concanavalin-a and Arthritis

Liver function abnormalities are common in patients with inflammatory arthritis. However, the precise mechanism is still unclear. In this study, inflammatory arthritis was established in mice by subcutaneous injection of complete Freund's adjuvant, and the intravenous injection of concanavalin A (Con A) was employed to induce acute immune-mediated hepatitis in mice. The result showed that the arthritis mice were more susceptible to ConA-induced hepatitis than the control mice, as evidenced by increased hepatic necrosis, elevated serum alanine aminotransferase activity, and raised inflammatory cytokines. Besides, the in vitro assay demonstrated that the T cells from arthritis mice were more sensitive to the Con A stimulation than those from control mice. Moreover, we determined that the level of leptin, a kind of adipokine, was significantly increased in the serum and hepatic T cells of arthritis mice. Interestingly, the data indicated that the enhanced expression of leptin in hepatic T cells is responsible for the hypersensitivity of arthritis mice-derived T cells to Con A challenge. Collectively, our findings demonstrate an unexpected role of leptin in the connection between inflammatory arthritis and acute immune-mediated hepatitis, thus providing new insight into the clinical therapy of arthritis-related liver dysfunction.

Topics: Acute Disease; Animals; Arthritis; Concanavalin A; Cytokines; Disease Susceptibility; Hepatitis; Hypersensitivity; Inflammation; Inflammation Mediators; Leptin; Liver; Lymphocyte Activation; Mice, Inbred C57BL; Signal Transduction; T-Lymphocytes

We have discovered a new class of colchicine-derived therapeutic agents for immune diseases including rejection of organ-transplantation and autoimmune disease. Compound 2, which had been developed to overcome poor pharmacokinetic properties of compound 1, a first-generation colchicine analog, turned out to show toxicity such as intestinal toxicity and loss of weight during in vivo tests. The deletion of 7-carboxamide group and middle ring-truncation in colchicine allowed us to have structurally simplified analogs with strong immunosuppressive activity. Herein, we report non-alkaloid tricyclic compound 7 and 12 as immunosuppressants which exhibited a strong immunosuppressive in vivo efficacy on the T-dependent antibody response, the Zymosan A-induced arthritis model and the Carrageenan-induced edema model. Compound 7 and 12 revealed less toxicity than the previous lead compound 2, and their minimum lethal doses (MLD) were proved to exceed 100 mg/kg.

Topics: Animals; Arthritis; B-Lymphocytes; Carrageenan; Cell Proliferation; Colchicine; Concanavalin A; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Discovery; Edema; Humans; Immunosuppressive Agents; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Molecular Structure; Structure-Activity Relationship; T-Lymphocytes; Zymosan

The chimeric cell surface receptor scC2Fv/CD8/zeta was constructed to engineer primary mouse T lymphocytes with antibody-type specificity to type II collagen (CII). Such cells could be used as gene carriers in the anti-inflammatory gene therapy of an autoimmune arthritis. This receptor includes the single chain Fv domain (scFv) of the anti-CII monoclonal antibody (mAb) C2, hinge region of CD8alpha and the transmembrane and cytoplasmic domains of TCRzeta. The scC2Fv/CD8/zeta gene was transduced into T cell hybridomas and primary mouse lymphocytes using retrovirus-mediated gene transfer. The chimeric receptor scC2Fv/CD8/zeta forms covalently bound homodimers, as demonstrated in T cell hybridomas and packaging fibroblasts. It does not associate with endogenous signalling subunits of the TCR complex. When scC2Fv/CD8/zeta-expressing clones of T cell hybridomas MD.45 and HCQ6 were stimulated with CII they produced IL-2. The level of their IL-2 response correlated with the expression level of the chimeric receptor on the cell surface. Splenocytes isolated from DBA/1 mice were stimulated with Con A in vitro to facilitate retrovirus-mediated transfer of the scC2Fv/CD8/zeta gene. As a result of transduction, approximately 4% of the Con A-activated splenocytes expressed the chimeric receptor scC2Fv/CD8/zeta on the cell surface. These cells proliferated in response to stimulation with CII.

Topics: Animals; Arthritis; Autoimmune Diseases; Collagen; Concanavalin A; Genetic Therapy; Hybridomas; Immunoglobulin Fragments; Interleukin-2; Membrane Proteins; Mice; Mice, Inbred DBA; Receptors, Antigen, T-Cell; Stimulation, Chemical; T-Lymphocytes; Transfection

Adjuvant arthritis (AA) and type II collagen (CII)-induced arthritis (CIA) in the rat serve as models of chronic human arthritis. Adoptive transfer of AA was observed in 21 of 25 Lewis rats given concanavalin A (Con A)-treated spleen cells prepared from animals immunized with Mycobacterium butyricum in mineral oil (complete Freund's adjuvant, CFA). No arthritic changes were noted in rats given spleen cells obtained from donors that had received incomplete Freund's adjuvant (IFA, 0/22), type I collagen in IFA (CI-IFA, 0/6) or CII-IFA (0/28). Administration of spleen cells from IFA, CI-IFA or CII-IFA-injected animals did not modify the development of CIA when these rats were subsequently challenged with CII-IFA. However, partial protection against induction of AA was provided by the transfer of spleen cells prepared from rats immunized with CII-IFA (6/11) but not by those obtained from rats injected with IFA (1/15) or CI-IFA (0/3). Rats that did not develop clinically evident arthritis following the administration of spleen cells prepared from CFA-injected rats were also resistant to AA induction by CFA. Pre-treatment of rats with a synthetic peptide, corresponding to amino acids 180-188 of the Mycobacterium 65 kD heat shock protein (65 kD HSP), significantly delayed the onset of AA, but not that of CIA. Disease-specific resistance to AA, provided by spleen cells prepared from rats injected with CII-IFA and by pre-treatment with the 65 kD HSP 180-188 peptide, may result from the induction of protective tolerance to arthritogenic epitopes present in the Mycobacterium and CII preparations.

Topics: Amino Acid Sequence; Animals; Arthritis; Arthritis, Experimental; Autoimmune Diseases; Bacterial Proteins; Chaperonin 60; Chaperonins; Collagen; Concanavalin A; Freund's Adjuvant; Heat-Shock Proteins; Immunotherapy, Adoptive; Lymphocyte Activation; Molecular Sequence Data; Nontuberculous Mycobacteria; Peptide Fragments; Rats; Rats, Inbred Lew; Spleen; T-Lymphocyte Subsets

Intra-articular injections of murine recombinant IL-1 (mrIL-1) during the chronic phase of antigen-induced arthritis (AIA) induced a flare-up of the smouldering inflammation. The exacerbation was characterized by acute and transient joint swelling and this coincided with the extravascular accumulation of neutrophils. IL-1 injected into arthritic joints of neutropenic mice demonstrated that joint swelling was independent of the neutrophil influx into the joint. Both phenomena were absent when IL-1 was injected into a naive joint. The IL-1-induced flare-up was not T cell mediated as in the antigen-induced flare-up, and suggestive evidence is presented that IL-1 sensitivity depended on the resident macrophage population. This explained why the hypersensitivity is not restricted to the immunologically mediated arthritis but reflects a more general hypersensitivity of previously injured joints, e.g. zymosan-induced arthritis and IL-1-affected joints. In addition, IL-1 could also potentiate the antigen-specific flare-up of chronic AIA and prolongs the duration of the exacerbation. Our data indicate that joints bearing a chronic infiltrate are at risk from exacerbations in two ways: a T cell mediated rechallenge with antigen, and a non-specific reactivation by systemic and local IL-1 generation.

Topics: Acute Disease; Animals; Antigens; Arthritis; Concanavalin A; Indomethacin; Interleukin-1; Joints; Macrophages; Male; Mice; Mice, Inbred C57BL; Neutrophils; Prostaglandins; Time Factors

SK&F 105685 (N,N-Dimethyl-8,8-dipropyl-2-azaspiro[4.5]decane-2-propanamine+ ++ dihydrochloride) is a novel azaspirane with beneficial activity in animal models of autoimmune diseases such as adjuvant-induced arthritis and experimental autoimmune encephalomyelitis in the Lewis rat and lupus-like disease in the MRL mouse. The effect of SK&F 105685 on the proliferation of rat lymphoid cells was examined in vitro. The compound inhibited the proliferative response of spleen, thymus and lymph node cells to the mitogen concanavalin A (Con A) in a dose-dependent manner but had little or no effect on the mitogenic response of peripheral blood lymphocytes. Although less potent than cyclosporin A, SK&F 105685 was able to inhibit the proliferation of spleen cells stimulated with PMA and ionomycin or the mitogens phytohemagglutinin (PHA), Con A and pokeweed mitogen (PWM). Relatively early event(s) in cell proliferation were affected by SK&F 105685 since delaying addition of the drug by 24 to 48 hours after Con A stimulation of rat spleen cells resulted in reduced levels of suppression. The mode of action of SK&F 105685 appeared to differ from that of cyclosporin A or rapamycin. Unlike cyclosporin A, SK&F 105685 did not affect IL-2 production by Con A-stimulated spleen cells or the IL-2-producing Jurkat cell line, but, like rapamycin, the compound significantly reduced the IL-2-induced proliferation of rat ConA blasts. These results suggest that inhibition of lymphocyte proliferation by SK&F 105685 may require the activity of an intermediate effector cell(s) present in susceptible populations such as cells from the spleen, thymus, lymph nodes and Con A blast preparations but absent or present in low numbers in resistant populations such as peripheral blood cells. Indomethacin and NG-monomethyl-L-arginine (NGMMA), a competitive inhibitor of nitric oxide synthase, were both unable to relieve SK&F 105685-induced suppression of splenic Con A responses thereby ruling out a role for the production of prostaglandins or nitric oxide by macrophages as an intermediate in drug-mediated suppression. In summary, SK&F 105685 was unable to inhibit lymphoproliferative responses by a mechanism distinct from that of cyclosporin A or rapamycin and which appears to involve regulation of cellular interactions rather than a direct effect on responding lymphocytes.

Topics: Animals; Arginine; Arthritis; Concanavalin A; Cyclosporine; Dose-Response Relationship, Drug; Humans; In Vitro Techniques; Indomethacin; Interleukin-2; Lymphocyte Activation; Male; omega-N-Methylarginine; Polyenes; Rats; Rats, Inbred Lew; Sirolimus; Spiro Compounds; Spleen; Time Factors; Tumor Cells, Cultured

Pristane-induced arthritis was investigated in DBA/1, DBA/2, and BALB/c mice, and F1 hybrid mice generated from inter-crosses between these strains. The incidence of disease in F1 hybrid mice was significantly lower than the susceptible parental strains (DBA/1 and BALB/c), and resistance to arthritis was observed in both DBA/2 mice and the (DBA/2 x BALB/c) F1 hybrid mice. Several cellular immune abnormalities were observed in pristane-injected DBA/1 mice. Con A mitogen responses were depressed following pristane injection, and a functional suppressor cell population was detected. Delayed type hypersensitivity responses to type II collagen were observed in pristane injected mice. The intraperitoneal injection of pristane appears to alter immune regulation and induce autoimmune responses to connective tissue components.

Topics: Animals; Arthritis; Carcinogens; Collagen; Concanavalin A; Disease Models, Animal; Hybridization, Genetic; Hypersensitivity, Delayed; Immune Tolerance; Lipopolysaccharides; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Terpenes

Effective T cell vaccination against experimental autoimmune diseases involves treatment with activated, autoimmune T lymphocytes. The present study was undertaken to learn whether antigen-specific T cells present in low frequency could be selected in vitro without using the specific antigen. The rat models of adjuvant arthritis and experimental autoimmune encephalomyelitis were investigated using proliferation assays and limiting dilution techniques to quantify the changes in reactivity of a heterogenous population of lymphocytes to the relevant antigen. Stimulation with concanavalin A for 2 d and then culture in IL-2-containing medium led to a substantial increase in the activity and frequency of the specific autoimmune T cells. Enrichment of antigen-specific T cells could be demonstrated using lymph node, spleen, or peripheral blood lymphocytes, from rats late in the course of disease. The effect was not evident in lymphocytes from the thymus. These results are relevant to the clinical application of T cell vaccination and to investigation of self-antigens in autoimmune disease.

Topics: Animals; Arthritis; Arthritis, Experimental; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Female; Interleukin-2; Lymph Nodes; Lymphocyte Activation; Male; Mycobacterium tuberculosis; Rats; Rats, Inbred Lew; T-Lymphocytes; Vaccination

The migration of intravenously administered adjuvant sensitized T lymphocytes to the knee synovium of recipient rats undergoing passive adjuvant arthritis has been followed. Using fluorescein isothiocyanate (FITC)-labeled adjuvant-sensitized T cells and anticollagen IgG, the present studies demonstrate the presence of fluorescent cells in the inflamed knee synovium of recipient rats undergoing passive arthritis. Proliferation studies indicate that synovial cells from these rats respond to Mycobacterium tuberculosis (MT). Since cross-reactivity between Mycobacterial antigens and cartilage proteoglycans has been previously demonstrated, it is suggested that adjuvant-sensitized T cells that are injected into naive rats migrate to the synovium, proliferate in response to cartilage proteoglycan, and initiate passive arthritis.

Topics: Animals; Antigens, Bacterial; Antigens, Differentiation, T-Lymphocyte; Arthritis; Arthritis, Experimental; Cell Movement; Collagen; Concanavalin A; Immunization, Passive; Lymphocyte Activation; Mycobacterium tuberculosis; Rats; T-Lymphocytes

The chronicity of the antigen-induced arthritis is characterized as dependent on the development of cell-mediated immunity to the antigen, but the exact mechanisms underlying are unclear. We have evidenced decreased suppressor and increased helper cell potential in the early phase of arthritis as result of the immunization procedure. In the late phase of arthritis proliferative responses of spleen lymphocytes to cartilage proteoglycans were revealed which were neither present in immunized animals without arthritis induction nor in the early phase of arthritis. The changes of the regulatory properties on the T-cell level are probably responsible for the transition of acute arthritis into the chronic stage. The deficiency of an effective suppression and/or the increased helper cell potential results in the activation of B- and T-lymphocytes with increased cell-mediated and humoral immune responsiveness to the antigen maintaining the inflammatory process for a long time. In this situation the release of cartilage proteoglycans during the acute joint reaction induces autoimmune responses against cartilage which could contribute to the chronification of inflammation and to cartilage degradation.

Topics: Animals; Antigens; Arthritis; Autoimmunity; Cartilage; Concanavalin A; Female; Immunity, Cellular; Knee Joint; Lymphocyte Activation; Male; Rabbits; T-Lymphocytes, Regulatory

Experimental immunotherapy of murine collagen-induced arthritis can be achieved by administration of specific T suppressor cell hybridomas. The present study examines the immunoregulation noted in this experimental immunotherapy by describing the immunomodulatory effects of a cytokine produced by a suppressor T cell line, T101N. The inhibition of the activation of splenic lymphocytes in response to T cell mitogens and the erythema and edema associated with arthritis were assayed. Mice given T101N ascites showed reduced inflammation (p less than 0.05). Lymphocytes derived from naive mice and cultured in the presence of T101N culture supernatant showed reduced response to concanavalin A. Therefore, this form of experimental immunotherapy of arthritis may be associated with cytokines secreted from T suppressor cells which modulate T cell activation.

Topics: Animals; Arthritis; Arthritis, Experimental; Biological Factors; Concanavalin A; Cytokines; Hybridomas; Lymphocyte Activation; Male; Mice; Mice, Inbred AKR; Mice, Inbred DBA; Spleen; T-Lymphocytes, Regulatory

We studied synoviocytes obtained from rats that had been injected with Freund's complete adjuvant 12 days prior to killing. We found that several activation parameters were affected in these synoviocytes, namely, the concanavalin A protein-binding pattern, the production of superoxide, and the appearance of the p77 polypeptide, which we have previously shown to be associated with the activation caused by the induction of polyarthritis. Our findings suggest that prior to the establishment of the inflammatory process, synoviocytes are in a state of activation, and may be a component of the molecular mechanism during the early stages of disease.

Topics: Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Fluorescein-5-isothiocyanate; Fluoresceins; Glycoproteins; Histocompatibility Antigens Class II; Iodine Radioisotopes; Superoxides; Synovial Membrane

Rats injected with peptidoglycan-polysaccharide polymers derived from group A streptococcal cell walls (PG-APS) develop a chronic, remittant, erosive synovitis. Spleen cells from injected rats failed to proliferate when stimulated in vitro by Con A or PHA, unless nylon wool adherent cells were first removed. The suppression could also be reversed by removing phagocytic cells which had ingested carbonyl iron. Cells from control rats were suppressed in vitro by co-culture with unfractionated or nylon wool-adherent cells from PG-APS injected rats, and the suppressor activity was still expressed after exposure of the suppressor cells to 3,000 rad of irradiation. Addition of catalase and indomethacin to cultures only partially reversed the suppression. T lymphocytes from rats given a single arthropathic dose of PG-APS remained suppressed for at least 86 days after injection. Cells from rats given a low, non-arthropathic dose of PG-APS did not become suppressed. Cells from the Buffalo rat, which is resistant to development of PG-APS-induced chronic arthritis, showed less suppression than cells from the susceptible Lewis and Sprague-Dawley rat strains.

Topics: Animals; Arthritis; Arthritis, Experimental; Catalase; Cell Adhesion; Cells, Cultured; Concanavalin A; Female; Immune Tolerance; Immunity, Cellular; Indomethacin; Mitosis; Peptidoglycan; Phagocytes; Rats; Rats, Inbred Lew; Spleen; Streptococcus pyogenes; T-Lymphocytes

Spirogermanium is a novel metal containing azaspirane compound with reported antitumor activity. The results of the present investigation demonstrate that spirogermanium also exhibits antiarthritic and immunoregulatory activities after p.o. administration to rats. Spirogermanium decreased hindleg inflammatory lesions of adjuvant arthritic rats when administered p.o. before or after the development of the arthritic lesions. After termination of spirogermanium administration, the adjuvant-injected hindleg lesions remained significantly suppressed for at least 2 weeks postdrug treatment; whereas, the uninjected, immune-mediated hindleg inflammation tended to increase postdrug treatment. In multiparameter ex vivo studies, untreated arthritic rats exhibited enhanced cyanine dye fluorescence in peripheral blood monocytes, enhanced interleukin (IL)-1 production by adherent spleen cells and depressed IL-2 and IL-3 production by splenic lymphocytes. Spirogermanium normalized these changes to various degrees, with the exception of the depressed IL-2 and IL-3 production. Spirogermanium administered to normal nonarthritic rats decreased mitogenic responses of spleen cells to Concanavalin A which was found to be caused, at least in part, by enhanced suppressor cell activity. The antiarthritic and immunoregulatory profile of spirogermanium appeared to be different from the profiles of the antiarthritic agents, auranofin and indomethacin.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents; Arthritis; Arthritis, Experimental; Auranofin; Aurothioglucose; Concanavalin A; Fluorescence; Germanium; Immunosuppressive Agents; Indomethacin; Interleukin-1; Interleukin-2; Macrophages; Male; Monocytes; Organometallic Compounds; Rats; Rats, Inbred Lew; Spiro Compounds; T-Lymphocytes, Regulatory

We have investigated the cellular composition of 108 consecutive samples of synovial fluid from patients with Juvenile Chronic Arthritis (JCA), Rheumatoid Arthritis (RA) and Osteoarthritis (OA). Particular emphasis was placed upon the enumeration of cells with dendritic morphology and the study of their in vitro function. Whilst the cellularity of the synovial fluids varied by a factor of greater than 100 within patient groups, the fluids obtained from patients with inflammatory arthritis (JCA & RA) were more cellular than those from patients with non-inflammatory arthritis (OA). This was also noted with respect to both the number and proportion of dendritic cells. The dendritic cells stimulated allogeneic mixed leucocyte reactions, and enhanced mitogenic responses of peripheral blood lymphocytes when present in numbers as low as 1% of the total mononuclear cells. Syngeneic stimulation of blood lymphocytes by similar numbers of dendritic cells was usually negative. However, occasionally there was a marked syngeneic stimulation, which may be evidence for the presentation of antigen by dendritic cells within the arthritic joint.

Topics: Arthritis; Arthritis, Juvenile; Concanavalin A; Humans; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoid Tissue; Synovial Fluid

We studied the in vivo effect of transfer of nonspecific suppressor spleen cells into syngeneic Lewis rats. The rats were immunized with Freund's adjuvant on day 0, and then given 5 X 10(7) cells on days--7, 0 and 7. These cells had been incubated for 3 days, with or without mitogenic stimulation. Transfer of the cultured cells markedly diminished the severity of adjuvant arthritis (AA). On the contrary, transfer of concanavalin A (Con A)-stimulated cells led to no suppressive activity. These results differed from findings in a prior in vitro experiment, in which the suppressive influence of previously cultured cells on T and B lymphocytes proliferation rates was examined and significant suppressive activity was detected in Con A-stimulated cells but not in cultured cells. Suppressor activity by transfer of solely cultured cells was identified in T cell fractions. Transfer of 5 X 10(7) spleen cells proved to be the optimal dose for suppressing AA, and transfer of less than 2 X 10(7) cells had no significant effect. The number and time of transfer were also important factors in the suppression. Each group of transfer on days--7, 0 and 7, days--7 and 0, and days 0 and 7 led to a similar reduction in the severity of AA, and was less prominent in the group injected on days 7 and 14. A single injection on days--7, 0, or 7 revealed minimal or no effects. These observations indicate that the transfer of in vitro cultured spleen cells nonspecifically modified the course of rat AA in vivo, thereby differing from the results of the suppressor activity seen in vitro. The transfer of cultured cells, as a potential tool for the treatment of clinical diseases warrants further attention.

Topics: Animals; Arthritis; Arthritis, Experimental; B-Lymphocytes; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Immunologic; Female; Immunization; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes; T-Lymphocytes, Regulatory; Time Factors

Intraperitoneal administration of group A streptococcal cell walls to Lewis rats induces a chronic arthritis, whereas the Fischer strain is resistant to the development of the lesion. Spleen cells from cell wall-treated rats (Lewis and Fischer) are deficient in the synthesis of IL-2. Using an mAb directed against the rat IL-2-R, the present studies indicate that the expression of IL-2-R on spleens of cell wall-treated rats is normal. However, the addition of exogenous IL-2 to spleen cells cultured with Con A does not stimulate the mitogenic response.

Topics: Animals; Arthritis; Cell Wall; Concanavalin A; Female; Immunologic Deficiency Syndromes; Interleukin-2; Lymphocyte Activation; Rats; Rats, Inbred F344; Rats, Inbred Lew; Receptors, Immunologic; Receptors, Interleukin-2; Spleen; Streptococcus pyogenes

Several pharmacological agents, some of which are known to have effects on the immune system, decrease the incidence of collagen II-induced arthritis when added to the antigen emulsion. Concanavalin A, which has been reported to exert suppressive effects on the immune system in vivo, consistently reduced the immune response to the collagen antigen. These effects were dose and time dependent. The suppressive effects of pokeweed mitogen, tilorone and carrageenan on anti-collagen II responses were somewhat variable. Suppressive activity could be observed with concanavalin A and levamisole when the drugs were injected at a site distant from the collagen emulsion. These studies indicate that local administration of drugs is an effective approach for demonstrating the activity of some agents that may alter the course of collagen II disease through an effect on the immune system.

Topics: Animals; Antigens; Arthritis; Arthritis, Rheumatoid; Collagen; Concanavalin A; Emulsions; Female; Hypersensitivity, Delayed; Levamisole; Rats

Nephelometry of serum acute-phase glycoproteins by binding to concanavalin A (con-A) was compared with assays for haptoglobin and alpha 1-acid glycoprotein, and for C-reactive protein. The cutoff points for positive reactions were determined on the basis of results for a random sample (n = 130) from a middle-aged population. The sensitivity of the con-A binding assay compared favorably with that of individual acute-phase glycoproteins in a follow-up cohort of 198 patients with inflammatory joint diseases. Unlike the case in many individual acute-phase glycoprotein assays, the distribution of con-A binding values in healthy subjects is remarkably symmetrical, allowing an easy distinction between abnormal and normal values.

Topics: Acute-Phase Proteins; Adolescent; Adult; Aged; Arthritis; Arthritis, Rheumatoid; Blood Proteins; C-Reactive Protein; Concanavalin A; Glycoproteins; Haptoglobins; Humans; Immunochemistry; Middle Aged; Nephelometry and Turbidimetry; Orosomucoid; Protein Binding

We have been studying the pathogenesis of adjuvant arthritis in rats using a long-term cell line of T lymphocytes, the A2 line, which can induce polyarthritis and can also be used to vaccinate rats against adjuvant arthritis. Although line A2 was selected for its proliferative response to mycobacteria, it also responded to collagen type II. To elucidate its role of responsiveness to collagen type II and the relationship between arthritogenicity and vaccination, we cloned A2 and selected a subline A2b. We now report that subline A2b, which bore a marker of helper/delayed hypersensitivity T lymphocytes, was strongly arthritogenic, but could not vaccinate against arthritis. Moreover, A2b showed no response to collagen type II. Therefore, reactivity to collagen type II is not a requisite for arthritogenicity, and mediation of arthritis and vaccination can be distinct properties of different populations of T lymphocytes.

Topics: Animals; Antigens, Bacterial; Arthritis; Arthritis, Experimental; Autoimmune Diseases; Clone Cells; Collagen; Concanavalin A; Immunity, Innate; Lymphocyte Activation; Mice; Mycobacterium tuberculosis; Phenotype; Rats; Rats, Inbred Lew; T-Lymphocytes; T-Lymphocytes, Helper-Inducer

Naproxen was evaluated for possible disease modifying effects in the Freund's adjuvant injected rat (AIR). Oral administration of the clinical dose, 7 mg/kg/day, lead to an almost complete inhibition of hindpaw swelling and cartilage and bone erosion. This was noted in animals maintained on drug as well as those in which therapy was discontinued. AIR, comparable to arthritic patients, demonstrate a reduced lymphocytic response to T cell mitogens. This response was normalized in naproxen-treated rats. These results suggest that naproxen has a disease modifying effect in the AIR.

Topics: Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Female; Lymphocyte Activation; Mitogens; Naproxen; Phytohemagglutinins; Rats; T-Lymphocytes

The role of macrophages and of macrophage-lymphocyte co-operation in the induction of T-suppressor cell activity in rats with adjuvant arthritis was investigated. Macrophages from arthritic rats had no direct effect on the induction of T-suppressor cells, but, in the presence of arthritic rats' lymphocytes or supernatants from such lymphocytes, macrophages inhibited the induction of T-suppressor cell activity. Addition of indomethacin during the induction of T-suppressor cells reduced the inhibitory effects, suggesting that prostaglandins released from lymphocyte-activated macrophages might be involved in the defective induction of T-suppressor cell activity in rats with adjuvant arthritis.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Adhesion; Cells, Cultured; Concanavalin A; Female; Indomethacin; Lymphocyte Activation; Lymphocytes; Macrophages; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes, Regulatory

Concanavalin-A-induced T-suppressor cell activity was decreased in spleen cell suspensions from Lewis rats with adjuvant arthritis. This decreased activity was evident on day 10 after the induction of the disease, just before the development of the polyarthritic lesions, and persisted during the period of active inflammatory and immunological disease. The extent of the impairment of T-suppressor cell activity was positively correlated with the severity of the arthritic lesions. Removal of phagocytic cells prior to induction of T-suppressor cells abolished the observed decrease in suppressor cell activity. It is suggested that this model may be of value for the investigation of suppressor cell function in immunologically mediated disorders.

Topics: Animals; Arthritis; Arthritis, Experimental; Cells, Cultured; Concanavalin A; Female; Lymphocyte Activation; Macrophages; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes, Regulatory

Two reliable systems for the cell-mediated passive transfer of adjuvant arthritis have been developed. Donor rats were sensitized with Mycobacterium butyricum in mineral oil. In the first system, intravenous injection of adjuvant-sensitized donor lymph node or spleen cells into adult-thymectomized, lethally irradiated, bone marrow cell-reconstituted syngeneic rats induced arthritis in the recipients. In the second system, adjuvant-sensitized donor lymph node or spleen cells were cultured in vitro with concanavalin A; these cells induced arthritis in normal recipients as well as in thymectomized, irradiated, bone marrow cell-reconstituted recipients. The passively transferred disease in both systems resembled classical adjuvant-induced arthritis clinically, radiographically, and histologically. Neither irradiated, adjuvant-sensitized donor cells nor cells from donors not injected with complete adjuvant could passively transfer arthritis.

Topics: Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Graft vs Host Reaction; Immune Tolerance; Immunity, Cellular; Immunization, Passive; Lymph Nodes; Lymphocyte Activation; Male; Mice; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes

It has previously been reported that lymph node or spleen cells from rats with adjuvant-induced arthritis can transfer the disease to normal recipients after being cultured with concanavalin A (Con A). In this report, it is shown that a subpopulation of cells that (1) lack surface Ig and the antigen reactive with the monoclonal antibody OX8, (2) are largely nonadherent and esterase negative, and (3) are predominantly marked by the monoclonal antibody W3/25 can transfer arthritis after stimulation with Con A. Adjuvant-sensitized lymph node or spleen cells stimulated with Con A but not PHA transfer arthritis, and this difference correlates with relatively higher levels of interleukin 2 secretion by Con A-stimulated cells. A synthetic adjuvant, CP-20961, a substituted propanediamine, induces arthritis that is passively transferable under the same conditions as arthritis induced by classical mycobacterium-containing adjuvant. The data support the hypothesis that adjuvant inoculation in the rat results in the induction of a unique subpopulation of T cells that initiate the inflammatory joint disease.

Topics: Animals; Arthritis; Arthritis, Experimental; Cells, Cultured; Concanavalin A; Diamines; Immunization, Passive; Interleukin-2; Lymph Nodes; Male; Rabbits; Rats; T-Lymphocytes

The relationship between cell mediated immunity to collagen and arthritis was studied with lymphocytes from arthritic and nonarthritic rats after immunisation with native bovine type II collagen. With the in-vivo radiometric ear assay arthritic rats gave a significantly higher response to native type II collagen than did nonarthritic rats. However, there was an overlap of values, and some arthritic rats gave no response to collagen even on the day of onset of arthritis. There was no difference in the response of lymphocytes from arthritic and nonarthritic rats with in-vitro transformation to native type II collagen, responses being found in both groups. All rats which developed arthritis had serum antibodies to native type II collagen, but not all responded to the tests for cell mediated immunity. These findings suggest that antibodies to collagen are more associated with the development of arthritis than is cell mediated immunity to collagen.

Topics: Animals; Antibody Formation; Arthritis; Cattle; Collagen; Concanavalin A; Female; Immunity, Cellular; Immunization; Immunoglobulin G; Lymphocyte Activation; Male; Rats; Rats, Inbred Strains; Time Factors

Fisher rats from a inbred colony, when fed on a salt-free high-protein diet, developed only a mild arthritis after adjuvant injection. Their spleen cells failed to respond in vitro to concanavalin A (a T-cell mitogen), although they possessed a B-cell function of plaque formation to sheep red blood cells. When a full salt supplement was included in the diet, or magnesium or copper or zinc was included in the drinking water, adjuvant-induced arthritis was severe and the response to the T-cell mitogen was restored. The above results suggest that these trace elements may stabilize or activate certain cell populations needed for some immune responses in rats.

Topics: Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Copper; Diet, Sodium-Restricted; Freund's Adjuvant; Lymphocyte Activation; Magnesium; Rats; Rats, Inbred F344; Spleen; Zinc

D-penicillamine (D-PA) has beneficial therapeutic effects for patients with rheumatoid arthritis but no convincing explanation has been offered for the mode of action. Experiments reported here were designed to gain an insight into the related mechanisms. Wistar rats were inoculated with various doses of Mycobacterium tuberculosis to induce adjuvant arthritis, and on the 21st day, the lesions of paws and ears were graded according to the extent of the erythema and swelling. Rats given D-PA simultaneously with the inoculation of M. tuberculosis developed a more severe arthritis than that seen in the control group, when they were inoculated with low doses of M. tuberculosis. To investigate the effect of D-PA on hemolytic plaque forming cells (PFC) in the spleen, BDF1 mice were immunized with various doses of sheep red blood cells (SRBC) and D-PA was injected in various doses and at various times. D-PA produced either enhancement or depression of the number of PFC, depending on the dose of antigenic stimulus of SRBC. Furthermore, D-PA slightly enhanced the concanavalin A-induced blastogenesis of the spleen cells in vitro, at a concentration of 1-50 microM, but at concentrations of 500 microM, inhibition was evident. These results indicate that D-PA may act as an immunomodulating agent.

Topics: Adjuvants, Immunologic; Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Dose-Response Relationship, Immunologic; Female; Hemolytic Plaque Technique; Injections, Intraperitoneal; Lymphocyte Activation; Mice; Mycobacterium tuberculosis; Penicillamine; Rats; Rats, Inbred Strains; Spleen

Nuclear refringence test (NRT) was studied in two inbred strains of rats: Lewis (LEW) and Wistar AG (WAG), the first develops a severe arthritis while the later only slight inflammation. Before adjuvant injection, a good NRT to ConA, PHA and Isoprinosine was observed in LEW but a poor one in WAG. After adjuvant injection a striking difference appears between the two strains concerning ConA response: in LEW the response is significantly lower on day 14, while in WAG the initial poor response is not modified. These data suggest that in WAG the suppressor T lymphocytes are already activated in vivo and are no longer responsive in vitro. The responses to PHA and Isoprinosine are parallel in both strains, the NRT response diminishes on day 14 in LEW and on day 21 in WAG.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Nucleus; Concanavalin A; Lymphocytes; Male; Rats; Rats, Inbred Lew; Rats, Inbred Strains

Adherent spleen cells from rats with adjuvant arthritis inhibit the incorporation of 3H-thymidine into DNA and 3H-leucine into protein in nonadherent spleen lymphocytes, stimulated by the mitogens Concanavalin A and E. coli Lipopolysaccharide. This suppressive activity was abolished by pretreatment of the adherent cells with the selective macrophage toxin silica. It is suggested that suppressor macrophages directly, or through interaction with suppressor T cells, inhibit the response of lymphocytes to mitogens by inhibition of cellular protein synthesis, followed by inhibition of DNA-synthesis and death of about 30% of the lymphocytes. Treatment of adjuvant arthritis rats with D-penicillamine resulted in significantly increased incorporation of 3H-thymidine in spleen lymphocytes, compared to cultures from untreated, arthritic rats. This approach may prove useful in the investigation of cellular interactions in a model of immunologically induced inflammation and provide a tool for the evaluation of the effects of immunoregulatory drugs.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Adhesion; Concanavalin A; Female; Leucine; Lipopolysaccharides; Penicillamine; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes, Regulatory; Thymidine

Administration of D-penicillamine (50 mg/kg/day orally) to rats with adjuvant arthritis for up to 42 days significantly modified the incorporation of 3H-thymidine (3H-TdR) in concanavalin A (Con A)-stimulated lymph node cells. Treatment with D-penicillamine abolished the ability of macrophages from arthritic rats to inhibit lymphocyte responsiveness to Con A and lipopolysaccharide (LPS) 14 days after the induction of the disease. Increased T-cell responsiveness to Con A was found from day 14 to 35 in cultures of unseparated and adherent-cell-depleted lymph node cells from D-penicillamine-treated arthritic rats. B-cell responsiveness to LPS was not affected. Experiments with bovine serum albumin gradient-separated lymph node cells confirmed these findings and indicated that treatment with D-penicillamine may specifically enhance T-helper cell responsiveness to Con A. It is suggested that administration of D-penicillamine may interfere with macrophage function during the course of an immunologically induced chronic inflammation, leading to an increased response of T-helper cells. The theoretical implications of these findings are discussed.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Adhesion; Cell Separation; Concanavalin A; Female; Indomethacin; Lipopolysaccharides; Lymphocytes; Macrophages; Penicillamine; Rats; Rats, Inbred Lew; Thymidine

Topics: Arthritis; Arthritis, Experimental; Concanavalin A; Glycoproteins; Humans; Inflammation; Lectins; Plant Lectins; Plants, Toxic; Protein Binding; Ricinus

Experimental model of human chronic inflammatory arthritis, adjuvant arthritis may be induced only in several strains of inbred Rats: it is well developed in LEW and practically absent in WAG. After adjuvant injection, the PHA-stimulable lymphocytes subpopulation quite disappears from the blood, if polyarthritis is well developed. These cells are probably capted in the tissues implicated in immunological conflict. On the contrary, the ConA-stimulable subpopulation is enhanced in both strains after adjuvant injection, earlier and more intense in WAG than in LEW and that phenomenon is probably linked to a stimulation of suppressor T lymphocytes. Treatment with prednisone or gamma rays inhibits partially and delays the appearance of arthritis in LEW, acting essentially on ConA-stimulable subpopulation.

Topics: Adjuvants, Immunologic; Animals; Arthritis; Arthritis, Experimental; Concanavalin A; Gamma Rays; Lectins; Lymphocyte Activation; Male; Prednisone; Rats; Rats, Inbred Lew; Rats, Inbred Strains; T-Lymphocytes

Serum from adjuvant arthritic rats inhibits the concanavalin A- (Con A) and lipopolysaccharide-induced stimulation of lymph node cells, leaving the basal and phytohemagglutinin-stimulated 3H-thymidine incorporation unaffected. Con A-stimulated 3H-thymidine uptake is also inhibited in rat spleen and peripheral blood lymphocytes and in dog peripheral blood lymphocytes. The intensity of the inhibitory activity in serum is positively correlated with the intensity of the secondary lesions of adjuvant arthritis. Inhibitory activity was not found in serum from rats bearing nystatin-induced inflammation. Serum fractionation studies indicated that the inhibitory activity cannot be attributed to low molecular weight alpha2-glycoproteins or to gamma-globulins and alpha2-macroglobulins, but it is present in a fraction migrating with beta-globulins. The inhibitory activity in arthritic rat serum is reduced by treatment with non-steroidal anti-inflammatory drugs, but is unaltered by D-penicillamine. It is suggested that this inhibitory activity is part of the systemic response to an immunologically mediated inflammation.

Topics: Animals; Arthritis; Arthritis, Experimental; Chemical Fractionation; Chromatography, Gel; Concanavalin A; Female; Ibuprofen; Indomethacin; Lymph Nodes; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Penicillamine; Phenylbutazone; Phytohemagglutinins; Rats; Spleen

Serum from adjuvant arthritic rats inhibits the mitogenic response of rat lymph node cells to concanavalin A, leaving unaffected the response to phytohemagglutinin. This activity is already evident 4 days after adjuvant administration and persists for at least 28 days.

Topics: Animals; Arthritis; Arthritis, Experimental; Concanavalin A; DNA; Female; Lymph Nodes; Lymphocytes; Rats; Rats, Inbred Lew; Thymidine

The criteria for selecting and establishing a animal model for arthritis were described. Rats are the most frequently used animals. Adjuvant and Myocobacterium induced arthritis provide a model of chronic joint inflammation, although significant differences exist when compared with human disease. A better model of arthritis in rats and mice can be induced by the injection of one strain of Mycoplasma arthritidis. An even better model is presented by rabbits first immunized against a protein such as heterologous fibrin or albumin, and then challenged by the same protein injected directly into a joint. This results in a localized chronic arthritis pathologically similar to that of man. Arithritis can also be induced in rabbits by the injection of polymers such as chitinor or Concanavalin A into the joint. Although there is no lack of arthritis animal models, there is no animal model which gives a true replication of rheumatoid arthritis.

Topics: Animals; Arthritis; Arthritis, Infectious; Arthritis, Rheumatoid; Concanavalin A; Disease Models, Animal; Dog Diseases; Dogs; Haplorhini; Mice; Mycobacterium; Mycobacterium tuberculosis; Mycoplasma Infections; Rabbits; Rats; Rodent Diseases

Diminished splenic T-cell function, as measured by their response to phytohemagglutinin and Concanavalin-A, was noted only during the active phase of adjuvant induced disease. This diminished function is due to at least two types of suppressor cell: (a) cells which adhere to glass but not plastic, and which are sensitive to methotrexate in vitro; (b) cells which adhere to plastic and glass, and which are relatively insensitive to methotrexate in vitro. Methotrexate treatment in vivo inhibits adjuvant induced disease, and is associated with the absence of both types of suppressor cells in the spleen.

Topics: Animals; Arthritis; Arthritis, Experimental; Cell Adhesion; Concanavalin A; Glass; Lectins; Leukocyte Count; Lipopolysaccharides; Lymphocyte Activation; Male; Methotrexate; Mitogens; Plastics; Rats; Spleen; T-Lymphocytes

Topics: Arthritis; Concanavalin A; Humans; Immunity, Cellular; Lectins; Lymphocyte Activation; Mitogens; Skin Tests