ABSTRACT
It is becoming increasingly clear that the clinical courses of a variety of autoimmune and infectious diseases are influenced by the balance of TH1 and TH2 cell subsets that are generated during the immune response. IL-10 is one of several cytokines which influences the differentiation of TH cell subsets and represents a target for therapeutic intervention. We have evaluated a variety of pharmacological agents for their ability to modulate IL-10 release by the murine D10.G4.1 TH2 cell line when stimulated with concanacavalin-A in the presence of IL-1 alpha. Several were inhibitory, and the concentrations which caused a 50% reduction in IL-10 production were 0.38 microM cyclosporin-A, 0.0073 microM dexamethasone, 0.045 microM prednisolone and 0.31 microM cycloheximide. Methotrexate and pentoxifyline caused a weak but statistically significant reduction in IL-10 production at a concentration of 10 microM (P < or = 0.05), whereas amrinone and azathioprine had no clear effect. The pharmacological agents tested are known to exert multiple effects and were evaluated with a view to their use as reference standards in an ongoing screening programme to identify novel compounds which specifically modulate Il-10 production.
Subject(s)
Interleukin-10/biosynthesis , T-Lymphocytes/metabolism , Animals , Anti-Inflammatory Agents/pharmacology , Antineoplastic Agents/pharmacology , Cell Line , Concanavalin A/pharmacology , Immunosuppressive Agents/pharmacology , Interleukin-1/physiology , Mice , Recombinant Proteins/pharmacology , Steroids , T-Lymphocytes/drug effectsABSTRACT
The effect of BTS 71 412, 4-acetyl-1-(4-chlorophenyl)-3-(2-methylthiophenyl)- 3-pyrazolin-5-one, has been determined on a variety of immune reactions in vitro in order to gain a further insight into the mechanisms whereby this novel immunosuppressive drug suppresses cell and antibody mediated immune responses in vivo. BTS 71 412 markedly inhibited [3H]thymidine incorporation by mouse splenocytes activated with concanavalin-A (IC50 = 20.1 microM), phytohaemagglutinin (IC50 = 4.6 microM), lipopolysaccharide (LPS) (IC50 = 3.2 microM), anti-IgM (mu-chain specific) (IC50 = 2.6 microM), or mixed lymphocyte culture (IC50 = 8.4 microM). Activity of BTS 71 412 was not associated with a reduction in cell viability. BTS 71 412 also prevented [3H]thymidine incorporation by the murine HT-2 helper T-cell clone when cultured with IL-2 (IC50 = 7.6 microM) or IL-4 (IC50 = 7.3 microM), enriched Thy-1+ T-lymphocytes stimulated with phorbol 12-myristate 13-acetate plus ionomycin (IC50 = 3.6 microM), and enriched B220- B-lymphocytes stimulated with LPS (IC50 = 3.0 microM). Splenocytes cultured with BTS 71 412 produced lower levels of interleukin (IL)-2, IL-10 and interferon-gamma when stimulated with concanavalin-A (IC50 values 42 microM, 22 microM and 60 microM, respectively). The compound suppressed in vitro antibody responses to keyhole limpet haemocyanin (IC50 = 2.3 microM), but did not reproducibly inhibit IL-6 or tumour necrosis factor alpha production by adherent peritoneal macrophages stimulated with LPS in vitro. These data indicate that BTS 71 412 specifically inhibits both B- and T-lymphocyte activation and proliferation but does not affect macrophage activation in vitro.
Subject(s)
Immunosuppressive Agents/pharmacology , Pyrazoles/pharmacology , Pyrazolones , Animals , Antibody Formation/drug effects , Antigens/immunology , Cell Line , Cells, Cultured , Cycloheximide/pharmacology , Cyclosporine/pharmacology , Cytokines/biosynthesis , Cytokines/pharmacology , Dexamethasone/pharmacology , Dose-Response Relationship, Immunologic , Female , Hemocyanins/immunology , In Vitro Techniques , Lymphocyte Activation/drug effects , Lymphocyte Culture Test, Mixed , Lymphocyte Subsets/immunology , Lymphocyte Subsets/metabolism , Lymphocytes/immunology , Lymphocytes/metabolism , Macrophage Activation/drug effects , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Pyrazoles/chemistry , Spleen/immunology , Spleen/metabolismABSTRACT
The effect of a number of drugs commonly used to treat the more severe exacerbations of the autoimmune disease systemic lupus erythematosus (SLE) in humans has been investigated in the murine chronic graft-versus-host (GVH) induced model of lupus. This was undertaken in order to determine the value of this model for the investigation of immunomodulant drugs, with particular regard to the reproducibility of disease induction and methods of monitoring disease progression. The drugs were azathioprine, cyclophosphamide, cyclosporin A and dexamethasone. All of these, except for azathioprine, reduced disease severity, assessed as the development of lupus nephritis. Anti-ssDNA autoantibodies were also reduced in titre in the dexamethasone-treated group. Overall, these findings, combined with the reproducible induction of disease seen in this model, support the use of chronic GVH disease as a model for SLE and show that the induced disease can be ameliorated by drugs effective in the treatment of SLE in humans.
Subject(s)
Disease Models, Animal , Graft vs Host Disease/drug therapy , Immunosuppressive Agents/therapeutic use , Lupus Erythematosus, Systemic/drug therapy , Animals , Azathioprine/therapeutic use , Cyclophosphamide/therapeutic use , Cyclosporins/therapeutic use , Deoxyguanosine , Dexamethasone/therapeutic use , Female , Lupus Nephritis/prevention & control , Mice , Proteinuria/prevention & control , Reproducibility of ResultsABSTRACT
A commercially available medium supplement, Nu-Serum, was compared with foetal calf serum for its ability to support the murine allogeneic mixed lymphocyte reaction. Nu-Serum supported proliferation whilst at the same time producing significantly lower counts in unstimulated cultures. There was considerably less batch-to-batch variation with Nu-Serum than with foetal calf serum, which obviated the need for pretesting in order to find a suitable batch.
