ovalbumin and Arthritis

Phosphoinositide-3 kinase (PI3K)-δ and PI3K-γ are preferentially expressed in immune cells, and inhibitors targeting these isoforms are hypothesized to have anti-inflammatory activity by affecting the adaptive and innate immune response. We report on a potent oral PI3K-δ and PI3K-γ inhibitor (IPI-145) and characterize this compound in biochemical, cellular, and in vivo assays. These studies demonstrate that IPI-145 exerts profound effects on adaptive and innate immunity by inhibiting B and T cell proliferation, blocking neutrophil migration, and inhibiting basophil activation. We explored the therapeutic value of combined PI3K-δ and PI3K-γ blockade, and IPI-145 showed potent activity in collagen-induced arthritis, ovalbumin-induced asthma, and systemic lupus erythematosus rodent models. These findings support the hypothesis that inhibition of immune function can be achieved through PI3K-δ and PI3K-γ blockade, potentially leading to significant therapeutic effects in multiple inflammatory, autoimmune, and hematologic diseases.

Topics: Animals; Arthritis; Asthma; Collagen Type II; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Humans; Isoquinolines; Lupus Erythematosus, Systemic; Molecular Structure; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Purines; Rats; Rats, Inbred Lew; Rats, Wistar; Structure-Activity Relationship

Previously, it has been shown that heat shock protein 70 (HSP70) can prevent inflammatory damage in experimental autoimmune disease models. Various possible underlying working mechanisms have been proposed. One possibility is that HSP70 induces a tolerogenic phenotype in dendritic cells (DCs) as a result of the direct interaction of the antigen with the DC. Tolerogenic DCs can induce antigen-specific regulatory T cells and dampen pathogenic T cell responses. We show that treatment of murine DCs with either mycobacterial (Mt) or mouse HSP70 and pulsed with the disease-inducing antigen induced suppression of proteoglycan-induced arthritis (PGIA), although mouse HSP70-treated DCs could ameliorate PGIA to a greater extent. In addition, while murine DCs treated with Mt- or mouse HSP70 had no significantly altered phenotype as compared to untreated DCs, HSP70-treated DCs pulsed with pOVA (ovalbumin peptide 323-339) induced a significantly increased production of IL-10 in pOVA-specific T cells. IL-10-producing T cells were earlier shown to be involved in Mt HSP70-induced suppression of PGIA. In conclusion, this study indicates that Mt- and mouse HSP70-treated BMDC can suppress PGIA via an IL-10-producing T cell-dependent manner.

Topics: Animals; Arthritis; Bacterial Proteins; Bone Marrow Cells; CD4-Positive T-Lymphocytes; Cytokines; Dendritic Cells; Endopeptidase K; Female; HSP70 Heat-Shock Proteins; Humans; Interleukin-10; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Transgenic; Mycobacterium; Ovalbumin; Peptide Fragments; Phenotype; Proteoglycans

B cell tolerance to self-antigen is critical to preventing antibody-mediated autoimmunity. Previous work using B cell antigen receptor transgenic animals suggested that self-antigen-specific B cells are either deleted from the repertoire, enter a state of diminished function termed anergy, or are ignorant to the presence of self-antigen. These mechanisms have not been assessed in a normal polyclonal repertoire because of an inability to detect rare antigen-specific B cells. Using a novel detection and enrichment strategy to assess polyclonal self-antigen-specific B cells, we find no evidence of deletion or anergy of cells specific for antigen not bound to membrane, and tolerance to these types of antigens appears to be largely maintained by the absence of T cell help. In contrast, a combination of deleting cells expressing receptors with high affinity for antigen with anergy of the undeleted lower affinity cells maintains tolerance to ubiquitous membrane-bound self-antigens.

Topics: Adoptive Transfer; Animals; Arthritis; Autoantigens; B-Lymphocytes; Cell Membrane; Clonal Anergy; Clonal Deletion; Clone Cells; Female; Flow Cytometry; Glycosylphosphatidylinositols; Lymphocyte Count; Male; Mice; Mice, 129 Strain; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, Knockout; Ovalbumin; Protein Multimerization; Receptors, Antigen, B-Cell

The aim of this study was to evaluate the value of contrast-enhanced gray-scale sonography in quantitative assessment of synovial pannus angiogenesis in antigen-induced arthritis in rabbits.. Twenty-four adult New Zealand White rabbits were divided into two groups. Inflammatory arthritis was induced by intra-articular injection of ovalbumin into right knee joints with 4 mg in low-dose group (LD, n = 12) and 8 mg in high-dose group (HD, n = 12). The left side of the knee on each rabbit was used as normal control. Contrast-enhanced gray-scale sonography with time intensity curve (TIC) was performed on the synovia of suprapatellar bursa and posterior capsules 4 weeks after the injection. Immunohistochemical examinations of synovia were applied to assess the microvessel density and the expression of vascular endothelial growth factor. Correlation analysis between sonographic and immunohistochemical findings was performed.. Contrast-enhanced gray-scale sonography of all right knees demonstrated intra-articular hypoechoic lesions with enhanced blood flow and no abnormal findings on all left knees. Parameters of TIC, including ascending curve (A), derived peak intensity (DPI), and area under curve (AUC) on arthritis joints were significantly higher in HD group compared to LD group (P < .05). Positive correlation was found between immunohistochemical findings and parameters of A, DPI, and AUC (P < .05). However, no correlation was found between other parameters (slope of descending rate, time to peak, time to rise, and initial intensity) and immunohistochemical findings.. Contrast-enhanced gray-scale sonography with TIC measurement could provide reliable method for noninvasive quantitative assessment of synovial pannus angiogenesis of arthritis in clinical settings.

Topics: Animals; Antigens; Arthritis; Contrast Media; Image Enhancement; Image Interpretation, Computer-Assisted; Neovascularization, Pathologic; Ovalbumin; Phospholipids; Rabbits; Reproducibility of Results; Sensitivity and Specificity; Sulfur Hexafluoride; Synovial Membrane; Ultrasonography

To evaluate the thermo-responsive poly(N-isopropylacrylamide) (PNiPAAm) polymer as an adjuvant, we synthesized PNiPAAm through free radical polymerization and characterized it both in vitro and in vivo. The polymer when mixed with collagen type II (CII) induced antigen-specific autoimmunity and arthritis. Mice immunized with PNiPAAm-CII developed significant levels of CII-specific IgG response comprising major IgG subclasses. Antigen-specific cellular recall response was also enhanced in these mice, while negligible level of IFN-γ was detected in splenocyte cultures, in vitro. PNiPAAm-CII-immunized arthritic mouse paws showed massive infiltration of immune cells and extensive damage to cartilage and bone. As determined by immunostaining, most of the CII protein retained its native configuration after injecting it with PNiPAAm in naive mice. Physical adsorption of CII and the high-molecular-weight form of moderately hydrophobic PNiPAAm induced a significant anti-CII antibody response. Similar to CII, mice immunized with PNiPAAm and ovalbumin (PNiPAAm-Ova) induced significant anti-ovalbumin antibody response. Comparable levels of serum IFN-γ, IL-1β and IL-17 were observed in ovalbumin-immunized mice with complete Freund, incomplete Freund (CFA and IFA) or PNiPAAm adjuvants. However, serum IL-4 levels were significantly higher in PNiPAAm-Ova and CFA-Ova groups compared with the IFA-Ova group. Thus, we show for the first time, biocompatible and biodegradable thermo-responsive PNiPAAm can be used as an adjuvant in several immunological applications as well as in better understanding of the autoimmune responses against self-proteins.

Topics: Acrylamides; Acrylic Resins; Adjuvants, Immunologic; Animals; Arthritis; Autoimmunity; Biocompatible Materials; Female; Freund's Adjuvant; Immunoglobulin G; Interferon-gamma; Interleukin-17; Interleukin-1beta; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Polymers; Temperature

Methotrexate (MTX) encapsulated microspheres release MTX in the joint in a slow, controlled manner following intra-articular injection in healthy rabbits. The objective of this study was to determine the pharmacokinetics of MTX and to evaluate the efficacy following intra-articular treatment of MTX-loaded microspheres in an antigen-induced inflammatory arthritis rabbit model.. Arthritis was induced in both knee joints of rabbits using ovalbumin. Rabbits were intra-articularly treated with MTX solution or MTX microspheres and plasma concentrations of MTX were determined in the first 8 h following intra-articular treatment. Rabbits were killed 14 days following treatment and histological analysis of rabbit joints was conducted to determine efficacy.. Arthritis was successfully induced in the joints of rabbits with the observation of histopathological features resembling rheumatoid arthritis. No significant differences were detected between MTX solution and MTX microspheres treated groups compared to phosphate buffered saline (control) animals.. MTX microspheres effectively delivered the drug to the intra-articular space. However, a high degree of inter-animal variability, the severity of the disease induced and insufficient length of the observation period were suggested to be possible causes for the lack of therapeutic responses to MTX-loaded microspheres treatment.

Topics: Animals; Antirheumatic Agents; Arthritis; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Edema; Female; Hindlimb; Injections, Intra-Articular; Joints; Methotrexate; Microspheres; Ovalbumin; Rabbits

Intravenous Ig (IVIg) mediates protection from the effects of immune thrombocytopenic purpura (ITP) as well as numerous other autoimmune states; however, the active antibodies within IVIg are unknown. There is some evidence that antibodies specific for a cell-associated antigen on erythrocytes are responsible, at least in part, for the therapeutic effect of IVIg in ITP. Yet whether an IVIg directed to a soluble antigen can likewise be beneficial in ITP or other autoimmune diseases is also unknown. A murine model of ITP was used to determine the effectiveness of IgG specific to soluble antigens in treating immune thrombocytopenic purpura. Mice experimentally treated with soluble OVA + anti-OVA versus mice treated with OVA conjugated to rbcs (OVA-rbcs) + anti-OVA were compared. In both situations, mice were protected from ITP. Both these experimental therapeutic regimes acted in a complement-independent fashion and both also blocked reticuloendothelial function. In contrast to OVA-rbcs + anti-OVA, soluble OVA + anti-OVA (as well as IVIg) did not have any effect on thrombocytopenia in mice lacking the inhibitory receptor FcgammaRIIB (FcgammaRIIB(-/-) mice). Similarly, antibodies reactive with the endogenous soluble antigens albumin and transferrin also ameliorated ITP in an FcgammaRIIB-dependent manner. Finally, broadening the significance of these experiments was the finding that anti-albumin was protective in a K/BxN serum-induced arthritis model. We conclude that IgG antibodies directed to soluble antigens ameliorated 2 disparate IVIg-treatable autoimmune diseases.

Topics: Animals; Antibodies; Antibody Specificity; Antigens; Antigens, CD; Arthritis; Immunoglobulin G; Immunoglobulins, Intravenous; Immunotherapy; Inflammation; Mice; Mice, Knockout; Ovalbumin; Purpura, Thrombocytopenic, Idiopathic; Receptors, IgG; Solubility

The present study was designed to investigate the effect of the phosphodiesterase IV inhibitor rolipram on Th1 and Th2 immune responses in mice. Mice were immunized subcutaneously at the base of the tail with ovalbumin (OVA) emulsified with complete Freund's adjuvant (day 0) and were treated daily with oral administration of various doses of rolipram from days 0 to 20. On day 21, production of anti-OVA IgG and proliferative responses to the antigen were determined. Anti-OVA IgG2a and interferon-gamma (IFN-gamma), as indicators of Th1 responses, and anti-OVA IgG1 and interleukin-10 (IL-10), as indicators of Th2 responses, were also measured. The results showed that treatment with rolipram failed to affect the production of OVA-specific IgG but decreased the proliferation of spleen cells to the antigen. Its inhibitory effect on these immune responses was correlated with a marked decrease in IFN-gamma but not IL-10 production, although neither anti-OVA IgG2a nor IgG1 production was affected by rolipram. These results suggest that rolipram may preferentially inhibit Th1 responses more effectively than Th2 responses. Administration of rolipram resulted in suppression of antigen (OVA)-induced arthritis in mice. The suppression of joint inflammation by rolipram was associated with the inhibition of the OVA-specific proliferative responses of spleen cells and IFN-gamma secretion. These results indicate that rolipram may be effective in regulating Th1-mediated diseases such as rheumatoid arthritis.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Animals; Arthritis; Cyclic Nucleotide Phosphodiesterases, Type 4; Female; Freund's Adjuvant; Immunoglobulin G; Interferon-gamma; Interleukin-10; Mice; Ovalbumin; Phosphodiesterase Inhibitors; Rolipram; Th1 Cells; Th2 Cells

Most inflammatory disorders are becoming more prevalent, especially in Western countries. The pro-inflammatory cytokine tumor necrosis factor-alpha (TNF) plays a prominent role in many of these inflammatory disorders. We have previously shown that SPRET/Ei mice exhibit an extreme and dominant resistance to high doses of TNF. In this report, we investigate the response of heterozygous (C57BL/6xSPRET/Ei)F1 mice in different models of inflammatory diseases. Compared with C57BL/6 mice, (B x S)F1 mice are protected against TNF-induced arthritis and are partially protected against allergic asthma in an ovalbumin-induced model. However, these mice display complete susceptibility to TNF-induced inflammatory bowel disease. These results indicate that the SPRET/Ei genome harbors potent dominant antiinflammatory genes that might be relevant for the treatment of certain chronic inflammatory diseases. It is very well possible that different genes are implicated in the different models.

Topics: Animals; Arthritis; Asthma; Bronchoalveolar Lavage Fluid; Crosses, Genetic; Disease Models, Animal; Female; Genetic Predisposition to Disease; Genome; Heterozygote; Inflammation; Inflammatory Bowel Diseases; Joints; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Tumor Necrosis Factor-alpha

Photodynamic therapy (PDT) using the photosensitizer BPD-Verteporfin (liposomal benzoporphyrin derivative-monoacid ring A) has been shown in previous studies to be effective in the amelioration of inflammatory arthritis in both the MRL-lpr mouse and the New Zealand White (NZW) rabbit models, and could potentially offer alleviation of certain inflammation-related symptoms of rheumatoid arthritis. Time and dose dependency of BPD-MA tissue uptake was carried out in the inflamed synovium and other articular and peri-articular tissues following intravenous and intra-articular administration in the NZW rabbit model. As some articular and peri-articular tissues are difficult to extract, this study uses a rapid fluorimetric sampling of tissues following dissolution in Soluene 350. Our results showed that i.v. injected BPD-MA preferentially distributed in the inflamed synovium, and in tissues with a high degree of vascularization. Little or no association was found with avascular tissues such as cartilage and tendons. Clearance from the synovium was rapid, supporting earlier rather than late light treatment. Much higher association of BPD-MA with the synovium was achieved using intra-articular injection, and BPD-MA concentrations were maintained at relatively steady levels for several hours. These observations support the possibility that PDT could offer a safe, highly versatile clinical option for the management of inflamed joints in autoimmune disorders.

Topics: Animals; Antineoplastic Agents; Arthritis; Injections, Intra-Articular; Injections, Intravenous; Liposomes; Ovalbumin; Photochemotherapy; Photosensitizing Agents; Porphyrins; Rabbits; Synovial Fluid; Tissue Distribution; Verteporfin

The effects of oral administration of ovalbumin (OVA) on acute OVA-induced arthritis (OIA) in rats, which is mediated by Arthus reaction to the antigen in the joint space, were investigated. The oral administration of OVA before immunization with OVA significantly suppressed the development of acute OIA in a dose-dependent manner, in accordance with decreases in both the in vivo anti-OVA IgG antibody production and in vitro lymphocyte proliferative responses to OVA. These results were shown in both the single high-dose (200 mg x 1) or the multiple low-dose (200 microg x 5) feeding protocols. In vitro study showed that rat IL-2 could reverse the reduced OVA-specific lymphocyte proliferative responses. The spleen cells obtained from OVA-feeding, unprimed rats neither adoptively transferred the suppression to naive recipient rats nor suppressed the in vitro lymphocyte proliferation. These results demonstrate that the acute OIA can be suppressed by the induction of oral tolerance (OT) to OVA, and strongly suggest that the OT was due to clonal anergy of antigen-reactive T lymphocytes, not the active suppression by OVA-specific regulatory cells.

Topics: Acute Disease; Administration, Oral; Adoptive Transfer; Animals; Antigens; Arthritis; Clonal Anergy; Female; Immunization; Immunoglobulin G; In Vitro Techniques; Interleukin-2; Lymphocyte Activation; Male; Ovalbumin; Rats; Rats, Inbred Lew; T-Lymphocytes

To assess the utility of macromolecular contrast material-enhanced magnetic resonance (MR) imaging parameters for determining the histopathologic severity of temporomandibular joint (TMJ) arthritis.. Ovalbumin was used to induce arthritis in the TMJs of 10 previously sensitized adult white rabbits. Five rabbits composed the sham-treated control group. Dynamic spin-echo imaging was performed immediately before and for 30 minutes after injection of macromolecular contrast medium. Histologic specimens of TMJ were assessed quantitatively for arthritis. Changes in MR signal intensity were derived from the synovial and subsynovial tissues of the TMJ, and plasma volume (PV) and permeability surface area product (PS) were calculated. These MR parameters and the arthritic scores were compared between sham-treated and antigen-challenged TMJs. The relationships between MR parameters and histopathologic indexes were also determined.. Arthritic TMJs showed marked enhancement of the synovial and subsynovial tissues over the imaging period. PS and all histopathologic indexes of arthritis were significantly greater (P < .005) in antigen-challenged than in sham-treated TMJs. PS demonstrated strong positive relationships with all histologic parameters of arthritis, indicating its utility for assessing the severity of joint inflammation.. Macromolecular contrast-enhanced MR imaging enables quantification of PS and PV in inflamed joints. This technique may provide insights into the pathogenesis of joint inflammation and noninvasive monitoring of disease severity and treatment response in arthritis.

Topics: Albumins; Animals; Arthritis; Contrast Media; Gadolinium DTPA; Magnetic Resonance Imaging; Male; Molecular Weight; Observer Variation; Organometallic Compounds; Ovalbumin; Pentetic Acid; Rabbits; Temporomandibular Joint; Temporomandibular Joint Disorders

DA rats develop transient arthritis after subcutaneous immunization with adjuvant-oil, while chronic arthritis and collagen autoreactivity ensues when collagen is added to the oil. We show here that DA rats can be protected from oil-induced arthritis (OIA) and rat collagen-induced arthritis (rCIA) by addition of antigen to these arthritogenic inocula. We have investigated this remarkable phenomenon and demonstrate that both foreign and self antigens can be protective, apparently provided they are immunogenic; hence HSP-65kDa, ovalbumin, rat myelin basic protein, rat IgG and bovine albumin are effective while rat albumin is not. This protection is long-lasting and disease-specific because rats protected from rCIA resist a later attempt to induce arthritis, but not experimental autoimmune encephalomyelitis (EAE). Protection from rCIA depends neither on the blocking of humoral autoreactivity to collagen nor on a change in the isotype profile of anti-collagen antibodies. We demonstrate that immunogens can also be protective when injected intraperitoneally only a few days before onset of arthritis. Our results indicate that protection is mediated through bystander immune reactions towards the co-immunized antigen and that the arthritogenicity of a given provocation, be it adjuvants, microbes or autoantigens, may be a complex net result of arthritogenic and contra-arthritogenic immune reactions.

Topics: Amino Acid Sequence; Animals; Antigens; Arthritis; Bacterial Proteins; Chaperonin 60; Chaperonins; Collagen; Encephalomyelitis, Autoimmune, Experimental; Female; Immunization; Immunoglobulin G; Male; Molecular Sequence Data; Myelin Basic Protein; Oils; Ovalbumin; Rats; Rats, Inbred Strains; Serum Albumin, Bovine

To determine if systemic administration of human interleukin 1 receptor antagonist (IL-1ra) to rabbits during the induction phase of antigen induced arthritis (AIA) could block inflammation and cartilage proteoglycan loss.. Recombinant human IL-1ra was administered every 6 h to rabbits beginning 1 h before induction of arthritis. Joint swelling was monitored for 72 h and then animals were killed 6 h after the last dose of IL-1ra. Leukocyte accumulation in the joint space and synovial lining was determined and the proteoglycan content and capacity for synthesis was assessed in the articular cartilage of the control and arthritic joints.. Administration of IL-1ra had no detectable effect on the induction of arthritis. Swelling proceeded with a similar time course to untreated AIA animals and at 3 days the cellular infiltrate into synovial fluid (SF) was similarly high, the proteoglycan content of SF was also high and cartilage proteoglycan content was depleted. The biosynthesis of proteoglycan in cartilage was also similarly inhibited. No changes were detected in the cartilage and synovium or SF of the contralateral joints of animals receiving IL-1ra.. IL-1ra given at a dose shown to block synovitis and proteoglycan loss induced by a bolus injection of recombinant IL-1 in rabbits was unable to inhibit the induction of AIA. Our results suggest that the action of IL-1 is not the major factor responsible for the induction of arthritis in this animal model of inflammatory joint disease.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antigens; Arthritis; Cartilage, Articular; Female; Humans; Hyperplasia; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Ovalbumin; Proteoglycans; Rabbits; Receptors, Interleukin-1; Recombinant Proteins; Sialoglycoproteins; Synovial Membrane

Osteoarthritis was surgically induced in mature male Dutch Belted rabbits by sectioning the fibular collateral and sesamoid ligaments and removal of the anterior horn of the lateral meniscus. The site of surgical intervention was detectable by MRI. Histopathologic analysis revealed severe focal cartilage lesions on opposing surfaces of the tibia and femur. Histology of cartilage adjacent to the osteoarthritic lesions appeared normal. In another animal model, arthritis was induced by immunization against ovalbumin followed by intra-articular injection of ovalbumin. MRI of immune arthritic rabbit knees showed accumulation of synovial fluid and cartilage degradation. Histopathology was characterized by vascular necrosis of the synovium and depletion of cartilage proteoglycan. MRI can be used to non-invasively follow the therapeutic effects of drug treatment on synovial inflammation and cartilage degradation in rabbit knees.

Topics: Animals; Arthritis; Cartilage, Articular; Disease Models, Animal; Knee Joint; Magnetic Resonance Imaging; Male; Osteoarthritis; Ovalbumin; Rabbits

T-cell vaccination using antigen-specific lines or clones has been shown to be effective in down-regulating immunity in various experimental autoimmune models. Anti-idiotypic networks developing during differentiation of the immune system are considered to be a safeguard against autoimmunity and these pre-existing networks are supposed to be a prerequisite for successful vaccination. However, the interesting question of feasibility of T-cell vaccination beyond the area of autoimmunity remains to be answered. The present study is the first one providing evidence of successful T-cell vaccination in mice immunized against foreign protein antigens (in this system supposedly no pre-existing network exists). Intraperitoneal (i.p.) administration of hen egg lysozyme (HEL)- and chicken egg albumin (OVA)-specific lymph node cells (LNC) were shown to effectively down-regulate immunity (as measured in a delayed type of hypersensitivity) to HEL and OVA, respectively. In contrast, vaccination was unsuccessful with methylated bovine serum albumin (mBSA)-specific LNC in mBSA immunity. Suppression induced by HEL- and OVA-specific LNC was antigen specific. Unlike the greater part of other studies, in which antigen-specific lines or clones were used, we used draining LNC of immunized mice, which after activation were fixed with glutardialdehyde and injected i.p. 10 days before immunization. Finally, effects of T-cell vaccination were studied in a chronic HEL-induced arthritis. Joint swelling, cell influx and cartilage matrix depletion were significantly less in mice treated with antigen-specific cells. We conclude that successful vaccination is feasible in mice rendered immune to foreign protein antigens using a pool of LNC as source of vaccine, suggesting no necessity of a strong pre-existing network.

Topics: Animals; Antigens; Arthritis; Chronic Disease; Female; Immune Tolerance; Lymph Nodes; Mice; Mice, Inbred C57BL; Muramidase; Ovalbumin; Proteins; Serum Albumin, Bovine; T-Lymphocytes; Vaccination

Anti-inflammatory properties of zinc protoporphyrin disodium (Zn-PP-2Na) were studied. Zn-PP-2Na exhibits anti-allergic action against type III and IV reactions (passive Arthus reaction in rats and tuberculin-induced footpad reaction in mice), but does not affect type I and II reactions (homologous passive cutaneous anaphylaxis in mice and reversed cutaneous anaphylaxis in rats). Zn-PP-2Na also clearly inhibits type II collagen-induced arthritis in mice. The agent inhibits general arthritis symptoms, anti-type-II collagen antibody production and type II collagen-induced delayed type hypersensitivity (DTH) in arthritic mice. Zn-PP-2Na, however, did not affect carrageenin-induced paw edema and histamine- and serotonin-induced skin reactions in rats. Zn-PP-2Na inhibits IL-1-induced mouse lymphocyte proliferation, but does not affect PMA-induced O2- generation from guinea-pig neutrophil. These results indicate that Zn-PP-2Na inhibits type II collagen-induced arthritis in mice due to the antagonism of IL-1 activity and the inhibition of DTH against type II collagen.

Topics: Anaphylaxis; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis; Arthus Reaction; Ascaris; Collagen; Edema; Hypersensitivity; Interleukin-1; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Ovalbumin; Oxygen Consumption; Passive Cutaneous Anaphylaxis; Protoporphyrins; Rats; Rats, Wistar; Skin Tests

H2-c-fos transgenic (c-fos+) mice are characterized by the inability to raise specific IgG antibodies against immunizing Ag. To examine the contribution of Ag-specific IgG antibody to the development of arthritis, Ag-induced arthritis was produced in c-fos+ mice and their control littermates (c-fos- mice). Intra-articular injection of OVA into c-fos- mice hyperimmunized with OVA induced destructive arthritis with massive lymphocyte infiltration. The c-fos+ mice also developed destructive arthritis comparable in degree with that seen in c-fos- mice. However, joints from the c-fos+ mice had few or no infiltrating lymphocytes. The majority of cells invading the extensively eroded collagenous tissue in the c-fos+ mice had a mesenchymal appearance. These cells, producing excess amounts of c-Fos protein, adhered to and invaded the cartilage matrix when cultured on cartilage slices. These cells, thus, appear to directly cause joint destruction in c-fos+ mice.

Topics: Animals; Arthritis; Bone and Bones; Cartilage; Genes, fos; Immunoglobulin G; Lymphocytes; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Proto-Oncogene Proteins c-fos; Synovial Fluid

Chronic monarticular arthritis can be induced in ovalbumin-sensitized rabbits by intraarticular injection of ovalbumin (antigen-induced arthritis) or in naive rabbits by injecting hyaluronic acid mixed with the polycation poly-D-lysine (polycation-induced arthritis). Both models show some points of similarity, including joint swelling, the presence of inflammatory leukocytes and the inflammatory mediator prostaglandin E2, and the kinetics of cartilage proteoglycan loss. However, the assessment of the capacity of synovial lining and articular cartilage to synthesize and secrete neutral metalloproteinases reveals a difference between these models. We found that articular cartilage from the inflamed joints of rabbits with antigen-induced arthritis did not synthesize neutral metalloproteinases, although the synovial lining did. In contrast, both the synovial lining and the articular cartilage from the inflamed joints of rabbits with polycation-induced arthritis synthesized neutral metalloproteinases. These findings suggest that in inflammatory synovitis, different mechanisms can operate to produce damage to the matrix of articular cartilage.

Topics: Animals; Antigens; Arthritis; Arthritis, Experimental; Cartilage, Articular; Chronic Disease; Culture Media; Culture Techniques; Hyaluronic Acid; Injections, Intra-Articular; Lysine; Male; Metalloendopeptidases; Ovalbumin; Polyamines; Polyelectrolytes; Polymers; Prostaglandins E; Proteoglycans; Rabbits; Synovial Membrane; Time Factors

Homocytotropic antibody was stimulated in animals by administering protein antigens in a vaccine with B. pertussis adjuvant. The titers of the allergic antibody responses were judged by passive cutaneous anaphylaxis reactions. Sera or globulin fractions containing high titers of antibody activity were injected into the knee joints of experimental animals. After sufficient delay for unfixed proteins to be cleared from the knee joints, animals were challenged intravenously with the corresponding antigen. The resultant local reaction of swelling and warmth (passive synovial anaphylaxis) was judged visually and by scanning procedures. Histological studies showed evidence of mast cell degranulation concurrent with synovial reaction.

Topics: Animals; Antibodies; Antigens; Arthritis; Cell Count; Cell Degranulation; Evans Blue; Immunization; Joints; Mast Cells; Ovalbumin; Rabbits; Skin; Synovial Membrane

Joint inflammation was induced in C57B1/6 mice by injection of cloned MT4+, Lyt-2- T cells specific for the antigen methylated bovine serum albumin (mBSA), together with mBSA. In this model, after waning of the inflammation, flare reactions can be induced by a rechallenge with the specific antigen. Herein we show that such flare reactions can still be induced several weeks after waning of the joint inflammation, as was demonstrated both in normal C57B1/6 mice and in athymic C57B1 nude mice. The results in the latter group indicate that T cells of the recipient mice are not necessary for the elicitation of flare reactions. On histologic examination, the inflammatory infiltrates in the knee joints of the nude mice appeared to be mainly granulocytic. The cloned T cells persisted and remained functionally reactive in the knee joint for at least 2 weeks in the absence of the antigen, and thus, in the absence of inflammation. In view of the similarities between induced joint inflammation in mice and rheumatoid arthritis in humans, these data may be relevant to our understanding of the processes involved in the latter disease.

Topics: Animals; Antigens, Ly; Arthritis; Arthritis, Experimental; CD4-Positive T-Lymphocytes; Cells, Cultured; Clone Cells; Female; Hypersensitivity, Delayed; Mice; Mice, Inbred C57BL; Mice, Nude; Ovalbumin; Radionuclide Imaging; Serum Albumin, Bovine; Technetium

We examined the kinetics of proteoglycan (PG) depletion in rabbits with antigen-induced arthritis. There was a rapid loss of PG from arthritic cartilage, reaching 35-40% at day 7. Thereafter, the rate of PG depletion declined, and by day 42, the maximum loss was 55-60%. The initial loss of PG was accompanied by the appearance of large amounts of sulfated glycosaminoglycans (GAGs) in the joint fluid (measured as total sulfated GAGs by dye binding and as keratan sulfate by radioimmunoassay). However, by day 14, the levels of sulfated GAGs in arthritic joint fluid declined to control levels, even though the cartilage demonstrated a sustained depletion of PG. The cartilage PG degradation observed in antigen-induced arthritis could also be produced in normal animals by a single intraarticular injection of recombinant interleukin-1. The acute loss of cartilage PG occurred independently of neutrophil accumulation, both in the case of antigen-induced arthritis and after injection of interleukin-1.

Topics: Animals; Antigens; Arthritis; Arthritis, Experimental; Cartilage, Articular; Glycosaminoglycans; Immunization; Injections, Intra-Articular; Interleukin-1; Joints; Keratan Sulfate; Knee Joint; Leukopenia; Male; Mechlorethamine; Neutrophils; Ovalbumin; Proteoglycans; Rabbits

Dysprosium-165-ferric hydroxide macroaggregates (165Dy-FHMA) was used as an agent of radiation synovectomy in an antigen-induced arthritis model in New Zealand white rabbits. Animals were killed up to 6 months after treatment. 165Dy-FHMA was found to have a potent but temporary antiinflammatory effect on synovium for up to 3 months after treatment. Treated knees also showed significant preservation of articular cartilage architecture and proteoglycan content compared with untreated controls, but only during the first 3 months after treatment. In animals killed 3 and 6 months after treatment there were only minimal differences between the treated and untreated knees, indicating that the antiinflammatory effects on synovial tissue and articular cartilage preservation were not sustained.

Topics: Animals; Antigens; Arthritis; Arthritis, Experimental; Brachytherapy; Cartilage, Articular; Drug Carriers; Dysprosium; Ferric Compounds; Fibrosis; Hyperplasia; Knee Joint; Ovalbumin; Particle Size; Rabbits; Radiography; Radioisotopes; Synovial Membrane

Antigen-induced arthritis in the rabbit (AIAR) provides the closest experimental equivalent to human rheumatoid arthritis in terms of infiltration of synovial tissue by lymphoid cells. A method is described for quantitative histological analysis of AIAR. Measurements of total cell numbers, lymphocyte and polymorphonuclear leucocyte infiltration, and thickness of infiltrated synovium were obtained for ranges of antigen dosage and duration of arthritis. The method has been devised as part of a system for the analysis of joint swelling, synovial fluid biochemistry and cytology, cartilage proteoglycan chemistry and synovial histology on the same specimen.

Topics: Animals; Antigens; Arthritis; Cell Count; Dose-Response Relationship, Immunologic; Hindlimb; Leukocyte Count; Lymphocytes; Male; Neutrophils; Ovalbumin; Rabbits; Synovial Membrane; Time Factors

The relative contributions of cellular and humoral immunity to cartilage destruction in chronic arthritis has been investigated in a model of chronic synovitis in the rabbit. In this model, antigen-induced arthritis, immunization with ovalbumin in Freund's complete adjuvant (FCA) followed by intra-articular injection of this protein produces a chronic synovitis associated with loss of proteoglycan from articular cartilage. In addition, the synovial lining cell population is metabolically activated. Similar treatment of animals immunized with ovalbumin in Freund's incomplete adjuvant (FIA) produced a resolving arthritis which initially (over the first 7 days) appears to be identical to that in FCA-immunized animals, apart from the lack of activation of synovial lining cells. Following this initial synovitis the joints return to apparent normality apart from a persistent 'low grade' synovitis consisting mainly of a plasma cell infiltrate. The most striking finding in the FIA-immunized animals is the rapid loss (greater than 30% by day 7) and recovery of proteoglycan from the matrix of articular cartilage. These findings show that the perpetuation of chronic destructive synovitis in the rabbit requires the presence of active cellular immunity.

Topics: Acid Phosphatase; Animals; Antibodies; Arthritis; Arthritis, Experimental; Cartilage, Articular; Female; Guinea Pigs; Hypersensitivity, Delayed; Immunity, Cellular; Male; Ovalbumin; Proteoglycans; Rabbits; Synovial Fluid; Synovitis

The activity and distribution of the collagenolytic lysosomal enzyme, cathepsin L, has been assessed in the synovial lining of the rabbit during the initiation and development of chronic (antigen-induced) arthritis. Biochemical assay of homogenates of synovial lining revealed a marked increase in the activity of lysosomal enzymes, including cathepsin L, between 1 and 5 days following initiation of arthritis. Elevated levels of these enzymes were still present at 4 weeks duration of arthritis. Using a monospecific antibody to cathepsin L. enzyme was immunolocated only in synovial lining cells in normal joints. In arthritic joints the enzyme was found in synovial lining cells, synovial fibroblasts and in infiltrating macrophages. Measurement, by scanning and integrating microdensitometry, of the peroxidase-reaction product in synovial lining cells revealed an increased content of cathepsin L in these cells in the diseased tissues.

Topics: Acetylglucosaminidase; Animals; Arthritis; Arthritis, Experimental; Cathepsin L; Cathepsins; Cysteine Endopeptidases; Endopeptidases; Fibroblasts; Glucuronidase; Macrophages; Male; Ovalbumin; Rabbits; Synovial Membrane

We analyzed the histopathologic findings of arthritis in 3 rat models: adjuvant arthritis induced by active immunization to Mycobacterium tuberculosis (MT) antigens, arthritis produced by passive transfer of an intrinsically arthritogenic line of anti-MT T lymphocytes, and bystander arthritis produced by intraarticular injection of a foreign antigen, ovalbumin, into rats with T lymphocyte line cells specific for the ovalbumin antigen. The histopathology of the tibiotarsal and knee joints was studied by light microscopy and the articular surface of the cartilage by electron microscopy after labeling with cationized ferritin. The lesions in the 3 models of arthritis were compared. In active adjuvant arthritis, inflammatory lesions and cartilage destruction were found as early as 9 days after immunization, and persisted for as long as 11 months. Similar, but somewhat milder, lesions were found in arthritis produced by transfer of anti-MT T lymphocytes. Inflammatory signs were present at 4 days, when there was no evidence of joint edema. Severe inflammatory lesions were found in arthritis induced by transfer of anti-ovalbumin T lymphocytes that was followed by ovalbumin injection into the knee. Pathologic changes were found to be similar in all 3 models. Thus, the changes could be attributed to the action of T lymphocytes, irrespective of whether the target antigen was intrinsic to the joint.

Topics: Animals; Antigens, Bacterial; Arthritis; Arthritis, Experimental; Cartilage, Articular; Epitopes; Hindlimb; Immunization; Immunization, Passive; Joints; Microscopy, Electron; Mycobacterium tuberculosis; Ovalbumin; Rats; Rats, Inbred Lew; T-Lymphocytes

T-cell lines were established from the lymph node cells of syngeneic Louvain (LOU) rats previously immunized with native chick type II collagen (CII) emulsified in incomplete Freund's adjuvant. The CII lines proliferated in vitro to type II collagen but not to type I collagen, ovalbumin (OV), or PPD. Control lines, developed from LOU rats immunized with OV emulsified in complete Freund's adjuvant, were OV specific because they did not respond to other antigens in vitro. CII line cells could adoptively transfer delayed-type hypersensitivity (DTH) but did not induce IgG antibody production to collagen. Moreover, the intravenous administration of 2 X 10(7) CII line cells prevented the subsequent induction of collagen arthritis following immunization and suppressed DTH to collagen without affecting antibody responses in the recipients. Spleen cells, but not sera, from these resistant rats decreased CII line reactivity in vitro. OV or irradiated CII lines had no effect on clinical or immunologic parameters in this model. These findings demonstrate protection from arthritis afforded by T-cell line transfer and suggest that the phenomenon results from down-regulation of the recipients' cellular immunity to collagen.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Line; Collagen; Hypersensitivity, Delayed; Immunization, Passive; Immunoglobulin G; Lymphocyte Activation; Ovalbumin; Rats; T-Lymphocytes; Tuberculin

Chronicity of murine allergic arthritis depends on the charge-mediated retention of the cationic antigen in the joint. The authors examined whether arthritis induced with the positively charged antigen amidated bovine serum albumin (aBSA) could be modulated with a nonimmunogenic polycation by competition for anionic retention sites in the joint. Concomitant intraarticular injection of aBSA with the cationic protein protamine chloride (pI approximately 10) strongly reduced the retention of aBSA. Detailed analysis revealed that the retention of aBSA in the noncartilaginous tissues was significantly reduced by protamine, whereas the retention in the highly negatively charged cartilage was completely prevented. Joint inflammation was already significantly suppressed at Day 3 and suppression was still demonstrable at Day 28. Protamine treatment also caused a highly significant reduction in cartilage damage and bone apposition. Control experiments indicated that the suppressive effect of protamine was related to its interference with antigen retention in the joint and not to a mere antiinflammatory action.

Topics: Amides; Animals; Antibodies; Antigens; Arthritis; Arthritis, Experimental; Binding, Competitive; Cartilage; Cations; Hypersensitivity, Delayed; Knee; Metabolic Clearance Rate; Mice; Ovalbumin; Protamines; Serum Albumin, Bovine; Tissue Distribution; Zymosan

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

Acute phase protein levels have been measured during the induction and progression of antigen-induced mono-articular arthritis in rabbits and mice. In rabbits there was a short lived elevation in serum C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) immediately following intra-articular injection which returned to baseline levels 10-12 days after the injection. In BALB/c mice, serum amyloid P-component (SAP) and the third component of complement (C3) were elevated after intra-articular injection, returning towards baseline levels 6 weeks after the injection. The levels of CRP and SAP correlated with the inflammatory changes in the joints during the acute phase of the arthritic response (7 days after intra-articular injection). During the chronic phase the levels of these acute phase proteins bore no relationship to the degree of connective tissue destruction.

Topics: Acute-Phase Proteins; Animals; Arthritis; Blood Sedimentation; C-Reactive Protein; Complement C3; Female; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Rabbits; Serum Amyloid P-Component

Topics: Animals; Arthritis; Arthritis, Experimental; Catalase; Cations; Cattle; Mice; Ovalbumin; Rabbits; Serum Albumin, Bovine

Fibronectin (FN), a high molecular weight glycoprotein, is present in plasma and is a normal structural component of the synovium in the rabbit, as it is in man. FN is also involved in the sequence of changes seen in synovium in experimental antigen-induced arthritis. Its widespread distribution in inflamed synovia in the initial acute phase of induced arthritis probably merely reflects the presence of FN of plasma origin in serous exudates. In established experimental arthritis, FN co-distributes with fibrin, while in synovia undergoing organisation, FN is present intracellularly in several types of mesenchymal cells (suggesting local synthesis) and is deposited on immature collagen fibrils. However, it is no longer present when mature collagen is formed. The persistence of FN, along with fibrin, in inflamed joints, and its involvement in fibrosis, suggest that it may play a significant part in determining the chronicity of this form of experimental arthritis.

Topics: Animals; Antigens; Arthritis; Fibronectins; Fluorescent Antibody Technique; Ovalbumin; Rabbits; Synovial Membrane; Synovitis

We studied the ability of dimethyl sulfoxide (DMSO) to influence the course of an experimental model of inflammatory arthritis. A Dumonde-Glynn model of arthritis was induced in both tibio-femoral joints of 10 rabbits, using ovalbumin as the immunogen. At one month post induction of the arthritis, the right tibio-femoral joint of 6 animals was treated for 3 months with topical 80% DMSO--1 g/kg body weight applied to the shaved skin for 5 out of 7 days each week. In another 4 animals, the right tibio-femoral joint was injected with 0.5 ml of 80% DMSO at one month post induction of the arthritis. Joint radiographs were taken at monthly intervals. The rabbits were sequentially sacrificed and the joint tissues evaluated by a blinded observer. Neither the topical DMSO nor the intraarticular DMSO treated joints showed any favorable responses to therapy; in fact the topically treated joints exhibited somewhat more inflammatory and destructive changes than the untreated joints. However the repeated injection of DMSO into normal joints did not, of itself, produce any deleterious effects. This study indicates a need to assess more thoroughly a possible deleterious effect of DMSO on the course of untreated inflammatory arthritis.

Topics: Administration, Topical; Animals; Arthritis; Arthritis, Experimental; Cartilage, Articular; Dimethyl Sulfoxide; Female; Injections, Intra-Articular; Male; Ovalbumin; Rabbits; Radiography; Synovial Membrane

In an attempt to produce a superior model of rheumatoid arthritis, experiments have been performed to investigate the ease of induction of experimental arthritis in marmosets by immunological means. Marmosets were sensitised with the following combinations of antigen and adjuvant: ovalbumin in Freund's complete adjuvant (FCA), ovalbumin in FCA + Bordetella pertussis, methylated-BSA in FCA + B. pertussis or human fibrin in FCA + B. pertussis, and subsequently injected with the corresponding antigen in saline into one knee joint. Animals receiving ovalbumin, with or without B. pertussis, produced only a weak transient monoarticular synovitis. Animals receiving Met-BSA + B. pertussis produced a chronic synovitis but only mild erosive changes were apparent even 21 weeks after intraarticular injection. Animals receiving human fibrin produced a transient monoarticular synovitis of moderate intensity. These results indicate that the marmoset offers no obvious advantages over the rabbit for the induction of experimental rheumatoid arthritis.

Topics: Animals; Arthritis; Arthritis, Experimental; Callithrix; Fibrin; Freund's Adjuvant; Immunization; Injections, Intra-Articular; Knee Joint; Ovalbumin; Pertussis Vaccine; Rheumatoid Factor; Serum Albumin, Bovine

Antigen induced arthritis (AIA) was elicited in 28 adult rabbits using ovalbumin. All animals developed an intensive monarthritis manifested by local heat and swelling. Two, 7 and 21 days later, respectively, 0.3 ml of 1 per cent OsO4 was injected into the arthritic knees of three groups of 9 rabbits. The rabbits were killed 1, 3 and 6 months later. No modifying influence on the severe to complete cartilage destruction was evident. One group of 8 rabbits received only OsO4 and the cartilage of these animals remained intact after 6 months.

Topics: Animals; Arthritis; Cartilage; Knee Joint; Necrosis; Osmium; Osmium Tetroxide; Ovalbumin; Rabbits; Time Factors

Rabbits have been immunized with ovalbumin in Freund's incomplete adjuvant (FIA) followed by the intra-articular injection of ovalbumin, in order to follow the development of inflammation in the synovial lining. The kinetics of cell proliferation have been investigated using tritiated thymidine (3HTdR) autoradiography and Feulgen cytophotometry. Unexpectedly, marked histological changes were found in the synovium, with hyperplasia of synoviocytes, and of the connective-tissue cells of the subintima, being seen as early as 3 days after challenge. Large numbers of inflammatory cells, including many plasma cells, were found in the synovium at between 5 and 11 days. Labelling of synoviocytes and connective-tissue cells reached a maximum at 3 days and declined thereafter, reaching normal levels at 14 days. Three weeks after challenge the synovium was normal in appearance.

Topics: Animals; Arthritis; Arthritis, Experimental; Autoradiography; Cell Division; DNA; Female; Male; Neutrophils; Ovalbumin; Rabbits; Synovial Membrane; Time Factors

Topics: Animals; Arthritis; Autoantibodies; Chickens; Collagen; Female; Hypersensitivity, Delayed; Leukocyte Migration-Inhibitory Factors; Ovalbumin; Rats; Rats, Inbred BN; Rats, Inbred Lew; Spleen

Collagen-induced polyarthritis in rats is a new experimental model that shares clinical and histologic features with adjuvant arthritis. To determine whether collagen-induced arthritis is a form of adjuvant disease and to further exclude contamination of collagen with an adjuvant substance, native type II collagen was studied for adjuvant properties. IgM and IgG PFC activity and PBMC [3H]TdR incorporation were studied in rats after injection with TNP-OA combined with IFA, IFA and CII, or CFA. In general, humoral and CMI responses to TNP-OA were lower in rats injected with IFA/CII compared with those with IFA; the presence of CII during primary immunization failed to significantly enhance PFC activity to TNP after a boost. CFA-injected rats gave maximal values in both studies. Mice pretreated with BII in the absence of oil gave PFC responses below control after sensitization with SRC. Furthermore, CII was unable to replace mycobacteria in the induction of EAE in rats and was devoid of mitogenic or polyclonal stimulatory properties. It is concluded that collagen-induced arthritis is a distinct entity from adjuvant arthritis and is dependent upon the unique immunogenicity of type II collagen in rats rather than upon an adjuvant effect.

Topics: Animals; Arthritis; Arthritis, Experimental; Clone Cells; Collagen; Encephalomyelitis, Autoimmune, Experimental; Female; Hemagglutination Tests; Hemolytic Plaque Technique; Immunity, Cellular; Male; Mice; Mice, Inbred C57BL; Mitogens; Myelin Proteins; Oils; Ovalbumin; Rats; Rats, Inbred Lew; Trinitrobenzenes

Bee venom, administered subcutaneously, suppressed the development of carrageenan-induced paw edema and adjuvant arthritis in the rat in a dose-related manner. A single dose of bee venom administered subcutaneously the day before or on the day of injection of complete Freund's adjuvant (CFA) effectively suppressed the development of polyarthritis. This suppressive effect decreased progressively as dosing was delayed. Bee venom was found to be most effective when mixed and injected (sub-plantar) together with CFA, the disease-inducing agent. Similarly, antigens such as egg albumin, when incorporated into CFA, and injected into the hind paw, prevented the development of arthritis. These results suggest that at least two mechanisms are involved in the anti-arthritic action of bee venom: (1) alteration of the immune response, probably via antigen competition, and (2) an anti-inflammatory action via corticosteroids or through an as yet undetermined mechanism.

Topics: Animals; Anti-Inflammatory Agents; Arthritis; Arthritis, Experimental; Bee Venoms; Carrageenan; Cyclophosphamide; Edema; Hydrocortisone; Male; Ovalbumin; Rats

Cyclophosphamide is given intravenously in the high dose of 12 mg/kg/day during 2 months from the beginning of immunological systemic connective tissue disease in rabbits, according to the modified Glynn's model. The effects of this alkylating drug are studied after the termination of treatment. At short term (15 days), cyclophosphamide depresses the inflammation of synovia in stimulated and non-stimulated joints, the humoral and cellular immunities, but the drug does not modify the other systemic lesions. At long term (between 1 and 6 months), these effects of cyclophosphamide disappear completely.

Topics: Animals; Antigens, Bacterial; Arthritis; Arthritis, Experimental; Cyclophosphamide; Immunosuppression Therapy; Male; Ovalbumin; Rabbits; Time Factors

D-penicillamine given orally to rabbits at a dose of 15 mg/kg body weight, administered daily for long periods commencing prior to and at the time of immunization, reduced the concentration of specific IgG antibodies directed against ovalbumin. This effect was not observed when treatment with the drug commenced after immunization and the onset of a mono-articular arthritis. The concentration of serum IgA rose abruptly after antigen was injected intra-articularly to induce the arthritis. This increase was not affected by treatment with D-penicillamine. The concentration of serum IgA decreased markedly during the chronic phase of the mono-articular arthritis, falling below the lower limit of the normal range. Treatment with D-penicillamine for at least 50 days restored the serum IgA concentration to the middle of the normal range. D-penicillamine treatment had no significant effect on the concentration of IgG, or IgM in serum either before or after the onset of mono-articular arthritis.

Topics: Administration, Oral; Animals; Antibody Formation; Arthritis; Female; Immunoglobulins; Male; Ovalbumin; Penicillamine; Rabbits; Time Factors

Knee joint arthritis is induced among rabbits previously immunized by intradermal injection with egg albumin (EA) emulsified in adjuvant containing either M. tuberculosis or M. butyricum, then by intra-articular injection with EA. Arthritis evolution involves two phases, an early one during the first 2 months and a late one from 3 months to 1 year. During the early phase, arthritis intensities are similar no matter which Mycobacterium is used. However, during the late phase, only rabbits immunized with M. tuberculosis develop self-perpetuating arthritis. Among more than 50% of arthritic rabbits, immunological lesions of aortic artery and cardiac valvules are found. Among the rabbits immunized with M. tuberculosis, the humoral anti-EA antibody level remains constant during the whole arthritis evolution; but, among the rabbits immunized with M. butyricum, the arthritis intensity decreases from 3 months of evolution. The correlation between arthritis index (AI) and humoral antibody level is only significant among the rabbits with early arthritis. The intradermally immunized rabbits show a positive skin test with EA and tuberculin. The more intense the cutaneous reactions, the greater the chances of developing self-perpetuating arthritis after the EA intra-articular injection. The fluorescent anit-EA antibodies in the synovia and spleen are found only among the early arthritis. After 2 months of evolution, fluorescent antibodies disappear whatever the immunization may be. Among the immunized rabbits, it is probable that antigenic EA does not persist in the synovia. Indeed, the autologous inflamed synovia transplantation, from the donor-challenged knee joint, does not develop an inflammatory reaction in the non-challenged knee joint. The fluorescent immunoglobulins IgG and IgM in the synovium of arthritic rabbits are found with the same percentages as fluorescent anti-EA antibodies. The lymphocyte response to EA, PHA and PWM are positive whatever the immunization and arthritis evolution may be. There is no correlation between AIs and lymphocyte responses to specific and nonspecific mitogens. It is probable that self-perpetuation depends closely on M. tuberculosis whose adjuvant power is much superior than M. butyricum and not on antigenic EA whose essential role would be to trigger the inflammatory process.

Topics: Animals; Antibodies; Antigens, Bacterial; Arthritis; Arthus Reaction; Fluorescent Antibody Technique; Immunization Schedule; Immunoglobulin G; Immunoglobulin M; Injections, Intra-Articular; Injections, Intradermal; Lymphocyte Activation; Male; Mycobacterium; Ovalbumin; Rabbits; Skin Tests; Spleen; Synovial Membrane; Transplantation, Homologous; Tuberculin

The presence of antigen, IgG and C3 was shown by radioautography and immunofluorescence in the collagenous tissues of the joints of animals injected intra-articularly with antigen after having been previously immunized with that antigen in Freund's incomplete adjuvant. Since these joints were shown to be virtually free of inflammatory reactions, we suggest that the persistence of immune complexes activating complement cannot fully explain the chronicity of experimental allergic arthritis.

Topics: Animals; Antigens; Arthritis; Autoradiography; Complement C3; Fluorescent Antibody Technique; Hindlimb; Immunoglobulin G; Joints; Ovalbumin; Rabbits; Radiography

A water-soluble mycobacterial glycopeptide was obtained in large quantities from the culture supernatant fluid of M. tuberculosis strain DT. This glycopeptide was strongly adjuvant-active when injected, in a water-in-oil emulsion contianing ovalbumin, into guinea-pigs. In addition, it was devoid of cord factor toxicity in mice, polyarthritogenic activity in rats and cavity stimulating activity in rabbit lungs.

Topics: Adjuvants, Immunologic; Animals; Arthritis; Cord Factors; Cornea; Glycopeptides; Glycoproteins; Guinea Pigs; Hypersensitivity, Delayed; Immunoglobulin G; Mycobacterium tuberculosis; Ovalbumin; Precipitin Tests; Rabbits; Rats; Rats, Inbred Lew; Skin Tests; Solubility; Tuberculosis, Pulmonary

1 Old English (OE) rabbits produced more severe monoarticular arthritis (MAA) after sensitization and intra-articular challenge with ovalbumin than did either New Zealand White (NZW) or Dutch rabbits. NZW rabbits were better responders than Dutch rabbits.2 The swelling of the joint in all three strains of rabbits was triphasic. There was an initial acute swelling which appeared to peak at 2-4 days after challenge. This was followed by a decrease in joint size, and then a secondary increase in size beginning 1-2 weeks after challenge.3 An investigation of MAA in OE rabbits showed that there was an increase in E-type prostaglandins, total leucocyte counts and free acid phosphatase activity in the synovial fluid of the challenged joints at 6 h, 19 h, 47 h, 7 days and 46 days following challenge. There were also histopathological changes at these times. In addition, there was an increase in the surface temperature of both the challenged and non-challenged knees, and a rise in the body temperature.4 Prostaglandin levels peaked at 19 h and were equivalent to 19 ng E(2) per joint. In a separate experiment, the prostaglandin present at 18 h was shown to be mainly E(1). Maximum levels of prostaglandin appeared to coincide with maximum joint temperature, but preceded maximum joint swelling and a significant rise in both the number of inflammatory cells and the free acid phosphatase activity in the synovial fluid, all of which occurred at 47 hours.5 Indomethacin, 7.5 mg/kg orally twice daily, almost completely inhibited the increase in prostaglandin levels in the challenged joints and produced a moderate reduction in joint swelling. It also reduced the increased surface temperature of both knee joints and the raised body temperature. However, indomethacin had no effect on the number of leucocytes present, the free acid phosphatase levels, or the histopathological changes in the joint.6 The mean plasma level of indomethacin ranged from 0.5 to 3 mug/ml at the time when the animals were killed.7 Lysosomal enzymes may be more important than prostaglandins in rabbit MAA, and the lack of effect of indomethacin on joint histopathology may be due to its inability to prevent the release of these enzymes.

Topics: Acid Phosphatase; Animals; Arthritis; Body Temperature; Female; Immunization; Indomethacin; Knee Joint; Leukocyte Count; Lysosomes; Male; Mycobacterium; Ovalbumin; Prostaglandins; Rabbits; Species Specificity; Synovial Fluid; Time Factors

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

Digestion by lysozyme of delipidated cells of Mycobacterium smegmatis liberates a water-soluble immunoadjuvant fraction which is chemically very similar to the water-soluble adjuvant (WSA) obtained previously from purified cell walls, but which contains somewhat more non-peptidoglycan amino acids. The yield of peptidoglycan-arabinogalactan complex is about 10 times greater starting from whole cells than from cell walls. The main biological properties of this "neo-WSA" are described: it increases circulating antibodies to ovalbumin in guinea pigs, it does not produce polyarthritis in rats or induce hypersensitivity to tuberculin, it does not increase susceptibility to histamine or hyperreactivity to endotoxin, and does not produce spleen and liver hypertrophy. Analogous immunostimulant fractions have also been obtained from delipidated cells of Nocardia opaca by lysozyme treatment.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Arthritis; Cell Wall; Chromatography; Endotoxins; Guinea Pigs; Histamine; Lipid Metabolism; Liver; Mice; Muramidase; Mycobacterium; Nocardia; Ovalbumin; Rats; Spleen; Ultracentrifugation

Topics: Animals; Arthritis; Aspirin; Carrageenan; Croton Oil; Edema; Foot; Granuloma; Hindlimb; Hydrocortisone; Inflammation; Male; Methods; Mycobacterium; Ovalbumin; Phenylbutazone; Prostaglandins; Rats; Serotonin

Whole mycobacterial cells, which are used in Freund's complete adjuvant, besides inducing hypersensitivity to tuberculoprotein, also can elicit hyperreactivity to endotoxins, lymphoid hyperplasia, and allergic polyarthritis in rats. The data reported here demonstrate that a potent water-soluble adjuvant obtained from mycobacterial cell walls is also effective in increasing the immune response to viruses, and that it is free of the toxic effects observed with whole mycobacterial cells.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Antigen-Antibody Complex; Arthritis; Cell Wall; Encephalomyocarditis virus; Endotoxins; Freund's Adjuvant; Guinea Pigs; Hepatomegaly; Hypersensitivity, Delayed; Male; Mice; Mice, Inbred Strains; Mycobacterium; Orthomyxoviridae; Ovalbumin; Precipitins; Rats; Splenomegaly; Tuberculin

In an experimental arthritis induced by injection of bovine serum albumin or egg albumin into the joints of previously immunized animals, it has been demonstrated that the major portion of the radioactively labeled antigens injected was localized to avascular collagenous tissues in the joint, i.e., articular cartilage, menisci, and intra-articular ligaments. The antigens were partially eluted from the tissues with 5 M guanidine solution, but not with acid buffers or by 3 M magnesium chloride. The radioactive material eluted with guanidine was at least 80% precipitable by specific antisera. The radioactively labeled-inducing antigen was identified on the surface of articular collagenous tissues from arthritic joints by radioautography and immunofluorescence. Rabbit immunoglobulin and C3 were demonstrated in the same sites by immunofluorescence. The presence of specific antibody in collagenous tissues was demonstrated by the selective in vitro binding of (125)I-labeled-inducing antigen to menisci from arthritic joints of immunized animals. The evidence obtained indicates that in this model of chronic arthritis, the inducing antigen persists for long periods of time in the form of immune complexes in the surface layers of the intra-articular collagenous tissue. The antigen retained in this form may be responsible for the chronicity of the synovitis by serving as a direct stimulus for the maintenance of prolonged antibody synthesis in the synovium and by providing a source of complement-fixing antigen-antibody complexes for the mediation of joint inflammation.

Topics: Animals; Antibody Formation; Antigen-Antibody Complex; Antigens; Arthritis; Autoradiography; Cartilage, Articular; Chronic Disease; Collagen; Complement System Proteins; Disease Models, Animal; Fluorescent Antibody Technique; Guanidines; Immunoglobulins; Inflammation; Iodine Isotopes; Knee Joint; Ovalbumin; Rabbits; Serum Albumin, Bovine; Synovial Membrane; Synovitis

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Antigens; Arthritis; Benzoates; Bone Marrow; Erythrocytes; Immunoelectrophoresis; Immunosuppression Therapy; Lymph Nodes; Male; Mycobacterium tuberculosis; Ovalbumin; Procarbazine; Rats; Rats, Inbred Strains; Sheep; Spleen; Thymus Gland

Topics: Animals; Antigens; Arthritis; Autoradiography; Binding Sites; Chromatography, DEAE-Cellulose; Female; Immunoelectrophoresis; Immunoglobulin G; In Vitro Techniques; Injections, Intra-Articular; Iodine Isotopes; Macrophages; Ovalbumin; Rabbits; Synovial Fluid; Synovitis

Topics: Animals; Arthritis; Female; Freund's Adjuvant; gamma-Globulins; Hindlimb; Hyperplasia; Immunization; Inflammation; Injections, Intra-Articular; Iodine; Iodine Isotopes; Joints; Lymphocytes; Male; Ovalbumin; Patella; Rabbits; Staining and Labeling; Synovial Membrane

Topics: Animals; Arthritis; Cartilage, Articular; Chronic Disease; Disease Models, Animal; Female; Hindlimb; Leukocytes; Lymphocytes; Macrophages; Male; Ovalbumin; Prednisolone; Rabbits; Synovitis

Topics: Adrenalectomy; Animals; Anti-Inflammatory Agents; Arthritis; Calcium; Capillary Permeability; Carrageenan; Depression, Chemical; Dextrans; Edema; Erythema; Granuloma; Hemolysis; Hyaluronoglucosaminidase; Indicators and Reagents; Male; Malonates; Mice; Ovalbumin; Phenylbutazone; Phenylhydrazines; Prednisolone; Protein Denaturation; Radiation Injuries, Experimental; Rats; Serotonin; Ultraviolet Rays; Wound Healing

Topics: Animals; Arthritis; Cyclophosphamide; Disease Models, Animal; Guinea Pigs; Hindlimb; Hypersensitivity, Delayed; Ovalbumin; Tuberculin Test

Topics: Animals; Arthritis; Cyclophosphamide; Disease Models, Animal; Guinea Pigs; Hindlimb; Hypersensitivity, Delayed; Ovalbumin; Tuberculin Test

Topics: Animals; Arthritis; Arthus Reaction; Bradykinin; Capillary Permeability; Carrageenan; Depression, Chemical; Dextrans; Disease Models, Animal; Dogs; Edema; Exudates and Transudates; Female; Formaldehyde; Granuloma; Guinea Pigs; Histamine; Hyaluronoglucosaminidase; Kaolin; Lymph; Male; Mice; Mycobacterium Infections; Ovalbumin; p-Methoxy-N-methylphenethylamine; Phytotherapy; Plants, Medicinal; Rabbits; Rats; Serotonin; Species Specificity; Swine

Topics: Acetylcholine; Analgesics; Animals; Anti-Inflammatory Agents; Arthritis; Atropine; Croton Oil; Depression, Chemical; Edema; Exudates and Transudates; Female; Formaldehyde; Glycosides; Granuloma; Guinea Pigs; Histamine H1 Antagonists; In Vitro Techniques; Intestines; Male; Ovalbumin; Papaverine; Parasympatholytics; Phenylbutazone; Plant Extracts; Plants, Medicinal; Rabbits; Rats

Topics: Adjuvants, Immunologic; Adsorption; Animals; Antigens; Arthritis; Cattle; Female; Freund's Adjuvant; gamma-Globulins; Injections, Intradermal; Injections, Intramuscular; Ovalbumin; Proteins; Rats; Serum Albumin; Serum Albumin, Bovine; Waxes

Topics: Arthritis; Fibrinolysin; Glucose; Humans; Muramidase; Ovalbumin; Peptide Hydrolases; Phosphoric Monoester Hydrolases; Synovial Fluid; Synovial Membrane; Transaminases

Topics: Animals; Antigens; Arthritis; Freund's Adjuvant; Hypersensitivity; Ovalbumin; Rats; Skin Tests

Topics: Adjuvants, Immunologic; Animals; Arthritis; Biological Assay; Carrageenan; Female; In Vitro Techniques; Inflammation; Injections, Intramuscular; Injections, Intraperitoneal; Injections, Intravenous; Injections, Subcutaneous; Mineral Oil; Mycobacterium; Ovalbumin; Pregnancy; Pregnancy, Animal; Rats; Stress, Physiological