Subject(s)
Graft Survival/drug effects , Heart Transplantation/immunology , Immunosuppressive Agents/therapeutic use , Indolizines/therapeutic use , Intercellular Adhesion Molecule-1/biosynthesis , Lymphocyte Function-Associated Antigen-1/biosynthesis , Animals , Antigens, Differentiation, T-Lymphocyte/biosynthesis , Histocompatibility Antigens Class I/biosynthesis , Histocompatibility Antigens Class II/biosynthesis , Lymphoid Tissue/drug effects , Lymphoid Tissue/immunology , Rats , Rats, Inbred BN , Rats, Inbred Lew , Rats, Inbred Strains , Transplantation, HomologousSubject(s)
Graft Survival/drug effects , Heart Transplantation/immunology , Immunosuppressive Agents/therapeutic use , Indolizines/therapeutic use , Tacrolimus/therapeutic use , Animals , Drug Synergism , Immunosuppressive Agents/toxicity , Indolizines/toxicity , Lethal Dose 50 , Rats , Rats, Inbred Strains , Tacrolimus/toxicity , Time Factors , Transplantation, HomologousABSTRACT
The inhibition of intracellular oligosaccharide processing is a new approach to immunosuppression in allotransplantation. The net effect of such inhibition is reduction in the membrane expression of certain glycoproteins. Hence cell-cell interaction in allorejection may be impaired in the presence of glycoprotein processing inhibitors because the expression of key ligand-receptor pairs of N-linked glycoproteins including adhesion molecules is inhibited. The aims of this study were to measure the immunosuppressive ability of castanospermine (CAST) in a rat heart allograft model, to measure its effect on membrane expression of adhesion molecules (LFA-1 alpha, LFA-1 beta, ICAM-1), class I and class II MHC antigens and on other T cell associated molecules (CD4, CD8, CD39, CD45, W3/13), to test its tolerogenic potential and its toxicity. Membrane expression of these molecules was measured by flow cytometry for single cells and by immunoperoxidase staining for the allograft. In grafted rats CAST significantly reduced the expression of LFA-1 alpha on lymphoid cells in the thymus, lymph node, spleen and heart allografts. ICAM-1 expression on endothelial cells of the allograft vasculature, class I and class II MHC expression on lymphoid cells in the thymus, class II MHC expression on lymphoid cells in the allograft; and CD4, CD8, CD45 and W3/13 expression on lymphoid cells in some organs. By contrast, in non-grafted rats CAST significantly upregulated expression of class I MHC and CD45 in the thymus, lymph node and spleen, ICAM-1 and CD4 on lymphoid cells in the spleen, but reduced expression of LFA-1 alpha on lymphoid cells in the thymus. It also prolonged rat heart allograft survival in a dose-dependent manner and with limited testing was relatively non-toxic. In conclusion, CAST is an immunosuppressive molecule which may work by downregulation of the ligand-receptor adhesion molecule pair, LFA-1 alpha-ICAM-1 although subtle downregulation of class I and II MHC, CD4 and CD8 molecules could also contribute to its immunosuppressive activity. Hence, both lymphocyte-endothelial cell binding and lymphocyte activation may be inhibited by CAST. This work suggests that CAST may hold significant potential as a transplant immunosuppressant probably as an adjuvant agent to inhibitors of interleukin 2 secretion.
Subject(s)
Cell Adhesion Molecules/metabolism , Enzyme Inhibitors/therapeutic use , Glycoside Hydrolase Inhibitors , Graft Survival/drug effects , Heart Transplantation , Immunosuppressive Agents/therapeutic use , Indolizines/therapeutic use , Membrane Glycoproteins/metabolism , Oligosaccharides/metabolism , Protein Processing, Post-Translational/drug effects , Animals , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/toxicity , Glycosylation/drug effects , Heart Transplantation/immunology , Histocompatibility Antigens/metabolism , Immunosuppressive Agents/pharmacology , Immunosuppressive Agents/toxicity , Indolizines/pharmacology , Indolizines/toxicity , Intercellular Adhesion Molecule-1/metabolism , Liver/drug effects , Lymphocyte Function-Associated Antigen-1/metabolism , Lymphoid Tissue/drug effects , Lymphoid Tissue/metabolism , Rats , Rats, Inbred Strains , Transplantation, Heterotopic , Transplantation, Homologous/immunology , alpha-GlucosidasesSubject(s)
Heart Transplantation/immunology , Immunosuppressive Agents/pharmacology , Indolizines/pharmacology , Intercellular Adhesion Molecule-1/biosynthesis , Lymphocyte Function-Associated Antigen-1/biosynthesis , Animals , Cell Membrane/drug effects , Cell Membrane/immunology , Cell Membrane/metabolism , Cells, Cultured , Flow Cytometry , Gene Expression , Organ Specificity , Rats , Rats, Inbred Strains , Time Factors , Transplantation, Homologous , Transplantation, IsogeneicSubject(s)
Heart Transplantation/immunology , Immunosuppressive Agents/therapeutic use , Indolizines/therapeutic use , Tacrolimus/therapeutic use , Drug Synergism , Drug Therapy, Combination , Graft Survival/drug effects , Histocompatibility Testing , Indolizines/administration & dosage , Infusions, Intravenous , Lymphocyte Culture Test, Mixed , Major Histocompatibility Complex , Proportional Hazards Models , Transplantation, HomologousSubject(s)
Cell Adhesion Molecules/metabolism , Graft Rejection/prevention & control , Heart Transplantation/immunology , Indolizines/pharmacology , Animals , Down-Regulation , Glycoside Hydrolase Inhibitors , Graft Rejection/immunology , Intercellular Adhesion Molecule-1 , Lymphocyte Function-Associated Antigen-1/metabolism , Rats , Rats, Inbred Strains , Transplantation, Homologous , alpha-GlucosidasesSubject(s)
Carbohydrate Metabolism , Glucosidases/antagonists & inhibitors , Graft Survival/immunology , Heart Transplantation/immunology , Animals , Antibodies, Monoclonal , Dose-Response Relationship, Drug , Graft Survival/drug effects , Graft Survival/physiology , Heart Transplantation/physiology , Indolizines/pharmacology , Proportional Hazards Models , Rats , Rats, Inbred Strains , Skin Transplantation/immunology , Transplantation, Heterotopic , Transplantation, HomologousSubject(s)
Diabetes Mellitus, Experimental/surgery , Duodenum/transplantation , Glycoproteins/biosynthesis , Glycoside Hydrolases/antagonists & inhibitors , Graft Survival/drug effects , Indolizines/pharmacology , Pancreas Transplantation/immunology , Animals , Graft Rejection , Muscle, Smooth/transplantation , Pancreas Transplantation/physiology , Protein Processing, Post-Translational/drug effects , Rats , Rats, Inbred Strains , Time Factors , Transplantation, HomologousABSTRACT
Conditioned immunosuppression using a taste aversion paradigm has been demonstrated in a number of laboratory models but few reports have demonstrated changes in immunity sufficient to be of clinical relevance. The experiments reported here demonstrate that the survival of heart allografts in rats can be prolonged by behaviorally conditioned immunosuppression using cyclosporin A (CsA) as an unconditioned stimulus in taste aversion conditioning. Conditioned animals received saccharin as the conditioned stimulus paired with an injection of CsA at 10 and 6 days prior to transplantation. They were reexposed to saccharin alone 1 day prior to and 3 days after transplantation. On these occasions the conditioned group displayed taste aversion behavior when offered saccharin and a significant prolongation of heart graft survival was observed compared to the conditioned and nonconditioned control groups. These experiments suggest that behaviorally conditioned immunosuppression may have important clinical implications as an adjunct to drug treatments in transplantation medicine.
Subject(s)
Avoidance Learning , Conditioning, Operant , Cyclosporine/pharmacology , Heart Transplantation/immunology , Immune Tolerance , Abdomen , Animals , Cyclosporine/administration & dosage , Graft Survival/immunology , Psychoneuroimmunology , Rats , Rats, Inbred Lew/immunology , Rats, Inbred Strains/immunology , Saccharin , Taste , Transplantation, Heterotopic , Transplantation, HomologousSubject(s)
Pancreas Transplantation/pathology , Animals , Cell Adhesion , Cell Separation/methods , Collagen , Fetus , Gels , Graft Rejection , Islets of Langerhans/cytology , Pancreas/embryology , RatsSubject(s)
Blood Transfusion , Erythrocyte Transfusion , Hot Temperature , Immunization , Pancreas Transplantation , Animals , Fetus , Graft Rejection , Rats , Rats, Inbred WFSubject(s)
Blood Transfusion , Erythrocyte Transfusion , Graft Rejection , Pancreas Transplantation , Species Specificity , Tissue Donors , Animals , Fetus , Preoperative Care , Rats , Rats, Inbred WFSubject(s)
Graft Survival , Pancreas Transplantation/immunology , Animals , Fetus , Flow Cytometry , Immunosuppressive Agents/therapeutic use , Lymphocytes/immunology , Male , Mice , Mice, Inbred BALB C , Organ Preservation/methods , Oxygen , Pancreas Transplantation/pathology , Rats , Rats, Inbred Strains , Time Factors , Transplantation, HeterologousABSTRACT
A method for monitoring circulating lymphocytes subsets in the rat on an automated flow cytometer with monoclonal antibodies was used to ascertain in vivo effects of various doses of immunosuppressive agents. The agents tested were anti-lymphocyte serum (ALS), azathioprine (AZA), cyclophosphamide (CTX), cyclosporin A (CsA) and methylprednisolone (MP). Each immunosuppressive agent varied in its capacity to induce changes in T cell subsets and B cell numbers. The rapidity of onset of action of the agents varied considerably; with ALS and MP maximal effects were seen within hours whilst the effects with CsA, cyclophosphamide (CTX) and azathioprine (AZA) took several days to develop. ALS had marked anti-T cell activity but did not selectively affect the T cell subsets. AZA and CTX both exerted their major effect upon the B cell (OX4+) subpopulation. CsA administration was associated with the appearance of many circulating lymphocytes which expressed the pan-T marker (W3/13) but neither of the T cell subset markers (W3/25, OX8). With CsA there was no significant alteration in the W3/25:OX8 ratio, although a persistent decrease in the number of all T lymphocytes was observed after administration of this drug at a dose of 45 mg/kg had ceased. MP was the only drug which had a marked selective effect on a T cell subset. The numbers of circulating Class II major histocompatibility complex (MHC) reactive lymphocytes (W3/25+) were significantly more depressed than the Class I MHC reactive subset (OX8+). This effect persisted for up to 31 days after the single injection of a depot preparation of this drug, and was found to be associated with prolonged survival of precultured endocrine xenografts.