RESUMEN
The ratio of von Willebrand factor (VWF) to FVIII differs among available VWF/FVIII concentrates. Repeated infusions of concentrates with a low VWF:FVIII ratio may expose patients with von Willebrand disease to supranormal FVIII levels. The aim of this study was to determine the effects of repeated infusions with two VWF/FVIII concentrates differing in VWF:FVIII ratio on attained FVIII trough and peak levels as well as other pharmacokinetic parameters. Rabbits were randomized to receive multiple 150 IU kg⻹ VWF:RCo infusions at 4 h intervals with VWF/FVIII concentrates of a high (Haemate® P/Humate-P®) or low (Wilate®) VWF:FVIII ratio. Trough plasma FVIII and VWF levels were measured after each infusion. Pharmacokinetic analysis was performed using samples collected frequently after infusion. Mean FVIII trough level after the first Wilate infusion was 50.6 IU dL⻹ with a 95% confidence interval (CI) of 43.1-58.2 IU dL⻹, compared with 31.8 IU dL⻹ (CI, 24.4-39.1 IU dL⻹) for Haemate P (P<0.001). Trough levels progressively increased over the 24 h treatment period in both groups. After the final infusion, mean trough FVIII remained significantly higher (P = 0.002) in recipients of Wilate. Mean peak FVIII concentration after infusion was 67% higher in the Wilate group (167 vs. 100 IU dL⻹ , respectively; P = 0.002). Mean cumulative exposure to FVIII, as measured by area under the curve, was 84% greater in Wilate-treated animals. Half-life did not differ between the two concentrates. Animal model data suggest that exposure to elevated FVIII levels can be reduced through use of VWF/FVIII concentrates with higher VWF:FVIII ratios.
Asunto(s)
Coagulantes/administración & dosificación , Coagulantes/farmacocinética , Factor VIII/administración & dosificación , Factor VIII/farmacocinética , Factor de von Willebrand/administración & dosificación , Factor de von Willebrand/farmacocinética , Animales , Área Bajo la Curva , Combinación de Medicamentos , Femenino , Semivida , Infusiones Intravenosas , Modelos Animales , ConejosRESUMEN
BACKGROUND AND OBJECTIVES: In fetal alloimmune thrombocytopenia (FAIT), transplacental maternal antibodies cause destruction of fetal platelets. FAIT is similar to fetal Rhesus haemolytic disease, but half of the affected fetuses are born to primiparous women. In 10-20% of cases, prenatal and perinatal intracranial haemorrhages are reported. Different therapeutic approaches have been described, including maternally administered high-dose intravenous immunoglobulin (high dose IVIG) without or with steroids or intrauterine transfusion (IUT) of compatible platelets. For the latter, the use of plasma-free maternal and donor platelets has been described, but a comparison of these two sources of platelets has not been reported. MATERIALS AND METHODS: We retrospectively analyzed the clinical courses of cases with FAIT treated with IUT of either HPA-matched donor platelets or maternal platelets, done by a single team between 1990 and 1997. In 57 pregnancies, FAIT was treated by repeated IUT with either maternal (15 fetuses) or donor platelets (42 fetuses). RESULTS: There was no procedure-related fetal or neonatal loss. Platelets from both sources reliably raised the fetal platelet counts. Donor platelet preparations contained more platelets and yielded higher fetal post-transfusion platelet counts, but maternal platelets were clinically equally effective. CONCLUSIONS: Donor and maternal platelet concentrates are effective sources for the treatment of FAIT.
Asunto(s)
Antígenos de Plaqueta Humana/inmunología , Enfermedades Fetales/terapia , Histocompatibilidad Materno-Fetal/inmunología , Transfusión de Plaquetas/métodos , Trombocitopenia Neonatal Aloinmune/terapia , Adulto , Donantes de Sangre , Transfusión de Sangre Intrauterina , Cesárea , Femenino , Enfermedades Fetales/inmunología , Edad Gestacional , Hematoma/etiología , Humanos , Recién Nacido , Isoanticuerpos/sangre , Masculino , Madres , Embarazo , Púrpura/etiología , Estudios Retrospectivos , Trombocitopenia Neonatal Aloinmune/inmunologíaRESUMEN
Chronic lymphocytic leukemia (CLL) is a low-grade B-cell non-Hodgkin lymphoma. Skin involvement is observed in up to 25% of patients. A 70-year-old men with presented with prominent erythematous facial infiltrates. After the diagnosis of CLL was established, he showed a good hematologic response to the monoclonal antibody rituximab, but the skin infiltrates were therapy-resistant.
Asunto(s)
Resistencia a Medicamentos , Leucemia Linfocítica Crónica de Células B/patología , Infiltración Leucémica/tratamiento farmacológico , Piel/patología , Anciano , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Monoclonales de Origen Murino , Antineoplásicos/administración & dosificación , Antineoplásicos/uso terapéutico , Cara/patología , Humanos , Interferón-alfa/administración & dosificación , Interferón-alfa/uso terapéutico , Leucemia Linfocítica Crónica de Células B/clasificación , Leucemia Linfocítica Crónica de Células B/diagnóstico , Leucemia Linfocítica Crónica de Células B/tratamiento farmacológico , Infiltración Leucémica/patología , Masculino , Rituximab , Factores de TiempoRESUMEN
The association of cancer with thromboembolic events has been established for more than hundred years. While the thrombophilic diathesis of tumor patients and the neoplastic thrombogenesis have been elucidated pathophysiologically, at least in part, there is growing experimental and clinical evidence that factors of plasmic hemostasis promote tumor growth, angiogenesis, and metastasis. Thus, the rationale for antithrombotic prophylaxis and therapy of tumor patients might not only be based on the prevention of thromboembolic complications but also on the potentially antineoplastic and antimetastatic effects of pharmacological anticoagulation. Encouraging results of clinical studies indicate that prophylactic and therapeutic anticoagulation might improve the survival of selected patients with cancer.
Asunto(s)
Anticoagulantes/uso terapéutico , Neoplasias/tratamiento farmacológico , Tromboembolia/etiología , Humanos , Neoplasias/sangre , Neoplasias/complicaciones , Tromboembolia/tratamiento farmacológico , Tromboembolia/prevención & controlRESUMEN
To improve tissue regeneration of ischemic myocardium, autologous bone marrow-derived stem cells have been injected intramyocardially in five patients undergoing coronary artery bypass grafting and transmyocardial laser revascularization. An innovative method for the intraoperative isolation of CD133(+)-stem cells in less than 3 hours has been established. After induction of general anesthesia, approx. 60-240 ml of bone marrow were harvested from the posterior iliac crest and processed in the operating room under GMP conditions using the automated cell selection device Clini-MACS. Following standard CABG surgery, LASER channels were shot in predefined areas within the hibernating myocardium. Subsequently, autologous CD133(+)-stem cells (1.9-9.7 x 10(6) cells; purity up to 97%) were injected in a predefined pattern around the laser channels. Through the intraoperative isolation of CD133(+)-cells, this effective treatment of ischemic myocardium can be applied to patients scheduled both for elective and for emergency revascularisation procedures.
Asunto(s)
Células de la Médula Ósea/citología , Separación Celular/métodos , Glicoproteínas/análisis , Péptidos/análisis , Células Madre/citología , Antígeno AC133 , Antígenos CD , Humanos , Periodo Intraoperatorio , Células Madre/química , Factores de TiempoAsunto(s)
Enfermedades de von Willebrand , Humanos , Prevalencia , Enfermedades de von Willebrand/diagnóstico , Enfermedades de von Willebrand/epidemiología , Enfermedades de von Willebrand/etiología , Enfermedades de von Willebrand/terapia , Factor de von Willebrand/biosíntesis , Factor de von Willebrand/metabolismoRESUMEN
HISTORY: A 62-year-old man was admitted to hospital because of rectal bleeding of unknown cause. INVESTIGATIONS: Coloscopy showed a rectal tumour as cause of the bleeding and concomitant hypochromic microcytic anaemia. Histological examination established the diagnosis of anorectal malignant melanoma. Further staging examinations did not reveal metastatic disease. TREATMENT AND COURSE: An radical abdominoperineal rectal resection amputation was performed. The postsurgical course was uneventful. However, multiple lung metastases were detected 6 weeks later. CONCLUSION: Anorectal malignant melanoma is a rare cause in the differential diagnosis of anorectal tumours. Surgery remains the therapy of choice. Chemotherapy and immunotherapy are principally used in a palliative setting. Despite advances in therapy, the prognosis of this tumour entity remains unfavourable.
Asunto(s)
Melanoma/diagnóstico , Neoplasias del Recto/diagnóstico , Anemia Hipocrómica/etiología , Colonoscopía , Diagnóstico Diferencial , Hemorragia Gastrointestinal/etiología , Humanos , Neoplasias Pulmonares/secundario , Masculino , Melanoma/secundario , Melanoma/cirugía , Persona de Mediana Edad , Pronóstico , Neoplasias del Recto/patología , Neoplasias del Recto/cirugía , Recto/cirugíaRESUMEN
HISTORY AND CLINICAL FINDINGS: A 21-year-old patient was admitted to the hospital because of massively enlarged cervical lymph nodes. Additionally, a left-sided facial and brachial edema was visible. Auscultation of the left lung was remarkable for diminished breath sounds. EXAMINATIONS: Diagnostic imaging showed an extensive thoracic tumor and enlarged mediastinal and cervical lymph nodes. The diagnosis of neuroblastoma was established by biopsy. TREATMENT AND CLINICAL COURSE: The patient was treated with a polychemotherapy protocol according to the pediatric neuroblastoma study NB97. Subsequently, the patient underwent partial tumor resection, received two further chemotherapy courses and irradiation of the remaining tumor. Because of residual vital tumor cells, a second surgical tumour reduction followed by high-dose chemotherapy with autologous stem-cell support was performed. Two cycles of high-dose retinoic acid followed. Six months after the end of therapy, the patient is in a good condition despite of the presence of residual tumor. CONCLUSION: Neuroblastoma is a very rare tumor in adult patients. Therapy is multimodal and should follow pediatric guidelines for neuroblastoma treatment.
Asunto(s)
Neuroblastoma/diagnóstico , Neuroblastoma/terapia , Neoplasias Torácicas/diagnóstico , Neoplasias Torácicas/terapia , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Quimioterapia Adyuvante , Terapia Combinada , Diagnóstico Diferencial , Humanos , Imagen por Resonancia Magnética , Masculino , Neuroblastoma/cirugía , Radioterapia Adyuvante , Reoperación , Trasplante de Células Madre , Neoplasias Torácicas/cirugía , Trasplante Autólogo , Tretinoina/administración & dosificaciónRESUMEN
We report 2 cases in which patients with coronary heart disease not amenable for conventional revascularization underwent transmyocardial laser revascularization (TMLR) and implantation of AC133+ bone-marrow stem cells. The reason for using TMLR in combination with stem cell application is to take advantage of the synergistic angiogenic effect. The local inflammatory reaction induced by TMLR should serve as an informational platform for stem cells and may trigger their angiogenic differentiation. Functional analysis of myocardial performance after treatment in these 2 cases revealed dramatic improvement of the wall motion at the site of the TMLR and stem cell application. Because TMLR does not enhance myocardial contractility and there was no angiographic evidence of major collaterals to the ischemic region in either patient, we assume that the synergistic effect of stem cells and TMLR-induced angiogenesis occurred; however, our assumption is of a speculative nature. We think that TMLR in combination with stem cell transplantation might become a novel revascularization therapy for ischemic myocardium.
Asunto(s)
Trasplante de Médula Ósea/métodos , Enfermedad Coronaria/cirugía , Trasplante de Células Madre Hematopoyéticas/métodos , Terapia por Láser/métodos , Revascularización Miocárdica/métodos , Anciano , Terapia Combinada , Humanos , Trasplante Autólogo/métodos , Resultado del TratamientoRESUMEN
Recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood progenitor cells (PBPCs) from healthy individuals are a rapidly emerging alternative source to bone marrow for allogeneic transplantation. Although widely applied in the meantime, only limited information on feasibility and safety of mobilization and collection of PBPCs is currently available from prospective multicenter studies specifically designed to investigate this donation modality. This ongoing multicenter study on the performance as well as the short- and long-term safety profile of rhG-CSF-induced mobilization and collection of PBPCs was initiated in October 1999. The study is designed to recruit a total of 300 healthy family donors who will be followed regularly for a period of 5 years after donation. The first interim report presented here summarizes results obtained after enrollment of 150 donors from nine German institutions. The study protocol allowed the individual choice between two dose regimens of rh-CSF (10 micro g/kg per day vs 2x8 micro g/kg per day of donor body weight). The primary endpoint was defined as a yield of > or =5x10(6) CD34(+) cells/kg of recipient body weight in a single leukapheresis product. This endpoint was attained by 50% of donors receiving the lower rhG-CSF dose regimen and by 75% of donors with the higher dose regimen ( p<0.0009). A total of 478 acute adverse events attributable to the mobilization procedure were recorded and manifested predominantly as transient bone pain and headaches (80%). No persistent hematologic or nonhematologic adverse events have occurred in this study so far. Thus, the current experience in a prospective multicenter study supports previous single-center and retrospective registry reports in that the collection of PBPCs after rhG-CSF mobilization is feasible and associated with frequent, but generally mild and acceptable side effects.
Asunto(s)
Donantes de Sangre , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Movilización de Célula Madre Hematopoyética/efectos adversos , Leucaféresis/normas , Adolescente , Adulto , Antígenos CD34/análisis , Recuento de Células Sanguíneas , Femenino , Filgrastim , Movilización de Célula Madre Hematopoyética/métodos , Humanos , Sistema Inmunológico , Inmunofenotipificación , Subgrupos Linfocitarios , Masculino , Persona de Mediana Edad , Núcleo Familiar , Estudios Prospectivos , Proteínas RecombinantesAsunto(s)
Antimetabolitos Antineoplásicos/efectos adversos , Ataxia Cerebelosa/inducido químicamente , Fluorouracilo/efectos adversos , Enfermedades del Sistema Nervioso/inducido químicamente , Sistema Nervioso/efectos de los fármacos , Humanos , Leucoencefalopatía Multifocal Progresiva/inducido químicamenteAsunto(s)
Plasmacitoma/diagnóstico , Neoplasias de la Médula Ósea/diagnóstico , Neoplasias de la Médula Ósea/patología , Neoplasias de la Médula Ósea/terapia , Neoplasias Óseas/diagnóstico , Neoplasias Óseas/patología , Neoplasias Óseas/terapia , Diagnóstico Diferencial , Humanos , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/patología , Mieloma Múltiple/terapia , Estadificación de Neoplasias , Plasmacitoma/patología , Plasmacitoma/terapia , PronósticoRESUMEN
Monocytes/macrophages secrete various cytokines that induce proliferation of colony-forming unit granulocyte-macrophage (CFU-GM) in short-term assays. To determine whether macrophages also support proliferation of more primitive progenitors, i.e., cells that give rise to colony forming cells in a 5-week long-term culture (LTC), we established plastic-adherent macrophage layers from human peripheral blood (PB) and filgrastim (G-CSF)-mobilized progenitor cell collections in the presence of hydrocortisone, and compared these layers with bone marrow (BM) stroma regarding their suitability to support proliferation and differentiation of CD34+ BM and cord blood (CB) cells in 5-week LTCs. CD34+ cells were seeded onto irradiated macrophage and BM stromal layers, as well as without any preformed layer. After 5 weeks, colony formation (CFU-GM, BFU-E/CFU-E) and cell expansion were determined. CD34+ cells from BM and CB yielded more CFU-GM and total nucleated cells at 5 weeks in the presence of both types of adherent layer compared with cultures without a layer (p<0.05). For CD34+ BM cells, macrophage layers were superior to BM stroma in enhancing CFU-GM and CFU-E/BFU-E output (p < 0.05). In contrast, BM stroma was favorable compared with macrophages concerning nucleated cell expansion from CD34+ CB cells (p = 0.027). The macrophage nature of PB-derived adherent cells was confirmed immunocytochemically by positive staining for CD68, Ki-Mlp, CD31, CD54, inconstant staining for CD14, and negative staining for CD1a, CD3, CD15, CD34, and CD62E. Cytochemical reactions were positive for alpha-naphthyl acetate esterase and negative for peroxidase and periodic acid-Schiff, consistent with the immunophenotype. In conclusion, the results show that blood-derived macrophages support CFU-GM generation from CD34+ CB and BM progenitors for 5 weeks in vitro. Differential effects on proliferation and maturation of BM versus CB progenitors are discussed.
Asunto(s)
Antígenos CD34/análisis , Células de la Médula Ósea/inmunología , Sangre Fetal/inmunología , Células Madre Hematopoyéticas/citología , Macrófagos/citología , Células de la Médula Ósea/citología , Células Cultivadas , Sangre Fetal/citología , Humanos , Hidrocortisona , InmunohistoquímicaRESUMEN
A five-agent GVHD prophylaxis programme consisting of cyclosporin A, methotrexate, anti-thymocyte-globulin, pentaglobin and metronidazol was given to 48 recipients of unrelated donor marrow with chronic myelogenous leukemia, acute leukemia, myelodysplastic syndromes, and familiar lymphocytic hemophagocytosis of an average age of 33.5 (0.6-56) years. GVHD grades II-IV occurred in 18 patients (39%) and grades III-IV in five patients (11%). Chronic GVHD developed in nine patients (23%), three limited and six extensive. Fifteen patients died. Clinical relapse was detected in eight patients. Four patients died as a consequence of the underlying disease and subsequent treatment, 11 patients died of transplant-related causes. After a median follow-up of 19 months, the overall and disease-free survival are 67% and 62%, respectively. Survival by age is as follows: 0-19 years: 12/13 patients; 20-39 years: 14/25 patients; 40-59 years: 7/10 patients. The five-agent GVHD prophylaxis regimen is effective. Matched-unrelated donor transplants can be carried out safely in patients younger than 50 years of age. The results in patients younger than 20 years of age should encourage matched-unrelated donor transplants at earlier stages of the disease.
Asunto(s)
Antiinfecciosos/uso terapéutico , Trasplante de Médula Ósea , Enfermedad Injerto contra Huésped/prevención & control , Enfermedades Hematológicas/terapia , Inmunosupresores/uso terapéutico , Adolescente , Adulto , Antiinfecciosos/administración & dosificación , Suero Antilinfocítico/administración & dosificación , Suero Antilinfocítico/uso terapéutico , Niño , Preescolar , Ciclosporina/administración & dosificación , Ciclosporina/uso terapéutico , Quimioterapia Combinada , Femenino , Enfermedades Hematológicas/patología , Enfermedades Hematológicas/fisiopatología , Prueba de Histocompatibilidad , Humanos , Inmunoglobulina A/administración & dosificación , Inmunoglobulina A/uso terapéutico , Inmunoglobulina M/administración & dosificación , Inmunoglobulina M/uso terapéutico , Inmunosupresores/administración & dosificación , Lactante , Masculino , Metotrexato/administración & dosificación , Metotrexato/uso terapéutico , Metronidazol/administración & dosificación , Metronidazol/uso terapéutico , Persona de Mediana Edad , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Human lymphocytes remain among the most promising target cells for gene therapy. Gene-modified lymphocytes have been used successfully to treat adenosine deaminase (ADA)-deficient patients and to control GvHD after allogeneic BMT. Because activation and proliferation of T cells are necessary for efficient retrovirus-mediated gene transfer and subsequent selection of transduced cells, mononuclear cells (MNC) from steady-state and G-CSF-stimulated peripheral blood were activated by short exposure to the mitogen PHA, the anti-CD3 antibody OKT3, or both in the presence of different concentrations of recombinant IL-2. Using OKT3 (10 or 30 ng/ml) and IL-2 (100 U/ml), T cells expanded efficiently during a 14-day culture period. Cell expansion was similar under serum-free conditions. The immunophenotypic profile over time showed a marked increase in CD8+ cells, leading to a reversed CD4/CD8 ratio of 1:2 and a slight increase in CD56+ cells. Supernatant-based centrifugal transduction of primary human T lymphocytes was compared with supernatant transduction on the extracellular matrix protein fibronectin. Transduction with cell-free retrovirus-containing supernatant in tissue culture flasks coated with human plasma fibronectin led to significantly higher transduction efficiencies (20% +/- 7.5%) than centrifugal transduction in uncoated culture flasks (13.6% +/- 5.1%)(p = 0.041). To both rapidly characterize transduced cells and isolate these from residual nontransduced but biologically equivalent cells, an amphotropic Moloney murine leukemia virus (MoMuLV)-based retroviral vector containing the intracytoplasmically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene was used. FACS sorting of T cells after transduction resulted in >90% LNGFR+ cells and was much faster than enrichment of transduced cells through growth in G418-selection medium. These results show that supernatant-based retroviral gene transfer into primary human T lymphocytes can be enhanced by fibronectin. Ectopic expression of a cell surface protein can be used to rapidly and conveniently quantitate transduction efficiency through FACS analysis and to efficiently enrich transduced cells through FACS sorting.
Asunto(s)
Técnicas de Transferencia de Gen , Vectores Genéticos , Inmunoterapia Adoptiva , Activación de Linfocitos , Linfocitos T/inmunología , Factor Estimulante de Colonias de Granulocitos/inmunología , Humanos , Inmunofenotipificación , Muromonab-CD3/inmunología , RetroviridaeRESUMEN
The yield of CD34+ PBPC and colony-forming units-granulocyte-macrophage (CFU-GM) in leukapheresis products and the expression of the adhesion molecules CD11a, CD31, CD49d, CD49e, CD54, CD58, CD62L, c-kit (CD117), Thy-1 (CD90), CD33, CD38, and HLA-DR on CD34+ PBPC were analyzed in patients with cancer of the testis (n = 10), breast cancer (n = 10), Hodgkin's disease (n = 20), high-grade (n = 20) and low-grade (n = 20) non-Hodgkin's lymphoma, and healthy donors (n = 20) undergoing G-CSF (filgrastim)-stimulated PBPC mobilization. For each disease entity, G-CSF was administered in two different doses, 10 microg G-CSF/kg body weight (BW)/day s.c. vs. 24 microg G-CSF/kg BW s.c./day in steady-state condition. Data were compared for each dose group separately. Patients with cancer of the testis and breast cancer mobilized significantly more CD34+ cells than patients with high-grade and low-grade non-Hodgkin's lymphoma and Hodgkin's disease (p<0.05). Correspondingly, expression of CD49d on CD34+ PBPC was significantly lower in the same patients with cancer of the testis compared with high-grade and low-grade non-Hodgkin's lymphoma and Hodgkins' disease and in patients with breast cancer compared with high-grade and low-grade non-Hodgkin's lymphoma, Hodgkins's disease, and healthy donors. Similar results were obtained for CD49e. These data suggest that the expression of the adhesion molecules CD49d and CD49e on G-CSF-mobilized CD34+ cells of patients with solid tumors, non-Hodgkin's lymphoma, Hodgkin's disease, and healthy donors is inversely correlated with the amount of mobilized CD34+ cells.
Asunto(s)
Neoplasias de la Mama/patología , Moléculas de Adhesión Celular/análisis , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Movilización de Célula Madre Hematopoyética/métodos , Células Madre Hematopoyéticas/inmunología , Enfermedad de Hodgkin/patología , Linfoma no Hodgkin/patología , Neoplasias Testiculares/patología , Adolescente , Adulto , Antígenos CD/análisis , Antígenos CD34/análisis , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/terapia , Ensayo de Unidades Formadoras de Colonias , Femenino , Filgrastim , Factor Estimulante de Colonias de Granulocitos/farmacología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/patología , Enfermedad de Hodgkin/inmunología , Enfermedad de Hodgkin/terapia , Humanos , Integrina alfa4 , Integrina alfa5 , Integrinas/análisis , Linfoma no Hodgkin/inmunología , Linfoma no Hodgkin/terapia , Masculino , Persona de Mediana Edad , Proteínas Recombinantes , Valores de Referencia , Neoplasias Testiculares/inmunología , Neoplasias Testiculares/terapiaRESUMEN
Genetically modified lymphocytes have been successfully used for correction of ADA deficiency in children and in controlling graft-versus-host disease (GvHD) after allogeneic bone marrow transplantation. Low transduction efficiencies are, however, limiting for gene therapeutic strategies based on lymphocytes. In this study we compared protocols for highly efficient gene transfer into human T cells using retroviral vector-containing supernatant. We showed that infection of both human primary T cells and CD4+ Jurkat cells is most efficient on the matrix component fibronectin. Transduction was carried out with a retroviral vector encoding both the human intracytoplasmatically truncated low-affinity nerve growth factor receptor (deltaLNGFR) as a gene transfer marker and the Herpes simplex virus thymidine kinase for negative selection. Based on LNGFR expression genetically modified cells were enriched to near purity by magnetic cell sorting (MACS). Enriched cells could be shown to be highly sensitive to ganciclovir.
Asunto(s)
Fibronectinas/metabolismo , Técnicas de Transferencia de Gen , Vectores Genéticos/genética , Retroviridae/genética , Linfocitos T/virología , Antialérgicos/metabolismo , Ganciclovir/farmacología , Humanos , Integrina alfa4beta1 , Integrinas/metabolismo , Células Jurkat , Receptores de Fibronectina/metabolismo , Receptores Mensajeros de Linfocitos/metabolismo , Receptores de Factor de Crecimiento Nervioso/metabolismo , Timidina Quinasa/genéticaRESUMEN
Twenty-six patients received peripheral blood progenitor cells (PBPC) from unrelated donors at five European Centres. Twenty-five donors were HLA-A, -B and -DR identical and one had a one antigen mismatch. PBPC were mobilised by treatment with G-CSF for 4-5 days. Eleven patients receiving PBPC were compared to 11 patients receiving unrelated bone marrow with comparable prognostic factors. The median mononuclear cell count, the CD3+ cells and the CD56+ cells were seven to 10 times higher in PBPC, compared to bone marrow (P < 0.001). The median CD34+ cell content was 6.1 x 10(6)/kg recipient weight with PBPC, compared to 4.3 with bone marrow (NS). Time from transplantation to neutrophils >0.5 x 10(9)/l was a median of 11 (range 6-21) days using PBPC vs 15 (10-22) days after transplantation of bone marrow (P = 0.03). Transfusions and time to discharge did not differ between the two groups. All 26 patients receiving PBPC engrafted. Acute GVHD grades II-IV was seen in 8/26 (31%) and chronic GVHD in 8/18 (44%). Overall, 13/26 (50%) of the patients are alive and well with a median follow-up of 9 (2-35) months.
Asunto(s)
Trasplante de Médula Ósea , Supervivencia de Injerto , Trasplante de Células Madre Hematopoyéticas , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Prueba de Histocompatibilidad , Humanos , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Relapse after transplant for malignant lymphomas remains the main cause of treatment failure. Most conditioning regimens contain total body irradiation (TBI). We investigated the toxicity and efficacy of an intensified chemotherapy conditioning regimen without TBI in patients with relapsed or high-risk malignant lymphoma who had received prior radiation therapy and were therefore not eligible for TBI. Twenty patients with a median age of 38 (18-56) and relapsed or high-risk malignant non-Hodgkin's lymphoma (NHL, n = 16) or Hodgkin's disease (HD, n = 4) underwent high-dose chemotherapy consisting of busulfan (16 mg/kg), cyclophosphamide (120 mg/kg) and etoposide 30 mg/kg (n = 8) or 45 mg/kg (n = 12) followed by peripheral stem cell support (n = 14), autologous bone marrow (n = 3), allogeneic (n = 2) or syngeneic (n = 1) transplantation. All but two had chemosensitive disease before high-dose chemotherapy. The main toxicity -- according to the Bearman score -- was mucositis II in 18 (90%) patients; five patients (25%) suffered a grade I hepatic toxicity. GI toxicity I occurred in three (15%) and renal toxicity I in two patients (10%). Sixty percent of the patients developed transient dermatitis with erythema and three of them (15%) had skin desquamation; one patient experienced asymptomatic pancreatitis. Toxicity was slightly higher in patients treated with 45 mg/kg etoposide. One patient (5%) died of treatment-related venoocclusive disease. After a median follow-up of 50 months (24-84) the disease-free and overall survival were 50% and 55%. One of the nine relapsing patients developed secondary AML 18 months after transplant. High-dose busulfan, cyclophosphamide and etoposide is an effective regimen resulting in long-term disease-free survival in 50% of patients with relapsed malignant lymphoma and prior radiation therapy. The toxicity is moderate with a low treatment-related mortality (5%).