult; Aged; Antibody-Producing Cells; Arthritis, Rheumatoid; Cells, Cultured; Epitopes; Female; Hemolytic Plaque Technique; Humans; Male; Middle Aged; Ovalbumin; T-Lymphocytes, Regulatory

Topics: Administration, Oral; Animals; Antibody Formation; Arthritis, Rheumatoid; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Ovalbumin; Penicillamine; Rabbits

The corneo-scleral changes described are highly characteristic and may be the first signs of an underlying systemic disorder so that otherwise healthy patients who present with limbal guttering and scleral disease must be continuously monitored with this association in mind. The clinical and histological features of limbal guttering in connective tissue disorders strongly suggest that a local antigen-antibody reaction triggers off a number of biochemical and cellular responses which combine to produce lysis of scleral and corneal collagen, although immune complexes have not so far been demonstrated in these eyes. Modes of therapy aimed at one particular chain of events have varying degrees of success, as indeed does more blunderbuss treatment with steroids, anti-inflammatory drugs, or cytotoxic agents. The early stages of the ocular lesion in the rabbit are now being studied, as is the immunological basis for its production. It is hoped that further work with the animal model will lead to a deeper understanding of the pathogenesis of these conditions, which will turn provide both ophthalmologists and rheumatologists with more scientific guidelines for treatment.

Topics: Animals; Arthritis, Rheumatoid; Collagen Diseases; Cornea; Corneal Diseases; Disease Models, Animal; Granulomatosis with Polyangiitis; Humans; Lupus Erythematosus, Systemic; Ovalbumin; Polyarteritis Nodosa; Rabbits

An experiment was designed to compare the efficiency of lymph node injection for the induction of adjuvant arthritis (AA) with that of conventional footpad injection in the rat. Quantitative studies revealed that the minimal dose required for induction of AA by the lymph node route is one fifth of that by the footpad route. Thus, the lymph node route was found to be more efficient than the footpad method in terms of higher incidence and earlier onset of AA. PPD in Freund's incomplete adjuvant was able to produce tuberculin sensitization in the rat. The lymph node route again proved to be superior in terms of consistent appearance of the 24-hour reaction on days 8 and 14 and prolongation of the skin reaction over 48 h. These findings show that the lymph node method is so efficient in the rat that it will be especially useful for the trial induction of AA with various materials of unknown potency as well as for production of delayed hypersensitivity. In addition, this injection method appears to be a simple and efficient technique for assay of other immunological reactions.

Topics: Animals; Arthritis, Rheumatoid; Female; Foot; Freund's Adjuvant; Guinea Pigs; Hindlimb; Hypersensitivity, Delayed; Inguinal Canal; Injections, Intradermal; Injections, Intralymphatic; Lymph Nodes; Mycobacterium tuberculosis; Ovalbumin; Rats; Rats, Inbred Lew; Tuberculin; Waxes

Cell walls of Mycobacterium smegmatis were able to produce much more severe arthritis in rats than the delipidated cells, whereas cell envelope and cell membrane fractions were unable to produce the disease. The lysozyme-solubilized product was able to produce mild disease with only 30% of incidence with an optimum dose, whereas the higher and the lower doses did not produce the disease. The rats immunized with cell envelope, cell membrane fraction and nonarthritogenic doses of lysozyme-solubilized product were protected against the subsequent homologous and heterologous challenge of delipidated cells. It was discussed that this preventative effect can be the result of antigenic competition between the arthritogenic and nonarthritogenic components of M. smegmatis. On the other hand, all the fractions separated here were able to serve as an immunoadjuvant in terms of inducing delayed hypersensitivity to ovalbumin in guinea pigs.

Topics: Animals; Arthritis, Rheumatoid; Cell Fractionation; Cell Wall; Female; Freund's Adjuvant; Guinea Pigs; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Mycobacterium; Mycobacterium Infections; Ovalbumin; Rats; Rats, Inbred Lew; Tuberculin

Daily intraperitoneal doses of concanavalin A (Con A) produced a dose-related inhibition of adjuvant-induced arthritis in rats. Con A was also effective on established arthritis, markedly relieving the disease after only three doses. The inhibitory effect of Con A was neutralised by pre-incubation with ovalbumin, although this treatment did not modify the delayed phlogistic action of Con A in rat paws.

Topics: Animals; Arthritis, Rheumatoid; Concanavalin A; Dose-Response Relationship, Drug; Edema; Female; Foot; Hindlimb; Immunosuppressive Agents; Ovalbumin; Rats; Time Factors

Water-soluble substances have been extracted from two strains of Mycobacterium tuberculosis var. hominis: the native hydrosoluble part (polysaccharide and peptidoglycan), a substance in which the polysaccharide moiety is less abundant than in the latter, the acetylated peptidoglycan and, finally a tetrasaccharide-heptapeptide. All four types of substances, when they were injected together with Freund's incomplete adjuvant, exerted an adjuvant effect on the production of delayed-type hypersensitivity to ovalbumin in the guinea pig and on the production of anti-influenza virus antibodies in the rabbit. Injected intravenously in the mouse, they increased the number of antibody-producing cells in the spleen and enhanced the graft versus host reaction; no effect was seen on the phagocytic activity of the reticulo-endothelial system. By contrast with wax D, the water-soluble substances were devoid of arthritis-inducing activity in the rat. Altogether, these water-soluble substances seem to be endowed with at least some of the adjuvant activities of Freund's complete adjuvant and some of the immunostimulant activities of a live Mycobacterium like BCG.

Topics: Adjuvants, Immunologic; Animals; Antibodies, Viral; Antibody Formation; Antibody-Producing Cells; Arthritis, Rheumatoid; Graft vs Host Reaction; Guinea Pigs; Hypersensitivity, Delayed; Mice; Mycobacterium tuberculosis; Orthomyxoviridae; Ovalbumin; Peptidoglycan; Phagocytosis; Polysaccharides; Rabbits; Solubility; Spleen

Prolonged sensitization with emulsion of an autologous or isologous subcutaneous abscess of Arthus type induced by injection of hen egg-white was performed in 34 rabbits which were divided into (high responder and intermediate responder groups (H- and M-groups) according to individual difference of immune responses. The development of a rheumatoid factor-like substance (RFLS) was demonstrated after 30 experimental days and subsequently observed in 21 out of 33 rabbits. There was no significant difference in the incidence of RFLS between both groups. As to the relation between the development of RFLS and types of focal antigens, the group of the autologous W-substance showed a higher incidence of RFLS than the N-substance. Acute and/or chronic synovitis was demonstrated in 13 of 33 rabbits and inflammatory changes were more intensive and extensive in the later period of experiment. Presence of RFLS in the affected synovial tissues, chiefly in the cytoplasm of plasma cells and mononuclear cells, occasionally in a free state was revealed by immunofluorescent study, and depositions positive for IgG and beta 1C were observed in the wall of blood vessels and fibrinous thrombi in the affected synovial tissues.

Topics: Animals; Antibodies; Antigens; Arthritis, Rheumatoid; Autoantigens; Fluorescent Antibody Technique; gamma-Globulins; Isoantigens; Methods; Ovalbumin; Rabbits; Rheumatoid Factor; Synovial Membrane; Time Factors

Topics: Animals; Arthritis, Rheumatoid; Chronic Disease; Disease Models, Animal; Glycosaminoglycans; Injections, Intra-Articular; Ovalbumin; Rabbits; Species Specificity; Swine; Synovitis

Topics: Animals; Antibodies; Antigen-Antibody Complex; Arthritis; Arthritis, Rheumatoid; Disease Models, Animal; Female; Fluorescent Antibody Technique; Freund's Adjuvant; gamma-Globulins; Guinea Pigs; Immunization; Injections, Intra-Articular; Joints; Ovalbumin; Phagocytosis; Rabbits; Synovial Fluid

Topics: Animals; Antigen-Antibody Complex; Arthritis, Rheumatoid; Bone Marrow Cells; Cell Division; Cell Migration Inhibition; Complement System Proteins; Disease Models, Animal; Freund's Adjuvant; Immunity, Cellular; Immunoglobulins; Macrophages; Ovalbumin; Protein Binding; Rabbits; Rats; Rheumatoid Factor; Synovial Fluid; Synovial Membrane

Topics: Animals; Antigens; Arthritis, Rheumatoid; Disease Models, Animal; Fluorouracil; Freund's Adjuvant; Hindlimb; Injections, Intra-Articular; Ovalbumin; Rabbits; Synovial Membrane; Synovitis; Time Factors

Topics: Age Factors; Anaphylaxis; Animals; Arthritis, Rheumatoid; Freund's Adjuvant; Ovalbumin; Prostaglandins; Rabbits; Species Specificity; Synovial Fluid

Topics: Animals; Arthritis, Rheumatoid; Disease Models, Animal; Fluorouracil; Knee Joint; Ovalbumin; Rabbits; Synovial Membrane

Topics: Adjuvants, Immunologic; Animals; Antibodies; Arthritis, Infectious; Arthritis, Rheumatoid; Complement Fixation Tests; Corneal Opacity; Glycolipids; Guinea Pigs; Humans; Hypersensitivity, Delayed; Immunoglobulins; Joints; Microscopy, Electron; Mycobacterium; Nocardia; Ovalbumin

Topics: Animals; Arthritis, Rheumatoid; Blood Proteins; Erythrocytes; Formaldehyde; gamma-Globulins; Guinea Pigs; Hemagglutination Tests; Humans; Immune Sera; Ovalbumin; Rheumatoid Factor; Serum Albumin, Bovine; Sheep; Tannins