ABSTRACT
To further the insight in the immunomodulating properties of the anticonvulsant 5,5-diphenylhydantoin (DPH), C57BL/6 (B6), C57BL/6-lpr/lpr (B6-lpr/lpr) and MRL/MpJ- +/+ (MRL) mice received DPH orally for six months to determine weekly urinary biopterin levels, a potential T-cell activation marker, by high performance liquid chromatography. At the end of the experiment serum antibody levels were measured by ELISA and relative lymphoid organ weights determined. DPH treatment resulted in reduced body weight in all strains, reduced spleen weights in B6 and MRL mice, profoundly reduced popliteal lymph node weights in B6-lpr/lpr mice and increased thymus weights in MRL mice. DPH treatment decreased serum IgM, IgG and IgA as well as IgM and IgG anti-ssDNA levels in B6-lpr/lpr mice, but did not affect these parameters in other strains. Effects of DPH on IgM rheumatoid factor levels in B6-lpr/lpr mice were inconsistent. Urinary biopterin levels of untreated B6 and B6-lpr/lpr mice were about equal and lower than those of MRL mice. During the first three months of DPH treatment, persistently elevated biopterin levels were observed in B6 and to a lesser degree in MRL mice, and alternately elevated and control levels in B6-lpr/lpr mice. Thereafter, the effects faded in all strains. Results show that long-term DPH treatment causes only minor lymphoid organ weight changes in B6 and MRL mice, but causes a clear reduction of the lymphadenopathy and (auto)antibody formation in B6-lpr/lpr mice. Observed changes could not be related to altered biopterin excretion indicating that the latter is an inappropriate marker of murine autoimmune disease.
Subject(s)
Autoimmune Diseases/drug therapy , Lymphatic Diseases/drug therapy , Phenytoin/pharmacology , Animals , Antibodies, Antinuclear/drug effects , Autoimmune Diseases/immunology , Biopterins/urine , Body Weight/drug effects , Drug Administration Schedule , Female , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Lymphatic Diseases/immunology , Lymphoid Tissue/drug effects , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Organ Size/drug effects , Phenytoin/administration & dosage , Rheumatoid Factor/drug effectsABSTRACT
Based on evidence that urinary neopterin levels are useful markers of disordered cellular immunity in man, we investigated murine urinary biopterin excretion during acute and chronic graft-versus-host (GvH)-reactions as well as after oral exposure to drugs with documented immune disregulating potential in man. Biopterin levels were determined in urine spot samples by reversed-phase high performance liquid chromatography and expressed in relation to the urinary creatinine content. Similarly increased and decreased biopterin levels were observed during acute and chronic GvH-disease in (C57BL/6J x DBA/2J)F1 (B6D2F1) mice. Increased and/or decreased levels of urinary biopterin were observed during treatment with 5,5-diphenylhydantoin (DPH), methimazole, propylthiouracil and nitrofurantoin, but no consistent pattern could be distinguished. The DPH-induced alterations were similar in B6 and B6D2F1 mice, were dose-dependent, reversible and independent of mature T-cells, as judged by the pronounced biopterin excretion of B6-nu/nu mice in comparison with their T-cell competent litter mates. The results indicate that monitoring of urinary biopterin excretion in mice does not represent a useful biochemical marker for T-cell activation.
Subject(s)
Biopterins/urine , Graft vs Host Disease/urine , Phenytoin/pharmacology , Animals , Biopterins/analogs & derivatives , Body Weight , Circadian Rhythm , Creatine/urine , Female , Lymphocyte Activation , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Nude , Neopterin , Phenytoin/blood , Time FactorsABSTRACT
Inhibitors of oxidative phosphorylation such as several triorganotin compounds, oligomycin, 2,4-dinitrophenol and carbonylcyanide p-trifluoromethoxyphenylhydrazone suppress energy metabolism of isolated rat thymocytes as indicated by a reduction of ATP levels, an increase in glucose consumption and by a marked accumulation of lactate. Also these compounds effectively inhibit the incorporation of DNA, RNA and protein precursors into acid-precipitable material of thymocytes. Moreover, the prostaglandin E1-induced elevation of cAMP is markedly reduced by these inhibitors. A correlation is observed between the effects on energy metabolism, macromolecular synthesis and cAMP production, since from a series of trialkyltin chlorides, tri-n-propyltin, tri-n-butyltin and tri-n-hexyltin are very effective inhibitors of these functions, while trimethyltin and tri-n-octyltin affect neither of them; other inhibitors of oxidative phosphorylation, each of them with quite different mechanisms of action, also inhibit macromolecular synthesis and cAMP production. The finding that a rise in intracellular ATP concentrations leads to a reversion of the tri-n-butyltin-induced inhibition of cAMP production and uridine incorporation, indicates a regulating role for the cellular energy state in these aspects of cellular function.
