ABSTRACT
Direct reprogramming of differentiated cells to pluripotent stem cells has great potential to improve our understanding of developmental biology and disorders such as cancers, and has implications for regenerative medicine. In general, the effects of transcription factors (TFs) that are transduced into cells can be influenced by pre-existing transcriptional networks and epigenetic modifications. However, previous work has identified four key TFs, Oct4, Sox2, Klf4 and c-Myc, which can reprogram various differentiated cells to generate induced pluripotent stem cells. Here, we show that in the heart, the transduction of cardiac mesenchymal progenitors (CMPs) with Klf4 and c-Myc (KM) was sufficient to drive the differentiation of these cells into adipocytes without the use of adipogenic stimulation cocktail, that is, insulin, 3-isobutyl-1-methylxanthine (IBMX) and dexamethasone. KM-transduced CMPs exhibited a gradually increased expression of adipogenic-related genes, such as C/Ebpα, Pparγ and Fabp4, activation of the peroxisome proliferator-activated receptor (PPAR) signaling pathway, inactivation of the cell cycle-related pathway and formation of cytoplasmic lipid droplets within 10 days. In contrast, NIH3T3 fibroblasts, 3T3-L1 preadipocytes, and bone marrow-derived mesenchymal stem cells transduced with KM did not differentiate into adipocytes. Both in vitro and in vivo cardiac ischemia reperfusion injury models demonstrated that the expression of KM genes sharply increased following a reperfusion insult. These results suggest that ectopic adipose tissue formation in the heart following myocardial infarction results from CMPs that express KM following a stress response.
Subject(s)
Cell Differentiation , Kinesins/metabolism , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Myocardium/cytology , Proto-Oncogene Proteins c-myc/metabolism , 3T3-L1 Cells , Adipocytes/cytology , Animals , CCAAT-Enhancer-Binding Protein-alpha/genetics , CCAAT-Enhancer-Binding Protein-alpha/metabolism , Cells, Cultured , Fatty Acid-Binding Proteins/genetics , Fatty Acid-Binding Proteins/metabolism , Kinesins/genetics , Kruppel-Like Factor 4 , Mice , Mice, Inbred C57BL , Models, Biological , NIH 3T3 Cells , PPAR gamma/genetics , PPAR gamma/metabolism , Proto-Oncogene Proteins c-myc/genetics , Real-Time Polymerase Chain Reaction , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Signal Transduction/drug effects , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
BACKGROUND AND OBJECTIVE: Currently, cells for transplantation in regenerative medicine are derived from either autologous or allogeneic tissue. The former has the drawbacks that the quality of donor cells may depend on the condition of the patient, while the quantity of the cells may also be limited. To solve these problems, we investigated the potential of allogeneic cardiac mesenchymal progenitors (CMPs) derived from postmortem hearts, which may be immunologically privileged similar to bone marrow-derived mesenchymal progenitors. MATERIALS AND METHODS: We examined whether viable CMPs could be isolated from C57/B6 murine cardiac tissues harvested at 24 hours postmortem. After 2- to 3-week propagation with a high dose of basic fibroblast growth factor, we performed cellular characteristics analyses, which included proliferation and differentiation property flow cytometry and microarray analyses. RESULTS: Postmortem CMPs had a longer lag phase after seeding than CMPs obtained from living tissues, but otherwise had similar characteristics in all the analyses. In addition, global gene expression analysis by microarray showed that cells derived from postmortem and living tissues had similar characteristics. CONCLUSION: These results indicate that allogeneic postmortem CMPs have potential for cell transplantation because they circumvent the issue of both the quality and quantity of donor cells.
Subject(s)
Mesenchymal Stem Cells/physiology , Myocytes, Cardiac/physiology , Animals , Antigens, Surface/metabolism , Cell Differentiation , Cell Proliferation , Cell Separation , Cell Survival , Cells, Cultured , Fibroblast Growth Factor 2/pharmacology , Gene Expression Regulation , Genetic Markers , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/immunology , Mice, Inbred C57BL , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/immunology , Phenotype , Postmortem Changes , Time FactorsABSTRACT
Mitochondria play an essential role in eukaryotes, and mitochondrial dysfunction is implicated in several diseases. Therefore, intercellular mitochondrial transfer has been proposed as a mechanism for cell-based therapy. In addition, internalization of isolated mitochondria cells by simple coincubation was reported to improve mitochondrial function in the recipient cells. However, substantial evidence for internalization of isolated mitochondria is still lacking, and its precise mechanism remains elusive. We tested whether enriched mitochondria can be internalized into cultured human cells by simple coincubation using fluorescence microscopy and flow cytometry. Mitochondria were isolated from endometrial gland-derived mesenchymal cells (EMCs) or EMCs stably expressing mitochondrial-targeted red fluorescent protein (EMCs-DsRed-mito), and enriched by anti-mitochondrial antibody-conjugated microbeads. They were coincubated with isogeneic EMCs stably expressing green fluorescent protein (GFP). Live fluorescence imaging clearly showed that DsRed-labeled mitochondria accumulated in the cytoplasm of EMCs stably expressing GFP around the nucleus. Flow cytometry confirmed the presence of a distinct population of GFP and DsRed double-positive cells within the recipient cells. In addition, transfer efficiency depended on mitochondrial concentration, indicating that human cells may possess the inherent ability to internalize mitochondria. Therefore, this study supports the application of direct transfer of isogeneic mitochondria as a novel approach for the treatment of diseases associated with mitochondrial dysfunction.
Subject(s)
Endometrium/physiology , Mesoderm/physiology , Mitochondria/transplantation , Cell Line , Endometrium/cytology , Endometrium/metabolism , Female , Flow Cytometry , Green Fluorescent Proteins/biosynthesis , Green Fluorescent Proteins/genetics , Humans , Luminescent Proteins/biosynthesis , Luminescent Proteins/genetics , Mesoderm/cytology , Mesoderm/metabolism , Microscopy, Fluorescence , Mitochondria/metabolism , Mitochondria/ultrastructure , TransfectionABSTRACT
Introduction. Amniotic membrane contains a multipotential stem cell population and is expected to possess the machinery to regulate immunological reactions. We investigated the safety and efficacy of allogeneic amniotic membrane-derived mesenchymal stromal cell (AMSC) transplantation in a porcine model of chronic myocardial ischemia as a preclinical trial. Methods. Porcine AMSCs were isolated from amniotic membranes obtained by cesarean section just before delivery and were cultured to increase their numbers before transplantation. Chronic myocardial ischemia was induced by implantation of an ameroid constrictor around the left circumflex coronary artery. Four weeks after ischemia induction, nine swine were assigned to undergo either allogeneic AMSC transplantation or normal saline injection. Functional analysis was performed by echocardiography, and histological examinations were carried out by immunohistochemistry 4 weeks after AMSC transplantation. Results. Echocardiography demonstrated that left ventricular ejection fraction was significantly improved and left ventricular dilatation was well attenuated 4 weeks after AMSC transplantation. Histological assessment showed a significant reduction in percentage of fibrosis in the AMSC transplantation group. Injected allogeneic green fluorescent protein (GFP)-expressing AMSCs were identified in the immunocompetent host heart without the use of any immunosuppressants 4 weeks after transplantation. Immunohistochemistry revealed that GFP colocalized with cardiac troponin T and cardiac troponin I. Conclusions. We have demonstrated that allogeneic AMSC transplantation produced histological and functional improvement in the impaired myocardium in a porcine model of chronic myocardial ischemia. The transplanted allogeneic AMSCs survived without the use of any immunosuppressants and gained cardiac phenotype through either their transdifferentiation or cell fusion.
ABSTRACT
Coronary malperfusion due to acute type A aortic dissection (DAA) is a lethal complication. It is especially difficult to rescue the patients with left coronary malperfusion because of acute global myocardial infarction (AMI), even with successful surgical treatments, including the replacement of the ascending aorta and coronary artery bypass grafting (CABG). We review our experience and illustrate our approach to these critically ill patients. In addition, we classify the mechanism of malperfusion into 4 types based upon perioperative findings and discuss surgical management indivisually. From January 1990 to April 2005, a total of 260 patients were operated for DAA in our institution. Twenty (7.7%) patients, 11 men and 9 women were suffering from coronary malperfusion due to DAA. The mean age was 55 (range 28-72) years. The right coronary artery was involved in 9 patients, and the left in 11. All procedures such as graft replacement and CABG were done on an emergent or urgent basis. Hospital mortality rate of right coronary malperfusion was 22% (2/9 patients), and that related to left coronary malperfusion was 5/11 (45%). Assisting device was required in 9 cases, veno-arterial bypass (VAB) in 6 cases, left ventricular assist system (LVAS) in 1, left heart bypass (LHB) in 1, LHB+right heart bypass (RHB) in 1. We lost all patients using VAB. Only 3 patients supported with strong assist device survived. Aggressive myocardial resuscitation and early operation are the key factors in the management of these critically ill patients. But once severe myocardial infarction occurs, V-A bypass (percutaneous cardiopulmonary support) is useless in treating patients with DAA who develop severe heart failure. We recommend to implant stronger assist device including LVAS immediately before exacerbation of multiple organ failure. In conclusion, surgical management is not easy for emergency patients with DAA in association with myocardial ischemia. However, reasonable surgical results can be obtained with supplemental CABG and strong mechanical support of the left ventricle.
Subject(s)
Aortic Aneurysm, Thoracic/surgery , Aortic Dissection/surgery , Coronary Disease/surgery , Myocardial Infarction/surgery , Adult , Aged , Aortic Dissection/complications , Aortic Aneurysm, Thoracic/complications , Coronary Artery Bypass , Coronary Disease/etiology , Female , Heart Bypass, Left , Heart-Assist Devices , Humans , Male , Middle Aged , Myocardial Infarction/etiology , Prognosis , Treatment OutcomeABSTRACT
Catheter-mediated, percutaneous, transluminal delivery of naked plasmid DNA (pDNA) into myocardium may offer a valuable strategy to heart diseases. Here, we examined whether clinically available transthoracic direct current (DC) shock improves intracoronary naked DNA transfection into myocardium. Plasmid vector encoding the GL3 luciferase was infused retrogradely into the coronary veins of beagle dogs, whereas another pDNA solution was infused into the left coronary artery. During and after these procedures, the coronary venous sinus was occluded by balloon, and transthoracic DC shock of 200 J was applied immediately after the infusions. Without DC shock, no remarkable increase in luciferase activity was demonstrated in any part of the left ventricular myocardium. In the presence of DC pulsation, significant luciferase expression was detected in the regions that were supplied by left anterior descending coronary artery (LAD), whereas the gene expression in the right coronary artery (RCA) regions was much less drastic. X-gal (5-bromo-4-chloro-3-indolyl-beta-D-galactoside) staining of cardiac cross-sections also revealed regional expression of beta-galactosidase. Immunohistochemical examinations of heart cryosections revealed that cardiomyocytes in LAD regions successfully expressed transgene product. The present system may enable a new strategy for myocardial gene therapy, without any special device or technique other than cardiac catheterization and DC cardioversion that are generally performed in ordinary hospitals.
Subject(s)
DNA/administration & dosage , Genetic Therapy/methods , Heart Diseases/therapy , Myocardium/metabolism , Transfection/methods , Animals , Cardiac Catheterization , Coronary Vessels , Dogs , Electric Stimulation , Electrocardiography , Electromyography , Female , Gene Expression , Immunohistochemistry/methods , Luciferases/analysis , Luciferases/geneticsABSTRACT
Naked plasmid DNA (pDNA) vaccine expressing herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) was tested for protective activity against acute HSV-1 infection in mice. The pDNA was intravenously injected into Balb/c mice via their tail vein under high pressure, and the vaccination was performed two times at an interval of 7 days. The gB gene vaccination significantly protected the mice from subsequent intraperitoneal challenge with a lethal dose of HSV-1, which killed all the animals given control plasmid or saline. The protective activity was correlated with the dose of the plasmid inoculated, the survival rate reaching 83% in mice vaccinated with 5 microg of pDNA. The vaccinated mice were also protected from latent HSV infection. The immunized mice showed significant elevation in neutralizing antibody against HSV-1 as well as serum levels of interleukin-12 (IL-12) and interferon-gamma (IFN-gamma). When mice were immunized with 5 microg of an Epstein-Barr virus (EBV)-based plasmid vector harboring the gB, the cytotoxic T lymphocytes (CTLs) activity and proliferative response for HSV-1 were also induced. The results strongly suggest that intravenous immunization of naked pDNA may induce humoral and cellular immune responses against the virus, leading to a significant prophylactic outcome against HSV-1 infection in mice.
Subject(s)
Genetic Therapy/methods , Herpes Simplex/immunology , Herpes Simplex/therapy , Herpesvirus 1, Human , Vaccines, DNA/administration & dosage , Viral Envelope Proteins/genetics , Animals , Antibody Formation , Cytotoxicity Tests, Immunologic , Female , Genetic Therapy/adverse effects , Herpesvirus 4, Human/genetics , Immunity, Cellular , Injections, Intravenous , Interferon-gamma/blood , Interleukin-12/blood , Mice , Mice, Inbred BALB C , Vaccines, DNA/genetics , Virus LatencyABSTRACT
Galalpha1-3Gal (Gal) is the major epitope on pig tissues bound by human natural antibodies. Xenogeneic hematopoietic cell transplantation is being investigated to induce immunological tolerance to xenografts. We have investigated the level of Gal expression on pig hematopoietic cells. Cells were collected from pig fetal liver and bone marrow (BM), and also from adult BM and peripheral blood, before and after treatment with pig-specific hematopoietic growth factors. Fluorescent activated cell sorting (FACS) analysis was performed with the M86 monoclonal antibody (specific for Gal), lineage markers, and biotinylated stem cell factor (SCF) to detect c-kit expression. In fetal pig BM and liver, there was no significant difference in Gal expression between monocytes/macrophages (myeloid cells) and lymphocytes. In adult hematopoietic cells from all sources, Gal-positive subpopulations in T cells showed weak expression of Gal, whereas B cells demonstrated higher expression, and myeloid cells showed highest expression. Adult BM and mobilized peripheral blood progenitor cells contained small populations with very low or negligible expression of Gal. A very small population of c-kit-positive cells, indicating progenitor cells, were Gal-negative. The small Gal-negative population that exists in progenitor cells might explain why some pig colony forming units (CFU) can be resistant to human serum.
Subject(s)
Disaccharides/metabolism , Epitopes/biosynthesis , Hematopoietic Stem Cells/immunology , Stem Cell Transplantation , Transplantation, Heterologous/immunology , Animals , Antigens, CD1/immunology , Bone Marrow/embryology , Bone Marrow Cells/immunology , CD3 Complex/immunology , Fetus , Flow Cytometry , Immune Tolerance , Liver/embryology , Liver/immunology , Lymphocytes/immunology , Primates , SwineABSTRACT
Previous clinical results of left ventricular assist system (LVAS) therapy for cardiogenic shock due to acute myocardial infarction (AMI) are still unacceptable. Japanese LVAS was designed for left atrial inflow cannulation. However, to obtain higher initial LVAS flow and more decompression of left ventricular (LV) cavity and to avoid thromboembolic event, LV inflow cannulation is a preferable procedure. Therefore, LV inflow cannula for Japanese LVAS (Toyobo) was developed. We treated three AMI cases with Toyobo-LVAS using the new LV inflow cannula. All patients were in cardiogenic shock status since broad antero-septal AMI and treated with percutaneous cardio-pulmonary support before the LVAS installation. The LVAS was effective for recovery from cardiogenic shock status and multiple organ failure. Two patients died because of serious LVAS associated complications such as bleeding (case 1, 8 days) and cerebral thromboembolism (case 2, 45 days). One of them was assisted for 202 days and underwent second operation. Sixty days after removal of LVAS, the patient died due to sepsis. The technique of LV inflow cannulation is improved through our experiences. However, our result suggests that renovation of the regimen for anti-coagulation and anti-septic therapy are necessary.
Subject(s)
Heart-Assist Devices , Myocardial Infarction/complications , Shock, Cardiogenic/therapy , Aged , Humans , MaleABSTRACT
Thrombotic microangiopathy (TM) is a serious complication of bone marrow transplantation (BMT) that resembles thrombotic thrombocytopenic purpura (TTP). In attempting to achieve hematopoietic cell chimerism in the pig-to-baboon model, we have observed TM following infusion of high doses (>10(10) cells/kg) of porcine peripheral blood mobilized progenitor cells (PBPC) into baboons. We performed investigations to analyze the pathobiology of this TM and to test therapeutic interventions to ameliorate it. PBPC were obtained by leukapheresis of cytokine-stimulated swine. The initial observations were made in two baboons that underwent a non-myeloablative regimen (NMR) prior to PBPC transplantation (TX) (group 1). We then studied three experimental groups. Group 2 (n = 2) received NMR without PBPC TX. Group 3 (n = 2) received PBPC TX alone. Group 4 (n = 6) received NMR + PBPC TX combined with prostacyclin, low-dose heparin, methylprednisolone, and cyclosporine was replaced by anti-CD40L mAb in five cases. Baboons in groups 1 and 3 developed severe thrombocytopenia (<10,000/mm3), intravascular hemolysis with schistocytosis (>10/high powered field (hpf)), increase in plasma lactate dehydrogenase (LDH) (2500-9000 U/l), transient neurologic changes, renal insufficiency, and purpura. Autopsy on two baboons confirmed extensive platelet thrombi in the microcirculation, and, similar to clinical BMT-associated TM/TTP, no unusually large vWF multimers or changes in vWF protease activity were observed in the plasma of baboons with TM. In group 2, self-limited thrombocytopenia occurred for 10-15 days following NMR. Group 4 baboons developed thrombocytopenia (<20,000/mm3) rarely requiring platelet transfusion, minimal schistocytosis (<3/hpf), minor increase in LDH (<1000 U/l), with no clinical sequelae. We conclude that high-dose porcine PBPC infusion into baboons induces a microangiopathic state with vWF biochemical parameters resembling clinical BMT-associated TM/TTP and that administration of antithrombotic and anti-inflammatory agents can ameliorate this complication.
Subject(s)
Hematopoietic Stem Cell Transplantation/adverse effects , Purpura, Thrombotic Thrombocytopenic/etiology , Thrombosis/drug therapy , Transplantation, Heterologous/adverse effects , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/pharmacology , Blood Coagulation Factors/drug effects , Blood Coagulation Factors/metabolism , Drug Therapy, Combination , Fibrinolytic Agents/administration & dosage , Fibrinolytic Agents/pharmacology , Microcirculation/pathology , Models, Animal , Papio , Swine , Thrombocytopenia/blood , Thrombocytopenia/drug therapy , Thrombocytopenia/etiology , Thrombosis/blood , Thrombosis/etiology , Transplantation ConditioningABSTRACT
Bone marrow stromal cells are able to differentiate into adipogenic, chondrogenic, myogenic, osteogenic, and cardiomyogenic lineages, all of which are limited to a mesoderm-derived origin. In this study, we showed that neurons, which are of an ectoderm-origin, could be generated from marrow-derived stromal cells by specific inducers, fibronectin/ornithine coating, and neurosphere formation. The neurons generated from marrow stroma formed neurites, expressed neuron-specific markers and genes, and started to respond to depolarizing stimuli as functional mature neurons. Among stromal cells, isolated mature osteoblasts which had strong in vivo osteogenic activity could be efficiently converted into functional neurons. This transdifferentiation or meta-differentiation was enhanced by Noggin, an inhibitor of bone morphogenetic proteins, in comparison with 5-azacytidine, a demethylating agent capable of altering the gene expression pattern. Marrow stroma is therefore a potential source of cells for neural cell transplantation.
Subject(s)
Azacitidine/pharmacology , Bone Marrow Cells/physiology , Cell Differentiation/drug effects , Neurons/physiology , Osteoblasts/physiology , Proteins/pharmacology , Animals , Bone Morphogenetic Proteins/antagonists & inhibitors , Bone Morphogenetic Proteins/physiology , Carrier Proteins , Cell Lineage , Cells, Cultured , Female , Mice , Potassium Channels/physiology , Regeneration , Stromal Cells/physiologyABSTRACT
BACKGROUND: Efforts to achieve tolerance to transplanted pig organs in nonhuman primates by the induction of a state of mixed hematopoietic chimerism have been associated with disorders of coagulation and thrombosis. Activation of recipient vascular endothelium and platelets by porcine hematopoietic cells and/or activation of donor organ vascular endothelium and/or molecular differences between the species may play roles. Irradiation or drug therapy could possibly potentiate endothelial cell activation and/or injury. METHODS: We have investigated parameters of coagulation and platelet activation in nonhuman primates after (1) a regimen aimed at inducing mixed hematopoietic chimerism and tolerance (TIR that included total body irradiation, T cell depletion, and splenectomy; (2) pig bone marrow or pig peripheral blood mobilized progenitor cell transplantation (PCTx); and/or (3) pig organ transplantation (POTx). Five experimental groups were studied. Baboons were the recipient subjects in all groups except Group 1. Gp 1 Cynomolgus monkeys (n=6) underwent TIR + allotransplantation of hematopoietic cells and a kidney or heart or TIR + concordant xenotransplantation (using baboons as donors) of cells and a kidney; Gp 2 Baboons (n=4) underwent TIR with or without (+/-) autologous hematopoietic cell infusion; Gp 3 (n=12) PCTx+/-TIR; Gp 4 (n=5) POTx+/-TIR; Gp 5 (n=4) TIR + PCTx + POTx. Platelet counts, with plasma prothrombin time, partial thromboplastin time, fibrinogen levels, fibrin split products and/or D-dimer were measured. RESULTS: In the absence of a discordant (porcine) cellular or organ transplant (Groups 1 and 2), TIR resulted in transient thrombocytopenia only, in keeping with bone marrow depression from irradiation. PCTx alone (Group 3) was associated with the rapid development of a thrombotic thrombocytopenic (TTP)-like microangiopathic state, that persisted longer when PCTx was combined with TIR. POTx (+/-TIR) (Group 4) was associated with a gradual fall (over several days) in platelet counts and fibrinogen with disseminated intravascular coagulation (DIC); after graft excision, the DIC generally resolved. When TIR, PCTx and POTx were combined (Group 5), an initial TTP-like state was superseded by a consumptive picture of DIC within the first week, necessitating graft removal. CONCLUSIONS: Both PCTx and POTx lead to profound alterations in hemostasis and coagulation parameters that must be overcome if discordant xenotransplantation of hematopoietic cells and organs is to be fully successful. Disordered thromboregulation could exacerbate vascular damage and potentiate activation of coagulation pathways after exposure to xenogeneic cells or a vascularized xenograft.
Subject(s)
Blood Coagulation Disorders/etiology , Hematopoietic Stem Cell Transplantation , Organ Transplantation , Thrombosis/etiology , Transplantation, Heterologous , Animals , Blood Coagulation Disorders/complications , Female , Graft Rejection/complications , Graft Rejection/etiology , Humans , Macaca fascicularis , Male , Organ Transplantation/physiology , Papio , Swine , Thrombosis/complications , Transplantation Chimera , Transplantation Immunology/immunology , Transplantation ToleranceABSTRACT
BACKGROUND: Anti-Galalpha1-3Gal (Gal) antibodies (Gal Ab) contribute to the rejection of porcine organs transplanted into primates. Extracorporeal immunoadsorption (EIA) has been developed to eliminate Gal Ab from the circulation. METHODS: Between 1995 and 1999 we performed 320 EIAs in baboons using a COBE-Spectra apheresis unit incorporating a synthetic Gal immunoaffinity column. Three plasma volumes were immunoadsorbed on each occasion. The 221 consecutive EIAs performed in 41 immunosuppressed baboons between January 1997 and April 1999 form the basis of this review. Of these 41 baboons, 29 underwent a series of three or four EIAs at daily intervals, seven had multiple series of three EIAs, and the remainder underwent single or double EIAs. Serum Gal Ab levels were monitored by ELISA before and at intervals after the course of EIA. RESULTS: There were two fatal complications, one from a respiratory mishap (unrelated to the EIA) and one from persistent hypotension unresponsive to therapeutic interventions. Seven procedures (3%) were terminated early owing to technical difficulties and/or persistent hypotension. Mean pre-EIA Gal Ab levels in naive baboons were 33.1 microg/ml (IgM) and 14.5 microg/ml (IgG). Immediately after three consecutive EIAs, IgM was depleted by a mean of 97.3% and IgG by 99.4%. By 18 to 24 h later, Gal Ab was returning but depletion remained at 80.1% (IgM) and 84.7% (IgG). The subsequent rate of return of Gal Ab depended on the immunomodulatory protocol used. CONCLUSIONS: (1) With appropriate monitoring, EIA is an acceptably safe procedure, even in small (<10 kg) baboons. (2) Three consecutive EIAs are effective in removing >97% of Gal Ab. (3) In the majority of cases, return of Gal Ab begins within 24 h, irrespective of the immunomodulatory protocol.
Subject(s)
Disaccharides/immunology , Immunoglobulin G/blood , Immunoglobulin M/blood , Plasmapheresis/methods , Animals , Chromatography, Affinity , Enzyme-Linked Immunosorbent Assay , Epitopes/immunology , Extracorporeal Circulation , Graft Rejection/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Immunosorbent Techniques , Papio , Perfusion , Swine , Transplantation, Heterologous/immunologySubject(s)
Hematopoietic Stem Cell Transplantation , Metalloendopeptidases/metabolism , Transplantation, Heterologous/physiology , von Willebrand Factor/metabolism , ADAM Proteins , ADAMTS13 Protein , Animals , Granulocyte Colony-Stimulating Factor/pharmacology , Hematopoietic Stem Cell Mobilization , Humans , Immunosuppression Therapy/methods , Interleukin-3/pharmacology , L-Lactate Dehydrogenase/blood , Papio , Stem Cell Factor/pharmacology , Thrombocytopenia , Whole-Body IrradiationSubject(s)
Hematopoietic Stem Cell Mobilization , Hematopoietic Stem Cells/cytology , Interleukin-3/pharmacology , Stem Cell Factor/pharmacology , Animals , Antilymphocyte Serum/pharmacology , Hematopoietic Stem Cells/drug effects , Hematopoietic Stem Cells/immunology , Lymphocyte Depletion , Papio , Splenectomy , Swine , Thymus Gland/radiation effects , Whole-Body IrradiationSubject(s)
Immunosuppression Therapy/methods , Skin Transplantation/immunology , Thymus Gland/transplantation , Transplantation, Heterologous/immunology , Animals , Animals, Newborn , Fetal Tissue Transplantation/immunology , Histocompatibility Testing , Lymphocyte Depletion , Papio , Splenectomy , Swine , T-Lymphocytes/immunology , Thymectomy , Whole-Body IrradiationSubject(s)
Hematopoietic Stem Cell Transplantation , Immunosuppression Therapy/methods , Membrane Glycoproteins/immunology , Transplantation, Heterologous/immunology , Animals , Antibody Formation , CD40 Antigens/immunology , CD40 Ligand , Complement Inactivator Proteins/pharmacology , Cyclosporine/therapeutic use , Elapid Venoms/pharmacology , Immunity, Cellular , Leukapheresis , Lymphocyte Culture Test, Mixed , Lymphocyte Depletion , Mycophenolic Acid/analogs & derivatives , Mycophenolic Acid/pharmacology , Papio , Splenectomy , Swine , T-Lymphocytes/immunology , Whole-Body IrradiationSubject(s)
Hematopoietic Stem Cell Transplantation , Immunosuppression Therapy/methods , Transplantation Chimera , Transplantation, Heterologous/immunology , Animals , Antibodies, Monoclonal/pharmacology , CD40 Antigens/immunology , CD40 Ligand , Complement Inactivator Proteins/pharmacology , Elapid Venoms/pharmacology , Immunoglobulin G/analysis , Immunosuppressive Agents/pharmacology , Interleukin-3/pharmacology , Lymphocyte Depletion , Membrane Glycoproteins/immunology , Papio , Stem Cell Factor/pharmacology , Swine , T-Lymphocytes/immunology , Whole-Body IrradiationABSTRACT
Advances in molecular biology and in techniques of gene transfer have resulted in the development of practical approaches to human gene therapy. Many applications are of relevance to manipulation of the immune system and have potential in organ and cell transplantation. For example, gene therapy approaches may facilitate the induction of immunological tolerance to a donor organ or protect it locally against the host's immune response. Based on a comprehensive review of the world literature, examples of current research efforts in both allogeneic and xenogeneic transplantation are presented and discussed.