mycophenolic-acid and Disease-Models--Animal

Patients with POAG have lower corneal endothelial cell density than healthy controls of the same age. This may be attributed to mechanical damage from elevated IOP and toxicity of glaucoma medications.. Mycophenolic acid was detected in all cats. The dose 10 mg/kg given q12h for 1 week was tolerated (n = 3). The efficacy of MMF as an immunosuppressant and long-term safety in cats of this dosage regimen is unknown.. T

Topics: Acetylcholine; Acinetobacter baumannii; Actinobacteria; Action Potentials; Adalimumab; Adaptation, Physiological; Adipates; Administration, Oral; Adolescent; Adrenal Glands; Adsorption; Adult; Aged; Aged, 80 and over; Aging; AIDS-Related Opportunistic Infections; Aldosterone; Amino Acids; Ammonia; Amoxicillin; AMP-Activated Protein Kinases; Animals; Antacids; Anti-Bacterial Agents; Antineoplastic Agents; Antirheumatic Agents; Apgar Score; Area Under Curve; ARNTL Transcription Factors; Arterial Pressure; Arthritis, Juvenile; Athletes; Attention; Biodegradation, Environmental; Biofilms; Biofuels; Biological Therapy; Biomass; Biomimetic Materials; Bioreactors; Birth Weight; Bismuth; Blood Flow Velocity; Bone and Bones; Brain Injuries, Traumatic; Calcium; Calcium Channels; Capsaicin; Carbon; Carcinoma, Hepatocellular; Cardiomegaly, Exercise-Induced; Cartilage; Cartilage, Articular; Case-Control Studies; Catalysis; Cats; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Death; Cell Differentiation; Cell Line, Tumor; Cell Membrane; Charcoal; Chemokine CCL2; Child; Child, Preschool; Chondrogenesis; Chronic Disease; Circadian Clocks; Circadian Rhythm Signaling Peptides and Proteins; Clarithromycin; Coccidioides; Coccidioidomycosis; Cognitive Behavioral Therapy; Coinfection; Color; Coloring Agents; Computer Simulation; Computers, Molecular; Consensus; Corticosterone; Cyclic AMP Response Element-Binding Protein; Cytochrome P-450 Enzyme System; Death, Sudden, Cardiac; Density Functional Theory; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Dialysis Solutions; Disease Models, Animal; Dogs; Dopamine Agonists; Dose-Response Relationship, Drug; Doxorubicin; Drug Administration Schedule; Drug Resistance, Bacterial; Drug Therapy, Combination; Electrocardiography; Electrocardiography, Ambulatory; Electrolytes; Endocardium; Endocrine Disruptors; Endocytosis; Endoscopy, Gastrointestinal; Escherichia coli Proteins; Esters; Evolution, Molecular; Executive Function; Feasibility Studies; Female; Ferric Compounds; Fluorescence; Fluorescent Dyes; Fluorine Radioisotopes; Frailty; Free Radical Scavengers; Gabapentin; Geriatric Assessment; Glucaric Acid; Glucocorticoids; Glucose; Glucose Metabolism Disorders; Halogenated Diphenyl Ethers; Heart Rate; Heart Ventricles; HEK293 Cells; Helicobacter Infections; Helicobacter pylori; Hep G2 Cells; Hepatocytes; Humans; Hungary; Hydrogen Sulfide; Hydrogen-Ion Concentration; Immunologic Factors; Immunomodulation; Immunosuppressive Agents; Independent Living; Indocyanine Green; Infant; Infant Formula; Infant Mortality; Infant, Newborn; Infant, Newborn, Diseases; Inflorescence; Insulin Resistance; Insulins; International Agencies; Iron; Isotonic Solutions; Kidney Failure, Chronic; Kinetics; Lactones; Leukocytes, Mononuclear; Liver Neoplasms; Macular Edema; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Magnetosomes; Male; Medical Audit; Mesenchymal Stem Cells; Metabolic Syndrome; Metformin; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Nude; Middle Aged; Molecular Conformation; Molecular Targeted Therapy; Motor Activity; Multiple Sclerosis; Mycophenolic Acid; Netherlands; Neuropsychological Tests; Nuclear Energy; Organs at Risk; Osteoarthritis; Osteoarthritis, Hip; Oxidation-Reduction; Palladium; Pericardium; Perinatal Death; Peritoneal Dialysis; Phantoms, Imaging; Pharmaceutical Preparations; Phospholipids; Phosphorylation; Physical Conditioning, Human; Physical Endurance; Pilot Projects; Polyketides; Polymers; Positron-Emission Tomography; Postoperative Period; Potassium; Powders; Pramipexole; Predictive Value of Tests; Pregabalin; Pregnancy; Pregnancy Outcome; Protein Structure, Secondary; Proton Pump Inhibitors; Puberty; Pulmonary Circulation; Quality Assurance, Health Care; Quantum Dots; Radiometry; Radiotherapy Dosage; Radiotherapy Planning, Computer-Assisted; Radiotherapy, Intensity-Modulated; Rats, Sprague-Dawley; Receptors, CCR2; Receptors, Transferrin; Regeneration; Registries; Renal Insufficiency, Chronic; Reproducibility of Results; Research Design; Restless Legs Syndrome; Retina; Retinoid X Receptor alpha; Retrospective Studies; Rhenium; Risk Factors; RNA, Messenger; Severity of Illness Index; Sex Factors; Sodium; Sodium Fluoride; Solvents; Spectrometry, Fluorescence; Spectroscopy, Fourier Transform Infrared; Stereoisomerism; Stroke; Structure-Activity Relationship; Tachycardia, Ventricular; Tetracycline; Tetrahydrofolate Dehydrogenase; Tetrahydronaphthalenes; Thermodynamics; Thiophenes; Time Factors; Tinidazole; Tomography, Optical Coherence; Tomography, X-Ray Computed; Topiramate; Toxoplasma; Toxoplasmosis, Cerebral; Transferrin; Treatment Outcome; Up-Regulation; Upper Extremity; Uremia; Uveitis; Vascular Remodeling; Ventricular Fibrillation; Ventricular Function, Left; Ventricular Function, Right; Ventricular Remodeling; Verapamil; Veterans; Visual Acuity; Vitrectomy; Water Pollutants, Chemical; Zea mays; Zirconium

Mycophenolic acid (MPA) was introduced into clinical practice as immunosuppressive drug therapy to prevent allograft rejection. Since then, its clinical application has widened. Our goal was to review the lessons learned from experimental nontransplant glomerular disease models on the mechanisms of MPA therapy. T and B lymphocytes are preferentially dependent on de novo purine synthesis. By inhibiting the rate-limiting enzyme of de novo purine synthesis, MPA depletes the pool of deoxyguanosine triphosphate (dGTP) and inhibits proliferation of these immune cells. Furthermore, MPA can also induce apoptosis of immune cells and is known to inhibit synthesis of fucose- and mannose-containing membrane glycoproteins altering the surface expression and binding ability of adhesion molecules. However, MPA exerts a direct effect also on nonimmune cells. Mesangial cells are partially dependent on de novo purine biosynthesis and are thus susceptible to MPA treatment. Additionally, MPA can inhibit apoptosis in podocytes and seems to be beneficial in preserving the expression of nephrin and podocin, and by attenuation of urokinase receptor expression leads to decreased foot-process effacement. In summary, our manuscript sheds light on the molecular mechanisms underlying the antiproteinuric effect of MPA. Overall, MPA is an excellent treatment option in many immunologic glomerulopathies because it possesses immunosuppressive properties, has a remarkable effect on nonimmune cells and counteracts the proliferation of mesangial cells, expansion of mesangial matrix, and foot-process effacement of podocytes combined with a low systemic toxicity.

Topics: Animals; Cell Proliferation; Disease Models, Animal; Glomerular Basement Membrane; Glomerulonephritis; Humans; Immunosuppressive Agents; IMP Dehydrogenase; Intracellular Signaling Peptides and Proteins; Lymphocytes; Membrane Proteins; Mesangial Cells; Monocytes; Mycophenolic Acid; Nephrotic Syndrome; Podocytes; Purines; Rats

The success of solid-organ transplantation was made possible by recognizing that destruction of the graft is caused by an alloimmune-mediated process. For the past decade, immunosuppressive protocols have used a combination of drugs that significantly decreased the rate of acute organ rejection. Despite advances in surgical and medical care of recipients of solid-organ transplants, long-term graft survival and patient survival have not improved during the past 2 decades. Current immunosuppression protocols include a combination of calcineurin inhibitors, such as tacrolimus, and antiproliferative agents (most commonly mycophenolate mofetil), with or without different dosing regimens of corticosteroids. Mammalian target of rapamycin inhibitors were introduced to be used in combination with cyclosporine-based therapy, but they did not gain much acceptance because of their adverse event profile. Belatacept, a costimulatory inhibitor, is currently being studied in different regimens in an effort to replace the use of calcineurin inhibitors to induce tolerance and to improve long-term outcomes. Induction therapy is now being used in more than 90% of kidney transplants and more than 50% cases of other solid-organ transplantation such as lung, heart, and intestinal transplants. As a result of these combination immunosuppressive (IS) therapy protocols, not only the incidence but also the intensity of episodes of acute rejection have decreased markedly, and at present 1-year graft and patient survival is almost 98% for kidney transplant recipients and approximately greater than 80% for heart and lung transplants. Evolving concepts include the use of donor-derived bone marrow mesenchymal cells to induce tolerance, to minimize the use of maintenance IS agents, and to prevent the development of adverse events associated with long-term use of maintenance IS therapy.

Topics: Abatacept; Alemtuzumab; Animals; Antibodies, Monoclonal, Humanized; Azathioprine; Bone Marrow Transplantation; Calcineurin Inhibitors; Cyclosporine; Disease Models, Animal; Everolimus; Heart Transplantation; Humans; Immunosuppression Therapy; Immunosuppressive Agents; Kidney Transplantation; Lung Transplantation; Mycophenolic Acid; Piperidines; Pyrimidines; Pyrroles; Randomized Controlled Trials as Topic; Sirolimus; Tacrolimus

Substantial progress has been made over the last two decades in our understanding of the immunopathogenesis of antineutrophil cytoplasmic antibodies (ANCA) associated vasculitides. Compelling evidence from in vitro studies and experimental models in conjunction with clinical trials has confirmed that ANCA directly contribute to the evolution and progression of the disease process. Continuous development in our understanding of the mechanisms that drive the disease may ultimately allow us to tailor the multitude of novel therapies, which are rapidly becoming available, to the requirements of individual patients. In this review we endeavour to provide a brief overview of the recent advances in ANCA-associated vasculitides and outline basic principles for diagnosis and treatment of these complex multisystem diseases.

Topics: Animals; Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis; Antibodies, Antineutrophil Cytoplasmic; Azathioprine; Cyclophosphamide; Disease Models, Animal; Disease Progression; Humans; Immunoglobulin G; Immunosuppressive Agents; Methotrexate; Mycophenolic Acid

The immunosuppressive agent mycophenolate mofetil (MMF) has beneficial effects in cardiac transplant patients beyond the suppression of tissue rejection. Patients with regimens containing MMF experience diminished intimal thickening and cardiac allograft vasculopathy compared to patients treated with azathioprine. Studies have shown that diet-induced atherosclerosis (a related vasculopathy) is a chronic inflammatory process, and so MMF has also been used to reduce atherosclerosis in a rabbit model of hyperlipidemia. These data hold out the intriguing possibility that MMF might be a viable primary or secondary preventive agent in people at significant risk for atherosclerosis.

Topics: Animals; Atherosclerosis; Disease Models, Animal; Humans; Mycophenolic Acid

Acute renal failure (ARF) is a common cause of mortality and morbidity in hospitalized patients. Ischemia is an important cause of ARF, and ARF caused by ischemic injury is referred to as ischemic acute tubular necrosis (ATN). There is growing evidence from models that ischemic ATN is associated with intrarenal inflammation. Consequently, intrarenal inflammation is an attractive target for the development of novel drug therapies for ARF. This review outlines ischemic ATN models, the pathophysiological roles of inflammatory cells such as T and B cells in ischemic ATN models, and effective T and B cell therapeutic reagents.

Topics: Animals; Anti-Inflammatory Agents; B-Lymphocytes; Cells, Cultured; Disease Models, Animal; Drug Evaluation, Preclinical; Gene Expression Profiling; Gene Expression Regulation; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Inflammation; Ischemia; Kidney; Kidney Tubular Necrosis, Acute; Mycophenolic Acid; Oligonucleotide Array Sequence Analysis; T-Lymphocytes

Glomerulonephritis is a challenging complication of systemic lupus erythematosus that still results in kidney loss in up to 30% of patients. In this review we highlight the development of integrated efforts to link pathogenesis with disease definition and new therapeutics.. Immune complex deposition in the kidney initiates an inflammatory cascade that causes glomerular disease but there are many modulating factors including genetic predisposition, products of the innate immune system, cytokines, complement and activated cells (both renal and immune). Animal models can help dissect potential disease mechanisms but the study of multiple models will be required since there are multiple subsets of human disease. Recent therapeutic studies in humans address the distinction between therapies for remission induction and remission maintenance. Multiple studies confirm the therapeutic equivalence of mycophenolate mofetil and cyclophosphamide in induction of remission but results are still far from ideal. The next few years should see the testing of new biologic reagents in humans. Another area of interest is the search for noninvasive measures of disease and disease response.. Although there has been remarkable progress in our understanding of the immunology and phenotype of lupus nephritis current therapies have insufficient efficacy. As new therapies emerge, improved clinical design coupled with mechanistic studies will be needed to identify agents that may be effective only in some patient subpopulations.

Topics: Animals; Cyclophosphamide; Disease Models, Animal; Humans; Immunosuppressive Agents; Lupus Nephritis; Mice; Mycophenolic Acid; Remission Induction

Mycofenolate mofetil (MMF-Cellcept) is an immunomodulatory drug utilized extensively in transplant medicine. The efficacy of regimes including Cellcept in preventing allograft rejection, and in the treatment of rejection, is now firmly established. The immunosuppressive actions of this drug enabled the investigation for the beneficial effects in autoimmune diseases. We review the evidence for the contribution of MMF in autoimmunity in animal models of systemic lupus erythematosus (SLE), mercury induced autoimmune glomerulonephritis, diabetes mellitus, experimental autoimmune uveoretinitis, and experimental allergic encephalitis. MMF has an influence on the T and B cell pathways. It is immunosuppressive and anti-inflammatory.

Topics: Animals; Autoimmune Diseases; Diabetes Mellitus, Experimental; Disease Models, Animal; Immunosuppressive Agents; Inflammatory Bowel Diseases; Mycophenolic Acid; Retinitis; Uveitis

Glomerulonephritis is an important manifestation of small vessel vasculitides such as Wegener granulomatosis, microscopic polyangiitis, and Churg-Strauss syndrome. Renal involvement in these diseases is characterized by a pauci-immune segmental necrotizing and crescentic glomerulonephritis that is strongly associated with circulating antineutrophil cytoplasmic autoantibodies. We will review recent advances in understanding the pathogenesis of antineutrophil cytoplasmic autoantibody-related renal vasculitides and innovative approaches to their treatment.. An experimental milestone in antineutrophil cytoplasmic autoantibody research has been reached in the past year. Using an innovative mouse model, investigators from the University of North Carolina in Chapel Hill have recently acquired robust data supporting the pathogenic role of antineutrophil cytoplasmic autoantibodies in the glomerulonephritis and small vessel vasculitis, analogous to those seen in microscopic polyangiitis and Wegener granulomatosis. Novel immunosuppressive approaches have been examined including preliminary studies using biologic agents, such as antagonists of tumor necrosis factor and monoclonal antibodies to B lymphocytes.. Recent insights into the pathogenesis of antineutrophil cytoplasmic autoantibody-related vascular injury and the availability of new biologic, immune response modifiers to complement standard chemical immunosuppressive agents offer exciting new prospects for investigation in the management of patients with small vessel renal vasculitides.

Topics: Adjuvants, Immunologic; Animals; Antibodies, Antineutrophil Cytoplasmic; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Azathioprine; Cyclophosphamide; Disease Models, Animal; Humans; Kidney; Kidney Diseases; Methotrexate; Mice; Mycophenolic Acid; Rituximab; Vasculitis

The optimal treatment of atopic dermatitis requires regular medical supervision. The course of this chronic skin disease is influenced by multiple triggers which are relevant for the treatment. The mainstays of topical therapy include regular use of emollients coupled with antimicrobial substances, corticosteroids and immune modulators as required. Ultraviolet radiation and immunosuppressive regimens represent further options for the treatment of severe exacerbations and may lead to long term improvement. Data from experimental studies provide insight into possible future treatment methods.

Topics: Adjuvants, Immunologic; Adolescent; Adrenal Cortex Hormones; Adult; Age Factors; Animals; Anti-Bacterial Agents; Anti-Infective Agents, Local; Antifungal Agents; Azathioprine; Child; Child, Preschool; Cross-Over Studies; Cyclosporine; Dermatitis, Atopic; Dermatologic Agents; Desensitization, Immunologic; Disease Models, Animal; Double-Blind Method; Histamine Antagonists; Humans; Immunoglobulins, Intravenous; Immunosuppressive Agents; Infant; Leukotriene Antagonists; Medicine, Chinese Traditional; Mice; Mice, Nude; Mycophenolic Acid; Ointments; Photopheresis; Phototherapy; Placebos; Probiotics; PUVA Therapy; Randomized Controlled Trials as Topic; Ultraviolet Therapy

Renal infiltration with macrophages and monocytes is a well-recognized feature of not only immune, but also nonimmune kidney disease. This review focuses on the investigations that have shown accumulation of immunocompetent cells in experimental models of acute and chronic ischemia, protein overload, hypercholesterolemia, renal ablation, obstructive uropathy, polycystic kidney disease, diabetes, aging, murine hypertension, and nephrotoxicity. We examine the mechanisms of infiltration of immunocompetent cells and their participation in the self-perpetuating cycle of activation of the angiotensin system, generation of reactive oxygen species, and further recruitment of monocytes and lymphocytes. We also discuss the possibility of antigen-dependent and antigen-independent mechanisms of immune cell activation in these animal models. Finally, we review the recent studies in which suppression of cellular immunity with mycophenolate mofetil has proven beneficial in attenuating or preventing the progression of renal functional and histologic damage in experimental conditions of nonimmune nature.

Topics: Animals; Antigens; Disease Models, Animal; Humans; Immunocompetence; Immunosuppressive Agents; Kidney Diseases; Lymphocyte Activation; Lymphocytes; Macrophage Activation; Macrophages; Models, Biological; Mycophenolic Acid

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Humans; Kidney Diseases; Kidney Glomerulus; Mycophenolic Acid

Both mizoribine (MZR) and mycophenolate mofetil (MMF) are immunosuppressive agents that inhibit the proliferation of lymphocytes selectively, via inhibition of IMPDH. MZR is a nucleoside of the imidazole class, isolated from the culture medium of the mold Eupenicillium brefeldianum M-2166. Although this compound has been found to have weak antimicrobial activity against Candida albicans, it has proved ineffective against experimental candidiasis. Unlike azathioprine, this compound is not taken up by nucleic acids in the cell. Instead, after phosphorylation MZR-5 -monophosphate inhibits GMP synthesis by the antagonistic blocking of IMPDH (Ki = 10(-8)M) and GMP- synthetase (Ki =10(-5) M). The drug has been found to inhibit both humoral and cellular immunity, and on this basis it was developed in Japan as an immunosuppressant. MZR has been shown in animal experiments to lack oncogenicity, and has been shown clinically to be associated with a low incidence of severe adverse reactions. MZR has been registered in Japan for the prevention of rejection in renal transplantation, and for the treatment of lupus nephritis, rheumatoid arthritis and the nephrotic syndrome. MMF is the morpholinoethyl ester prodrug of mycophenolic acid (MPA), which was first isolated in 1896 from the culture media of several Penicillium species. MPA has been evaluated for its unique properties as an anticancer, antiviral, antifungal and antibacterial agent, as well as for its therapeutic use in psoriasis and rheumatoid arthritis. MMF was designed to enhance the oral bioavailability of the parent compound. After beneficial effects were observed in animals, the clinical efficacy of MMF as an immunosuppressant in renal transplantation was studied in the United States. In 1995 the US Food and Drug Administration (FDA) approved the use of MMF for the prevention of rejection in renal transplantation, the drug also available on a number of European markets.

Topics: Animals; Arthritis; Carbon-Nitrogen Ligases; Cell Cycle; Cell Division; Clinical Trials as Topic; Creatine; Disease Models, Animal; Dogs; Dose-Response Relationship, Drug; Flow Cytometry; Humans; Immunosuppressive Agents; IMP Dehydrogenase; Kinetics; Lupus Nephritis; Lymphocytes; Mice; Mitogens; Models, Biological; Models, Chemical; Mycophenolic Acid; Nephrotic Syndrome; Organ Transplantation; Ribonucleosides; Time Factors; Tumor Cells, Cultured

The immunosuppressive drug mycophenolate mofetil is a potential new treatment for autoimmune renal disease. In addition to its effects in modulating the autoimmune response, mycophenolate mofetil reduces adhesion molecule expression and delays the progression of renal failure. Data from experimental models of glomerulonephritis, especially of lupus nephritis, have demonstrated that mycophenolate mofetil reverses both inflammatory and autoimmune aspects of disease and influences outcome. As yet, clinical experience with mycophenolate mofetil remains anecdotal but provides a strong platform for the scientific evaluation of mycophenolate mofetil as a new therapeutic agent for these conditions in the future.

Topics: Animals; Autoimmune Diseases; Disease Models, Animal; Glomerulonephritis; Humans; Immunosuppressive Agents; Kidney Diseases; Lupus Nephritis; Mycophenolic Acid; Vasculitis

Mycophenolate mofetil (MMF) is a novel immunosuppressive drug that shows promise in preventing the rejection of organ allografts and in the treatment of ongoing rejection. Orally administered MMF is hydrolyzed by esterases in the intestine and blood to release mycophenolic acid (MPA), a potent, selective, noncompetitive inhibitor of the type 2 isoform of inosine monophosphate dehydroxygenase (IMPDH) expressed in activated human T and B lymphocytes. By inhibiting IMPDH, MPA depletes the pool of dGTP required for DNA synthesis. MPA has a more potent cytostatic effect on lymphocytes than on other cell types, and this is the principal mechanism by which immunosuppressive activity is exerted. MPA also depletes pools of GTP in human lymphocytes and monocytes, thereby inhibiting the synthesis of fucose- and mannose-containing saccharide components of membrane glycoproteins. These are recognized by the family of adhesion molecules termed selectins. By this mechanism, MPA could decrease the recruitment of lymphocytes and monocytes into sites of graft rejection. In addition to preventing allograft rejection, MMF suppresses graft-versus-host reactions in lethal and nonlethal murine models. MMF inhibits primary antibody responses more efficiently than secondary responses. MPA inhibits the proliferation of human B lymphocytes transformed by Epstein-Barr virus and is not mutagenic. Clinically attainable concentrations of MPA suppress the proliferation of human arterial smooth muscle cells. These two properties of MPA may decrease the risk of lymphoma development and proliferative arteriopathy in long-term recipients of MMF.

Topics: Animals; B-Lymphocytes; Disease Models, Animal; DNA; Enzyme Inhibitors; Graft Rejection; Graft vs Host Disease; Guanosine Triphosphate; Humans; Immunosuppressive Agents; IMP Dehydrogenase; Lymphocyte Activation; Membrane Glycoproteins; Mice; Muscle, Smooth, Vascular; Mycophenolic Acid; Organ Transplantation; Purines; Selectins; T-Lymphocytes; Transplantation, Homologous

In summary, many new modalities of immunosuppression after transplantation are being investigated (Fig. 1). These approaches include various new drugs or monoclonal antibodies that target different cell subsets, cellular activation pathways, cellular effector function or mediators (such as cytokines) of effector function, ligands that stabilize cellular interactions, or antimetabolites that preferentially affect lymphocytes (Tables 4 and 5). Because of the excellent early graft and patient survival results after liver transplantation under various current immunosuppressive protocols, future clinical trials using these various new modalities will require large numbers of patients to show statistically significant differences in graft or patient survival. Therefore, other criteria in addition to graft and patient survival must be analyzed to evaluate the importance of new immunosuppressive therapies. These criteria may include incidence of acute or chronic rejection, long-term graft function, incidence of infectious complications, length of hospitalization, drug toxicity, and patient tolerance and compliance with new therapies.

Topics: Alprostadil; Animals; Antibodies, Monoclonal; Biphenyl Compounds; Child; Disease Models, Animal; Dogs; Guanidines; Humans; Immunosuppressive Agents; Liver Transplantation; Mycophenolic Acid; Polyenes; Rats; Sirolimus; Tacrolimus

The effects of an immunosuppressive agent, mycophenolate mofetil (MM), were investigated and compared with those of methylprednisolone (MP) and dexamethasone (DXM) on the traumatic nerve function.. This is a randomized controlled animal study.. This experimental study was performed on 84 male Wistar albino rats. The rats were assigned to 12 groups each consisting of 7 animals. The groups were formed according to application of normal-dose DXM (group 1A-B), high-dose MP (group 2A-B), normal-dose MP (group 3A-B), MM (group 4A-B), and MM with high-dose MP combination therapies (group VA-B). Right sciatic nerve dissection was performed, and compound muscle action potential thresholds were recorded. The nerve was traumatized with the compression of a Jeweller forceps for 20 seconds. Posttraumatic thresholds were also recorded. The compound muscle action potential thresholds were recorded in the first and fourth weeks for the assigned groups. Then, the nerve was transected and prepared for electron microscopic and histopathologic examinations. Nitric oxide and malondialdehyde assessments were performed on both tissue and blood samples.. Only the MM and MP+MM groups had satisfactory electron microscopic findings and were about to reach the tissue characteristics of the control animals. Despite the electrophysiologic recovery, the DXM group was found to have poor electron microscopic scoring.. Mycophenolate mofetil has been found to be beneficial in the treatment of traumatic nerve paralysis. Although a complementary investigation is needed, this immunosuppressive agent may be an alternative to corticosteroids for the selected cases where steroid therapy is contraindicated.

Topics: Animals; Dexamethasone; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Synergism; Electromyography; Facial Paralysis; Male; Methylprednisolone; Microscopy, Electron; Mycophenolic Acid; Nerve Regeneration; Peripheral Nerve Injuries; Rats; Rats, Wistar; Sciatic Nerve; Sciatic Neuropathy

To assess the efficacy of paquinimod, a new immunomodulatory small molecule, in a murine lupus model, and to evaluate its pharmacokinetics and tolerability in systemic lupus erythematosus (SLE) patients at doses predicted to be efficacious and safe and determine the maximum tolerated dose.. The efficacy of paquinimod was studied in lupus-prone MRL-lpr/lpr mice and compared with that of established SLE treatments. Dose-response data and pharmacokinetic data were used to calculate effective and safe clinical doses of paquinimod. The pharmacokinetics and tolerability of paquinimod were evaluated in a phase Ib double-blind, placebo controlled, dose-ranging study in which cohorts of SLE patients received daily oral treatment for 12 weeks.. Paquinimod treatment resulted in disease inhibition in MRL-lpr/lpr mice, comparable to that obtained with prednisolone and mycophenolate mofetil; prominent effects on disease manifestations and serologic markers and a steroid-sparing effect were observed. In patients with SLE, the pharmacokinetic properties of paquinimod were linear and well suitable for once-daily oral treatment. The majority of the adverse events (AEs) were mild or moderate, and transient. The most frequent AEs were arthralgia and myalgia, reported with the highest dose levels of paquinimod (4.5 mg/day and 6.0 mg/day). At the 4.5 mg/day dose level and higher, some AEs of severe intensity and serious adverse events were reported.. Paquinimod effectively inhibited disease and had a steroid-sparing effect in experimental lupus. Results from preclinical models together with pharmacokinetic data were successfully translated into a safe clinical dose range, and doses of up to 3.0 mg/day were well tolerated in the SLE patients. Taken together, the promising combined data from a murine model and human SLE support the future clinical development of paquinimod.

Topics: Adult; Aged; Animals; Disease Models, Animal; Double-Blind Method; Female; Humans; Immunosuppressive Agents; Kidney; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred MRL lpr; Middle Aged; Mycophenolic Acid; Prednisolone; Quinolines; Severity of Illness Index; Treatment Outcome; Young Adult

Systemic lupus erythematosus (SLE) is an autoimmune disease involving multi-organ systems with a widely heterogeneous clinical presentation. Renal involvement, observed mainly in lupus nephritis (LN), is the most common organ lesion associated with SLE and a determinant of prognosis. However, treatment of LN remains controversial and challenging, prompting the need for novel therapeutic approaches. In particular, development of a clinically relevant LN animal model would greatly facilitate the development of new treatments. Here, we report a novel murine model for LN established by administering polyinosinic-polycytidylic acid (Poly (I:C)) to NZB/W F1 mice. We investigated the effectiveness of administering Poly (I:C) to NZB/W F1 mice for accelerating nephritis onset and explored the optimal conditions under which to enroll mice with nephritis with similar pathology for studying treatment candidates. Gene-expression analysis revealed that activation of macrophages, which are reported to be involved in the progression of LN in patients, was a unique characteristic in this accelerated nephritis model. Evaluation of the therapeutic effect of mycophenolate mofetil (MMF), a recommended first-choice agent for LN, in this novel LN model showed that MMF significantly reduced proteinuria. The cathepsin S (CatS) inhibitor ASP1617, which has been reported to prevent development of lupus-like glomerulonephritis in the spontaneous NZB/W F1 mouse model, also showed marked therapeutic effect in this model. Our novel Poly (I:C) accelerated LN model would thus be very useful for screening clinical candidates for LN, and CatS may be an attractive therapeutic target for the treatment of LN.

Topics: Animals; Disease Models, Animal; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Lupus Nephritis; Mice; Mice, Inbred NZB; Mycophenolic Acid; Poly I-C

Mycophenolate mofetil (MMF) is an immunosuppressive agent widely applied in various autoimmune diseases, including autoimmune uveitis, a sight-threatening autoimmune disease mainly affecting the eyes. However, the mechanisms of action are not comprehensively understood. To investigate the potential impact of MMF on uveitis, we generated single-cell RNA sequence data from normal, experimental autoimmune uveitis (EAU) and MMF-treated EAU mice. We observed that some EAU-induced transcriptional changes were reversed by MMF treatment. Transcriptional data indicated that MMF may have a general inhibitory effect on the activation of immune cells during EAU. Each immune cell type showed a different response to MMF treatment. Pseudotime analysis showed that MMF treatment partly reversed the increased differentiation tendency from naïve to effector phenotypes of T and B cells in EAU. The reduced proportion of T-helper (Th)1 and T-helper (Th)17 cells after MMF treatment was confirmed using flow cytometry. MMF treatment downregulated the EAU-associated upregulation of several molecules (such as Cebpd, Pim1, Furin, Bhlhe40, and Hif1a) that promote pathogenic cytokine production by T helper (Th)-1 and Th17 cells. Abnormally enhanced immunoglobulin production, antigen processing, and presentation ability of B cells may also be inhibited by MMF treatment. In addition to T and B cells, MMF treatment countered EAU-induced transcriptional changes in other immune cells to different degrees. Overall, our findings provide novel insights into the mechanisms underlying MMF treatment and indicate that the therapeutic effect of MMF is not driven by a single molecule.

Topics: Animals; Autoimmune Diseases; Disease Models, Animal; Immunosuppressive Agents; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Th17 Cells; Transcriptome; Uveitis

Mycophenolic acid (MPA) is the most widely used immunosuppressive drug in transplantation and for autoimmune diseases. Unfortunately, more than 30% of patients experience a typical gastrointestinal adverse effect also referred to as mycophenolate-induced enteropathy. Due to its antibacterial, antifungal, and antiviral properties, MPA exposure is associated with intestinal dysbiosis characterized by a decrease in density and diversity of the microbiome regarding the main bacterial phyla (Firmicutes and Bacteroidetes). These bacterial phyla are known for their metabolic role in maintaining the homeostasis of the digestive tract, particularly through the production of short-chain fatty acids (SCFA) that could contribute to the pathophysiology of mycophenolate-induced enteropathy. Our study aimed at deciphering short-chain fatty acids (SCFA) profile alterations associated with gastrointestinal toxicity of MPA at the digestive and systemic levels in a mouse model.. Ten-week old C57BL/6 (SOPF) mice were randomly assigned in 2 groups of 9 subjects: control, and mycophenolate mofetil (MMF, 900 mg/kg/day). All mice were daily treated by oral gavage for 7 days. Individual faecal pellets were collected at days 0, 4 and 8 as well as plasma at day 8 for SCFA profiling. Additionally, after the sacrifice on day 8, the caecum was weighted, and colon length was measured. The proximal colon was cut for histological analysis.. MMF treatment induced around 10% weight loss at the end of the protocol associated with a significant decrease in caecum weight and a slight reduction in colon length. Histological analysis showed significant architectural changes in colon epithelium. Moreover, we observed an overall decrease in SCFA concentrations in faecal samples, especially regarding acetate (at day 8, control 1040.6 ± 278.161 μM versus MMF 384.7 ± 80.5 μM, p < 0.01) and propionate (at day 8, control 185.94 ± 51.96 μM versus MMF 44.07 ± 14.66 μM, p < 0.001), and in plasma samples for butyrate (at day 8, control 0.91 ± 0.1 μM versus MMF 0.46 ± 0.1 μM, p < 0.01).. These results are consistent with functional impairment of the gut microbiome linked with digestive or systemic defects during MMF treatment.

Topics: Animals; Anti-Bacterial Agents; Colon; Disease Models, Animal; Fatty Acids, Volatile; Feces; Female; Gastrointestinal Microbiome; Immunosuppressive Agents; Intestinal Diseases; Mice, Inbred C57BL; Mycophenolic Acid

Combination therapy with mycophenolate mofetil, tacrolimus and steroids are effective in achieving complete remission in lupus nephritis (LN). Combination therapy uniquely downregulated caspase-1 compared with monotherapies, which can cleave gasdermin D (GSDMD) and was recently identified as the pyroptosis executioner. We therefore investigated whether combination therapy enabled the suppression of caspase-1/GSDMD-mediated pyroptosis in LN.. Expression and activation of GSDMD were detected in kidney specimens of the human and mouse with LN using immunohistochemical staining and immunoblotting. Primary podocytes isolated from MRL/lpr mice were incubated with LPS+ATP, and pretreated with monotherapy or combination therapy. Inhibition of caspase-1/GSDMD-induced pyroptosis by combination therapy were assessed in MRL/lpr mice and human specimens. Pyroptosis was examined using a FAM caspase-1 kit and flow cytometry. The correlation between pyroptosis in peripheral blood and the systemic lupus erythematosus disease activity index (SLEDAI) was analyzed.. Kidney tissue specimens from LN patients and mice exhibited greatly increased expression levels and cleavage of GSDMD. In cultured podocytes, combination treatment significantly suppressed the activation of NLRP3 and caspase-1 and reduced GSDMD N-terminal levels. Combination therapy repressed disease progression through inhibition of caspase-1/GSDMD-mediated pyroptosis in both humans and MRL/lpr mice. Caspase-1/PI positive cell numbers in peripheral blood were positively correlated with SLE-DAI. LN patients with complete remission and partial remission had remarkably reduced caspase-1/PI positive cell numbers compared to baseline. Ac-FLTD-CMK, a GSDMD-derived inhibitor, prevented the development of LN.. Combination therapy suppressed caspase-1/GSDMD-mediated pyroptosis

Topics: Adolescent; Adult; Aged; Animals; Calcineurin Inhibitors; Caspase 1; Caspase Inhibitors; Cells, Cultured; Cohort Studies; Disease Models, Animal; Drug Therapy, Combination; Female; Humans; Intracellular Signaling Peptides and Proteins; Kidney; Lupus Nephritis; Mice; Mice, Inbred C57BL; Mice, Inbred MRL lpr; Middle Aged; Mycophenolic Acid; Phosphate-Binding Proteins; Podocytes; Prednisone; Pyroptosis; Tacrolimus; Young Adult

Microbiota has a role in the host blood pressure (BP) regulation. The immunosuppressive drug mofetil mycophenolate (MMF) ameliorates hypertension. The present study analyzes whether MMF improves dysbiosis in a genetic model of hypertension. Twenty weeks old male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were randomly divided into three groups: untreated WKY, untreated SHR, and SHR treated with MMF for 5 weeks. MMF treatment restored gut bacteria from the phyla Firmicutes and Bacteroidetes, and acetate- and lactate-producing bacteria to levels similar to those found in WKY, increasing butyrate-producing bacteria. MMF increased the percentage of anaerobic bacteria in the gut. The improvement of gut dysbiosis was associated with an enhanced colonic integrity and a decreased sympathetic drive in the gut. MMF inhibited neuroinflammation in the paraventricular nuclei in the hypothalamus. MMF increased the lower regulatory T cells proportion in mesenteric lymph nodes and Th17 and Th1 infiltration in aorta, improved aortic endothelial function and reduced systolic BP. This study demonstrates for the first time that MMF reduces gut dysbiosis in SHR. This effect could be related to its capability to improve gut integrity due to reduced sympathetic drive in the gut associated to the reduced brain neuroinflammation.

Topics: Animals; Antihypertensive Agents; Aorta, Thoracic; Bacteria; Blood Pressure; Cells, Cultured; Colon; Cytokines; Disease Models, Animal; Dysbiosis; Gastrointestinal Microbiome; Hypertension; Inflammation Mediators; Mycophenolic Acid; Neuroimmunomodulation; Paraventricular Hypothalamic Nucleus; Rats, Inbred SHR; Rats, Inbred WKY; Sympathetic Nervous System; T-Lymphocytes

Topics: Actins; Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Line, Tumor; Collagen; Disease Models, Animal; Drug Compounding; Drug Delivery Systems; Endopeptidases; Fibroblasts; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Linoleic Acid; Liver Neoplasms; Membrane Proteins; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Nanoparticles; NIH 3T3 Cells; Polymers; Xenograft Model Antitumor Assays

When Zika virus emerged as a public health emergency there were no drugs or vaccines approved for its prevention or treatment. We used a high-throughput screen for Zika virus protease inhibitors to identify several inhibitors of Zika virus infection. We expressed the NS2B-NS3 Zika virus protease and conducted a biochemical screen for small-molecule inhibitors. A quantitative structure-activity relationship model was employed to virtually screen ∼138,000 compounds, which increased the identification of active compounds, while decreasing screening time and resources. Candidate inhibitors were validated in several viral infection assays. Small molecules with favorable clinical profiles, especially the five-lipoxygenase-activating protein inhibitor, MK-591, inhibited the Zika virus protease and infection in neural stem cells. Members of the tetracycline family of antibiotics were more potent inhibitors of Zika virus infection than the protease, suggesting they may have multiple mechanisms of action. The most potent tetracycline, methacycline, reduced the amount of Zika virus present in the brain and the severity of Zika virus-induced motor deficits in an immunocompetent mouse model. As Food and Drug Administration-approved drugs, the tetracyclines could be quickly translated to the clinic. The compounds identified through our screening paradigm have the potential to be used as prophylactics for patients traveling to endemic regions or for the treatment of the neurological complications of Zika virus infection.

Topics: Animals; Antiviral Agents; Artificial Intelligence; Chlorocebus aethiops; Disease Models, Animal; Drug Evaluation, Preclinical; High-Throughput Screening Assays; Immunocompetence; Inhibitory Concentration 50; Methacycline; Mice, Inbred C57BL; Protease Inhibitors; Quantitative Structure-Activity Relationship; Small Molecule Libraries; Vero Cells; Zika Virus; Zika Virus Infection

To determine the effect of mycophenolate mofetil (MMF) on retinal degeneration on two mouse models of retinitis pigmentosa.. Intraperitoneal injections of MMF were administered daily in rd10 and c57 mice starting at postoperative day 12 (P12) and rd1 mice starting at P8. The effect of MMF was assessed with optical coherence tomography, immunohistochemistry, electroretinography, and OptoMotry. Whole retinal cyclic guanosine monophosphate (cGMP) and mycophenolic acid levels were quantified with mass spectrometry. Photoreceptor cGMP cytotoxicity was evaluated with cell counts of cGMP immunostaining.. MMF treatment significantly delays the onset of retinal degeneration and cGMP-dependent photoreceptor cytotoxicity in rd10 and rd1 mice, albeit a more modest effect in the latter. In rd10 mice, treatment with MMF showed robust preservation of the photoreceptors up to P22 with associated suppression of cGMP immunostaining and microglial activation; The neuroprotective effect diminished after P22, but outer retinal thickness was still significantly thicker by P35 and OptoMotry response was significantly better up to P60. Whereas cGMP immunostaining of the photoreceptors were present in rd10 and rd1 mice, hyperphysiological whole retinal cGMP levels were observed only in rd1 mice.. Early treatment with MMF confers potent neuroprotection in two animal models of RP by suppressing the cGMP-dependent common pathway for photoreceptor cell death. The neuroprotective effect of MMF on cGMP-dependent cytotoxicity occurs independently of the presence of hyperphysiological whole retinal cGMP levels. Thus our data suggest that MMF may be an important new class of neuroprotective agent that could be useful in the treatment of patients with RP.

Topics: Animals; Cyclic GMP; Disease Models, Animal; Electroretinography; Mass Spectrometry; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Neuroprotective Agents; Photoreceptor Cells, Vertebrate; Retina; Retinitis Pigmentosa; Tomography, Optical Coherence

The aim of the present study was to evaluate the histological, histochemical, and stereological changes caused by mycophenolate mofetil (MMF) on the tunica albuginea of rat penises submitted to an injection of transforming growth factor beta (TGF-β) for the induction of Peyronie's disease (PD). Twenty adult male Wistar rats were divided into four groups: Control group; TGF-β group (TGF-β injection); MMF-7d group (treated with MMF 7 days after induction with TGF-β); and MMF-30d group (treated with MMF 30 days after induction with TGF-β). The steorological evaluation included the relative volume of different types of connective fibres of the tunica albuginea. The histochemical analysis revealed the fragmentation and degradation of elastin in the tunica albuginea. This process was partially reversed in the MMF-7d group and a situation very close to normality was observed in the MMF-30d group. In the collagen III/collagen I ratio it was observed increase in this ratio in the TGF-β (59.4 ± 5.53) and MMF-7d (49 ± 18.2) groups and a decrease in the MMF-30d group (28.7 ± 4), approaching normality. The injection of TGF-β promoted fibrotic alterations in the penile tunica albuginea in Wistar rats corresponding to PD. In this model, MMF acts as a regenerating anti-fibrotic agent.

Topics: Animals; Disease Models, Animal; Fibrosis; Male; Mycophenolic Acid; Penile Erection; Penile Induration; Penis; Rats; Rats, Wistar; Transforming Growth Factor beta

Mycophenolate mofetil (MMF) is commonly prescribed and has proven advantages over other immunosuppressive drugs. However, frequent gastrointestinal side effects through an unknown mechanism limit its use. We have found that consumption of MMF alters the composition of the gut microbiota, selecting for bacteria expressing the enzyme β-glucuronidase (GUS) and leading to an up-regulation of GUS activity in the gut of mice and symptomatic humans. In the mouse, vancomycin eliminated GUS-expressing bacteria and prevented MMF-induced weight loss and colonic inflammation. Our work provides a mechanism for the toxicity associated with MMF and a future direction for the development of therapeutics.

Topics: Animals; Bacteria; Bacterial Proteins; Body Weight; Colitis; Disease Models, Animal; Female; Gastrointestinal Microbiome; Gastrointestinal Tract; Glucuronidase; Immunosuppressive Agents; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Up-Regulation; Vancomycin

The role of neuroinflammatory mediators has been well established in the pathogenesis of temporal lobe epilepsy (TLE) and associated neurobehavioral comorbidities. Mycophenolate mofetil (MMF) is commonly used as an immunosuppressant in organ transplantations. Its neuroprotective effect is well explored in different preclinical and clinical studies. The present study was designed to investigate the effect of MMF in rat model of lithium pilocarpine (LiPc)-induced spontaneous recurrent seizures and its associated neurobehavioral comorbidities. MMF treatment showed a dose-dependent decrease in seizure severity and reduced aggression in epileptic rats. There was marked improvement in spatial and recognition memory functions, along with substantial decrease in depression-like behavior in MMF treated epileptic rats. There was considerable decrease in mossy fiber sprouting in the dentate gyrus and the cornu ammonis 3 regions of the hippocampus, along with reduction in neuronal death in the treated groups. Furthermore, the hippocampal mRNA level of IL-1β, IL-2, PI3K, AKT, HIF-1α, RAPTOR, mTOR, Rps6kb1 and Rps6 was found to be decreased in MMF treated animals. mTOR, S6, pS6 and GFAP protein expression was decreased, whereas NeuN was increased in the rat hippocampus of the treated animals. The results concluded that MMF suppress recurrent seizures, and improves its associated behavioral impairments and cognitive deficit in rat model of TLE. The observed effects of MMF be correlated with the inhibition of IL-2 and IL-1β linked PI3K/AKT/mTOR signaling pathway hyperactivation.

Topics: Animals; Comorbidity; Disease Models, Animal; Epilepsy, Temporal Lobe; Hippocampus; Interleukin-1beta; Interleukin-2; Male; Mycophenolic Acid; Neurons; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; Seizures; Signal Transduction; Temporal Lobe; TOR Serine-Threonine Kinases

Tumor recurrence is a major complication following liver transplantation (LT) as treatment for hepatocellular carcinoma (HCC). Immunosuppression is an important risk factor for HCC recurrence, but conceivably may depend on the type of immunosuppressive medication. Mycophenolic acid (MPA) is a currently widely used immunosuppressant. This study investigated the effects of MPA on HCC.. Three human HCC cell lines and organoids from mouse primary liver tumor were used as experimental models. MTT, Alamar Blue assay, cell cycle analysis, colony formation, and [3H]-thymidine assays were performed. An LT database was used for retrospective analysis of the effect of mycophenolate mofetil, the prodrug of MPA, on HCC recurrence.. With clinically achievable concentrations, MPA effectively inhibited HCC cell proliferation and single-cell colony-forming unit. In short-term experiments, MPA effectively elicited S phase arrest in HCC cell lines. In addition, the initiation and growth of liver tumor organoids were effectively inhibited by MPA. Most importantly, the use of mycophenolate mofetil in patients with HCC-related LT was significantly associated with less tumor recurrence and improved patient survival.. MPA can specifically counteract HCC growth in vitro and tumor recurrence in LT patients. These results warrant prospective clinical trials into the role of MPA-mediated immunosuppression following LT of patients with HCC.

Topics: Adult; Aged; Animals; Carcinoma, Hepatocellular; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Immunosuppressive Agents; Kaplan-Meier Estimate; Liver Neoplasms; Liver Transplantation; Male; Mice; Middle Aged; Mycophenolic Acid; Neoplasm Recurrence, Local; Postoperative Complications; Primary Cell Culture; Prospective Studies; Retrospective Studies; Treatment Outcome; Young Adult

A recent clinical study showed that combination therapy consisting of mycophenolate mofetil, tacrolimus and steroids was shown to be more effective in achieving complete remission in patients with severe forms of lupus nephritis than conventional therapy consisting of intravenous cyclophosphamide and steroids. To explore the underlying molecular and cellular mechanisms of increased efficacy of the combination therapy regimen, we employed a mouse model of lupus nephritis, MRL/lpr mice, and treated them with monotherapies of prednisone, mycophenolate mofetil, or tacrolimus, or with their combination. Consistent with previous clinical findings, combination therapy markedly improved renal outcome compared to the monotherapies in mice with lupus nephritis. Transcriptomic analysis of their kidneys revealed distinct molecular pathways that were differentially regulated in combination therapy versus monotherapies. Combination therapy not only provided additive immunosuppressive effects, but also induced gene expression and molecular pathways to confer enhanced renoprotection. Specifically, combination therapy inhibited TLR7 expression in the kidneys of mice with lupus nephritis; combination of tacrolimus and mycophenolate mofetil led to better stabilization of the podocyte actin cytoskeleton through the reciprocal regulation of RhoA and Rac1 activities. Combination therapy strongly suppressed the IL-6/Stat3 pathway. These findings were further validated in renal biopsy samples from patients with lupus nephritis before and after treatments with mycophenolate mofetil, tacrolimus or combination therapy. Thus, our study further supports the earlier clinical finding and further provides insights into the molecular basis for increased efficacy of combination therapy.

Topics: Animals; Cytoskeleton; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Female; Gene Expression Profiling; Gene Expression Regulation; Glucocorticoids; Humans; Immunosuppressive Agents; Kidney; Lupus Nephritis; Mice, Inbred MRL lpr; Mycophenolic Acid; Podocytes; Prednisone; Signal Transduction; Tacrolimus; Transcriptome

To design colon-targeted codrugs of mycophenolic acid (MPA) and aminosugars as a safer option to mycophenolate mofetil (MMF) in the management of inflammatory bowel disease.. Codrugs were synthesized by coupling MPA with aminosugars (D-glucosamine and D-galactosamine) using EDCI coupling. The structures were confirmed by infrared radiation, nuclear magnetic resonance, mass spectroscopy and elemental analysis. The release profile of codrugs was extensively studied in aqueous buffers, upper gastrointestinal homogenates, faecal matter and caecal homogenates (. The prodrugs resisted activation in HCl buffer (pH 1.2) and stomach homogenates of rats with negligible hydrolysis in phosphate buffer (pH 7.4) and intestinal homogenates. Incubation with colon homogenates (. The significant attenuating effect of prodrugs and individual aminosugars on colonic inflammation proved that the rationale of the codrug approach is valid.

Topics: Animals; Colitis, Ulcerative; Colon; Disease Models, Animal; Drug Liberation; Drug Synergism; Galactosamine; Gastric Mucosa; Glucosamine; Humans; Immunosuppressive Agents; Male; Mycophenolic Acid; Prodrugs; Rats; Rats, Wistar; Severity of Illness Index; Treatment Outcome; Trinitrobenzenesulfonic Acid

CXC motif chemokine receptor 4 (CXCR4) blockade is pursued as an alternative to mesenchymal stem cell treatment in transplantation based on our previous report that burixafor, through CXCR4 antagonism, mobilizes immunomodulatory mesenchymal stem cells. Here, we explored the efficacy of combining mycophenolate mofetil (MMF)-based immunosuppressants with repetitive burixafor administration.. Swine heterotopic cardiac allograft recipients received MMF and corticosteroids (control, n = 10) combined with burixafor as a 2-dose (burixafor2D, n = 7) or 2-dose plus booster injections (burixafor2D + B, n = 5) regimen. The efficacy endpoints were graft survival, freedom from first acute rejection, and the severity of intimal hyperplasia. Each specimen was sacrificed either at its first graft arrest or after 150 days.. After 150 days, all specimens in the control group had died, but 28.5% of the burixafor2D group survived, and 60% of the burixafor2D + B group survived (P = 0.0088). Although the control group demonstrated acute rejection at a median of 33.5 days, the burixafor2D + B group survived without acute rejection for a median of 136 days (P = 0.0209). Burixafor administration significantly attenuated the incidence rate of acute rejection (P = 0.002) and the severity of intimal hyperplasia (P = 0.0097) at end point relative to the controls. These findings were associated with reduced cell infiltrates in the allografts, and modulation of C-reactive protein profiles in the circulation.. The augmentation of conventional MMF plus corticosteroids with a CXCR4 antagonist is potentially effective in improving outcomes after heart transplantation in minipigs. Future studies are warranted into optimizing the therapeutic regimens for humans.

Topics: Acute Disease; Allografts; Animals; Coronary Artery Disease; Disease Models, Animal; Drug Therapy, Combination; Female; Graft Rejection; Graft Survival; Heart Transplantation; Immunosuppressive Agents; Male; Mycophenolic Acid; Receptors, CXCR4; Swine; Swine, Miniature; Time Factors

Mycophenolate mofetil (MMF) is commonly prescribed after transplantation and has major advantages over other immunosuppressive drugs, but frequent gastrointestinal (GI) side-effects limit its use. The mechanism(s) underlying MMF-related GI toxicity have yet to be elucidated.. To investigate MMF-related GI toxicity, experimental mice were fed chow containing MMF (0.563%) and multiple indices of toxicity, including weight loss and colonic inflammation, were measured. Changes in intestinal microbial composition were detected using 16S rRNA Illumina sequencing, and downstream PICRUSt analysis was used to predict metagenomic pathways involved. Germ-free (GF) mice and mice treated with orally administered broad-spectrum antibiotics (ABX) were utilized to interrogate the importance of the microbiota in MMF-induced GI toxicity.. Mice treated with MMF exhibited significant weight loss, related to loss of body fat and muscle, and marked colonic inflammation. MMF exposure was associated with changes in gut microbial composition, as demonstrated by a loss of overall diversity, expansion of Proteobacteria (specifically Escherichia/Shigella), and enrichment of genes involved in lipopolysaccharide (LPS) biosynthesis, which paralleled increased levels of LPS in the feces and serum. MMF-related GI toxicity was dependent on the intestinal microbiota, as MMF did not induce weight loss or colonic inflammation in GF mice. Furthermore, ABX prevented and reversed MMF-induced weight loss and colonic inflammation.. An intact intestinal microbiota is required to initiate and sustain the GI toxicity of MMF. MMF treatment causes dynamic changes in the composition of the intestinal microbiota that may be a targetable driver of the GI side-effects of MMF.

Topics: Animals; Colon; Disease Models, Animal; Gastrointestinal Tract; Germ-Free Life; High-Throughput Nucleotide Sequencing; Humans; Immunosuppressive Agents; Male; Mice; Mice, Inbred Strains; Microbiota; Mycophenolic Acid; Proteobacteria; RNA, Ribosomal, 16S; Sequence Analysis, RNA; Weight Loss

Purpose To investigate the effect of CellCept nanoliposomes on Adriamycin-induced nephrotic syndrome in rats. Methods To model nephrotic syndrome, rats were injected with 6.5 mg/kg of Adriamycin in the tail vein. The rats were randomly divided into three groups, including a control group, a free mycophenolate mofetil (MMF)-treated group, and a liposome-encapsulated MMF-treated group. Five weeks after the Adriamycin treatment, the free MMF-treated group received CellCept while the liposome-encapsulated MMF-treated group received the CellCept nanoliposomes for 2 weeks. The general condition of the animals was observed, which included urine volume over 24 h, urine protein levels, and serum biochemical indexes. Renal morphology was also observed. Results The level of urine protein over 24 h was increased in the control group, while plasma albumin (ALB) was decreased. The total cholesterol (TC) and triacylglycerol (TG) levels increased significantly (P < 0.05, P < 0.01). The pathological examination of the kidneys showed some abnormalities. In contrast, these parameters were improved significantly in the free mycophenolate mofetil (MMF)-treated and liposome-contained mycophenolate mofetil (MMF)-treated groups. Conclusion The CellCept nanoliposomes have a good therapeutic effect on Adriamycin-induced nephrotic syndrome in rats.

Topics: Animals; Disease Models, Animal; Doxorubicin; Liposomes; Mycophenolic Acid; Nanoparticles; Nephrotic Syndrome; Rats

Ischaemia/reperfusion (I/R) is one of the main causes of acute kidney injury (AKI), which is characterized by sterile inflammation and oxidative stress. Immune cell activation can provoke overproduction of inflammatory mediators and reactive oxygen species (ROS), leading to perturbation of the microcirculation and tissue oxygenation associated with local and remote tissue injury. This study investigated whether the clinically employed immunosuppressant mycophenolate mofetil (MMF) was able to reduce I/R-induced renal oxygenation defects and oxidative stress by preventing sterile inflammation. Rats were divided into three groups (n=6/group): (1) a sham-operated control group; (2) a group subjected to renal I/R alone (I/R); and (3) a group subjected to I/R and MMF treatment (20 mg/kg prior to I/R) (I/R+MMF). Ischaemia was induced by a vascular occluder placed on the abdominal aorta for 30 minutes, followed by 120 minutes of reperfusion. Renal I/R deteriorated renal oxygenation (P<.001) and oxygen delivery (P<.01), reduced creatinine clearance (P<.01) and tubular sodium reabsorption (P<.001), and increased iNOS, renal tissue injury markers (P<.001), and IL-6 (P<.001). Oral MMF administration prior to insult restored renal cortical oxygenation (P<.05) and iNOS, renal injury markers, and inflammation parameters (P<.001) to near-baseline levels without affecting renal function. MMF exerted a prophylactic effect on renal microvascular oxygenation and abrogated tissue inflammation and renal injury following lower body I/R-induced AKI. These findings may have clinical implications during major vascular or renal transplant surgery.

Topics: Acute Kidney Injury; Animals; Aorta, Abdominal; Disease Models, Animal; Hemodynamics; Immunosuppressive Agents; Ischemia; Kidney; Male; Microcirculation; Microvessels; Mycophenolic Acid; Oxygen Consumption; Rats, Wistar; Renal Circulation; Reperfusion Injury

This study evaluated the effect of mycophenolate mofetil (MMF) on uterine tissue preservation following ischaemia/reperfusion (I/R) injury. Uterine I/R injury was induced in rats by clamping the lower abdominal aorta and ovarian arteries for 30 min. Group I/R + V (n = 7) received vehicle alone while Group I/R + M (n = 7) received 20 mg/kg/day MMF. Control groups underwent sham surgery and received vehicle (Group C) or 20 mg/kg/day MMF (Group M) (n = 7 for both). Four hours after detorsion, uterine tissue 8-hydroxy-2'-deoxyguanosine (8-OHdG), glutathione, malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and serum ischaemia modified albumin (IMA) concentrations were measured. Histopathological analyses were performed. The I/R + M group showed significant reduction in serum IMA and uterine tissue 8-OHdG, MDA and MPO and significant increase in SOD concentrations compared with the I/R + V group, indicating a protective effect against I/R oxidative damage (P = 0.009, P = 0.006, P = 0.002, P = 0.003 and P = 0.009, respectively). Histopathological evaluation revealed MMF treatment resulted in significantly less tissue and cellular damage and apoptosis compared with the I/R + V group. These results indicate MMF is effective in attenuating uterine tissue damage and preventing apoptosis following uterine I/R injury, probably via anti-inflammatory and anti-oxidative action.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Albumins; Animals; Antibiotics, Antineoplastic; Antioxidants; Aorta, Abdominal; Arteries; Deoxyguanosine; Disease Models, Animal; Female; Glutathione; Immunosuppressive Agents; Mycophenolic Acid; Ovary; Peroxidase; Rats; Rats, Wistar; Reperfusion Injury; Superoxide Dismutase; Uterus

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that predominantly affects women and is associated with prevalent hypertension, renal injury, and cardiovascular disease. Immune system dysfunction is recognized as an important factor in the pathogenesis of hypertension. We recently showed that preventing autoimmunity prevents the development of hypertension in an experimental model of SLE (female NZBWF1 mice). The present study tests the hypothesis that mycophenolate mofetil (MMF), an immunosuppressive therapy used clinically to treat SLE by depleting proliferating B and T lymphocytes, can improve blood pressure control.. Female SLE and control (NZW/LacJ) mice were treated daily for 8 weeks with 60 mg/kg MMF. Circulating CD45R. These data suggest that MMF selectively depleted CD45R

Topics: Albuminuria; Animals; Antibodies, Antinuclear; Arterial Pressure; B-Lymphocytes; Blood Pressure; Diabetic Nephropathies; Disease Models, Animal; DNA; Female; Hypertension; Immunoglobulin G; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Mycophenolic Acid; T-Lymphocytes

The transplant vasculopathy as a sign of chronic graft rejection affects both the epicardial and the intramyocardial arteries of the graft. This is at least partially mediated by NO synthases. The aim of this study was to assess possible protective effects of cyclosporine A (CsA), tacrolimus (FK506), and mycophenolate mofetil (MMF) on the expression of NO synthases in an experimental transplant rat model.. Heart transplantation was performed in 322 rats. These were randomly assigned to four equal groups (control, CsA, FK506, MMF). Recipients were monitored up to 60 days after transplantation, while transplanted hearts were recovered at certain time points for analysis. Expression and staining intensity for endothelial nitric oxide synthases (e-nos) and inducible nitric oxide synthases (i-nos) were analyzed in epicardial and intramyocardial vessels in each group.. All employed drugs led to a significant reduction of expression or staining intensity of i-nos and e-nos. MMF was most effective in reduction in expression of both NO synthases.. These results imply that all described drugs prevent endothelial impairment induced by toxicity of NO and thereby prevent transplant vasculopathy. MMF seems to be the most effective drug.

Topics: Allografts; Animals; Coronary Artery Disease; Coronary Vessels; Cyclosporine; Disease Models, Animal; Down-Regulation; Graft Rejection; Graft Survival; Heart Transplantation; Immunosuppressive Agents; Mycophenolic Acid; Nitric Oxide; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Rats, Inbred Lew; Tacrolimus; Time Factors

Hyperammonemia syndrome is an often fatal complication of lung transplantation which has been recently associated with Ureaplasma infection. It has not been definitely established that Ureaplasma species can cause hyperammonemia. We established a novel immunocompromised murine model of Ureaplasma urealyticum infection and used it to confirm that U. urealyticum can cause hyperammonemia. Male C3H mice were pharmacologically immunosuppressed with mycophenolate mofetil, tacrolimus and oral prednisone for seven days, and then challenged intratracheally (IT) and/or intraperitoneally (IP) with 107 CFU U. urealyticum over six days, while continuing immunosuppression. Spent U. urealyticum-free U9 broth was used as a negative control, with uninfected immunocompetent mice, uninfected immunosuppressed mice, and infected immunocompetent mice serving as additional controls. Plasma ammonia concentrations were compared using Wilcoxon ranks sum tests. Plasma ammonia concentrations of immunosuppressed mice challenged IT/IP with spent U9 broth (n = 14) (range 155-330 μmol/L) were similar to those of normal mice (n = 5), uninfected immunosuppressed mice (n = 5), and U. urealyticum IT/IP challenged immunocompetent mice (n = 5) [range 99-340 μmol/L, p = 0.60]. However, immunosuppressed mice challenged with U. urealyticum IT/IP (n = 20) or IP (n = 15) had higher plasma ammonia concentrations (range 225-945 μmol/L and 276-687 μmol/L, respectively) than those challenged IT/IP with spent U9 broth (p<0.001). U. urealyticum administered IT/IP or IP causes hyperammonemia in mice pharmacologically immunosuppressed with a regimen similar to that administered to lung transplant recipients.

Topics: Ammonia; Animals; Disease Models, Animal; Hyperammonemia; Immunocompromised Host; Immunosuppressive Agents; Male; Mice; Mice, Inbred C3H; Mycophenolic Acid; Prednisone; Real-Time Polymerase Chain Reaction; Tacrolimus; Ureaplasma Infections; Ureaplasma urealyticum

Mycophenolic acid (MPA), the active metabolite of mycophenolate mofetil, has anti-inflammatory effects, and is widely used as an immunomodulatory agent. However, the beneficial effect of MPA in hair-loss disorders is not fully understood.. To investigate the direct effect of MPA on dermal papilla cells (DPCs), and to examine the hair growth-stimulating effects of MPA topically applied to mouse skin.. Cultured DPCs were treated with various concentrations of MPA and analysed by MTT assay. Expressions of hair growth-related genes, including Wnt/β-catenin pathway-related genes and cellular apoptosis-regulating genes, such as Bcl-2, Bax and caspase-9, were examined using reverse transcription (RT)-PCR and western blotting. The Wnt/extracellular signal-regulated kinase (ERK) pathway was analysed by western blotting. The effect of topically applied MPA on anagen hair follicle induction after microneedle (MN) treatment with or without minoxidil (MXD) was evaluated by histopathological examination and RT-PCR.. MPA showed a promoting effect on DPC proliferation, which was associated with increased Axin2 transcription levels. In addition, phospho-ERK protein was detected in the MPA-treated DPCs. An increased Bcl-2/Bax transcript ratio contributed to cellular proliferation, and this was maintained in the MPA-treated environment. Topically applied MPA promoted anagen hair follicle induction in mice. The effect of MPA on hair follicles was compatible with that of MXD, and this effect was accelerated by MN treatment.. MPA promotes proliferation of DPCs and induction of anagen hair follicles in mice. This finding raises the possibility that MPA could be used as a treatment option for hair-loss disorders.

Topics: Alopecia; Animals; Apoptosis; Blotting, Western; Cell Proliferation; Cells, Cultured; Dermis; Disease Models, Animal; Female; Gene Expression Regulation; Growth Substances; Hair Follicle; Immunosuppressive Agents; Mice; Mice, Inbred C3H; Mitogen-Activated Protein Kinases; Mycophenolic Acid; Proto-Oncogene Proteins; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Transforming Growth Factor beta; Wnt Signaling Pathway

To analyse the effects of mycophenolate mofetil (MMF) on pulmonary artery pressure (PAP) and the expression of cytokines and their receptors in a rat model of pulmonary arterial hypertension (PAH).. Thirty-eight healthy male inbred Sprague-Dawley rats were divided randomly into four groups: a control group, a monocrotaline (MCT) group, an MMF20 group (MCT+20 mg/kg/day MMF), and an MMF40 group (MCT+40 mg/kg/day MMF). Systolic PAP (SPAP), the right ventricular hypertrophy index (RVI), and the levels of expression of cytokines and their receptors were measured and analysed.. SPAP, RVI, levels of expression of basic fibroblast growth factor (bFGF) in serum and lung homogenates, alveolar arterial wall thickness, and the number of muscular arteries in the MMF20 and MMF40 groups were decreased in comparison with the MCT group.. MMF inhibits the formation of vascular muscle and decreases SPAP and RVI by inhibition of the expression of bFGF, endothelin-1 (ET-1), and their receptors, resulting in the inhibition of smooth muscle proliferation and amelioration of PAH.

Topics: Animals; Blood Pressure; Cell Proliferation; Cytokines; Disease Models, Animal; Endothelin-1; Fibroblast Growth Factor 2; Hypertension, Pulmonary; Lung; Male; Monocrotaline; Muscle, Smooth, Vascular; Mycophenolic Acid; Rats; Rats, Sprague-Dawley; Receptor, Endothelin A; Receptors, Cytokine; Receptors, Fibroblast Growth Factor

To evaluate whether xenografts derived from hysterectomized patients would implant successfully and lead to uterine leiomyoma in Wistar rats.. This experimental study examined six female Wistar rats implanted with uterine leiomyoma obtained from patients who underwent hysterectomies at the gynecological surgery service of the HUUFMA. The rats were divided into two groups. Group I consisted of three rats in which the uterine leiomyoma had been implanted in the parietal peritoneum, and group II consisted of three rats in which the uterine leiomyoma was implanted in the subcutaneous tissue. The immunosuppressant mycophenolate mofetil (MMF) was administered orally by gavage (at a dose of 40 mg/kg of body weight) to prevent transplant rejection starting 15 days before the transplant and continuing throughout the entire experiment. After four weeks, necrosis and neovascularization were evaluated histologically in both groups and were classified as either absent or present. Lymphocytic inflammatory infiltration was also examined and classified as mild, moderate or intense (by hematoxylin and eosin staining), and fibrosis was classified as grade I-III (by Masson's trichrome staining).. Necrosis was absent from all three rats in group I and was observed in only one rat from group II. Neovascularization was present in two rats from group I and in only one rat from group II. The lymphocytic inflammatory infiltrate was mild in two rats and moderate in one rat from group I, and it was moderate in two rats and intense in one rat from group II. Two rats from group 1 exhibited grade III fibrosis, and one rat presented grade I fibrosis. In group II, two rats presented grade I fibrosis and one rat had grade II fibrosis. When necrosis and neovascularization were evaluated as variables, group I demonstrated greater evidence of successful implantation when compared to group II, indicating that the peritoneal implantation technique produces better results than the subcutaneous approach (p=0.039).. This study demonstrates that the xenotransplantation of uterine leiomyoma into the parietal peritoneum is more effective than the xenotransplantation of uterine leiomyoma into the subcutaneous tissue, and it describes a promising new model for the study of leiomyoma in vivo.

Topics: Animals; Disease Models, Animal; Female; Fibrosis; Graft Rejection; Humans; Immunosuppressive Agents; Leiomyoma; Mycophenolic Acid; Necrosis; Neoplasm Transplantation; Neovascularization, Pathologic; Peritoneum; Pilot Projects; Rats; Rats, Wistar; Subcutaneous Tissue; Transplantation, Heterologous; Uterine Neoplasms

Progression to chronic renal failure varies between patients with lupus nephritis. We compared the effects of mycophenolate mofetil (MMF) and cyclophosphamide (CTX), on renal histology and cellular pathways of fibrosis in murine lupus nephritis. Female NZBWF1/J mice were randomized to treatment with vehicle, methylprednisolone (MP) alone, MMF + MP or CTX + MP for up to 12 weeks, and the effects on clinical parameters, renal histology, and fibrotic processes were investigated. Treatment with MMF + MP or CTX + MP both improved survival, renal function, and decreased anti-dsDNA antibody level and immune complex deposition in kidneys of mice with active nephritis. Vehicle-treated mice showed progressive increase in mesangial proliferation, inflammatory cell infiltration and renal tubular atrophy, associated with PKC-α activation, increased TGF-β1 expression and increased matrix protein deposition. MP treatment alone did not have any significant effect. MMF + MP or CTX + MP treatment for 12 weeks reduced these abnormalities. MMF + MP was more effective than CTX + MP in suppressing fibrotic mediators, histological fibrosis score and expression of TGF-β1, fibronectin and collagen I in the kidney. Results from in vitro experiments on human mesangial cells (HMC) showed that mycophenolic acid (MPA) was more effective than CTX in suppressing PKC-α activation and TGF-β1 secretion induced by human polyclonal anti-dsDNA antibodies. While both MPA and CTX decreased TGF-β1- and TNF-α-induced fibronectin synthesis, only MPA decreased IL-6 induced fibronectin synthesis. MPA and CTX show distinct effects on fibrotic and inflammatory processes in NZBWF1/J murine lupus nephritis, suggesting that MMF + MP may be more effective than CTX + MP in preserving normal renal histology in lupus nephritis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Proliferation; Cyclophosphamide; Disease Models, Animal; Disease Progression; Female; Fibronectins; Fibrosis; Gene Expression Regulation; Humans; Immunosuppressive Agents; Interleukin-6; Kidney; Kidney Function Tests; Lupus Nephritis; Male; Mesangial Cells; Methylprednisolone; Mice; Mycophenolic Acid; Protein Kinase C-alpha; Survival Analysis; Transforming Growth Factor beta1

Encapsulated peritoneal sclerosis (EPS) is a rare complication of long-term peritoneal dialysis usually associated with the inadequacy and early termination of dialysis modality. Adequate treatment of peritoneal fibrosis has not been achieved by medical intervention so far. Mycophenolate mofetil (MMF), which inhibits inosine monophosphate dehydrogenase reversibly and highly selectively, is the most widely used drug for maintenance immunosupression in renal transplantation. Recent studies have shown that MMF has also antifibrotic effects. In this study, we evaluated the effects of MMF on EPS model in rats based on antifibrotic effects.. Twenty-four Wistar albino rat have been randomly divided into four groups. Group I (control group) received isotonic saline intraperitoneally (i.p) 2 ml/day for (0-3rd weeks). Group II (chlorhexidine (CG) group) received CG 2 ml/day i.p. for (0-3rd weeks). Group III (chlorhexidine + MMF group) received CG (2 ml/day) i.p. for (0-3rd weeks) plus MMF 30 mg/kg/day peroral (4th-6th weeks). Group IV (resting group) received CG 2 ml/day) i.p. (0-3rd weeks) plus peritoneal resting without any treatment (4th-6th weeks) At the end of the sixth weeks, all of the rats were killed. All of the groups were analyzed in terms of peritoneal thickness, degree of inflammation, vasculopathy, neovascularization and fibrosis. Also, the parietal peritoneal tissue samples were evaluated for matrix metalloproteinase 2 (MMP-2) by using the immunohistochemical analysis.. When the CG group was compared with the MMF group, the medication resulted in a statistically significant reduction in peritoneal thickness, inflammation and fibrosis score (53.23 ± 16.24 vs. 17.22 ± 3.62, 1 ± 1.225 vs. 1 ± 0, 1.6 ± 0.548 vs. 0.2 ± 0.447, respectively, all p < 0.05). In the resting group, no beneficial effects on morphological abnormality of the peritoneum were observed as compared with MMF group. However, according to immunohistochemical analysis of the expression of MMP-2 on peritoneal samples, the highest expression of MMP-2 was observed in the MMF group.. MMF was effective for the treatment of encapsulating peritoneal fibrosis in our rat model. Most recently, MMF may be first choice for EPS due to antifibrotic effect.

Topics: Animals; Disease Models, Animal; Immunosuppressive Agents; IMP Dehydrogenase; Mycophenolic Acid; Peritoneal Fibrosis; Rats; Rats, Wistar; Treatment Outcome

The aim of this study was to elucidate functional and molecular effects of mycophenolic acid (MPA) on non-lymphatic, kidney epithelial cells treated with transforming growth factor (TGF). MPA effects were studied using HK2 cells incubated with EGF and TGF. The reversibility of these effects was verified using guanosine and 8-aminoguanosine. The following assays were applied: cell proliferation, viability, collagen matrix contraction, scratch wound closure, spindle index, FACS with anti-CD29 and anti-CD326, promoter demethylation of RAS protein activator like 1 (RASAL1), as well as gene expression of RASAL1, integrin 1ß (ITGB1) (CD29) and epithelial cell adhesion molecule (EpCam) (CD326). Cell proliferation was inhibited by increasing concentrations of MPA, whereas neither apoptosis nor cytotoxicity was detected. Stimulation with EGF and/or TGF led to a significant collagen matrix contraction that was successfully inhibited by MPA. In addition, scratch wound closure was inhibited by incubation with TGF alone or with EGF. Under the same conditions, cell morphology (spindle shape) and molecular phenotype (ITGB1(High)EpCam(Low)/ITGB1(Low)EpCam(High)) were both significantly changed, suggesting an epithelial to mesenchymal transformation. Cell morphology and motility, as well as molecular phenotype, were reversible after MPA treatment with TGF transformation in both presence/absence of EGF, thereby suggesting a correlation with the previously described antifibrotic effects of MPA. Dysregulation of TGF signal transduction appears to be related to progression of fibrosis. A TGF-transformed kidney epithelial cell line derived from human proximal tubules was used to study whether the immunosuppressive drug: MPA possesses any functional or molecular antifibrotic effects. Functional and morphological in vitro changes induced by both the TGF and epithelial-growth-factor were reversible by treatment with MPA. An inhibitory effect of MPA on the TGF pathway appears to be responsible for the previously described antifibrotic effects of the MPA in the COL4A3-deficient mouse model of renal fibrosis.

Topics: Animals; Autoantigens; Cell Line; Cell Movement; Cell Proliferation; Cell Survival; Collagen Type IV; Disease Models, Animal; Epidermal Growth Factor; Fibrosis; Humans; Kidney; Mice; Mycophenolic Acid; Signal Transduction; Transforming Growth Factor beta

Mycophenolate mofetil (MMF) is an alternative immunosuppressive agent that has been reported to be effective and well tolerated for the treatment of refractory inflammatory bowel disease (IBD). The aim of this study was to investigate the therapeutic effect of MMF on intestinal injury and tissue inflammation, which were caused by Crohn's disease (CD). Here, trinitrobenzene sulfonic acid-relapsing (TNBS) colitis was induced in mice; then, we measured the differentiation of Th1/Th2 cells in mouse splenocytes by flow cytometry and the secretion of cytokines in mice with TNBS-induced colitis by real-time polymerase chain reaction and/or enzyme-linked immunosorbent assay (RT-PCR/ELISA). The results show that MMF significantly inhibited mRNA expression of pro-inflammatory cytokines IFN-γ, TNF-α, IL-12, IL-6, and IL-1β in mice with TNBS-induced colitis; however, MMF did not inhibit the expression of IL-10 mRNA. Additionally, ELISA showed that the serum levels of IFN-γ, TNF-α, IL-12, IL-6, and IL-1β were down-regulated in a TNBS model of colitis. Flow cytometric analysis showed MMF markedly reduced the percentages of Th1 and Th2 splenocytes in the CD mouse model. Mycophenolic acid (MPA) also significantly decreased the percentages of splenic Th1 and Th2 cells in vitro. Furthermore, MMF treatment not only significantly ameliorated diarrhea, and loss of body weight but also abrogated the histopathologic severity and inflammatory response of inflammatory colitis, and increased the survival rate of TNBS-induced colitic mice. These results suggest that treatment with MMF may improve experimental colitis and induce inflammatory response remission of CD by down-regulation of pro-inflammatory cytokines via modulation of the differentiation of Th1/Th2 cells.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Differentiation; Colitis; Crohn Disease; Disease Models, Animal; Female; Gene Expression; Immunosuppressive Agents; Interferon-gamma; Interleukin-12; Interleukin-1beta; Interleukin-6; Mice; Mice, Inbred BALB C; Mycophenolic Acid; Spleen; Th1 Cells; Th1-Th2 Balance; Th2 Cells; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

This study aimed to examine the pharmacokinetics and pharmacodynamics of mycophenolic acid (MPA) in Nagase analbuminemic rats (NARs) to evaluate the effect of protein binding on the associated inosine-5'-monophosphate dehydrogenase (IMPDH) activity. Free fractions of MPA in the control rats and NARs were 2.09 and 24.8%, respectively. Pharmacokinetic and pharmacodynamic parameters simultaneously obtained by the nonlinear mixed effects modeling program NONMEM explained reasonably well the concentrations of MPA and MPA glucuronide as well as IMPDH activity in both rats. NARs showed a higher clearance and a smaller volume of distribution based on the free MPA concentration than the controls did, besides the increase in free fraction. The half-maximal inhibitory concentration based on free MPA was estimated as 163 ng/mL for both rats. Simulations based on the obtained pharmacokinetic and pharmacodynamic parameters showed that the area under the IMPDH activity-time curve decreased non-linearly according to the increase in free fraction of MPA. In conclusion, the experimental data obtained from NARs followed by the modeling and simulation approach quantitatively clarified that the free MPA concentration was suitable for the biomarker of immunosuppressive effect of MPA. Dose adjustments based on the total MPA may cause unnecessary overexposure to MPA in patients with hypoalbuminemia.

Topics: Acetylglucosaminidase; Animals; Computer Simulation; Disease Models, Animal; Drug Monitoring; Genetic Predisposition to Disease; Glucuronides; Hypoalbuminemia; Immunosuppressive Agents; IMP Dehydrogenase; Liver; Models, Biological; Mycophenolic Acid; Nonlinear Dynamics; Phenotype; Protein Binding; Rats, Transgenic

This study evaluated the clinical and pathological effects of the immunosuppressive agent mycophenolate mofetil (MMF) in rats with experimental autoimmune encephalomyelitis (EAE; a model of multiple sclerosis [MS]).. EAE rats were randomly divided into 4 groups: model alone (n = 7); low- or high-dose MMF (20 and 30 mg/kg per day, respectively, n = 6 each) orally for 14 days; methylprednisolone (20 mg/kg per day, n = 6) injected once daily for 3 days. Six normal Wistar rats served as controls. Clinical signs and histopathological findings were evaluated 14 days after treatment started.. Oral administration of high-dose MMF significantly ameliorated the course of EAE in rats: cumulative clinical scores were lower and weight loss was less than in rats receiving methylprednisolone. The ameliorated disease course was associated with alleviation of histopathological signs of EAE. Treatment increased the blood proportion of CD8(+), CD4(+)CD25(+) and CD4(+)CD45RA(+) T cells, with a concomitant reduced proportion of CD4(+) T cells and ratio of CD4(+) to CD8(+) T cells, compared with EAE model alone rats.. MMF may have pharmacological potential in MS treatment and these findings may help in understanding the pathophysiological mechanism of MS.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; CD4-CD8 Ratio; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Enzyme Inhibitors; Female; Guinea Pigs; Immunosuppressive Agents; IMP Dehydrogenase; Interleukin-2 Receptor alpha Subunit; Leukocyte Common Antigens; Lymphocyte Count; Methylprednisolone; Multiple Sclerosis; Mycophenolic Acid; Random Allocation; Rats; Rats, Wistar; T-Lymphocyte Subsets

Hepatocyte transplantation is proposed as a solution for liver failure. The allotransplantation of hepatocytes has been studied extensively, however, graft rejection remains a major problem. The aim of the present study was to evaluate the immunosuppressive activity of mycophenolate mofetil (MMF), sirolimus, and their combination in an experimental model of hepatocyte allotransplantation in rats with acute liver failure.. Five male Wistar rats were used as hepatocyte donors and 60 male Lewis rats as recipients. The recipients were divided into five groups of 12 animals each. Group 1: no treatment. Group 2: cyclosporine. Group 3: sirolimus. Group 4: MMF. Group 5: MMF and sirolimus. All surviving animals were preserved for 15 days. For the induction of acute liver failure the recipients were injected with N-dimethyl-nitrosamine 24 hr before transplantation. The isolated hepatocytes were transplanted intrasplenically.. Analysis of the results showed a statistically significant prolongation of animal survival for groups 3, 4, and 5. More animals in group 5 survived than those in groups 3 and 4, although the difference was not statistically significant. Transplant hepatocyte survival was significantly better in groups 3, 4, and 5. Hepatocytes transplanted in the spleen of animals of group 5 showed better survival compared with those of groups 3 and 4.. Use of MMF and sirolimus, as monotherapy or in combination, is both effective and safe as immunosuppressive treatment in hepatocyte transplantation, as was proven in this experimental protocol.

Topics: Animals; Disease Models, Animal; Graft Rejection; Hepatocytes; Immunosuppressive Agents; Liver Failure, Acute; Male; Mycophenolic Acid; Rats, Inbred Lew; Rats, Wistar; Sirolimus; Spleen

Mycophenolate mofetil (MMF) is an effective immunosuppressive agent that has been frequently used in laboratory animals including swine; however, the pharmacokinetic properties of MMF in swine have not been studied. This short-term study was designed to evaluate the feasibility and the pharmacokinetic profiles of MMF therapy in neonatal swine.. Twelve neonatal pigs were randomized into four groups including one control and three treated groups with oral MMF administered at 0.5, 1, and 2 g/m(2)/d for 4 days, divided by 2 half-doses at 9:00 and 17:00 (except day 4 during which MMF was not administered at 17:00). Blood samples were collected at 9:00 on days 0, 2, 3 and 4 for complete blood count and hepatic/renal function examination; the trough concentration of plasma mycophenolic acid (MPA) was also determined. On days 2 and 4, blood was collected to determine the area under the curve (AUC) of plasma MPA concentration. Animal body-weight growth and manifestations of MMF side-effects such as anorexia, vomiting, and diarrhea were also observed.. MMF has no acute hepatic/renal toxicity in newborn pigs; however, less body-weight growth was observed in treated groups. In the control group, a spontaneous increase of lymphocyte count was observed; in contrast, MMF therapy with doses of 1 and 2 g/m(2)/d reduced both lymphocyte and monocyte counts of piglets. Oral MMF had high bioavailability in neonatal swine. MPA-AUC0-12h of doses 0.5, 1, and 2 g/m(2)/d was 22.00 ± 3.32, 57.57 ± 34.30, and 140.00 ± 19.70 μg × h/mL, respectively. Neither MPA trough concentration (MPA-C0), nor MPA maximum concentration (MPA-Cmax) or MPA-AUC0-6h had high correlation with MMF-dose. For surveillance of MPA exposure, MPA-C0 had significant correlation with MPA-AUC0-12h (Spearman's ρ = 0.933, AUC0-12h = 17.882 × C0 + 14.479, r(2) = 0.966).. To reach adequate drug exposure and to reduce dose-dependent side effects, an MMF dose of 1 g/m(2)/d is recommended to be used as an initial dose for immunosuppressive therapy in piglets, and MPA-C0 monitoring is the most practical strategy for experimental transplantation study.

Topics: Animals; Animals, Newborn; Disease Models, Animal; Follow-Up Studies; Graft Rejection; Immunosuppressive Agents; Mycophenolic Acid; Organ Transplantation; Swine

In human, autoimmune uveitis is a leading cause of visual disability and ranks with diabetic retinopathy as a major source of blind registrations in developed countries. Since most cases of non-infectious uveitis are considered to be autoimmune or at least immune-mediated, the management of such patients has rested on appropriate immunosuppression. Some patients, however, despite maximal immunotherapy, fail to respond or are seriously intolerant of the drug therapies. Since its establishment 20 years ago, the model of experimental autoimmune uveoretinitis has served as a useful template for novel therapeutic approaches. The aim of our study was to compare the efficacy of mycophenolate mofetil and cyclophosphamide and golimumab treatment in the mouse model of experimental autoimmune uveitis. The intensity of intraocular inflammation was evaluated histologically in the treatment and control groups. Experimental autoimmune uveitis has been induced in mouse strain C57BL/6 by subcutaneous application of interphotoreceptor retinoid binding protein in complete Freund's adjuvant and pertussis toxin. The treatment was commenced on the day of uveitis induction. Cyclophosphamide was applied intraperitoneally in a single dose (100 mg/kg), mycophenolate mofetil intraperitoneally daily (30 mg/kg or 50 mg/kg), golimumab subcutaneously weekly (70 mg/kg). Sham intraperitoneal injection of a placebo (aqua pro injectione) and untreated mice with experimental autoimmune uveitis served as controls. The results show statistically significant suppression of experimental uveitis both with mycophenolate mofetil and with cyclophosphamide, and thus support its use in human medicine.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antibodies, Monoclonal; Cyclophosphamide; Disease Models, Animal; Inflammation; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Uveitis

Lupus nephritis (LN) is usually associated with widespread effacement of the podocytes' foot processes leading to proteinuria. Induction of urokinase receptor (uPAR) signaling in podocytes leads to foot process effacement and urinary protein loss via promoting podocytes' motility and kidney permeability in the glomerulus. Very little is known about uPAR signaling in LN. Mycophenolate mofetil (MMF), an immunosuppressive agent, efficiently modulates the development of LN in humans and mice, but there are no data concerning the direct uPAR involvement on podocytes in LN. The MMF efficiency and uPAR involvement signaling in NZB×NZW F1 lupus-prone mice were examined by proteinuria, renal function and pathology, immune complex deposits, and uPAR expression of podocytes by immunofluorescence staining and quantitative RT-PCR. After MMF treatment, the proteinuria (p < 0.01), BUN level (p < 0.05) and immunodeposition in glomeruli (p < 0.001) were significantly improved. Most important, the renal uPAR mRNA levels (p < 0.001) and uPAR protein level of podocytes (p < 0.001) were significantly reduced. The beneficial effect of MMF on LN could be attributed, at least in part, to the inhibition of uPAR expression in podocytes. These findings demonstrated uPAR could have potential as a predictive index for response to LN therapeutics.

Topics: Animals; Blood Urea Nitrogen; Disease Models, Animal; Female; Fluorescent Antibody Technique; Humans; Immunosuppressive Agents; Lupus Nephritis; Mice; Mice, Inbred NZB; Mycophenolic Acid; Podocytes; Proteinuria; Receptors, Urokinase Plasminogen Activator; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction

The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression.. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively.. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression.. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.

Topics: Animals; Biomarkers; Cardiotonic Agents; Cecum; Cyclosporine; Disease Models, Animal; Hemodynamics; Immunocompromised Host; Immunosuppressive Agents; Inflammation Mediators; Ligation; Methylprednisolone; Mycophenolic Acid; Norepinephrine; Punctures; Sepsis; Swine; Swine, Miniature; Time Factors

To investigate the efficacy of different doses of the soluble form of the receptor for advanced glycation end products (sRAGE) (conjugated to the Fc portion of immunoglobulin) in the treatment of nephritis in lupus-prone mice, in comparison with the efficacy of combination therapy with mycophenolate mofetil plus prednisolone.. Twenty-eight female (NZB/NZW)F1 mice were divided into 5 groups (untreated, sRAGE [dose groups of 0.5, 1, or 2 μg], or mycophenolate mofetil plus prednisolone). Proteinuria and histologic damage were evaluated. Immune complex deposition and the nuclear translocation of NF-κB in the kidney tissue were assessed by immunofluorescence staining. Serum concentrations of anti-double-stranded DNA (anti-dsDNA) and IgG subclasses were also measured. The population of T cells was evaluated using a fluorescence-activated cell sorter, and expression of intracellular adhesion molecule 1 and vascular cell adhesion molecule 1 in the kidney tissue was assessed by immunohistochemical staining.. In comparison with untreated mice, mice treated with 1 or 2 μg sRAGE showed significantly reduced proteinuria and attenuated histologic renal damage, with efficacy comparable to that of combination therapy. Treatment with 2 μg sRAGE significantly reduced immune complex deposition and decreased the serum concentrations of anti-dsDNA, IgG2a, IgG2b, and IgG3. In addition, sRAGE interrupted the nuclear translocation of NF-κB in the kidney, resulting in reduction in the expression of downstream genes of NF-κB in vivo and in vitro. Furthermore, sRAGE effectively modified T cell populations.. Treatment with sRAGE significantly improved nephritis in lupus-prone mice, with efficacy comparable to that of standard induction treatment for lupus nephritis. These data suggest that sRAGE has antiinflammatory effects on the pathophysiology of lupus nephritis and could serve as a potent new therapy for this disease.

Topics: Animals; Anti-Inflammatory Agents; Antigen-Antibody Complex; Disease Models, Animal; Drug Therapy, Combination; Female; Kidney; Lupus Nephritis; Mice; Mice, Inbred NZB; Mycophenolic Acid; NF-kappa B; Prednisolone; Proteinuria; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Treatment Outcome

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease that is characterized by autoantibody production and inflammatory disease involving multiple organs. Premature atherosclerosis is a common complication of SLE and results in substantial morbidity and mortality from cardiovascular disease (CVD). The reasons for the premature atherosclerosis in SLE are incompletely understood, although chronic inflammation is thought to play an important role. There is currently no known preventative treatment of premature atherosclerosis in SLE. Mycophenolate mofetil (MMF) is an immunosuppressive agent that is commonly used for treatment of patients with SLE. In order to study the impact of this drug on murine lupus disease including premature atherosclerosis development, we treated gld.apoE(-/-) mice, a model of SLE and accelerated atherosclerosis, with MMF. We maintained seven-week old gld.apoE(-/-) mice on a high cholesterol Western diet with or without MMF. After 12 weeks on diet, mice receiving MMF showed decreased atherosclerotic lesion area compared to the control group. MMF treatment also improved the lupus phenotype, indicated by a significant decrease circulating autoantibody levels and ameliorating lupus nephritis associated with this model. This data suggests that the effects of MMF on the immune system may not only be beneficial for lupus, but also for inflammation driving lupus-associated atherosclerosis.

Topics: Animals; Apolipoproteins E; Atherosclerosis; Autoantibodies; Body Weight; Disease Models, Animal; Disease Progression; Eating; Fas Ligand Protein; Humans; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Lymphatic Diseases; Mice; Mycophenolic Acid; Nephritis; Splenomegaly

Hirschsprung disease (HSCR) is a partially penetrant oligogenic birth defect that occurs when enteric nervous system (ENS) precursors fail to colonize the distal bowel during early pregnancy. Genetic defects underlie HSCR, but much of the variability in the occurrence and severity of the birth defect remain unexplained. We hypothesized that nongenetic factors might contribute to disease development. Here we found that mycophenolate, an inhibitor of de novo guanine nucleotide biosynthesis, and 8 other drugs identified in a zebrafish screen impaired ENS development. In mice, mycophenolate treatment selectively impaired ENS precursor proliferation, delayed precursor migration, and induced bowel aganglionosis. In 2 different mouse models of HSCR, addition of mycophenolate increased the penetrance and severity of Hirschsprung-like pathology. Mycophenolate treatment also reduced ENS precursor migration as well as lamellipodia formation, proliferation, and survival in cultured enteric neural crest–derived cells. Using X-inactivation mosaicism for the purine salvage gene Hprt, we found that reduced ENS precursor proliferation most likely causes mycophenolate-induced migration defects and aganglionosis. To the best of our knowledge, mycophenolate is the first medicine identified that causes major ENS malformations and Hirschsprung-like pathology in a mammalian model. These studies demonstrate a critical role for de novo guanine nucleotide biosynthesis in ENS development and suggest that some cases of HSCR may be preventable.

Topics: Animals; Cell Movement; Cell Proliferation; Cells, Cultured; Disease Models, Animal; DNA; Drug Evaluation, Preclinical; Enteric Nervous System; Female; Guanosine Monophosphate; Hirschsprung Disease; Humans; Male; Mice; Mice, 129 Strain; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Mycophenolic Acid; Pregnancy; Zebrafish

Is it possible to perform allogeneic uterus transplantation (UTx) with a donation from a live donor in a non-human primate species and what immunosuppression is needed to prevent rejection?. Allogeneic UTx in the baboon is a donor- and recipient-safe surgical procedure; immunosuppression with induction therapy and a triple protocol should be used.. UTx may become a treatment for absolute uterine factor infertility. Autologous UTx models have been developed in non-human primates with reports on long-term survival of the uterine grafts. STUDY DESIGN, SIZEAND DURATION: This experimental study included 18 female baboons as uterus donors and 18 female baboons as uterus recipients. The follow-up time was 5-8 weeks.. Uterus retrieval was performed with extended hysterectomy including bilateral uterine and internal iliac arteries and ovarian veins. After UTx, with vascular anastomoses unilateral to the internal iliac artery and the external iliac vein, the uterus recipients received one of the following: no immunosuppression (n = 4); monotherapy (oral slow release tacrolimus) (n = 4) or induction therapy (antithymocyte globulin) followed by triple therapy (tacrolimus, mycophenolate, corticosteroids; n = 10). Surgical parameters, survival, immunosuppression and rejection patterns were evaluated.. The durations of uterus retrieval and recipient surgery were around 3 and 3.5 h, respectively. The total ischemic time was around 3 h. All the recipients and the donors survived the surgery. All the recipients presented rejection to some extent within the first weeks following UTx. In one recipient, the uterus was of normal appearance at the end of the study period. In spite of occasional high (>60 ng/ml) blood levels of tacrolimus, there was no evidence of nephrotoxicity.. This initial non-human primate allogeneic UTx study indicates that further research is needed to optimize immunosuppression protocols in order to avoid uterine rejection.. The findings suggest that allogeneic UTx in primate species is feasible but continued work on this issue is needed.. The study was funded by the Swedish Research Council, ALF University of Gothenburg, Hjalmar Svensson Foundation and by Jane and Dan Olsson Research Foundation. The authors do not have any competing interest.

Topics: Adrenal Cortex Hormones; Animals; Antilymphocyte Serum; Disease Models, Animal; Drug Therapy, Combination; Feasibility Studies; Female; Graft Rejection; Graft Survival; Immunosuppression Therapy; Induction Chemotherapy; Infertility, Female; Living Donors; Maintenance Chemotherapy; Mycophenolic Acid; Papio; Tacrolimus; Transplantation, Homologous; Uterine Diseases; Uterus

Septic shock is a systemic inflammatory response syndrome, and it is the leading cause of death in intensive care units. Mycophenolate mofetil (MMF) is an immunosuppressant that has been shown to be effective in the treatment of various inflammatory diseases. In this study, the anti-inflammatory effect of MMF in a mouse model of acute lung injury (ALI) induced by lipopolysaccharide (LPS) was evaluated. ALI was induced by intrapleural injection of LPS (250 ng/cavity). The leukocyte migration, exudation, myeloperoxidase and adenosine deaminase activities, nitric oxide products, tumor necrosis factor alpha (TNF-α), and interleukin 1 beta (IL-1β) levels, as well as mRNA expression of TNF-α and IL-1β, were evaluated. This study showed that MMF significantly decreased all parameters studied in a manner comparable to treatment with dexamethasone. In conclusion, MMF has important anti-inflammatory effects that may be useful as an auxiliary treatment for septic shock.

Topics: Acute Lung Injury; Adenosine Deaminase; Animals; Anti-Inflammatory Agents; Cell Movement; Dexamethasone; Disease Models, Animal; Immunosuppressive Agents; Inflammation; Interleukin-1beta; Leukocytes; Lipopolysaccharides; Lung; Mice; Mycophenolic Acid; Nitric Oxide; Peroxidase; RNA, Messenger; Shock, Septic; Tumor Necrosis Factor-alpha

Recent clinical and preclinical studies have demonstrated that systemic lupus erythematosus (SLE) is associated with an increased risk for cardiovascular disease (CVD). However, unlike in the general population, little is known regarding the efficacy of atheroprotective interventions in patients with SLE. The current study aims to determine the benefit of lymphocyte inhibition on reducing the atherosclerotic burden in SLE-susceptible LDLr-deficient mice.. Female LDLr(-/-) mice were lethally irradiated and reconstituted with bone marrow from C57Bl/6 mice (LDLr.B6) or the SLE-susceptible B6.Sle1.2.3 mice (LDLr.Sle). At 16 weeks post transplant, mice were treated with atorvastatin (10 mg/kg), mycophenolate mofetil (MMF; 40 mg/kg), or both (MMF-A) for 8 weeks, after which the extent of atherosclerosis and the presence of SLE were assessed.. Following 8 weeks of treatment, we observed that atorvastatin-mediated reduction in cholesterol levels attenuated atherogenesis in LDLr.B6 mice but failed to significantly reduce atherosclerotic lesion size in LDLr.Sle mice, in spite of a significant reduction in serum cholesterol levels. Treatment with MMF and MMF-A attenuated atherogenesis in LDLr.B6 and LDLr.Sle mice. In addition, MMF-containing regimens inhibited recruitment of CD4+ T cells to atherosclerotic lesions in LDLr.Sle mice. In these mice, MMF also reduced the proportion of activated splenic T cells, as well as interleukin 10 secretion by T cells. With regard to lupus activity, MMF had no overt effect on anti-double-stranded DNA (dsDNA) antibody titres or kidney function and pathology.. The current study demonstrates that reduction of cholesterol levels alone is not atheroprotective in lupus-mediated atherogenesis. This is the first study to demonstrate that MMF reduces the atherosclerotic burden in a model of lupus-accelerated atherosclerosis. Our results suggest that MMF treatment may prove beneficial in preventing CVD in patients with SLE.

Topics: Animals; Atherosclerosis; Atorvastatin; CD4-Positive T-Lymphocytes; Cholesterol; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Therapy, Combination; Female; Genetic Predisposition to Disease; Heptanoic Acids; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Immunity, Cellular; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Mice; Mice, Inbred C57BL; Mycophenolic Acid; Pyrroles

Highly abbreviated micro-dystrophin genes have been intensively studied for Duchenne muscular dystrophy (DMD) gene therapy. Following adeno-associated virus (AAV) gene transfer, robust microgene expression is achieved in murine DMD models in the absence of immune suppression. Interestingly, a recent study suggests that AAV gene transfer in dystrophic dogs may require up to 18 weeks' immune suppression using a combination of three different immune-suppressive drugs (cyclosporine, mycophenolate mofetil, and anti-dog thymocyte globulin). Continued immune suppression is not only costly but also may cause untoward reactions. Further, some of the drugs (such as anti-dog thymocyte globulin) are not readily available. To overcome these limitations, we developed a novel 5-week immune suppression scheme using only cyclosporine and mycophenolate mofetil. AAV vectors (either AV.RSV.AP that expresses the heat-resistant human alkaline phosphatase gene, or AV.CMV.μDys that expresses the canine R16-17/H3/ΔC microgene) at 2.85×10(12) vg particles were injected into adult dystrophic dog limb muscles under the new immune suppression protocol. Sustained transduction was observed for nearly half year (the end of the study). The simplified immune suppression strategy described here may facilitate preclinical studies in the dog model.

Topics: Animals; Antilymphocyte Serum; Cyclosporine; Dependovirus; Disease Models, Animal; Dogs; Dystrophin; Female; Gene Expression; Genetic Therapy; Genetic Vectors; Humans; Immunosuppression Therapy; Immunosuppressive Agents; Injections, Intramuscular; Male; Mice; Muscle, Skeletal; Muscular Dystrophy, Duchenne; Mycophenolic Acid; T-Lymphocytes; Transgenes

T lymphocytes induce the transformation of fibroblasts into myofibroblasts, the main mediators of fibrogenesis. The inosine 5'-monophosphate dehydrogenase inhibitor mycophenolate mofetil (MMF) and the anti-CD25 monoclonal antibody daclizumab (DCZ) have been reported to suppress the proliferation of T lymphocytes.. To evaluate the preventive effects of MMF and DCZ in early stages of bleomycin (BLM)-induced scleroderma.. This study involved five groups of Balb/c mice (n = 10 per group). Mice in four of the groups were injected subcutaneously (SC) with BLM [100 μg/day in 100 μL phosphate-buffered saline (PBS)] for 4 weeks; the remaining (control) group received only 100 μL PBS. Three of the BLM-treated groups also received either intraperitoneal MMF 50 or 150 mg/kg/day, or SC DCZ 100 μg/week. At the end of the fourth week, all mice were killed, and blood and tissue samples were obtained for further analysis.. In the BLM-treated group, increases were seen in inflammatory-cell infiltration, α-smooth muscle actin-positive (α-SMA+) fibroblastic cell count, tissue hydroxyproline content, and dermal thickness. Dermal fibrosis was histopathologically prominent. In BLM-treated mice also given MMF or DCZ, inflammatory-cell infiltration, tissue hydroxyproline content and dermal thickness were decreased. In the MMF groups, decreases were also noted in α-SMA+ fibroblastic cell count.. In this BLM-induced dermal fibrosis model, MMF and DCZ treatments prevented the development of dermal fibrosis. Further studies are needed to evaluate whether targeting T lymphocytes is effective in resolving pre-existing fibrosis in human scleroderma.

Topics: Animals; Antibiotics, Antineoplastic; Antibodies, Monoclonal, Humanized; Bleomycin; Cytokines; Daclizumab; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hydroxyproline; Immunoglobulin G; Immunohistochemistry; Immunosuppressive Agents; Injections, Subcutaneous; Mice; Mice, Inbred BALB C; Mycophenolic Acid; Scleroderma, Localized; Skin

Encapsulated peritoneal sclerosis (EPS) is a devastating complication of peritoneal dialysis. We aimed to investigate the effects of mycophenolate mofetil (MMF) treatment in experimental EPS in rats.. 40 nonuremic Wistar albino rats were divided equally into 4 groups: control rats received 2 ml isotonic saline intraperitoneally daily for 3 weeks without any other treatment. The chlorhexidine gluconate group received intraperitoneally 2 ml/200 g injection of chlorhexidine gluconate and ethanol dissolved in saline for 3 weeks. The resting group received chlorhexidine gluconate (0 - 3rd week) + peritoneal resting (4th - 6th week). The MMF group received chlorhexidine gluconate (0 - 3rd week) + 125 mg/l MMF in drinking water (4th - 6th week). Dialysate cytokine levels, leukocyte count, peritoneal thickness, inflammation and fibroblast activities were evaluated.. Although the MMF and resting groups showed beneficial effects on ultrafiltration and D1/D0 glucose compared to the chlorhexidine gluconate group, only MMF treatment improved dialysate TGFβ1, VEGF and MCP-1 levels compared to the resting group. Inflammatory activity and vascularity observed in a tissue biopsy, including capillaries number per mm2 of submesothelial area, decreased in the treatment group.. MMF treatment has beneficial effects on EPS via inhibiting inflammation and neovascularisation by reducing dialysate VEGF overexpression.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Chemokine CCL2; Disease Models, Animal; Gene Expression; Male; Mycophenolic Acid; Neovascularization, Pathologic; Peritoneal Dialysis; Peritoneal Fibrosis; Peritoneum; Rats; Rats, Wistar; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

Immunosuppressive (IS) medication is needed to avoid graft rejection in porcine transplantation models. An ideal IS therapy should have no side-effects, but increased susceptibility to infections, disturbed intestinal microflora and toxic effects on organs and tissues are commonly reported. The aim of the present study was to design an IS protocol with tacrolimus and mycophenolic acid to be used for maintenance therapy in the post-transplant period. An eligible whole blood trough value for tacrolimus was 5-15 μg/L. Conventional specific pathogen-free pigs were fitted with an indwelling catheter under general anaesthesia, and after the acclimatization period three groups were formed: group A (n= 4) received 0.15 mg/kg body weight (BW) twice daily tacrolimus and 500 mg twice daily mycophenolic acid; group B (n= 4) received 0.3 mg/kg BW twice daily tacrolimus and 500 mg twice daily mycophenolic acid; group C (n= 2) did not receive any medication. Daily clinical examinations and analyses of blood concentrations of tacrolimus and glucose were performed. Total and differential white blood cell counts, enzyme activities, bilirubin and electrolyte concentrations were measured every fourth day. At the end of the experiment, the pigs were killed with an overdose of pentobarbital intravenously and a necropsy was performed immediately. All animals seemed to tolerate the IS treatment well. No alterations in their clinical state of health were observed throughout the study and daily weight gain was similar for the three groups. The necropsy did not reveal any pathological findings related to medication. The study showed that 0.25 mg/kg BW twice daily tacrolimus and 500 mg twice daily mycophenolic acid would be an appropriate maintenance dosage for conventional pigs.

Topics: Administration, Oral; Animals; Clinical Protocols; Disease Models, Animal; Drug Administration Schedule; Graft Rejection; Immunosuppressive Agents; Islets of Langerhans Transplantation; Mycophenolic Acid; Postoperative Complications; Specific Pathogen-Free Organisms; Swine; Tacrolimus; Toxicity Tests

The treatment of some inflammatory diseases, such as rheumatoid arthritis, remains an important target for studies because some patients are refractory to conventional treatment. Mycophenolate mofetil (MMF), an immunosuppressive drug, has been shown to have a beneficial effect on the therapy of inflammatory and autoimmune diseases. In the present study, we aimed to analyse the anti-inflammatory effect of MMF administered by oral route in the mouse carrageenan-induced air pouch model.. MMF significantly inhibited the influx of leukocytes, exudate concentrations (P<0.01), activities of myeloperoxidase (MPO) and adenosine deaminase (ADA), levels of nitrite/nitrate (NO(x)) and inducible nitric oxide synthase (iNOS) mRNA expression, as well as the levels of mRNA expression and proteins of tumor necrosis factor-alpha (TNF-α), Interleukin-beta (IL-1β) and vascular endothelial growth factor-alpha (VEGF-α) (P<0.05). These results provide evidence that MMF has an important anti-inflammatory effect in reducing the influx of leukocytes and exudate concentrations. These inhibitory effects are correlated with the inhibition of specific pro-inflammatory enzymes (MPO, ADA and iNOS), and the levels of mRNA expression and proteins of TNF-α, IL-1β and VEGF-α.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Carrageenan; Cell Movement; Cytokines; Disease Models, Animal; Humans; Immunosuppression Therapy; Inflammation Mediators; Leukocytes; Mice; Mycophenolic Acid; Nitric Oxide; Peroxidase; Vascular Endothelial Growth Factor A

The aim of this study was to investigate the immunosuppressive activity of a mycophenolate mofetil (MMF) intraocular-implantable drug delivery system (IDDS) in a rabbit model of high-risk penetrating keratoplasty and to determine the biocompatibility of such a device when implanted in the anterior chamber.. Corneal vascularization was induced in New Zealand white rabbits by passing 5-0 silk sutures through the corneal stroma in each quadrant. The corneal neovascularized rabbits received a unilateral 7-mm-diameter central-penetrating keratoplasty. New Zealand white rabbits were used as donors and were divided into 4 treatment groups: the control group, which received no therapy; the 1% MMF eye drop group; the 1.0 mg cyclosporin A (CsA)-IDDS-implanted group; and the 1.0 mg MMF-IDDS-implanted group. Animals were followed up for 150 days, which involved examination of the corneal allografts (opacity, edema, and neovascularization) by slit-lamp biomicroscopy. The survival time of corneal allografts of these animal models was recorded in 4 groups. Histopathologic studies were carried out on the procured specimens of corneal allografts. The biocompatibility of MMF-IDDS in the anterior chamber in rabbits was also investigated.. The mean survival time of corneal allografts in the control and MMF eye drop groups was 18.7±3.0 and 37.5±6.2 days, respectively (P=0.005). Allografts from the CsA-IDDS-implanted group were transparent, except 1 allograft, which showed immune rejection after 130 days. Allografts from the MMF-IDDS-implanted group were transparent throughout the entire observation period. The incidence of allograft rejection was 100% in the control and MMF eye drop groups, respectively. The rejected allografts were much more edematous and more heavily infiltrated with leukocytes than the nonrejected allografts. MMF-IDDS was tolerated well in the anterior chamber, even with 3 MMF-IDDS implanted in the anterior chamber at 1 time.. MMF-IDDS was able to prolong high-risk allograft survival time and significantly inhibited corneal immune rejection in the rabbit model of penetrating keratoplasty. The device could safely be implanted in the anterior chamber without adverse effects.

Topics: Animals; Anterior Chamber; Corneal Neovascularization; Corneal Transplantation; Disease Models, Animal; Drug Delivery Systems; Drug Implants; Female; Graft Rejection; Graft Survival; Immunosuppressive Agents; Incidence; Male; Mycophenolic Acid; Rabbits; Risk; Time Factors; Treatment Outcome

Autoantibodies to brain proteins are present in Juvenile Neuronal Ceroid Lipofuscinosis (Batten disease) patients and in the Cln3-/- mouse model of this disease, suggesting an autoimmune component to pathogenesis. Using genetic or pharmaceutical approaches to attenuate this immune response in Cln3-/- mice, we demonstrate decreased neuroinflammation, decreased deposition of immunoglobulin G in the brain and protection of vulnerable neuron populations. Moreover, immune suppression results in a significant improvement in motor performance providing for the first plausible therapeutic approach for juvenile Batten disease.

Topics: Animals; Autoantibodies; Blotting, Western; Brain; Cell Count; Disease Models, Animal; Immunohistochemistry; Immunosuppression Therapy; Immunosuppressive Agents; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Chaperones; Motor Skills; Mycophenolic Acid; Neuronal Ceroid-Lipofuscinoses; Neurons

The aim of this study was to investigate the effect of mycophenolate mofetil (MMF) using differential kidney proteome profiling of COL4A3-deficient mice as a model of progressive renal disease.. Histological evaluation of kidney sections was performed. Total protein lysate from kidneys of placebo- and MMF-treated COL4A3-deficient mice was studied for significant differences in protein abundance using 2-dimensional electrophoresis and mass spectrometry.. While tubulointerstitial fibrosis in COL4A3-deficient mice was inhibited by MMF, 19 proteins in the kidneys were regulated: 12 with lower (ATPO, TAGL2, CAH1, TPD52, VA0D1, SERPH, GNAL, PSB6, EF1D, OTUB1, NDUS8, and NAPSA) and 7 with higher (ACADM, ACY3, CK054, ACTB/G, ACTB, UBP5, and ACY1) spot intensity. Nine differentially expressed proteins showed interaction potential (ATPO, TPD52, PSB6, EF1D, OTUB1, NAPSA, ACTB, ACTG, and UBP5).. The identified proteins take part in different signaling pathways. With the highest probability, the VEGF signaling pathway (TAGL2, EF1D, and ACTB) and hypoxia (CAH1, PSB6, and ACTG) were influenced by MMF in fibrotic conditions. Moreover, MMF may modulate antifibrotic and apoptotic pathways as well as epithelial-mesenchymal transition (EMT). Different signaling pathways may be influenced by MMF therapy. These results suggest an inhibitory effect of MMF on renal EMT in COL4A3-deficient mice. Further studies are necessary to validate these findings.

Topics: Animals; Autoantigens; Collagen Type IV; Databases, Protein; Disease Models, Animal; Disease Progression; Down-Regulation; Electrophoresis, Gel, Two-Dimensional; Epithelial-Mesenchymal Transition; Fibrosis; Kidney; Kidney Diseases; Mice; Mice, Knockout; Mycophenolic Acid; Proteomics; Random Allocation; Signal Transduction; Up-Regulation

Pulmonary arterial hypertension (PAH) is a severe clinical and pathophysiologic syndrome with no effective treatment at present. Mycophenolate mofetil (MMF), a prodrug of mycophenolic acid (MPA), has been applied widely to the treatment of connective tissue diseases with the effect of immunosuppressant. Its anti-proliferation has been found recently. Thereby, we tried to examine the effects of MMF on rats with PAH which was induced by monocrotaline injection, so as to investigate the mechanisms of treatment on PAH by MMF. The results substantiated that MMF therapy can alleviate thickening of pulmonary arterial walls and inhibit abnormal vascular remodeling, and the MPA concentrations which demonstrated efficacy in this study are within clinical applicable range, suggesting huge potentiality of MMF in the treatment of human PAH.

Topics: Animals; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Hypertension, Pulmonary; Hypertrophy; Immunosuppressive Agents; Male; Monocrotaline; Muscle, Smooth, Vascular; Mycophenolic Acid; Pulmonary Artery; Rats; Rats, Sprague-Dawley; Treatment Outcome

To evaluate the efficacy and safety of topically applied mycophenolate mofetil (MMF) for the prophylaxis of corneal graft rejection in an experimental keratoplasty model.. A total of 12 female Lewis rats received 3.5-mm MHC I/II-incompatible corneal grafts from DA donors. Recipients were randomly assigned to receive either topical MMF + beta-cyclodextrin therapy (1%), beta-cyclodextrin therapy alone or to remain untreated. Therapy was applied every 2 h (over 24 h) during the first 3 postoperative days, then twice hourly during daytime. Grafts were graded every day based on a rejection score including the parameters transplant clarity and edema.. The mean survival time (MST) of the grafts in the MMF-treated group was 12 days, the MST in the vehicle-treated group was 14.3 days and the MST in the untreated group was 13.3 days. So, the survival curves of the 3 treatment groups did not differ significantly.. Topical MMF is ineffective for prophylaxis of corneal graft rejection.

Topics: Administration, Topical; Animals; beta-Cyclodextrins; Corneal Transplantation; Disease Models, Animal; Female; Graft Rejection; Graft Survival; Immunosuppressive Agents; Mycophenolic Acid; Ophthalmic Solutions; Rats; Rats, Inbred Lew; Transplantation, Homologous; Treatment Failure

Within the scope of this study, the potential antifibrotic effect of mycophenolate mofetil (MMF) on COL4A3-deficient mice as an animal model for progressive renal fibrosis was investigated regarding kidney function and survival. Thirty-five animals were randomly assigned to one of five groups and treated with doses of 0, 10, 50, 100, or 150 mg/kg MMF per day, respectively. When increasing somnolence was observed, indicating end-stage renal disease, the mice were euthanized and blood was obtained. Serum concentrations of creatinine, urea nitrogen, total protein, mycophenolic acid (MPA), and mycophenolic acid glucuronide (MPAG) were quantified. The kidney histology was examined using hematoxylin and eosin as well as trichrome staining. The mean overall survival was 65.9 (+/-6.1) days with no significant difference between the treatment groups (P > 0.05, Mantel-Cox test). Serum predose concentrations of MPA and MPAG showed considerable interindividual variability. There was no correlation between survival time and MPA or MPAG concentrations (P > 0.05, Spearman rank correlation). However, an apparent decrease in serum creatinine and urea nitrogen concentrations was observed at higher doses of MMF, eg, -54% for creatinine in the 150-mg/kg/day group compared with placebo. A highly significant reciprocal correlation between MPA concentrations and serum creatinine was demonstrated (P < 0.01, r = -0.655, Spearman rank correlation). In conclusion, MMF may be a candidate drug for preserving kidney function in progressive renal fibrosis.

Topics: Animals; Autoantigens; Collagen Type IV; Disease Models, Animal; Disease Progression; Dose-Response Relationship, Drug; Fibrosis; Immunosuppressive Agents; Kidney Diseases; Kidney Function Tests; Mice; Mice, Knockout; Mycophenolic Acid; Survival Rate

Although the therapeutic potential of regulatory T lymphocytes (Tregs) in preventing allograft rejection has been well documented, accumulating evidence indicates that supplemental measures, such as concomitant use of immunosuppressive agents, are essential for effective application of Treg cell therapy in clinical transplantation. Thus, it is important to know the effect of immunosuppressive agents on Treg cell therapy.. We examined the impact of various immunosuppressive agents on the in vivo proliferation and therapeutic efficacy of in vitro-expanded Tregs using the murine graft-versus-host reaction and skin allograft model (BDF1 [H-2] to C57BL/6 [H-2]), respectively.. All six immunosuppressive agents tested inhibited the alloantigen-stimulated proliferation of Tregs as efficiently as they inhibited the proliferative response of conventional CD3 T cells. We further show that blockade of the CD40-CD40L interaction by treatment with a MR-1 antibody significantly increased the therapeutic efficacy of Tregs, a synergistic effect that seemed to be related to the strong regulatory activity of adoptively transferred Tregs together with effector T-cell hyporesponsiveness. Although concomitant use of rapamycin marginally augmented the therapeutic effectiveness of Tregs, mycophenolate mofetil and cyclosporine A at their full therapeutic doses exerted an antagonistic effect on Treg cell therapy.. These results demonstrate that inhibition of CD40-CD40L interaction or treatment with rapamycin could be successfully combined with in vitro-expanded Treg cell therapy, but the concomitant use of mycophenolate mofetil or cyclosporine A in this type of Treg cell therapy should be carefully considered.

Topics: Adoptive Transfer; Animals; Antibodies; CD40 Antigens; CD40 Ligand; Cell Proliferation; Cells, Cultured; Cyclosporine; Disease Models, Animal; Forkhead Transcription Factors; Graft Rejection; Graft vs Host Disease; Immunosuppressive Agents; Interleukin-2 Receptor alpha Subunit; Isoantigens; Male; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred DBA; Mycophenolic Acid; Sirolimus; Skin Transplantation; T-Lymphocytes, Regulatory; Time Factors; Transplantation Tolerance; Transplantation, Homologous

Lymphocytes participate in the early pathogenesis of ischemia-reperfusion injury (IRI) in kidney; however, their role during repair is largely unknown. Recent data have shown that Foxp3(+) regulatory T cells (Tregs) traffic into kidney during healing from IRI and directly participate in repair. Since lymphocyte-targeting therapy is currently administered to prevent rejection during recovery from IRI in renal transplants, we hypothesized that mycophenolate mofetil (MMF) would alter Treg trafficking and kidney repair. C57BL/6J and T cell deficient mice underwent unilateral clamping of renal pedicle for 45 min, followed by reperfusion, and were sacrificed at day 10. Mice were treated with saline (C) or MMF (100mg/kg) i.p. daily starting at day 2 until sacrifice (n=5-12/group). MMF worsened kidney tubular damage compared to C at 10 days (cortex and outer medulla: p<0.05) in wild-type mice; tubular apoptotic index was increased in cortex in MMF group as well (p=0.01). MMF reduced the total number of kidney-infiltrating mononuclear cells (p<0.001 versus C) and the percentages of TCRbeta(+)CD4(+) and TCRbeta(+)CD8(+) T cells (p<0.01), but not natural killer (NK), NKT or B lymphocytes. MMF specifically reduced kidney Foxp3(+) Tregs (0.82+/-0.11% versus 1.75+/-0.17%, p<0.05). Tubular proliferative index and tissue levels of basic FGF were increased in MMF group (p<0.05), IL-10 and IL-6 were decreased (p<0.05). To evaluate if MMF effect occurred through non-lymphocytic cells, T cell deficient mice were treated with MMF. Tubular injury in T cell deficient mice was not affected by MMF treatment, though MMF-treated animals had increased VEGF and decreased PDGF-BB protein tissue levels compared to controls (p<0.05). Thus, MMF modifies the structural, epithelial proliferative and inflammatory response during healing, likely through effects on T cells and possibly Tregs. Kidney repair after IRI can be altered by agents that target lymphocytes.

Topics: Animals; Disease Models, Animal; Forkhead Transcription Factors; Immunosuppressive Agents; Kidney; Kidney Tubular Necrosis, Acute; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mycophenolic Acid; Reperfusion Injury; T-Lymphocytes, Regulatory

The aim of this study was to develop and characterize a rat glomerulonephritis model, which progresses to renal fibrosis and renal failure. A single immunization of female WKY rats with more than 10 microg of recombinant alpha3(IV)NC1 protein caused severe proteinuria followed by progressive increases in plasma creatinine and blood urea nitrogen (BUN) level within 42 days. Sequential histopathological evaluation revealed crescent formation in glomeruli followed by tubular dilation and interstitial fibrosis. Hydroxyproline content and expression of type I collagen and smooth muscle actin genes in the renal cortex increased as renal dysfunction progressed. Furthermore, the TGF-beta1 level in the renal cortex also increased. In the evaluation of antinephritic agents in this model, prednisolone and mycophenolate mofetil (MMF) treatment significantly decreased plasma creatinine and BUN, and suppressed renal fibrosis and histological changes involving crescent formation, compared with the vehicle-treated nephritic rats, whereas lisinopril treatment failed to improve renal function and histology. We demonstrated that immunization of female WKY rats with a sufficient dose of recombinant alpha3(IV)NC1 induces end-stage kidney disease accompanied by renal fibrosis. The relatively short period needed to induce the disease and the high incidence of functional and structural changes were considered a great advantage of this model for clarifying the mechanisms of progressive glomerulonephritis and for evaluating agents used to treat renal failure.

Topics: Actins; Animals; Autoantigens; Collagen Type I; Collagen Type IV; Creatinine; Disease Models, Animal; Female; Gene Expression; Glomerulonephritis; Hydroxyproline; Immunosuppressive Agents; Kidney Failure, Chronic; Kidney Function Tests; Lisinopril; Mycophenolic Acid; Prednisolone; Rats; Rats, Inbred WKY; Recombinant Proteins; Transforming Growth Factor beta1

We evaluated the role of renal tubular Nox-2 in the pathogenesis of epithelial-to-mesenchymal transition (EMT) in kidney allografts.. We examined this question in the human kidney allografts with interstitial fibrosis and tubular atrophy not otherwise specified (IFTANOS), in the Fisher to Lewis rat transplant model, and in the in vitro model of transforming growth factor-beta1-induced EMT in normal rat kidney epithelial cells (NRK52E).. We first demonstrated that Nox-2 and alpha-smooth muscle actin (SMA) were increased in renal tubules from kidney transplant recipients on calcineurin inhibitors, mycophenolic acid (MPA), and prednisone with IFTANOS, suggestive of EMT (n=6). Next, we examined Nox-2 expression and fibrogenesis in syngeneic transplants, allogeneic transplants treated with MPA 40 mg/kg per 24 hr, and untreated allogeneic transplants for 6 months (n=14 in each group). Immunofluorescent and immunohistochemical studies for Nox-2, alpha-SMA, and E-cadherin showed that similar to patients with IFTANOS, rat allografts had greater tubulointerstitial staining for Nox-2 and alpha-SMA. MPA therapy prevented these changes. Immunoblot analyses examining Nox-2 signaling (phospho-nuclear factor [NF]-kappaB), redox signaling (phospho-smad2), and fibrosis (alpha-SMA and fibronectin) demonstrated that MPA treatment prevented the up-regulation of Nox-2, inhibited p-NF-kappaB and p-smad2, and down-regulated alpha-SMA and fibronectin levels. Finally, we examined Nox-2 signaling in vitro and confirmed that MPA inhibited phospho-NF-kappaB, Nox-2, phospho-smad2, and alpha-SMA during transforming growth factor-beta1-induced EMT of NRK52E cells while reducing Nox-2, vimentin, and fibronectin mRNA levels.. MPA may down-regulate Nox-2 activation and EMT through the NF-kappaB pathway in the tubular epithelial cells, suggesting a novel role for this drug independent of its immunosuppressive properties.

Topics: Adult; Animals; Anti-Bacterial Agents; Atrophy; Biopsy; Creatinine; Disease Models, Animal; Fibrosis; Humans; Immunosuppressive Agents; Kidney Glomerulus; Kidney Transplantation; Kidney Tubules; Mycophenolic Acid; NF-kappa B; Oxidative Stress; Rats; Rats, Inbred Lew; Reactive Oxygen Species; Transforming Growth Factor beta1; Transplantation, Homologous

Inflammatory events antecede established renal injury in rats with 5/6 renal ablation (Nx), as indicated by the beneficial effects of early, uninterrupted treatment with mycophenolate mofetil (MMF). Angiotensin II also exerts a major pathogenic role at this initial phase. We investigated whether losartan (L) or L+MMF treatment, started early, and L+MMF treatment, started late, would exert lasting renoprotection in Nx even after being discontinued.. Adult male Munich-Wistar rats underwent Nx and were divided into three groups: Nx (untreated), Nx(L) (given L), and Nx(LMMF) (given L and MMF). Protocol 1: treatments began on day 1, and ceased on day 30, after Nx. Protocol 2: L+MMF treatment began on day 30 and ceased on day 60.. Protocol 1: on day 30, hypertension, albuminuria and renal injury were strongly attenuated in Groups Nx(L) and Nx(LMMF). On day 120, these abnormalities were still attenuated in group Nx(LMMF). Protocol 2: on day 120, all parameters were similar between this late Nx(LMMF) group and untreated Nx.. In Nx, temporary suppression of early, transitory hemodynamic/inflammatory phenomena affords relatively durable renoprotection even after treatment discontinuation. This effect is not obtained with similar temporary treatment initiated later in the course of renal disease.

Topics: Albuminuria; Angiotensin II Type 1 Receptor Blockers; Animals; Anti-Inflammatory Agents; Blood Pressure; Chronic Disease; Disease Models, Animal; Drug Therapy, Combination; Kidney Diseases; Losartan; Macrophages; Male; Mycophenolic Acid; Nephrectomy; Protective Agents; Rats; Rats, Inbred Strains; Time Factors

To examine fibronectin (FN) expression in human lupus nephritis and the effect of anti-DNA antibodies on transforming growth factor beta1 (TGFbeta1) and FN synthesis in cultured human mesangial cells. The effects of mycophenolic acid (MPA) on this pathway, and the effects of mycophenolate mofetil (MMF) treatment in (NZB x NZW)F(1)/J mice were also studied.. Immunohistochemical analyses of renal biopsy samples from patients with active diffuse proliferative lupus nephritis were performed. Cultured human mesangial cells were incubated with human polyclonal anti-DNA antibodies, with or without MPA. (NZB x NZW)F(1)/J mice with active nephritis were randomized to receive either MMF (100 mg/kg/day) or vehicle treatment for 12 weeks.. Glomerular FN expression was increased in patients with lupus nephritis, and it colocalized with IgG deposition. Anti-DNA antibodies induced protein kinase Calpha (PKCalpha), PKCbetaI, and PKCbetaII activation, increased levels of bioactive TGFbeta1, and increased FN synthesis in human mesangial cells (P < 0.001 for each comparison versus control conditions). Pretreatment of anti-DNA antibodies with exogenous DNA reduced their cellular binding and abrogated their induction of TGFbeta1 and FN synthesis. Inhibition of PKC activation in human mesangial cells prior to anti-DNA antibody stimulation had no effect on cell proliferation, but resulted in significantly reduced antibody-mediated TGFbeta1 secretion and FN synthesis. MPA treatment down-regulated PKCalpha, PKCbetaI, and PKCbetaII phosphorylation, reduced levels of TGFbeta1 bioactivation, and decreased FN synthesis and deposition into the extracellular matrix. MMF treatment in (NZB x NZW)F(1)/J mice resulted in a reduction in glomerular IgG deposition, PKC activation, and FN expression, as well as an amelioration of proteinuria.. Human polyclonal anti-DNA antibodies induce TGFbeta1 and FN synthesis in human mesangial cells through PKC activation, which is inhibited by MPA.

Topics: Adult; Animals; Antibodies, Antinuclear; Biopsy; Cells, Cultured; Disease Models, Animal; Enzyme Inhibitors; Female; Fibronectins; Humans; Immunoglobulin G; Kidney; Lupus Nephritis; Male; Mesangial Cells; Mice; Mice, Inbred NZB; Middle Aged; Mitogen-Activated Protein Kinase Kinases; Mycophenolic Acid; Phosphorylation; Protein Kinase C; Transforming Growth Factor beta1

Hypertensive disorders of pregnancy, including preeclampsia (PE), affect approximately 7-10% of pregnancies in the US. Clinical and experimental studies strongly suggest that the maternal immune system plays a role in the development of these disorders; however, few therapeutic options exist aside from delivery.. Using a deoxycorticosterone acetate (DOCA)/salt-low renin rat model, which exhibits hypertension, proteinuria, endothelial dysfunction, and intrauterine growth restriction (IUGR), we measured serum cytokine levels as an indication of immune system activation. In addition, we suppressed the immune system with either azathioprine (Aza) or mycophenolate mofetil (MMF) during the second half of pregnancy to determine whether the these symptoms could be ameliorated.. Our results demonstrate that serum T helper-1 (Th1)-type inflammatory cytokines interleukin (IL)-2, IL-12, interferon-gamma (IFNgamma), and RANTES were significantly elevated in hypertensive pregnant rats while the Th2-type cytokine IL-4 was elevated in normal pregnant animals. Either Aza or MMF significantly attenuated the hypertension, proteinuria, and endothelial dysfunction as well as the increased proinflammatory Th1 cytokine profile in pregnant rats treated with DOCA/salt, and had no effect on these parameters in normal pregnant rats.. These data strongly suggest that maternal immune system activation plays a role in the development of pregnancy-induced hypertension (PIH).

Topics: Animals; Azathioprine; Blood Pressure; Cytokines; Desoxycorticosterone; Disease Models, Animal; Endothelium, Vascular; Female; Fetal Growth Retardation; Hypertension, Pregnancy-Induced; Immunosuppression Therapy; Immunosuppressive Agents; Mycophenolic Acid; Pregnancy; Proteinuria; Rats; Rats, Sprague-Dawley

Generation of oxidative stress induced by reactive oxygen species (ROS) and nitrogen (RNS) is believed to be a primary factor in the etiology of various inflammatory diseases. Although, the process of generation of oxygen species is a physiological event, in the inflammatory process this event is increased and produces large amounts of reactive species that leads to lipid peroxidation and to cell death. Mycophenolate mofetil (MMF) is a drug effective in protecting against chronic allograft failure and recently was introduced as an alternative for the treatment of various inflammatory diseases such as glomerulopathies, systemic lupus erythematosus and systemic vasculitis. Based on studies of the anti-inflammatory effect of MMF the aim of this study was to evaluate the effects of MMF on the inhibition of leukocytes and exudation, as well as myeloperoxidase and some antioxidant enzyme activities using carrageenan-induced pleurisy in mice. Our results showed that MMF significantly decreased leukocyte influx (P<0.01), exudation (P<0.01), superoxide dismutase (P<0.05), catalase (P<0.05), glutathione peroxidase (P<0.01), glutathione S-transferase (P<0.01) activities, levels of lipid peroxidation (P<0.05), as well as myeloperoxidase activity (P<0.05) on both phases (4h and 48h) of the inflammatory response induced by carrageenan into the mice pleural cavity. In conclusion, the anti-inflammatory effect of MMF may be, at least in part, via inhibition of ROS and/or NRS overgeneration, and consequently, attenuating the related oxidative stress.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Catalase; Cell Count; Cell Movement; Dexamethasone; Disease Models, Animal; Glutathione Peroxidase; Glutathione Transferase; Indomethacin; Leukocytes; Leukocytes, Mononuclear; Lipid Peroxidation; Mice; Mice, Inbred Strains; Mycophenolic Acid; Neutrophils; Oxidative Stress; Peroxidase; Pleural Effusion; Pleurisy; Superoxide Dismutase; Thiobarbituric Acid Reactive Substances

We aimed to explore the effect of Mycophenolate mofetil (MMF) on loss of renal function and cyst progression compared to rapamycin in Han: SPRD rats. We also sought to assess whether the effect of combination therapy of MMF plus rapamycin was better than that of monotherapy.. Sixty heterozygous (Cy/+) and littermate control (+/+) male Han: SPRD rats were weaned at 4 weeks of age, then divided into four groups randomly to receive different treatments by intragastric administration for 2 months: vehicle-treated group as control, MMF-treated group (20mg/kg/day), rapamycin-treated group (2mg/kg/day), and MMF+Rapa- treated group (MMF 20mg/kg/day plus Rapamycin 2mg/kg/day). RESULLS: After 2 months of treatment, rapamycin caused a 22 % decrease in body weight in comparison to the control group, whereas MMF had no significant effect on weight gain. The steady increase of BUN in Cy/+ rats was reduced by 15% in MMF-treated Cy/+ rats. However, rapamycin and combination therapy reduced BUN by 42% and 43%, respectively. CCr was 0.93+/-0.11ml/min in vehicle-treated Cy/+ rats, 1.67+/-0.23 ml/min in MMF-treated Cy/+ rats (P<0.05), 1.72+/-0.44 ml/min and 1.83+/-0.21 ml/min in rapamycin- and MMF+Rapa-treated Cy/+ rats, respectively (.P<0.01). Cyst volume density was 57.1 % in vehicle-treated Cy/+ rats, 45.2% in MMF-treated Cy/+ rats (P<0.05), 32.9% and 37.7% in rapamycin- and MMF+Rapa-treated Cy/+ rats, respectively (P<0.01). MMF markedly ameliorated interstitial inflammation and fibrosis. Rapamycin showed a similar effect on interstitial inflammation and fibrosis, but to a lesser degree.. MMF is more tolerable than rapamycin and can retard deterioration of renal function in Han: SPRD rats, though its effect is weaker than that of rapamycin. Combination therapy does not exert more favorable effect than monotherapy.

Topics: Animals; Disease Models, Animal; Drug Therapy, Combination; Immunosuppressive Agents; Male; Mycophenolic Acid; Polycystic Kidney Diseases; Rats; Rats, Sprague-Dawley; Sirolimus; Time Factors

To evaluate the expression of TNF-related apoptosis inducing ligand (TRAIL) and nuclear factor (NF)-kappaB in the kidney tissues of diabetic rats and the effects of valsartan, mycophenolate mofetil (MMF), and their combined application on the renal TRAIL and NF-kappaB expression.. Eighty uninephrectomized male Wistar rats were randomly divided into 2 groups: normal control (NC) group (n = 28), undergoing intraperitoneal injection of citric acid buffer, and diabetes mellitus (DM) group, undergoing intraperitoneal injection of streptozotocin (STZ) to establish DM models. The 52 DM rats were randomly divided into 7 equal subgroups: DM without treatment for 4 weeks (DM4), DM without treatment for 8 weeks (DM8) DM without treatment for 12 weeks (DM12) , and DM without treatment for 16 weeks (DM16), valsartan treatment (DM + V), MMF treatment (DM + M), and combined treatment (DM + V + M). The treatment subgroups were treated for 8 weeks immediately after the diabetic models ere established. Twenty-four hour urine was collected to measure the amount of protein 4, 8, 12, and 16 weeks after the induction of DM respectively. The rats were sacrificed. Blood samples were collected from the abdominal aorta to detect the blood urea nitrogen (BUN), serum creatinine (sCr), albumin, and glucose. The kidneys were taken out. Hypertrophy index (left kidney weight/body weight) was determined. Quantitative real time RT-PCR was performed to detect the expression of TRAIL and NF-kappaB mRNA. Immunohistochemistry was used to detect the protein expression of TRAIL and NF-kappaB.. 1) The 24 h urine protein levels and hypertrophy indexes of all DM subgroups were significantly higher than those of the NC groups (all P <0.05). 24 h urine protein and hypertrophy index increased gradually and peaked in the l2th week; blood albumin gradually decreased since the 8th week(P <0.01), and BUN and sCr began to decrease only since the 16th week (both P <0.01). Compared with the DM 8 subgroup, the hypertrophy index and 24 h urine protein of the different treatment subgroups, especially the DM + V + M subgroup, were significantly lower (all P <0.05). 2) Quantitative real time RT-PCR showed that compared with the NC group, the TRAIL expression levels of the DM subgroups were significantly lower before the 12th week after induction of DM model (all P < 0.01), and then significantly higher in the 16th week (all P < 0.01). The TRAIL expression of the treatment groups, especially that of the DM + M subgroup, were significantly higher than that of the DM8 group (all P <0.05). Compared with the NC group, the NF-kappaB expression levels of the DM subgroups were significantly higher time-dependently (all P <0.01). Compared with the DM8 group, the NF-kappaB expression levels of the treatment subgroups, especially that of the DM + V + M subgroup were significantly lower (all P <0.05). 3) The expression of TRAIL was mainly located in the convoluted tubule of kidney, and no TRAIL protein expression was detected in the glomeruli or renal vasculature. The levels of NF-kappaB protein expression, shown in glomeruli and convoluted tubules, of all DM subgroups were all higher than that of the NC group. The NF-kappaB protein expression level of the DM + V + M subgroup was significantly lower. The number of NF-kappaB positive cells was significantly related to the mononuclear macrophage infiltration , kidney function, and structural lesion.. An important monitoring factor in the autoimmune system, TRAIL closely participates in the pathogenesis of diabetic nephropathy, possibly controlled by NF-kappaB. In the early stage combination of valsartan and MMF may upregulate the expression of TRAIL, thus protecting the kidney function.

Topics: Animals; Antihypertensive Agents; Apoptosis; Apoptosis Regulatory Proteins; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Disease Models, Animal; Drug Therapy, Combination; Immunosuppressive Agents; Kidney; Male; Mycophenolic Acid; NF-kappa B; Random Allocation; Rats; Rats, Wistar; Tetrazoles; TNF-Related Apoptosis-Inducing Ligand; Treatment Outcome; Valine; Valsartan

Currently used non-specific immunosuppressive drugs often require intervention in myasthenia gravis (MG) and clinical improvement varies widely. To analyze the therapeutic effect of mycophenolate mofetil (MMF) in experimental autoimmune MG (EAMG), rats were immunized with acetylcholine receptors (AChRs) and subsequently treated with MMF or vehicle. MMF treatment resulted in a significant suppression of anti-rat AChR antibody titers. Interestingly, no abnormalities of neuromuscular transmission and adverse side effects were detected in MMF-treated EAMG animals. Moreover, anti-rat AChR antibody titers correlated to an improvement of clinical outcome. In conclusion, our data suggest that MMF acts as a potent immunosuppressant drug in EAMG.

Topics: Action Potentials; Analysis of Variance; Animals; Antibodies; Disease Models, Animal; Electromyography; Female; Immunosuppressive Agents; Muscle Proteins; Muscle, Skeletal; Myasthenia Gravis, Autoimmune, Experimental; Mycophenolic Acid; Rats; Rats, Inbred Lew; Receptors, Cholinergic; Statistics, Nonparametric; Time Factors; Vesicular Acetylcholine Transport Proteins

In this study, we used a murine intestinal inflammation model that mimics immunologic characteristics of human Crohn's disease (CD) to investigate the anti-inflammatory effects of mycophenolate mofetil (MMF) on intestinal injury and tissue inflammation. When these colitic mice were pretreated with MMF, we observed a significant decrease in mortality rates and body weight loss as well as an improvement in both wasting and histopathologic signs of colonic inflammation, relative to untreated colitic mice. To determine the mechanisms of action of MMF, we compared various immunological characteristics of the untreated and MMF-pretreated colitic mice. MMF-pretreated colitic mice showed an 18% decrease in the proportion of CD19+ B cells compared with untreated colitic mice 3 days. As a result, MMF pretreatment increases proportion of apoptotic T and B cells, especially CD19+ B cells. Also, down-regulation of Th1 cytokines (TNF-alpha, IFN-gamma) and augmentation of CD4+CD45RB(low) regulatory T (Treg) cells were observed in MMF-pretreated colitic mice compared with untreated colitic mice. Furthermore, mycophenolic acid (MPA) reduced TNF-alpha-stimulated NF-kappaB activation in HT-29 colon epithelial cells. Also, MMF-pretreated colitic mice significantly reduced expression of MD-1 compared with untreated colitic mice on B cells and dendritic cells (DCs). These studies show that MMF pretreatment can improve experimental colitis by down-regulation of expanded B cells population through apoptosis and augmentation of Treg cells. Through these mechanisms, MMF might also be an effective agent for the treatment of other diseases characterized by mucosal inflammation.

Topics: Animals; Antigens, CD19; Antigens, Surface; Apoptosis; B-Lymphocytes; Colitis; Disease Models, Animal; Down-Regulation; Female; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mycophenolic Acid; NF-kappa B; T-Lymphocytes, Regulatory; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

Mycophenolate mofetil (MMF) has been used successfully in solid organ transplantation (SOT) and more recently in nonmyeloablative hematopoietic stem cell transplantation (HSCT) for prophylaxis of graft rejection and acute graft-versus-host disease. However, the pharmacokinetics of MMF seem to differ when applied in HSCT compared to SOT. Here, we analyzed pharmacokinetics of mycophenolic acid (MPA), the active metabolite of MMF, in a nonmyeloablative canine HSCT model. Dogs received nonmyeloablative TBI for conditioning followed by leukocyte antigen-identical littermate HSCT and immunosuppression containing cyclosporin A (CsA) and different doses of MMF. Pharmacokinetics were performed on days 2, 14 and 27. Dose escalation of MMF from 10 to 30 mg/kg tended to increase area under the curve (AUC) and the apparent oral clearance by 45 and 110%, respectively. Doses applied had no linear association with MPA concentration or blood trough level. No significant drug accumulation occurred over time. Using a twice daily MMF regimen, we conclude that an AUC of 30-60 mug/ml h as recommended for SOT cannot be reached in HSCT. Toxicities did not permit single doses higher than 30 mg/kg. Thus, if larger AUCs are desired in order to assure sufficient immunosuppression in HSCT, MMF might have to be administered at least three times daily.

Topics: Animals; Cyclosporine; Disease Models, Animal; Dogs; Dose-Response Relationship, Drug; Drug Therapy, Combination; Graft Rejection; Hematopoietic Stem Cell Transplantation; Immunosuppressive Agents; Mycophenolic Acid

The present study determined the effect of immune suppression with mycophenolate mofetil (MMF) on sodium-sensitive hypertension following recovery from ischemia reperfusion (I/R)-induced acute renal failure. Male Sprague-Dawley rats fed 0.4% NaCl chow were subjected to 40 min bilateral I/R or control sham surgery. After 35 days of recovery, when plasma creatinine levels had returned to normal, the rats were switched to 4.0% NaCl chow for 28 days and administered vehicle or MMF (20 mg.kg(-1).day(-1) ip). High-salt mean arterial pressure was significantly higher in I/R rats (144 +/- 16 mmHg) compared with vehicle-treated sham rats (122 +/- 2 mmHg). Treatment of I/R rats with MMF during the period of high salt intake prevented the salt-induced increase in arterial pressure (114 +/- 3 mmHg). Conscious creatinine clearance was lower in I/R rats (0.27 +/- 0.07 ml.min(-1).100 g body wt(-1)) compared with vehicle-treated sham rats (0.58 +/- 0.04 ml.min(-1).100 g body wt(-1)); MMF treatment prevented the decrease in creatinine clearance in I/R rats (0.64 +/- 0.07 ml.min(-1).100 g body wt(-1)). I/R injury also significantly increased glomerular tissue damage and increased the presence of ED-1 positive (macrophages) and S100A4 positive cells (fibroblasts) in the renal interstitium. The I/R rats treated with MMF exhibited a significant reduction in infiltrating macrophages and fibroblasts and decreased histological damage. The present data indicate that infiltrating immune cells mediate or participate in the development of sodium-sensitive hypertension and renal damage in rats apparently recovered from renal I/R injury.

Topics: Acute Kidney Injury; Animals; Blood Pressure; Cell Movement; Creatinine; Disease Models, Animal; Fibroblasts; Hypertension; Immunosuppressive Agents; Ischemia; Kidney; Macrophages; Male; Mycophenolic Acid; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Sodium Chloride, Dietary; Time Factors

Topics: Amyotrophic Lateral Sclerosis; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Carcinoma, Small Cell; Disease Models, Animal; Humans; Immunologic Factors; Immunosuppressive Agents; Lambert-Eaton Myasthenic Syndrome; Lung Neoplasms; Mycophenolic Acid; Myelin-Associated Glycoprotein; Neuromuscular Diseases; Polyneuropathies; Rats; Rituximab; Superoxide Dismutase; Superoxide Dismutase-1

Activation of the renin-angiotensin system contributes to the progression of chronic kidney disease. Based on the known cellular effects of ANG II to promote inflammation, we posited that stimulation of lymphocyte responses by ANG II might contribute to the pathogenesis of hypertensive kidney injury. We therefore examined the effects of the immunosuppressive agent mycophenolate mofetil (MMF) on the course of hypertension and kidney disease induced by chronic infusion of ANG II in 129/SvEv mice. Although it had no effect on the severity of hypertension or cardiac hypertrophy, treatment with MMF significantly reduced albuminuria and ameliorated kidney injury, decreasing glomerulosclerosis and reducing lymphocyte infiltration into the renal interstitium. Attenuation of renal pathology with MMF was associated with reduced expression of mRNAs for the proinflammatory cytokines interferon-gamma and tumor necrosis factor-alpha and the profibrotic cytokine transforming growth factor-beta. As infiltration of the kidney by T lymphocytes was a prominent feature of ANG II-dependent renal injury, we carried out experiments examining the effects of ANG II on lymphocytes in vitro. We find that exposure of splenic lymphocytes to ANG II causes prominent rearrangements of the actin cytoskeleton. These actions require the activity of Rho kinase. Thus, ANG II exaggerates hypertensive kidney injury by stimulating lymphocyte responses. These proinflammatory actions of ANG II seem to have a proclivity for inducing kidney injury while having negligible actions in the pathogenesis of cardiac hypertrophy.

Topics: Albuminuria; Angiotensin II; Animals; Cardiomegaly; Cell Proliferation; Cytoskeleton; Disease Models, Animal; Hypertension; Immunosuppressive Agents; Interferon-gamma; Kidney Diseases; Male; Mice; Mice, Knockout; Mycophenolic Acid; Sodium Chloride, Dietary; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Vasoconstrictor Agents

The goal of this study was to test the hypothesis that renal infiltration of immune cells in Dahl S rats on increased dietary sodium intake contributes to the progression of renal damage, decreases in renal hemodynamics, and development of hypertension. We specifically studied whether anti-immune therapy, using mycophenolate mofetil (MMF), could help prevent increases in renal NF-kappaB activation, renal infiltration of monocytes/macrophages, renal damage, decreases in glomerular filtration rate (GFR) and renal plasma flow, and increases in arterial pressure. Seventy-four 7-to 8-wk-old Dahl S, Rapp strain rats were maintained on an 8% Na, 8% Na + MMF (20 mg.kg(-1).day(-1)), 0.3% Na, or 0.3% Na + MMF diet for 5 wk. Arterial and venous catheters were implanted at day 21. By day 35, renal NF-kappaB in 8% Na rats was 47% higher than in 0.3% Na rats and renal NF-kappaB was 41% lower in 8% Na + MMF rats compared with the 8% Na group. MMF treatment significantly decreased renal monocyte/macrophage infiltration and renal damage and increased GFR and renal plasma flow. In high-NA Dahl S rats mean arterial pressure increased to 182 +/- 5 mmHg, and MMF reduced this arterial pressure to 124 +/- 3 mmHg. In summary, in Dahl S rats on high sodium intake, treatment with MMF decreases renal NF-kappaB and renal monocyte/macrophage infiltration and improves renal function, lessens renal injury, and decreases arterial pressure. This suggests that renal infiltration of immune cells is associated with increased arterial pressure and renal damage and decreasing GFR and renal plasma flow in Dahl salt-sensitive hypertension.

Topics: Animals; Blood Pressure; Disease Models, Animal; Dose-Response Relationship, Drug; Glomerular Filtration Rate; Glutathione; Heart Rate; Hypertension; Immunosuppressive Agents; Kidney; Kidney Diseases; Macrophages; Male; Mycophenolic Acid; NF-kappa B; Proteinuria; Rats; Rats, Inbred Dahl; Renal Circulation; Sodium Chloride, Dietary; Superoxide Dismutase; Superoxides; Time Factors; Vascular Resistance

Aim of this study was to prove the efficacy and safety of the new malononitrilamide immunosuppressive FK778 in prolonging clear graft survival following allogeneic orthotopic keratoplasty in rats.. Sixty-seven penetrating keratoplasties were performed using Fisher and Lewis rats as donors and recipients, respectively: group 1 (n=11), allogeneic control without therapy; group 2 (n=12), syngeneic control; group 3 (n=11), mycophenolate mofetil (MMF) 40 mg/kg bodyweight; group 4 (n=12), FK778 5 mg/kg bodyweight; group 5 (n=12), FK778 10 mg/kg bodyweight; and group 6 (n=9), FK778 20 mg/kg bodyweight. Four animals in each group were killed for immunohistological evaluation on day 14. Therapy was administered orally for 18 days. The grafts were evaluated every three days by means of a scoring system including opacity, oedema, and vascularization. Time to rejection was analysed with the Kaplan-Meier survival analysis and compared with the log-rank test. The densities of infiltrating immune cells were compared statistically using the non-parametric Mann-Whitney test.. Mean rejection-free graft survival was 11.4 days in group 1 (allogeneic control), 100 days (total follow-up time) in group 2 (syngeneic control), 24.0 days in group 3 (MMF 40 mg/kg), 15.7 days in group 4 (FK778 5 mg/kg), 19.1 days in group 5 (FK778 10 mg/kg), and 25.4 days in group 6 (FK778 20 mg/kg) (P<0.005).. Systemic immunosuppression with FK778 prolongs graft survival in the rat keratoplasty model. FK778's efficacy is comparable with that of MMF in preventing immunologic graft rejection.

Topics: Alkynes; Animals; Cornea; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Graft Rejection; Graft Survival; Immunosuppressive Agents; Isoxazoles; Keratoplasty, Penetrating; Mycophenolic Acid; Nitriles; Postoperative Care; Postoperative Period; Rats; Rats, Inbred F344; Rats, Inbred Lew; Survival Analysis; Treatment Outcome

Finding UDP-glucuronosyltransferases (UGT) require protein kinase C-mediated phosphorylation is important information that allows manipulation of this critical system. UGTs glucuronidate numerous aromatic-like chemicals derived from metabolites, diet, environment and, inadvertently, therapeutics to reduce toxicities. As UGTs are inactivated by downregulating PKCs with reversibly-acting dietary curcumin, we determined the impact of gastro-intestinal glucuronidation on free-drug uptake and efficacy using immunosuppressant, mycophenolic acid (MPA), in mice. Expressed in COS-1 cells, mouse GI-distributed Ugt1a1 glucuronidates curcumin and MPA and undergoes irreversibly and reversibly dephosphorylation by PKC-specific inhibitor calphostin-C and general-kinase inhibitor curcumin, respectively, with parallel effects on activity. Moreover, oral curcumin-administration to mice reversibly inhibited glucuronidation in GI-tissues. Finally, successive oral administration of curcumin and MPA to antigen-treated mice increased serum free MPA and immunosuppression up to 9-fold. Results indicate targeted inhibition of GI glucuronidation in mice markedly improved free-chemical uptake and efficacy using MPA as a model.

Topics: Animals; Chlorocebus aethiops; COS Cells; Curcumin; Disease Models, Animal; Drug Delivery Systems; Duodenitis; Glucuronosyltransferase; Immunosuppressive Agents; Mice; Mycophenolic Acid; Protein Kinase C; Treatment Outcome

Immunosuppressive therapies allow long-term patient and transplant survival, but are associated with increased development of UV-induced skin cancers, particularly squamous cell carcinomas. The mechanisms by which CsA, MMF, tacrolimus (TAC) or sirolimus (SRL), alone or in dual combinations, influence tumor development and progression are not completely understood. In the current study, chronically UV-exposed mice treated with SRL alone or in combination with CsA or TAC developed more tumors than mice treated with vehicle or other immunosuppressants, but the tumors were significantly smaller and less advanced. Mice treated with CsA or TAC developed significantly larger tumors than vehicle-treated mice, and a larger percentage in the CsA group were malignant. The addition of MMF to CsA, but not to TAC, significantly reduced tumor size. Immunosuppressant effects on UVB-induced inflammation and tumor angiogenesis may explain these findings. CsA enhanced both UVB-induced inflammation and tumor blood vessel density, while MMF reduced inflammation. Addition of MMF to CsA reduced tumor size and vascularity. SRL did not affect inflammation, but significantly reduced tumor vascularity. Thus the choice of immunosuppressants has important implications for tumor number, size and progression, likely due to the influence of immunosuppressants on UVB-induced inflammation and angiogenesis.

Topics: Animals; Blood Vessels; Carcinoma, Squamous Cell; Cyclosporine; Disease Models, Animal; Drug Therapy, Combination; Female; Immunosuppressive Agents; Inflammation; Mice; Mice, Hairless; Mycophenolic Acid; Neoplasms, Radiation-Induced; Neovascularization, Pathologic; Sirolimus; Skin Neoplasms; Tacrolimus; Ultraviolet Rays

Administration of the NO inhibitor N(wdelta)-nitro-L-arginine methyl ester (NAME) and a high-salt diet (HS) promotes severe albuminuria and renal injury, which regresses upon discontinuation of treatments.. We investigated whether these changes reappear after reinstitution of HS, and whether they are prevented by treatment with the antilymphocyte agent mycophenolate mofetil (MMF) or the AT-1 receptor blocker losartan (L). Adult male Munich-Wistar rats received NAME and HS. A control Group (C) received only HS. After 20 days, rats receiving HS and NAME exhibited severe hypertension and albuminuria. After a 30-day recovery period, hypertension was attenuated and albuminuria had virtually disappeared.. Rats were then distributed among the following groups: HS, receiving HS; NS, receiving a normal salt (NS) diet; HS-MMF, receiving HS and MMF; HS-LOS, receiving HS and L; HS-HDZ, receiving HS and hydralazine (HDZ). Sixty days later, NS rats showed only slight albuminuria and renal damage or inflammation. In contrast, HS rats developed severe hypertension, marked glomerulosclerosis with interstitial expansion and renal infiltration by macrophages and angiotensin II-positive cells. The group treated with losartan had lowered blood pressure and a lack of albuminuria or renal injury. MMF provided similar protection without altering blood pressure, suggesting a nonhemodynamic effect, a hypothesis reinforced by the finding that HDZ lowered blood pressure without preventing renal injury.. These results indicate that treatment with HS and NAME predisposes to the development of hypertension and renal injury upon salt overload, characterizing a new model of chronic nephropathy.. The response to MMF or L, but not HDZ, suggests a key role for inflammatory rather than hemodynamic factors.

Topics: Angiotensin II Type 1 Receptor Blockers; Animals; Antihypertensive Agents; Disease Models, Animal; Drug Evaluation, Preclinical; Hydralazine; Hypertension; Immunohistochemistry; Immunosuppressive Agents; Kidney; Kidney Failure, Chronic; Losartan; Male; Mycophenolic Acid; Nitric Oxide; Rats; Rats, Wistar; Sodium Chloride, Dietary

Here we compare the efficacy of cyclophosphamide (CYC) for treatment of crescentic nephritis (CGN) with the newer immunosuppressants 15-deoxyspergualin (DSG) and mycophenolate mofetil (MMF) in SCG/Kj mice, an inbred mouse strain that spontaneously develops CGN, systemic necrotizing vasculitis and antineutrophil cytoplasmic antibodies (ANCAs).. Mice were randomly assigned to intraperitoneal treatment with either DSG (2 mg/kg/day), CYC (50 mg/kg/week), MMF (60 or 100 mg/kg/day) or vehicle (VEH, dextrose 5% 0.3 ml/day) beginning at the 10th week of life. ANCA, blood urea nitrogen (BUN) and proteinuria were determined in all animals regularly, and survival was calculated. Renal histology was obtained in the 18th week of life in the MMF- or VEH-treated groups and in the 24th week in DSG- or CYC-treated animals.. Mean survival in VEH-treated animals was 123 days. At that point, survival was 100% in the CYC- or DSG-treated animals (P<0.001). Survival in the MMF group (pooled data) was not significantly different from the VEH-treated animals [MMF, 117 days (95% CI 108-127)]. BUN (18th week, CYC 43+/-9 mg/dl and DSG 36+/-6 mg/dl vs VEH 73+/-28 mg/dl, P<0.001, MMF 66+/-26 mg/dl), 24 h proteinuria (18th week, CYC 0.4+/-0.2 mg and DSG 0.7+/-0.6 mg vs VEH 2.7+/-3 mg, P<0.001, MMF 2.2+/-3 mg) crescent formation (18th week, VEH 42+/-9%, MMF 39+/-11%; CYC 5+/-2% and DSG 22+/-7% vs VEH, P<0.05), glomerular immune complex deposition, and ANCA formation were significantly improved in CYC- and DSG- but not in MMF-treated animals when compared with controls.. DSG and CYC, but not MMF, prolong life, limit renal damage and prevent autoantibody formation in SCG/Kj mice.

Topics: Animals; Biopsy, Needle; Cyclophosphamide; Disease Models, Animal; Guanidines; Immunohistochemistry; Immunosuppression Therapy; Immunosuppressive Agents; Mice; Mice, Inbred Strains; Mycophenolic Acid; Nephritis; Random Allocation; Risk Factors; Sensitivity and Specificity; Survival Rate

The most common complication after allogenic islet transplantation is rejection. This study was to evaluate the effect of anti-rejection of glucocorticoid-free immunosuppressive regimen on allogenic islet transplantation.. Tacrolimus (FK506) + mycophenolate mofetil (MMF) and FK506 + MMF + prednisone (Pred) were administered respectively for 2 weeks to inhibit rejection after allogenic islet transplantation in rats, which were compared with the control group. The concentrations of blood glucose, insulin and C-peptide were determined dynamically in recipients and the sites of transplantation were observed morphologically.. As compared with the control group without immunosuppressive agents, FK506 +MMF and FK506 + MMF + Pred could prolong the survival time of grafts significantly. There were many morphologically intact islets in the liver of recipients 2 months after transplantation. Group FK506 + MMF kept normal levels of blood glucose, insulin and C-peptide beyond 60 days after transplantation. In contrast, group FK506 + MMF + Pred secreted less C-peptide (P<0.05) and maintained a higher level of blood glucose concentration (P<0.01) after the operation. There was no significant difference in insulin concentrations between the two groups. The level of blood glucose beyond the first 2 weeks after drug withdrawal in group FK506 + MMF + Pred decreased obviously (P<0.05), and the secretion of insulin and C-peptide increased. These results were compared with those the first 2 weeks after transplantation and the first 2 weeks after drug withdrawal.. Both regimens of FK506 + MMF and FK506 + MMF + Pred could provide effective immunosuppression. Moreover the combined glucocorticoid-free immunosuppressive strategy of low-dose FK506 and MMF could protect islet grafts in islet transplantation without diabetogenic side-effects.

Topics: Animals; Blood Glucose; C-Peptide; Diabetes Mellitus; Disease Models, Animal; Drug Therapy, Combination; Female; Glucocorticoids; Graft Rejection; Immunosuppression Therapy; Immunosuppressive Agents; Insulin; Islets of Langerhans Transplantation; Male; Mycophenolic Acid; Prodrugs; Rats; Rats, Sprague-Dawley; Rats, Wistar; Tacrolimus; Transplantation, Homologous

Renal ischemia and hypertension have been suggested to be involved in the progression of renal diseases. Recently, we developed a model of accelerated major histocompatibility complex-independent renal injury, where high-renin hypertension aggravates functional and morphological changes induced by ischemia/reperfusion (I/R). In this model, we evaluated the effect of immunosuppressant mycophenolate mofetil (MMF) to test its capability to slow the progression of accelerated nephropathy.. 34 anesthetized uninephrectomized hypertensive transgenic (mREN2)27 rats (TGR) received a clamp on the renal pedicle for 45 min. Animals were treated with MMF 10 mg/kg/day (n = 10), 20 mg/kg/day (n = 10) or placebo (n = 10) orally via gavage for 12 weeks. Four animals were sham operated and not treated. Proteinuria and blood pressure were evaluated throughout the experiment. At the end of the experiment, kidney function was evaluated and kidneys harvested for morphological analysis and immunohistochemistry (CD4+, CD8+ lymphocytes and specific rat monocyte/macrophage marker ED-1+ cells).. At week 12, both MMF-treated groups had lower proteinuria as compared to the placebo group (MMF 10: 22.4 +/- 9.8, MMF 20: 20.9 +/- 5.6 vs. 126.7 +/- 35.8; p < 0.01; sham 28.1 +/- 1.4 mg/day) and reduced glomerulosclerosis (MMF 10: 11.4 +/- 7.8, MMF 20: 5.2 +/- 2.7 vs. 20.9 +/- 10.9; p < 0.05; sham 15.7 +/- 9.2%). There were no differences in systolic blood pressure among groups. MMF-treated rats had lower CD4+ (MMF 10: 61.2 +/- 46.4, MMF 20: 29.3 +/- 18.2 vs. 125.3 +/- 42.8; p < 0.01, sham 84.9 +/- 6.1 cells/field of view) and CD8+ (MMF 10: 13.7 +/- 10.2, MMF 20: 10.0 +/- 8.1 vs. 37.8 +/- 14.3; p < 0.01; sham: 31.8 +/- 7.6 cells/field of view) lymphocytes infiltration and ED-1 macrophages infiltration (MMF 10: 5.5 +/- 6.4, MMF 20: 2.5 +/- 2.8 vs. 16.7 +/- 4.1; p < 0.01; sham 12.2 +/- 4.6 cells/field of view) than placebo-treated rats.. Our results thus support the hypothesis about the key role of immune mechanisms in progression of chronic nephropathies.

Topics: Animals; Animals, Genetically Modified; Disease Models, Animal; Disease Progression; Hypertension, Renal; Immunosuppressive Agents; Kidney; Kidney Function Tests; Male; Mycophenolic Acid; Rats; Reperfusion Injury

The purpose of our study was to compare the effectiveness of immunosuppressive drugs on the prevention of allograft rejection in a murine model of low-risk and high-risk keratoplasty. The therapy included FK 506 (tacrolimus; 0.2 mg/kg), mycophenolate mofetil (30 mg/kg), aminoguanidine (0.1 g/kg), and combination of FK506 + mycophenolate mofetil or FK506 + aminoguanidine. The results obtained from the Gray's survival model stratified according to the type of subjects suggest that a major rejection risk reduction was achieved using FK506; good results also were obtained for mycophenolate mofetil. Although the point estimates of both the survival and relative risk of rejection suggest a deferred effect of the combination FK506 + mycophenolate mofetil, this finding did not prove statistically significant.

Topics: Animals; Corneal Diseases; Corneal Transplantation; Disease Models, Animal; Drug Combinations; Female; Graft Rejection; Graft Survival; Guanidines; Immunosuppressive Agents; Male; Mice; Mycophenolic Acid; Risk Factors; Tacrolimus; Transplantation, Homologous

Cytoprotective genes have shown to display potent anti-inflammatory and antiapoptotic functions in endothelial and smooth muscle cells. We investigated whether cytoprotective genes, especially A20, were involved in mycophenolate mofetil (MMF)'s ability to ameliorate transplant arteriosclerosis in an experimental chronic rejection model.. Sprague-Dawley rat renal grafts were orthotopically transplanted into Wistar rats following the procedure of Kamada with our modification. The recipients were divided into three oral treatment groups: (1) vehicle group (cyclosporine [CsA] 10 mg/kg.d x 10 d followed by vehicle), (2) CsA group (CsA 10 mg/kg.d x 10 d followed by CsA 6 mg/kg.d), (3) MMF group (converted from CsA 10 mg/kg.d x 10 d to MMF 20 mg/kg.d on day 11). At the same time points after transplantation, the rats were sacrificed to harvest the renal allografts. The expression of four cytoprotective genes, A20, heme oxygenase (HO)-1, Bcl-2, and Bcl-XL, was analyzed by quantitative reverse-transcriptase polymerase chain reaction and immunohistochemistry.. The four-cytoprotective genes were all detected in rat kidney allografts undergoing chronic allograft nephropathy. The expression of A20 in grafted kidneys was significantly higher in the MMF than in the CsA or the vehicle group (P < .01). There was no significant difference between the CsA and the MMF groups in the expression of HO-1, Bcl-2, and Bcl-XL.. We demonstrated that MMF improved the expression of A20 in rat kidney allografts undergoing chronic allograft nephropathy. The correlation between MMF and A20 provide an explanation for the mechanism by which MMF ameliorates transplant arteriosclerosis in an experimental animal model of chronic rejection.

Topics: Animals; bcl-X Protein; Chronic Disease; Cyclosporine; Disease Models, Animal; DNA Primers; Gene Expression Regulation; Graft Rejection; Heme Oxygenase-1; Kidney Transplantation; Male; Mycophenolic Acid; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

To assess the mechanism of gastrointestinal disorders by mycophenolate mofetil (MMF), the intestinal absorption and enterohepatic circulation of mycophenolic acid (MPA), an active metabolite of MMF, and its 7-O-glucuronide (MPAG) were investigated using rat intestinal loops and a linked-rat model. The stability of MPAG in the intestinal fluids, the toxicity of MPA and MPAG to intestinal mucosa, and biliary excretion of MPAG in rats with acute renal failure (ARF) were also characterized. MPA was rapidly and extensively absorbed from the rat intestine whereas MPAG was much less absorbable. When MPA was administered intravenously to bile-donor rats, 1.2% of dose was excreted in bile of receiver rats exclusively as MPAG during 4 h. MPAG was minimally deconjugated in the intestinal fluids. MPAG, but not MPA, significantly enhanced the release of lactate dehydrogenase from intestinal mucosa. When MPA was intravenously administered to ARF rats, the biliary excretion of MPAG significantly increased, compared with that in normal rats. These results demonstrated that MPAG accumulated in the intestinal lumen following biliary excretion and exerted some toxic effect on the intestinal mucosa. It was also suggested that enterohepatic circulation of MPAG under renal dysfunction increased the risk of gastrointestinal disorders due to MPAG.

Topics: Acute Kidney Injury; Animals; Bile; Blood Urea Nitrogen; Chromatography, High Pressure Liquid; Colon; Creatinine; Disease Models, Animal; Enterohepatic Circulation; Glucuronides; Injections, Intravenous; Intestinal Absorption; Intestinal Mucosa; Intestine, Small; L-Lactate Dehydrogenase; Male; Membrane Transport Proteins; Molecular Structure; Multidrug Resistance-Associated Protein 2; Multidrug Resistance-Associated Proteins; Mycophenolic Acid; Rats; Rats, Wistar

The authors have investigated whether cyclosporine decreases the serum concentration of mycophenolic acid, the active principle of the immunosuppressant mycophenolate mofetil, and increases that of the inactive metabolite 7-O-mycophenolic acid glucuronide by reducing their enterohepatic recirculation. Rats were treated daily with methylcellulose (1.66 mL/kg PO) plus 0.9% NaCl (6 mL/kg IP), mycophenolate mofetil (20 mg/kg PO) plus 0.9% NaCl (6 mL/kg IP), methylcellulose (1.66 mL/kg PO) plus cyclosporine (5 mg/kg IP), and mycophenolate mofetil (20 mg/kg PO) plus cyclosporine (5 mg/kg IP). After 14 days a bile fistula was installed to measure the biliary excretion of the immunosuppressants and their metabolites. After 90 minutes blood was taken to determine their concentrations in blood or serum by HPLC. Cyclosporine significantly decreased the serum concentration of mycophenolic acid by 39% and increased, not significantly, that of 7-O-mycophenolic acid glucuronide by 53%. The biliary excretion of 7-O-mycophenolic acid glucuronide was significantly reduced by cyclosporine by 57%, whereas that of mycophenolic acid was not affected. Mycophenolate mofetil did not show a significant effect on either the blood concentration or the biliary excretion of cyclosporine and its metabolites AM1, AM9, AM1c, and AM4N. Cyclosporine significantly decreased the serum concentration of active mycophenolate acid and increased, not significantly, the serum concentration of inactive 7-O-mycophenolic acid glucuronide, presumably by reducing the biliary excretion of this inactive metabolite.

Topics: Animals; Bile; Bile Acids and Salts; Biliary Fistula; Bilirubin; Body Weight; Cyclosporine; Disease Models, Animal; Drug Administration Schedule; Drug Therapy, Combination; Injections, Intraperitoneal; Male; Mycophenolic Acid; Organ Size; Rats; Rats, Wistar

Experimental autoimmune myocarditis (EAM) is characterized by the appearance of multinucleated giant cells. EAM leads to severe myocardial damage and is a useful model of human giant cell myocarditis. We investigated whether mycophenolate mofetil (MMF), which is a potent immunosuppressant, prevents the development of myocarditis in a rat EAM model, and focused on the role of osteopontin (OPN) in the pathogenesis of this disorder. Adult Lewis rats were immunized with porcine cardiac myosin to establish EAM. The early MMF treatment completely prevented the development of EAM, and the late MMF treatment was also effective even against established EAM. Echocardiogram demonstrated that left ventricular function was also improved by the treatment with MMF. Real-time RT-PCR analysis showed that both early and late MMF treatments significantly inhibited myocarditis-induced OPN mRNA expression in the heart. Immunohistochemistry revealed that OPN expression was prominent in the myocardium on day 14, whereas expression was observed in the infiltrated macrophages on day 21. Mycophenolic acid (MPA) did inhibit agonist-induced OPN expression in cultured cardiomyocytes. These results show the therapeutic potential of MMF for autoimmune myocarditis and provide new insights into the pathogenesis of this disease.

Topics: Animals; Animals, Newborn; Autoimmune Diseases; Body Weight; Cells, Cultured; Disease Models, Animal; Echocardiography; Heart Ventricles; Immunohistochemistry; Immunosuppressive Agents; Male; Mycophenolic Acid; Myocarditis; Myocardium; Myocytes, Cardiac; Organ Size; Osteopontin; Rats; Rats, Inbred Lew; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sialoglycoproteins; Time Factors

Cellophane wrapping of the kidneys (Page kidney) induces perinephrits and hypertension, assumed to be due to renal ischemia resulting from parenchymal compression by the fibrous hull surrounding the kidneys. We investigated if interstitial nephritis, rather than plasma angiotensin activity, played a role in the development of hypertension in the Page kidney model.. We followed for 7 weeks rats with bilateral cellophane wrapping of the kidneys that received 20 mg/kg/day of the immunosuppressive antiproliferative drug mycophenolate mofetil (MMF) (two-kidney wrap/MMF) (N = 10) or vehicle (two-kidney wrap) (N = 10), and sham-operated rats (N = 10).. The two-kidney wrap group had progressive increment in blood pressure, inflammatory damage occupying 25% to 50% of the renal tubulointerstitial region and increased number of angiotensin II-positive cells, angiotensin II content, and oxidative stress in the kidney. MMF treatment prevented the development of hypertension and renal inflammation without modifying the perinephritic hull or the increment it induced in the intrarenal pressure. The plasma levels of angiotensin II were similar in the two-kidney wrap group, the two-kidney wrap/MMF group and the sham-operated animals and unchanged from baseline, despite the blood pressure increase in the two-kidney wrap group.. Our results indicate that renal wrap hypertension is unrelated to plasma angiotensin II levels and related to the inflammatory damage caused by the external compression of the kidney.

Topics: Angiotensin II; Animals; Blood Pressure; Body Weight; Cellophane; Disease Models, Animal; Glutathione; Hypertension, Renal; Immunosuppressive Agents; Ischemia; Kidney; Kidney Function Tests; Male; Malondialdehyde; Mycophenolic Acid; Nephritis, Interstitial; Pressure; Rats; Rats, Sprague-Dawley

Tacrolimus (FK506)/mycophenolate mofetil (MMF)/prednisone combination immunosuppression therapy has been found to effectively prevent composite tissue allograft (CTA) rejection with minimal toxicity in a preclinical porcine model. These findings have been reproduced in 24 human hands transplanted in 18 patients. In CTAs containing bone, adequate bone quality and healing are essential for long-term functional success. The purpose of this study was to determine the effect FK506/MMF/prednisone immunotherapy has on bone quality and healing.. Forelimb CTA-flaps were transplanted in nine pigs. Recipient animals received FK506/MMF/prednisone therapy for 3 months. Bone quality was studied pre- and posttransplant by measuring acoustic velocity and density and by calculating elastic coefficients. Additional bone quality analyses were performed on unoperated limbs, and in bone grafts from two pigs that had autograft procedures performed. Bone healing was assessed using radiographic analysis.. Three animals were lost to immunosuppression-related complications before the endpoint of the study. The bone component of all six CTA-flaps showed normal healing. Although results of the bone density measurements were not significantly different when comparing pre- to posttransplant values, acoustic velocity and elastic coefficient measurements showed a significant decrease posttransplant indicating a decrease in bone quality.. FK506/MMF/prednisone combination therapy prevented rejection, did not adversely affect bone quality, and showed normal bone healing. The transplant procedure itself decreased bone quality more than the immunosuppression regimen did over the observation period in this study. Based on these findings, we conclude to prevent CTA failure it is important to monitor bone quality posttransplant.

Topics: Animals; Bone Density; Disease Models, Animal; Drug Therapy, Combination; Forelimb; Glucocorticoids; Graft Rejection; Graft Survival; Immunosuppressive Agents; Immunotherapy; Mycophenolic Acid; Prednisone; Radiography; Swine; Tacrolimus; Transplantation, Homologous; Ultrasonography; Wound Healing

To determine the effectiveness of treatment with immunosuppressive drugs and monoclonal antibodies (mAb) after penetrating keratoplasty in two different models of high risk mouse recipients.. Corneas were grafted orthotopically in mouse models of high risk recipients with either neovascularisation of the graft bed or presensitisation to graft donor antigens. Recipients were treated with mAb against CD4(+) or CD8(+) cells or against T cells, or were treated with cyclosporin A (CsA) or mycophenolate mofetil (MMF), or a combination of both drugs.. Control untreated recipients with neovascularised graft bed or presensitised to the graft donor antigens rejected corneal allografts in 12.5 (SD 2.3) and 9.9 (1.6) days, respectively. Treatment of graft recipients with a neovascularised graft bed with mAb anti-CD4 or anti-T cells, but not with mAb anti-CD8 or with immunosuppressive drugs, resulted in a significant prolongation of graft survival; 75% and 28.5%, respectively, of grafts survived for more than 45 days after grafting. However, none of the treatments were successful in presensitised recipients.. Treatment of high risk recipients with mAb anti-CD4 is more effective in preventing corneal allograft rejection than the treatment with mAb anti-CD8 or the immunosuppressive drugs MMF and CsA. However, the effectiveness of the treatment depends on the recipients' pretransplantation risk type.

Topics: Animals; Antibodies, Monoclonal; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cyclosporine; Disease Models, Animal; Female; Graft Rejection; Graft Survival; Immunosuppression Therapy; Immunosuppressive Agents; Keratoplasty, Penetrating; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mycophenolic Acid; Postoperative Care

The inability to reconstruct extensive and often life-threatening tracheal defects is a clinical dilemma. The objective of this study was to achieve microvascular revascularization and transplantation of long-segment circumferential tracheal allografts in a canine model. Fifteen mongrel dogs were randomly assigned to 5 treatment groups. Twelve dogs underwent an excision of an 8-cm tracheal segment followed by transplantation and microvascular revascularization of an 8-cm cervical trachea allograft. Group 1 (n = 4) was treated with 10 mg/kg per day of cyclosporin A (CsA) and 7.5 mg/kg per day of mycophenolate mofetil (MM). Group 2 (n = 4) was treated with 5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. Group 3 (n = 4) was treated with 2.5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. Group 4 (n = 2) underwent an autograft tracheal transplant and received postoperative 2.5 mg/kg perday of CsA and 7.5 mg/kg per day of MM. Group 5 (n = 1) did not undergo surgery, but received postoperative 2.5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. The animals were maintained for a duration of 30 days, during which time the graft was assessed by routine endoscopic examination and tracheal biopsies. Ex vivo, tracheal autografts were examined grossly for graft healing and microscopically for histologic architecture. The mean survival times were 13.25 days (group 1), 16 days (group 2), and 20 days (group 3). There was 1 early allograft failure secondary to microvascular thrombosis, and there were 4 delayed failures secondary to postoperative wound infections. Five dogs were euthanized before the end of the 30-day observation period because of failure to thrive or hypocalcemic tetany. None of the dogs in the study demonstrated endoscopic or histologic evidence of rejection before euthanasia. Postmortem examination of the surviving dogs demonstrated normal histologic architecture without evidence of rejection. For the first time, we have achieved allotransplantation of long tracheal segments based on the cranial thyroid artery and internal jugular vein. Minimal systemic immunosuppression appears to be associated with a higher survival rate and a lower complication rate.

Topics: Animals; Cyclosporine; Disease Models, Animal; Dogs; Dose-Response Relationship, Drug; Drug Administration Schedule; Graft Rejection; Graft Survival; Immunosuppressive Agents; Microscopy, Electron; Microsurgery; Mycophenolic Acid; Random Allocation; Trachea; Transplantation, Autologous; Transplantation, Homologous; Vascular Surgical Procedures

We and others have shown that mixed allogeneic chimerism induces donor-specific tolerance to composite tissue allografts across major histocompatibility complex barriers without the need for immunosuppression. However, a delay period between bone marrow transplantation and limb allotransplantation is required, making such protocols impractical for clinical application. This study eliminates this delay period in a rat hind limb allotransplantation model by performing mixed allogeneic chimerism induction and transplantation "simultaneously.". Group 1 included controls in which naïve Wistar Furth (WF) hosts received ACI hind limbs. Group 2 included (ACI-->WF) chimeras that received limbs from third-party donors (Fisher), and group 3 included chimeras that received irradiated (1,050 cGy) ACI limbs. In group 4, WF hosts conditioned with 950 cGy received irradiated (1,050 cGy) ACI limbs followed by infusion of 100 x 10(6) ACI T-cell-depleted bone marrow cells and immunotherapy (tacrolimus and mycophenolate mofetil) for 28 days. Group 5 animals received the same treatment as group 4 animals without immunotherapy.. The rats in groups 1 and 2 rejected their limbs within 10 days. Only one rat in group 4 survived to the end of the study. Groups 3 and 5 demonstrated long-term limb survival without rejection or graft-versus-host disease. High levels of donor chimerism (>80%) were achieved and maintained throughout the study. Mixed lymphocyte reaction assays in both groups revealed donor-specific hyporesponsiveness with vigorous third-party reactivity.. This study demonstrated that infusion of donor bone marrow cells into conditioned hosts immediately after limb transplantation results in stable mixed chimerism, robust tolerance, and reliable limb allograft survival.

Topics: Animals; Disease Models, Animal; Drug Therapy, Combination; Graft Survival; Hindlimb; Immunosuppressive Agents; Lymphocyte Depletion; Major Histocompatibility Complex; Male; Mycophenolic Acid; Rats; Rats, Inbred ACI; Rats, Inbred WF; T-Lymphocytes; Transplantation Chimera; Transplantation Conditioning; Transplantation, Homologous; Whole-Body Irradiation

Topics: Animals; Disease Models, Animal; Drug Therapy, Combination; Fingolimod Hydrochloride; Graft Survival; Heart Transplantation; Immunosuppressive Agents; Mycophenolic Acid; Propylene Glycols; Rats; Rats, Inbred Lew; Rats, Inbred Strains; Sphingosine; Time Factors; Transplantation, Homologous

Topics: Animals; Apoptosis; Cytokines; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Disease Models, Animal; Immunosuppressive Agents; Male; Mice; Mice, Inbred CBA; Mycophenolic Acid; Rats; Rats, Inbred Strains; Th1 Cells

Interstitial mononuclear cell infiltration is a feature of experimental models of salt-sensitive hypertension (SSHTN). Since several products of these cells are capable of modifying local vascular reactivity and sodium reabsorption, we investigated whether mycophenolate mofetil (MMF), a drug known to inhibit infiltration and proliferation of immune cells, would modify the SSHTN induced by angiotensin II (Ang II) infusion.. Sprague-Dawley rats received Ang II for two weeks using subcutaneous minipumps. A high-sodium (4% NaCl) diet was started on the third week and was maintained until the eighth week. MMF (30 mg/kg, N = 15), an immunosuppressive drug, or vehicle (N = 15) was given daily by gastric gavage during the initial three weeks. Sham-operated rats (N = 9) were used as controls. Body weight, blood pressure (tail-cuff plethysmography), and serum creatinine were determined weekly. Urinary malondialdehyde (MDA) excretion, renal histology, and immunohistology, including the presence of Ang II and superoxide-producing cells, were analyzed at the end of Ang II infusion and at eight weeks.. MMF treatment did not modify hypertension induced during exogenous Ang II infusion, but prevented the subsequent SSHTN. Tubulointerstitial injury resulting from Ang II infusion was significantly reduced by MMF treatment, as were proliferative activity, T-cell infiltration and activation (interleukin-2 receptor expression), superoxide-producing cells, and urinary MDA excretion. Ang II-producing cells were present in the renal tubulointerstitium of rats with SSHTN (60 +/- 30 Ang II-positive cells/mm(2) at 8 weeks) and were reduced by two thirds in the MMF-treated group. Forty percent of lymphocytes infiltrating the tubulointerstitium stained positive for Ang II. The expression of Ang II receptors in the kidney was unmodified.. SSHTN resulting from Ang II infusion is associated with infiltration and activation of immune cells that produce Ang II. MMF treatment reduces these features and prevents the development of SSHTN.

Topics: Angiotensin II; Animals; Anti-Inflammatory Agents, Non-Steroidal; Blood Pressure; Body Weight; Cell Division; Creatinine; Disease Models, Animal; Fibronectins; Hypertension, Renal; Kidney; Leukocytes, Mononuclear; Lipid Peroxidation; Male; Malondialdehyde; Mycophenolic Acid; Osteopontin; Rats; Rats, Sprague-Dawley; Receptor, Angiotensin, Type 1; Receptor, Angiotensin, Type 2; Receptors, Angiotensin; Sialoglycoproteins; Superoxides; Vasoconstrictor Agents

Approaches to the treatment of lupus nephritis include immunosuppressants associated with anti-inflammatory drugs, mainly steroids, which, however, cause major side effects. Mycophenolate mofetil (MMF) has been described as being less toxic than conventional immunosuppressants, and it was effective in preventing progressive nephritis in lupus mice. Our study evaluated the therapeutic effect of MMF in NZB/W F1 hybrid mice with established disease. We also examined the combination of MMF with a selective cyclooxygenase-2 (COX-2) inhibitor, DFU, based on previous findings of excessive renal production of COX-2-derived thromboxane A2 (TXA2) in lupus nephritis.. Four groups of NZB/W mice (N = 30 each group), starting at five months of age, were given daily by gavage the following: vehicle, MMF 60 mg/kg, DFU 3 mg/kg, or MMF + DFU. Fifteen mice for each group were used for the survival studies, and the remaining mice were sacrificed at nine months.. MMF or DFU alone partially delayed the onset of proteinuria compared with vehicle. Combined therapy was significantly (P < 0.05) more effective than single drugs. Animal survival was partially ameliorated by MMF and significantly improved by the drug combination in comparison with the vehicle (P = 0.005) and DFU alone (P < 0.03). At nine months, serum blood urea nitrogen (BUN) levels were lower in all of the treated groups than in the vehicle group. Renal damage was also limited, but to a greater extent in mice given the combined therapy. In untreated mice, renal COX-2 mRNA expression was up-regulated, and generation of TXB(2), the stable breakdown product of TXA(2), increased. DFU prevented the abnormal renal TXB(2) production, confirming the COX-2 origin of this eicosanoid, whereas renal 6-keto-PGF(1 alpha) and prostaglandin E(2) (PGE(2)) were not affected substantially.. These results offer a strong case for exploring the possibility that in humans MMF combined with COX-2 inhibitors has a role in the treatment options for lupus nephritis. This combined drug therapy may be at least as effective as steroids but without the obvious nephrotoxicity of the latter.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Antibodies, Antinuclear; Blood Urea Nitrogen; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Disease Models, Animal; Drug Therapy, Combination; Female; Furans; Gene Expression Regulation, Enzymologic; Immunosuppressive Agents; Isoenzymes; Kidney; Lupus Nephritis; Lymphocyte Count; Lymphocyte Subsets; Membrane Proteins; Mice; Mice, Inbred NZB; Mycophenolic Acid; Prostaglandin-Endoperoxide Synthases; Proteinuria; Spleen; Survival Rate; Thromboxane B2

GB virus B (GBV-B) is the closest relative of hepatitis C virus (HCV) and is an attractive surrogate model for HCV antiviral studies. GBV-B induces an acute, resolving hepatitis in tamarins. Utilizing primary cultures of tamarin hepatocytes, we have previously developed a tissue culture system that exhibits high levels of GBV-B replication. In this report, we have extended the utility of this system for testing antiviral compounds. Treatment with human interferon provided only a marginal antiviral effect, while poly(I-C) yielded >3 and 4 log units of reduction of cell-associated and secreted viral RNA, respectively. Interestingly, treatment of GBV-B-infected hepatocytes with ribavirin resulted in an approximately 4-log decrease in viral RNA levels. Guanosine blocked the antiviral effect of ribavirin, suggesting that inhibition of IMP dehydrogenase (IMPDH) and reduction of intracellular GTP levels were essential for the antiviral effect. However, mycophenolic acid, another IMPDH inhibitor, had no antiviral effect. Virions harvested from ribavirin-treated cultures exhibited a dramatically reduced specific infectivity. These data suggest that incorporation of ribavirin triphosphate induces error-prone replication with concomitant reduction in infectivity and that reduction of GTP pools may be required for incorporation of ribavirin triphosphate. In contrast to the in vitro studies, no significant reduction in viremia was observed in vivo following treatment of tamarins with ribavirin during acute infection with GBV-B. These findings are consistent with the observation that ribavirin monotherapy for HCV infection decreases liver disease without a significant reduction in viremia. Our data suggest that nucleoside analogues that induce error-prone replication could be an attractive approach for the treatment of HCV infection if administered at sufficient levels to result in efficient incorporation by the viral polymerase.

Topics: Animals; Antiviral Agents; Cells, Cultured; Disease Models, Animal; Flaviviridae; Guanosine; Hepatitis, Viral, Animal; Hepatocytes; Mycophenolic Acid; Polymerase Chain Reaction; Ribavirin; RNA, Viral; Saguinus; Virus Replication

The aim of this study was to compare the effects of cyclosporine, tacrolimus, mycophenolate mofetil and SDZ RAD on an animal model of transplant arteriosclerosis involving alloantigen dependent and independent mechanisms.

Topics: Animals; Aorta, Abdominal; Arteriosclerosis; Chronic Disease; Cyclosporine; Disease Models, Animal; Everolimus; Graft Rejection; Immunosuppressive Agents; Isoantigens; Male; Mycophenolic Acid; Rats; Rats, Inbred F344; Rats, Inbred Lew; Sirolimus; Tacrolimus

In order to assess the effects of mycophenolate mofetil (MMF) on the development of adriamycin-induced nephropathy, the development of this nephropathy in rats treated with MMF was compared to that in non-treated animals (group ADR + V) over 28 weeks. At weeks 8, 16 and 20, 24-h proteinuria of the treated group statistically differed from that of the non-treated group. However, no significant difference in proteinuria was observed thereafter between the groups. At the end of the experiment, there was no significant difference between both groups regarding the frequency of glomerular lesion (Group ADR + V: Md = 35%, P25 = 20%, P75 = 68%; Group ADR + MMF: Md = 27%, P25 = 9.5%, P75 = 54%); tubulointerstitial lesion index (Group ADR + V: Md = 7, P25 = 1.5, P75 = 9; Group ADR + MMF: Md = 8, P25 = 2, P75 = 9); glomerulosclerosis area (group ADR + V = 2779 microm2, P25 = 751.8 microm2, P75 = 3115 microm2; Group ADR + MMF = 1147 microm2, P25 = 3969.7 microm2, P75 = 1560 microm2); and, interstitial fibrosis area (Group ADR + V: Md = 218200 microm2, P25 = 78670 microm2, P75 = 282700 microm2 group ADR + MMF: Md = 136000, P25 = 25010, P75 = 255800 microm2). In conclusion, MMF caused a temporary reduction in proteinuria but did not change the severity of the renal lesion observed after 28 weeks.

Topics: Animals; Disease Models, Animal; Doxorubicin; Immunohistochemistry; Kidney; Kidney Function Tests; Male; Mycophenolic Acid; Nephritis, Interstitial; Probability; Rats; Rats, Wistar; Reference Values; Treatment Outcome

Immunosuppressive therapy has been shown to either improve or, more frequently, enhance the development of atherosclerosis. We tested the effect of mycophenolate mofetil (MMF), an inhibitor of nucleotide synthesis widely used in transplant therapy, in diet-induced atherosclerosis in the rabbit. Two groups (n=10 each) of New Zealand White (NZW) rabbits were fed a 1% cholesterol diet for 12 weeks. One group received MMF (CHOL+MMF group) by gastric gavage (30 mg/kg daily) and the other group (CHOL) received the same volume of saline by the same route. There were no differences in the serum cholesterol (mean values > or =30 mmol/l in both groups after 2 weeks) or in the triglyceride, blood sugar, total protein, and albumin serum levels and weight gain in both groups of animals. The cholesterol-fed untreated rabbits had atherosclerotic plaques covering 43.9.1+/-SD 16.40% of their thoracic aorta and 41.9+/-22. 59% of their abdominal aorta, while the MMF treated group had 18. 5+/-7.17% and 17.7+/-9.71%, respectively (P<0.01). The cholesterol content of the aorta (mg/g) in the cholesterol-fed untreated group was 4.61+/-SD 1.21 in the thoracic aorta and 4.54+/-2.07 in the abdominal aorta, whereas the MMF treated group had and 2.83+/-0.84 and 2.77+/-1.44, respectively (P<0.01). Infiltrating macrophages (RAM 11 positive cells/100 nuclei) in the intimal layer of the aorta were 58.4+/-SD26.16 in the CHOL group and 8.5+/-5.51 in the CHOL+MMF group: (P<0.001). CD18 positive cells/100 nuclei were 27.4+/-17.6 in the CHOL group and 5.3+/-3.82 in the CHOL+MMF group (P<0.01), and the intima/media ratio was 0.66+/-0.11 in the CHOL group and 0. 30+/-0.09 in the MMF treated rabbits (P<0.001). MMF also reduced proliferating smooth muscle cells (HHF35 positive) infiltrating between the macrophages. These results indicate that MMF ameliorates importantly the atherogenic potential of a high cholesterol diet and this effect is associated with a reduction in macrophage and foam cell infiltration and smooth muscle cell proliferation and infiltration. Since chronic treatment with this drug is given routinely in various clinical conditions with relatively minor side effects, consideration may be given to its use as adjuvant therapy in atherosclerotic cardiovascular disease.

Topics: Analysis of Variance; Animals; Aorta, Abdominal; Aorta, Thoracic; Arteriosclerosis; Cholesterol; Diet, Atherogenic; Disease Models, Animal; Enzyme Inhibitors; Immunohistochemistry; Male; Muscle, Smooth, Vascular; Mycophenolic Acid; Rabbits; Reference Values; Statistics, Nonparametric

Topics: Animals; Clinical Trials as Topic; Disease Models, Animal; Female; Humans; Immunosuppressive Agents; Lupus Nephritis; Male; Mycophenolic Acid; Prodrugs

Extensive renal ablation is associated with progressive sclerosis of the remnant kidney. Because lymphocytes and monocytes accumulate in the remnant kidney, it is likely that they play a role in the renal scarring. Therefore, we treated rats with 5/6 nephrectomy (5/6Nx) with mycophenolate mofetil (MMF), a drug that has an antiproliferative effect and that suppresses the expression of intercellular adhesion molecules.. Sprague-Dawley rats with 5/6Nx received MMF (30 mg. kg-1. day-1 by daily gastric gavage, N = 15) or vehicle (N = 16). Ten additional rats were sham operated. All rats were fed a 30% protein diet. Body weight, serum creatinine, and urinary protein excretion were determined weekly. Lipid peroxidation, as a measure of oxidative stress observed by urinary malondialdehyde determinations, was performed every two weeks. Histologic studies were done in the remnant kidney four weeks (9 rats from the vehicle-treated group, 7 rats from the MMF group, and 5 sham-operated rats) and eight weeks after surgery (the remaining rats). Glomerular volume, sclerosis in glomeruli (segmental and global) and interstitium (semiquantitative scale), infiltrating lymphocytes and macrophages (CD43- and ED1-positive cells), and expression of adhesion molecules (CD54, CD18, and CD11b) were analyzed.. MMF treatment prevented the progressive increment in serum creatinine and the proteinuria observed in the 5/6 nephrectomized rats during the eight weeks of observation (P < 0.01). Weight gain was comparable in the MMF-treated and sham-operated rats, whereas weight gain was decreased in untreated 5/6 nephrectomized rats. Excretion of malondialdehyde increased after surgery but returned sooner to control levels in the MMF-treated rats. Increments in glomerular size and mean arterial blood pressure induced by renal ablation were not modified by MMF treatment. Eight weeks after surgery, segmental sclerosis was present in 48.4 +/- 8.35% (+/- sd) glomeruli in the vehicle-treated group versus 25 +/- 10.5% in the MMF-treated group (P < 0.001). Interstitial fibrosis was reduced significantly with MMF treatment (P < 0.001). Infiltration with CD43- and ED1-positive cells in glomeruli and interstitium was two to five times lower in MMF-treated rats (P < 0.01). Expression of adhesion molecules CD18 and CD11b was similarly reduced.. MMF ameliorates the progressive renal damage in the remnant kidney after 5/6Nx. This effect is associated with a reduction in the infiltration of lymphocytes and monocytes, whereas glomerular hypertrophy and systemic hypertension are unchanged.

Topics: Animals; Blood Pressure; Cell Adhesion Molecules; Creatinine; Disease Models, Animal; Hypertension, Renal; Hypertrophy; Immunosuppressive Agents; Kidney Failure, Chronic; Kidney Glomerulus; Lymphocytes; Male; Monocytes; Mycophenolic Acid; Nephrectomy; Proteinuria; Rats; Rats, Sprague-Dawley

The aim of the present study was to evaluate the therapeutic effect of mycophenolate mofetil (MMF) on the course of disease in SLE-prone MRLlpr/lpr mice. Three-months-old mice displaying clinical symptoms of glomerulonephritis were given MMF (100 mg/kg per day) orally via the drinking water. Control mice received i.p. injections of cyclophosphamide (CYC) (1.8 mg/mouse per week) or saline. Survival, albuminuria and haematuria, immunoglobulin levels and anti-dsDNA antibodies in serum, frequencies of immunoglobulin-producing B lymphocytes and glomerular deposits of immunoglobulin and C3 were analysed. The results showed that MMF treatment significantly prolonged survival and reduced the occurrence of albuminuria and haematuria in MRLlpr/lpr mice. In addition, the number of immunoglobulin-producing B cells and serum levels of IgG and IgG anti-dsDNA antibodies were reduced after MMF and CYC treatment. MMF treatment significantly reduced the extent of deposition of C3 in glomeruli. We conclude that the reduced severity of glomerulonephritis following treatment of lupus-prone mice with MMF was as efficacious as that of CYC. These results warrant clinical trials of MMF in SLE patients with glomerulonephritis.

Topics: Alkylating Agents; Animals; Antibodies, Antinuclear; Cyclophosphamide; Disease Models, Animal; Enzyme Inhibitors; Female; Immunoglobulins; IMP Dehydrogenase; Lupus Erythematosus, Systemic; Lupus Nephritis; Lymphocyte Activation; Male; Mice; Mice, Inbred MRL lpr; Mycophenolic Acid; Spleen

Chronic renal diseases progress to organ insufficiency, which may require replacement therapy within one to three decades even independently of the type of initial insults. In the majority of cases, the degrees of proteinuria and interstitial leukocyte infiltration and scarring are strictly correlated with the rate of disease progression. This study tests the hypothesis that excess intrarenal protein traffic may cause lymphocyte-dependent interstitial injury that, while not fully controlled by antiproteinuric therapy, can be further inhibited by concomitant immunosuppression. A primarily nonimmune model was used to reproduce progressive renal disease due to a critical loss of nephron mass. Angiotensin-converting enzyme (ACE) inhibitor limited proteinuria, interstitial inflammation, MHC class II antigen expression, and severe lesions. Combined treatment with ACE inhibitor and a specific antilymphocyte agent, mycophenolate mofetil, dramatically attenuated macrophage and T cell infiltration, MHC-class II overexpression, dendritic cells, and all manifestations of the disease. Evidence of lymphocyte-mediated renal injury in the setting of excess protein traffic provides the basis for combining ACE inhibition and immunosuppression to halt progression of proteinuric kidney disease and minimize the need for dialysis or transplantation.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Blood Pressure; Disease Models, Animal; Immunosuppressive Agents; Kidney; Kidney Failure, Chronic; Lisinopril; Lymphocytes; Male; Mycophenolic Acid; Proteinuria; Rats; Rats, Sprague-Dawley

Effects on T lymphocyte mediated pathology, phenotypes, and functions in MRLlpr/lpr mice given mycophenolate mofetil (MMF) (100 mg/kg/day) via drinking water or controls given ip cyclophosphamide (CYC) injections (1.8 mg/mouse/week) or water were described. Both MMF and CYC treatment diminished kidney and large salivary gland perivascular cell infiltrates, reduced profoundly double-negative (DN) T cell frequencies, decreased total lymphocyte number in spleen, and increased in vitro proliferative response to Con A. IFN-gamma and IL-10 in supernatants from Con A stimulated spleen cells were augmented after MMF but not CYC treatment. MMF treatment increased whereas CYC reduced IL-12 in serum. Kidney expressions of IFN-gamma, IL-10, and IL-12 mRNA were unaffected by MMF but decreased by CYC. Our results demonstrate that MMF and CYC suppress perivascular T lymphocyte inflammation by reducing the DN T cell population and by amelioration of T cell function. The varying cytokine patterns suggest different mechanisms of the two drugs.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; B-Lymphocytes; Cell Count; Cell Division; Concanavalin A; Cyclophosphamide; Cytokines; Disease Models, Animal; Female; Hypersensitivity, Delayed; Immunophenotyping; Immunosuppressive Agents; Kidney; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred MRL lpr; Mycophenolic Acid; RNA, Messenger; Salivary Glands; Spleen; T-Lymphocytes; Vasculitis

Mycophenolate mofetil (MMF) is the morpholinoethyl ester of mycophenolic acid, which is its active metabolite. MMF is effective in prolonging survival of allografts and xenografts. However, little is known about the effects and the main mechanism of action of MMF in autoimmune diseases. In this study, the effect of MMF on the spontaneous disease progression in the MRL/lpr mouse model of lupus was examined. Eight-week-old MRL/lpr mice (n=18) were orally treated with MMF dissolved in a vehicle (90 mg/kg) once a day. Control animals received vehicle alone (n=17). The incidence of albuminuria (>300 microg/18 h) was significantly reduced by MMF treatment compared with vehicle-treated controls (cumulative incidence of albuminuria at 23 wk in MMF-treated mice; 22% versus 88% in controls; P=0.0001). The glomerulonephritis was histologically less severe in MMF-treated mice than in control mice (P=0.005). Furthermore, in immunofluorescence studies the amount of immunoglobulin and C3 deposits in the glomerular capillary wall was significantly less in MMF-treated mice (P < or = 0.002). Surprisingly, in vivo no clear-cut immune-modulating effects were observed because there were no differences between MMF-treated and control animals with regard to autoantibody formation. Also, spleen enlargement and numbers of CD3+, CD4+, and CD8+ T cells in spleen, lymph nodes, and peripheral blood were not different between both groups. Furthermore, no immunosuppressive properties of 90 mg/kg MMF were found in BALB/c mice on delayed-type hypersensitivity and primary antibody response to methylated bovine serum albumin. Interestingly, renal perfusion experiments revealed that binding of nucleosome/antinucleosome complexes to the glomerular basement membrane is decreased in MMF-treated mice compared with control mice. It is concluded that MMF suppresses the development of lupus glomerulonephritis and albuminuria in MRL/ lpr mice. The observed reduction of glomerular immunoglobulin deposits in MMF-treated mice and the renal perfusion studies indicate that MMF treatment leads to a decreased binding of immune complexes in the glomerular capillary wall in lupus nephritis.

Topics: Albuminuria; Animals; Antibodies, Monoclonal; Antigen-Antibody Complex; Autoantibodies; Cattle; Disease Models, Animal; Hypersensitivity, Delayed; Immunosuppressive Agents; In Vitro Techniques; Lupus Nephritis; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred MRL lpr; Mycophenolic Acid; Serum Albumin, Bovine; Spleen

The effect of mycophenolate mofetil (MMF) was examined in active Heymann nephritis (HN), an animal model of human membranous nephropathy. HN was induced in Lewis rats with Fx1A/complete Freund's adjuvant (CFA), and controls only received CFA. The induction of HN was prevented by MMF (30 mg/kg per d) from 0 to 4 wk after immunization. Proteinuria was not different in CFA controls up to 16 wk, and was significantly less than in untreated HN from 6 wk onward. Serum anti-Fx1A antibody (Ab) levels and glomerular Ig deposition were suppressed during therapy. The interstitial infiltrate of alphabetaTCR+, CD4+ and CD8+ T cells, natural killer cells, and macrophages (mphi) observed in untreated HN at 8 wk was absent from rats treated from 0 to 4 wk with MMF. Semiquantitative reverse transcription-PCR for renal mononuclear cell cytokine mRNA at 8 wk demonstrated that MMF from 0 to 4 wk prevented the increased expression of Th1 (interferon-gamma, lymphotoxin), Th2 (interleukin-4), and mphi (tumor necrosis factor-alpha) cytokines identified in untreated HN. In lymph node draining sites of immunization, MMF limited both enlargement and the increased proportion of CD3+, CD4+, and CD8+ T cells observed in untreated HN and CFA controls. MMF suppressed Th2 (interleukin-4) but not Th1 (interferon-gamma, lymphotoxin) cytokine mRNA expression in lymph nodes. MMF from 4 to 8, 6 to 12, or 10 to 14 wk did not prevent proteinuria, serum anti-Fx1A Ab, or glomerular IgG deposition when compared with untreated HN. This study showed that MMF from 0 to 4 wk prevented the induction of HN and was associated with preferential suppression of Th2 cytokines. This therapy may prove useful in human idiopathic membranous nephropathy.

Topics: Animals; Disease Models, Animal; Drug Evaluation, Preclinical; Glomerulonephritis; Glomerulonephritis, Membranous; Humans; Interferon-gamma; Interleukins; Kidney Cortex; Lymphokines; Male; Mycophenolic Acid; Proteinuria; Rats; Rats, Inbred Lew; RNA, Messenger; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

To examine the effects of the immunosuppressant, mycophenolate mofetil (MM), on autoimmunity, glomerulonephritis, and mortality in the female NZB x NZW F1 (B/W) mouse model of systemic lupus erythematosus (SLE).. The development of murine lupus was assessed during the lifespan of 10 female B/W mice given 200 mg/kg/day of MM compared to 10 female B/W mice given vehicle. At 6 week intervals, mice were examined for weight change, albuminuria, antibodies to DNA, and IgG immunoglobulin levels. Morbidity and mortality were assessed daily. In a parallel study, MM treated and control B/W mice were examined at 18 weeks of age for splenocyte phenotype and adhesion molecule expression, as well as antibody titers and in vitro cytokine production in response to immunization with dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH).. The administration of MM was well tolerated without apparent side effects. Weight gain in MM treated and control mice was identical through 36 weeks of age. In the treatment group, MM suppressed the development of albuminuria and anti-DNA antibodies compared to the control animals. There were no significant differences between groups in serum concentrations of total IgG. At 60 weeks of age survival in the MM treated group was 100% compared to 10% in the control group. MM did not alter the percentages of CD4, CD8, or IgM positive splenocytes; however, the percentage of CD4+ T lymphocytes expressing very late antigen 4 and intercellular adhesion molecule 1 was reduced. MM inhibited the antibody response to DNP-KLH immunization in vivo; however, in vitro cytokine production in response to KLH was not suppressed.. MM suppressed the development of autoimmunity and prolonged lifespan in the female B/W mouse model of SLE. Suppression of autoimmunity was achieved without obvious side effects or altered CD4:CD8 T cell ratios. MM may be a useful primary or adjunctive therapy in human SLE.

Topics: Albuminuria; Animals; Antibodies, Antinuclear; Autoimmunity; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Female; Fluorometry; Hemocyanins; Immunization; Immunoglobulin G; Immunosuppressive Agents; Kidney; Kidney Glomerulus; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Mice, Inbred Strains; Mycophenolic Acid; Spleen; Survival Analysis; Treatment Outcome

We demonstrate that the novel immunosuppressive agent mycophenolate mofetil (MMF), that has been approved for use in kidney transplant recipients, strongly potentiates the antiviral activity of acyclovir in murine models for herpesvirus infections. Hairless mice that were infected intracutaneously with herpes simplex virus type 1 were treated systemically with ACV (20 mg/kg per day) and topically with 5% MMF. Combined use of both drugs resulted in an almost complete protection, whereas single use of either compound had virtually no effect. When athymic-nude mice were infected with an ACV-resistant (ACVr)-thymidine kinase-deficient (TK-) HSV-2 strain, combined use of systemically administered ACV (100 mg/kg per day) and topically applied MMF (5%) protected 60% of the animals against the infection, whereas all mice treated with either drug alone succumbed. Since transplant recipients under MMF therapy may develop opportunistic herpesvirus infections, requiring treatment with acyclovir (or valaciclovir), our findings have important implications for the treatment of these herpesvirus infections.

Topics: Acyclovir; Animals; Antiviral Agents; Disease Models, Animal; Drug Synergism; Herpes Genitalis; Herpes Simplex; Herpesvirus 1, Human; Herpesvirus 2, Human; Humans; Mice; Mice, Nude; Mycophenolic Acid

Graft-versus-host disease (GVHD) remains the most significant obstacle to the use of allogeneic bone marrow transplantation as a treatment for leukemia and other hematological malignancies. Because current GVHD treatment regimens such as cyclosporine and methotrexate are only partially effective, there is a need for new immunosuppressive drugs for the treatment of this condition.. A recently developed immunosuppressive drug, mycophenolate mofetil (MM), was tested in a fully mismatched (C57BL/6 donors to BALB/c recipients) murine model of acute GVHD after bone marrow transplantation.. A dose regimen of 30 mg/kg/day given by oral gavage and begun at 1 day before transplant had no positive effect on survival and was found to retard the rate of marrow engraftment as measured by absolute blood neutrophil counts. In all subsequent experiments, treatment was begun on day 5 after transplant. Three different doses (30, 60, and 90 mg/kg/day) were tested, but no significant improvement in mean survival time (MST) was observed for the first two doses (P=0.412 and 0.100, respectively). The highest dose (90 mg/kg/day) reduced MST (P=0.059), and no further dose increases were attempted. MM in combination with cyclosporine also failed to improve MST compared with animals treated with cyclosporine alone or controls.. These results suggest that MM given orally is not effective in this murine model of GVHD and may not have a role in the treatment and prevention of acute GVHD arising from bone marrow transplantation in the clinical setting.

Topics: Animals; Bone Marrow Transplantation; Disease Models, Animal; Dose-Response Relationship, Drug; Graft vs Host Disease; Immunosuppressive Agents; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mycophenolic Acid; Survival Analysis; Time Factors

The effects of a topical preparation of mycophenolic acid on the experimental allergic contact dermatitis induced by dinitrofluorobenzene was investigated. Visual assessment of skin reactions showed significant efficacy of a topical preparation of mycophenolic acid. This efficacy appeared from the early stage and endured up to 3 days. Morphological changes in the epidermis and dermis layers of animals treated with a mycophenolic acid cream were moderate compared with that in animals treated with vehicle only. In particular, hyperkeratosis was strongly suppressed. Since suppression of inflammatory cell infiltration was also observed, this efficacy might reach to the epidermis and dermis layer.

Topics: Administration, Topical; Animals; Butadienes; Decanoic Acids; Dermatitis, Allergic Contact; Dinitrofluorobenzene; Disease Models, Animal; Epidermis; Ethanol; Fatty Alcohols; Female; Guinea Pigs; Hemiterpenes; Mycophenolic Acid; Myristates; Myristic Acids; Palmitates; Paraffin Embedding; Pentanes; Polyethylene Glycols; Polymers; Skin; Stearic Acids

The requirement for effective, minimally toxic immunosuppression remains a major obstacle to performing human composite tissue allotransplantation. The skin component of composite tissue (e.g., limb) allografts is especially antigenic, necessitating toxic immunosuppressant doses to prevent or reverse acute rejection. In previous experiments, RS-61443, an experimental mycophenolic acid ester that inhibits lymphocyte proliferation with minimal toxicity, prevented acute limb allograft rejection in rats for more than 8 months when started on the day of transplantation. In this study, the ability of RS-61443 to reverse established acute rejection was tested in a rat model of hindlimb allotransplantation. Brown-Norway donors and Fischer 344 recipients provided a MHC mismatch for orthotopic midfemur limb transplants that were performed with microsurgical repair of femoral vessels and sciatic nerve. Three groups were studied: untreated allografts (n = 6); allografts receiving RS-61443 at 30 mg/kg/day, started on postoperative day 7 (n = 11); and allografts receiving RS-61443 at 30 mg/kg/day, started on postoperative day 9 (n = 9). Skin and soft tissues were biopsied periodically to assess rejection. Untreated allografts had complete acute rejection within 12-13 days. Animals in both the 7- and 9-day groups developed moderate to severe rejection clinically and histologically before initiation of immunosuppressive therapy. In both groups, RS-61443 was able to reverse rejection completely in all animals from which biopsies were obtained at the time of death at 9-16 weeks after transplantation (P < 0.007). RS-61443 was highly effective as a primary immunosuppressant for reversing established acute rejection in rat hindlimb allografts.

Topics: Animals; Disease Models, Animal; Graft Rejection; Hindlimb; Immunosuppressive Agents; Mycophenolic Acid; Rats; Rats, Inbred BN; Rats, Inbred F344

Cultured tumor cell lines, tumor xenografts grown in athymic nude mice, and a murine experimental metastasis model were used to assess the in vitro and in vivo anti-tumor activity of the potent IMP dehydrogenase (IMPDH) inhibitor, mycophenolic acid (MPA), and its morpholinoethyl ester pro-drug, mycophenolate mofetil (MM). The growth of all the cell lines tested was inhibited by MPA in vitro, with EC50 values ranging from less than 0.1 microM to 3.9 microM. Mice were monitored for s.c. tumor outgrowth in the case of human tumor xenograft models or survival time for the murine experimental metastasis model. Treatment with MM p.o. was started 24 hr after tumor challenge or after tumors became palpable. Treatment of athymic nude mice bearing A3.01 (T-lymphoblast), Molt-4 (T-cell leukemia), CaPan-2 (pancreatic adenocarcinoma), CaLu-3 (non-small-cell lung adenocarcinoma), LS174T and T84 (colon adenocarcinoma), and Daudi (B-cell lymphoma) human tumor xenografts with MM significantly inhibited s.c. tumor growth. Treatment of BALB/c mice with MM after i.v. injection of murine RAW117-H10 lymphoma cells in an experimental metastasis assay resulted in increased survival time for treated animals. No significant inhibitory effect on s.c. tumor outgrowth was seen with MM treatment of SK-Hep-1, a human hepatic endothelioma, or Hep-3B, a liver adenocarcinoma, at any of the doses tested.

Topics: Animals; Antineoplastic Agents; Cell Division; Disease Models, Animal; Drug Screening Assays, Antitumor; Female; Humans; IMP Dehydrogenase; Leukemia; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Nude; Mycophenolic Acid; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms; Neoplasms, Experimental; Transplantation, Heterologous

Topics: Animals; Disease Models, Animal; Graft vs Host Disease; Immunosuppressive Agents; Intestine, Small; Mice; Mycophenolic Acid; Rats; Spleen

These studies with wild-type and mutant cells defective in IMP dehydrogenase and the previous data with the adenylosuccinate synthetase-deficient cell line suggest that among the clinical population with dominantly inherited hyperuricemia, patients with partial deficiencies in these enzymes exist. It is hoped that these pharmacogenetic cell culture models for overproduction hyperuricemia will lead to the initiation of a search for hyperuricemia patients with either of these deficiencies. If such patients are found it may be possible to design chemotherapeutic regimens by which effectors (inhibitors) of purine synthesis might ameliorate the overproduction of purines by the de novo pathway.

Topics: Animals; Cell Line; Disease Models, Animal; Gout; Humans; IMP Dehydrogenase; Ketone Oxidoreductases; Lymphoma; Mice; Mutation; Mycophenolic Acid; Ribonucleotides; Uric Acid