RESUMO
Hematopoietic gene therapy has markedly progressed during the last 15 years both in terms of safety and efficacy. While a number of serious adverse events (SAE) were initially generated as a consequence of genotoxic insertions of gamma-retroviral vectors in the cell genome, no SAEs and excellent outcomes have been reported in patients infused with autologous hematopoietic stem cells (HSCs) transduced with self-inactivated lentiviral and gammaretroviral vectors. Advances in the field of HSC gene therapy have extended the number of monogenic diseases that can be treated with these approaches. Nowadays, evidence of clinical efficacy has been shown not only in primary immunodeficiencies, but also in other hematopoietic diseases, including beta-thalassemia and sickle cell anemia. In addition to the rapid progression of non-targeted gene therapies in the clinic, new approaches based on gene editing have been developed thanks to the discovery of designed nucleases and improved non-integrative vectors, which have markedly increased the efficacy and specificity of gene targeting to levels compatible with its clinical application. Based on advances achieved in the field of gene therapy, it can be envisaged that these therapies will soon be part of the therapeutic approaches used to treat life-threatening diseases of the hematopoietic system.
Assuntos
Anemia Falciforme/terapia , Terapia Genética/tendências , Doenças Hematológicas/terapia , Talassemia beta/terapia , Anemia Falciforme/sangue , Células Sanguíneas/patologia , Células Sanguíneas/transplante , Vetores Genéticos/efeitos adversos , Doenças Hematológicas/sangue , Doenças Hematológicas/patologia , Transplante de Células-Tronco Hematopoéticas/tendências , Células-Tronco Hematopoéticas/citologia , Humanos , Talassemia beta/sangueRESUMO
In vitro cultured blood cells for transfusion purposes provide a safe alternative to donor blood, particularly for patients who require recurrent transfusions, and can be used as carriers of therapeutic molecules. In vitro derivation of hematopoietic cell types from human-induced pluripotent stem cells (iPSCs) allows for a constant, well-defined production pipeline for such advanced therapeutic and medicinal products. Application of selected iPSC-derived hematopoietic stem cells and hematopoietic effector cells in transplantation/transfusions would avoid the risk of alloimmunization and blood-borne diseases, as well as enable the production of enhanced blood cells expressing molecules that enforce blood cell function or endow novel therapeutic properties. Here, we discuss the state of the art approaches to produce erythroid, megakaryoid and myeloid cells from iPSCs and the biological and technical hurdles that we need to overcome prior to therapeutic application.
Assuntos
Células Eritroides/citologia , Células-Tronco Hematopoéticas/citologia , Células-Tronco Pluripotentes Induzidas/citologia , Células Mieloides/citologia , Células Sanguíneas/citologia , Células Sanguíneas/transplante , Doadores de Sangue , Transfusão de Sangue/métodos , Diferenciação Celular/genética , Humanos , Trombopoese/genéticaRESUMO
POI is characterized by "absent not abnormal" menstruation with hormonal disorders in woman younger than 40 years of age, and etiological and pathophysiological mechanisms underlying the POI development have not been clearly defined. Recently, due to advantages such as abundant sources and non-invasive methods of harvest, MenSCs have been emerging as a promising treatment strategy for the recovery of female reproductive damage. Here, we demonstrated that MenSCs graft in POI mice after CTX treatment could restore ovarian function by regulating normal follicle development and estrous cycle, reducing apoptosis in ovaries to maintain homeostasis of microenvironment and modulating serum sex hormones to a relatively normal status. Moreover, MenSCs participated in the activation of ovarian transcriptional expression in ECM-dependent FAK/AKT signaling pathway and thus restored ovarian function to a certain extent. MenSCs transplantation was proved to be an effective way to repair ovarian function with low immunogenicity, suggesting its great potential for POI treatment.
Assuntos
Células Sanguíneas/transplante , Charibdotoxina/toxicidade , Matriz Extracelular/metabolismo , Quinase 1 de Adesão Focal/metabolismo , Insuficiência Ovariana Primária/terapia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células Estromais/transplante , Adulto , Animais , Células Sanguíneas/citologia , Feminino , Humanos , Menstruação , Camundongos , Camundongos Endogâmicos C57BL , Neurotoxinas/toxicidade , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/metabolismo , Insuficiência Ovariana Primária/patologia , Células Estromais/citologiaRESUMO
Hematopoietic stem and progenitor cells (HSPC) are endowed with the role of generating and maintaining lifelong the extremely diverse pool of blood cells1. Clinically, transplantation of human HSPC from an allogeneic healthy donor or infusion of autologous gene-corrected HSPC can effectively replenish defective blood cell production caused by congenital or acquired disorders2-9. However, due to methodological and ethical constraints that have limited the study of human HSPC primarily to in vitro assays10 or xenotransplantation models11,12, the in vivo activity of HSPC has to date remained relatively unexplored in humans13-16. Here we report a comprehensive study of the frequencies, dynamics and output of seven HSPC subtypes in humans that was performed by tracking 148,093 individual clones in six patients treated with lentiviral gene therapy using autologous HSPC transplantation and followed for up to 5 years. We discovered that primitive multipotent progenitor and hematopoietic stem cell (HSC) populations have distinct roles during the initial reconstitution after transplant, compared with subsequent steady-state phases. Furthermore, we showed that a fraction of in vitro-activated HSC are resilient and undergo a defined delayed activation period upon transplant. Finally, our data support the concept that early lymphoid-biased progenitors might be capable of long-term survival, such that they can be maintained independently of their continuous production from HSC. Overall, this study provides comprehensive data on HSPC dynamics after autologous transplantation and gene therapy in humans.
Assuntos
Engenharia Genética , Terapia Genética , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Células Sanguíneas/citologia , Células Sanguíneas/transplante , Linhagem da Célula/genética , Vetores Genéticos/uso terapêutico , Células-Tronco Hematopoéticas/metabolismo , Humanos , Lentivirus/genética , Células-Tronco/citologia , Transplante Autólogo/efeitos adversosRESUMO
Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non-SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non-SP cells. From a cell regulation point of view, we showed that SP activity depended on O2 concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia-induced factors HIF-1α and HIF-2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non-SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy.
Assuntos
Células Sanguíneas/metabolismo , Metabolismo Energético , Células-Tronco Hematopoéticas/metabolismo , Células da Side Population/metabolismo , Animais , Antígenos CD34/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Biomarcadores/metabolismo , Células Sanguíneas/transplante , Hipóxia Celular , Proliferação de Células , Células Cultivadas , Feminino , Sangue Fetal/citologia , Glucose/metabolismo , Transplante de Células-Tronco Hematopoéticas , Xenoenxertos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Camundongos , Mitocôndrias/metabolismo , Fenótipo , Interferência de RNA , Células da Side Population/transplante , TransfecçãoRESUMO
OBJECTIVE: To analyze whether application of autologous blood cell salvage can reduce the transfusion volume of allogeneic blood and complications of blood transfusion in off-pump coronary artery bypass operations. Methods: We randomly divided 120 patients into autologous blood cell salvage group (experimental group, n = 60) and non-autologous blood cell salvage group (control group, n = 60). Volume of perioperative allogeneic blood transfusion of each patient was recorded. Moreover, complications and ICU retention times (H) of each patient were also recorded. The data were analyzed using t tests. Results: The volume of allogeneic blood transfusion was significantly less in the experimental group than in the control group. Conclusion: Application of autologous blood cell salvage in off-pump coronary artery bypass graft operation can reduce the volume of allogeneic blood transfusion, alleviate blood shortage, and reduce the incidence of postoperative complications, leading to medical, economic, and social benefits.
Assuntos
Células Sanguíneas/transplante , Perda Sanguínea Cirúrgica/prevenção & controle , Ponte de Artéria Coronária sem Circulação Extracorpórea/métodos , Doença da Artéria Coronariana/cirurgia , Transfusão de Eritrócitos/estatística & dados numéricos , Terapia de Salvação/métodos , Adulto , Perda Sanguínea Cirúrgica/mortalidade , China/epidemiologia , Doença da Artéria Coronariana/mortalidade , Feminino , Humanos , Masculino , Complicações Pós-Operatórias , Estudos Retrospectivos , Transplante AutólogoRESUMO
AIM: Pilot clinical trial of NY-ESO-1 (ESO) protein in ISCOMATRIX™ adjuvant pulsed onto peripheral blood dendritic cells (PBDC), to ascertain feasibility, evaluate toxicity and assess induction of ESO-specific immune responses. PATIENTS & METHODS: Eligible participants had resected cancers expressing ESO or LAGE-1 and were at high risk of relapse. PBDC were produced using CliniMACS®plus, with initial depletion of CD1c+ B cells followed by positive selection of CD1c+ PBDC. Patients received three intradermal vaccinations of ESO/IMX-pulsed PBDC at 4-week intervals. RESULTS: The process was feasible and safe. No vaccine-induced immune responses were detected. Assays of immunomodulatory cells did not correlate with outcomes. One patient had a long lasting complete remission. CONCLUSION: This method was feasible and safe but was minimally immunogenic.
Assuntos
Células Sanguíneas/fisiologia , Vacinas Anticâncer/imunologia , Carcinoma Basocelular/terapia , Células Dendríticas/fisiologia , Imunoterapia/métodos , Neoplasias Cutâneas/terapia , Linfócitos T/imunologia , Idoso , Apresentação de Antígeno , Antígenos CD1/metabolismo , Antígenos de Neoplasias/metabolismo , Células Sanguíneas/transplante , Carcinoma Basocelular/imunologia , Carcinoma Basocelular/patologia , Células Cultivadas , Colesterol/metabolismo , Células Dendríticas/transplante , Combinação de Medicamentos , Feminino , Seguimentos , Glicoproteínas/metabolismo , Humanos , Imunidade Humoral , Ativação Linfocitária , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias , Fosfolipídeos/metabolismo , Projetos Piloto , Saponinas/metabolismo , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Resultado do TratamentoRESUMO
Genomic studies have identified recurrent somatic mutations in acute leukemias. However, current murine models do not sufficiently encompass the genomic complexity of human leukemias. To develop preclinical models, we transplanted 160 samples from patients with acute leukemia (acute myeloid leukemia, mixed lineage leukemia, B-cell acute lymphoblastic leukemia, T-cell ALL) into immunodeficient mice. Of these, 119 engrafted with expected immunophenotype. Targeted sequencing of 374 genes and 265 frequently rearranged RNAs detected recurrent and novel genetic lesions in 48 paired primary tumor (PT) and patient-derived xenotransplant (PDX) samples. Overall, the frequencies of 274 somatic variant alleles correlated between PT and PDX samples, although the data were highly variable for variant alleles present at 0-10%. Seventeen percent of variant alleles were detected in either PT or PDX samples only. Based on variant allele frequency changes, 24 PT-PDX pairs were classified as concordant while the other 24 pairs showed various degree of clonal discordance. There was no correlation of clonal concordance with clinical parameters of diseases. Significantly more bone marrow samples than peripheral blood samples engrafted discordantly. These data demonstrate the utility of developing PDX banks for modeling human leukemia, and emphasize the importance of genomic profiling of PDX and patient samples to ensure concordance before performing mechanistic or therapeutic studies.
Assuntos
Xenoenxertos/patologia , Leucemia/genética , Doença Aguda , Adolescente , Adulto , Animais , Células Sanguíneas/transplante , Transplante de Medula Óssea , Bovinos , Criança , Perfilação da Expressão Gênica , Humanos , Imunofenotipagem , Leucemia/patologia , Camundongos , Pessoa de Meia-Idade , Adulto JovemRESUMO
Allogeneic hematopoietic stem cell transplantation and blood cell transfusions are performed commonly in patients with a variety of blood disorders. Unfortunately, these donor-derived cell therapies are constrained due to limited supplies, infectious risk factors, a lack of appropriately matched donors, and the risk of immunologic complications from such products. The use of autologous cell therapies has been proposed to overcome these shortcomings. One can derive such therapies directly from hematopoietic stem and progenitor cells of individuals, which can then be manipulated ex vivo to produce the desired modifications or differentiated to produce a particular target population. Alternatively, pluripotent stem cells, which have a theoretically unlimited self-renewal capacity and an ability to differentiate into any desired cell type, can be used as an autologous starting source for such manipulation and differentiation approaches. Such cell products can also be used as a delivery vehicle for therapeutics. In this review, we highlight recent advances and discuss ongoing challenges for the in vitro generation of autologous hematopoietic cells that can be used for cell therapy.
Assuntos
Células Sanguíneas/citologia , Células Sanguíneas/transplante , Transfusão de Sangue Autóloga , Terapia Baseada em Transplante de Células e Tecidos , Animais , Células Sanguíneas/metabolismo , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Terapia Baseada em Transplante de Células e Tecidos/métodos , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Humanos , Transplante AutólogoRESUMO
Systemic sclerosis is an autoimmune disease characterized by sclerosis (hardening) of the skin and deep viscera associated with microvascular functional and structural alteration, which leads to chronic ischemia. In the hands of patients, ischemic and fibrotic damages lead to both pain and functional impairment. Hand disability creates a large burden in professional and daily activities, with social and psychological consequences. Currently, the proposed therapeutic options for hands rely mainly on hygienic measures, vasodilatator drugs and physiotherapy, but have many constraints and limited effects. Developing an innovative therapeutic approach is crucial to reduce symptoms and improve the quality of life. The discovery of adult stem cells from adipose tissue has increased the interest to use adipose tissue in plastic and regenerative surgery. Prepared as freshly isolated cells for immediate autologous transplantation, adipose tissue-derived stem cell therapy has emerged as a therapeutic alternative for the regeneration and repair of damaged tissues. We aim to update literature in the interest of autologous fat graft or adipose derived from stromal vascular fraction cell-based therapy for the hands of patients who suffer from systemic sclerosis.
Assuntos
Tecido Adiposo/transplante , Células-Tronco Adultas/transplante , Deformidades Adquiridas da Mão/terapia , Transplante de Células-Tronco Mesenquimais , Escleroderma Sistêmico/terapia , Células-Tronco Adultas/metabolismo , Células Sanguíneas/transplante , Ensaios Clínicos como Assunto , Citocinas/metabolismo , Células Endoteliais/transplante , Deformidades Adquiridas da Mão/etiologia , Humanos , Injeções , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Lipectomia , Macrófagos/transplante , Pericitos/transplante , Escleroderma Sistêmico/complicações , Células Estromais/metabolismo , Células Estromais/transplante , Coleta de Tecidos e Órgãos , Transplante Autólogo , Resultado do TratamentoRESUMO
Multiple myeloma is a plasma cell neoplasm with an extremely variable clinical course. Animal models are needed to better understand its pathophysiology and for preclinical testing of potential therapeutic agents. Hematopoietic cells expressing the hypermorphic Rad50(s) allele show hematopoietic failure, which can be mitigated by the lack of a transcription factor, Mef/Elf4. However, we find that 70% of Mef(-/-)Rad50(s/s) mice die from multiple myeloma or other plasma cell neoplasms. These mice initially show an abnormal plasma cell proliferation and monoclonal protein production, and then develop anemia and a decreased bone mineral density. Tumor cells can be serially transplanted and according to array CGH and whole exome sequencing, the pathogenesis of plasma cell neoplasms in these mice is not linked to activation of a specific oncogene, or inactivation of a specific tumor suppressor. This model recapitulates the systemic manifestations of human plasma cell neoplasms, and implicates cooperativity between the Rad50(s) and Mef/Elf4 pathways in initiating myelomagenic mutations that promote plasma cell transformation.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Proteínas de Ligação a DNA/genética , Mieloma Múltiplo/genética , Fenótipo , Fatores de Transcrição/genética , Hidrolases Anidrido Ácido , Animais , Células Sanguíneas/metabolismo , Células Sanguíneas/patologia , Células Sanguíneas/fisiologia , Células Sanguíneas/transplante , Densidade Óssea , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Exoma , Camundongos , Camundongos Endogâmicos C57BL , Mieloma Múltiplo/patologiaRESUMO
Mesenchymal stem cells (MSCs) have been successfully used to treat multiple diseases in animal studies and clinical trials. Currently, the commonly used MSCs are derived from bone marrow and adipose tissue. Alternative approaches include differentiation of induced pluripotent stem cells (iPSCs) into MSCs, or direct reprogramming of blood cells into MSCs. This review summarizes recent progresses concerning how to generate MSCs by blood cell reprogramming and how studies in cellular reprogramming may help identify new factors to expand or even rejuvenate adult MSCs.
Assuntos
Células Sanguíneas/transplante , Reprogramação Celular/genética , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Tecido Adiposo/citologia , Células Sanguíneas/citologia , Diferenciação Celular/genética , Proliferação de Células/genética , Humanos , Células-Tronco Pluripotentes InduzidasRESUMO
STUDY DESIGN: Experimental study. OBJECTIVE: Several neuro-degenerative disorders such as Alzheimer's dementia, Parkinson's disease and amyotrophic lateral sclerosis (ALS) are associated with genetic mutations, and replacing or disrupting defective sequences might offer therapeutic benefits. Single gene delivery has so far failed to achieve significant clinical improvements in humans, leading to the advent of co-expression of multiple therapeutic genes. Co-transfection using two or more individual constructs might inadvertently result in disproportionate delivery of the products into the cells. To prevent this, and in order to rule out interference among the many promoters with varying strength, expressing multiple proteins in equimolar amounts can be achieved by linking open reading frames under the control of only one promoter. SETTING: Kazan, Russian Federation. METHODS: Here we describe a strategy for adeno-viral co-expression of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) interconnected through picorna-viral 2A-amino-acid sequence in transfected human umbilical cord blood mono-nuclear cells (hUCB-MCs). RESULTS: Presence of both growth factors, as well as absence of immune response to 2A-antigen, was demonstrated after 28-52 days. Following injection of hUCB-MCs into ALS transgenic mice, co-expression of VEGF and FGF2, as well as viable xeno-transplanted cells, were observed in the spinal cord after 1 month. CONCLUSION: These results suggest that recombinant adeno-virus containing 2A-sequences could serve as a promising alternative in regenerative medicine for the delivery of therapeutic molecules to treat neurodegenerative diseases, such as ALS.
Assuntos
Esclerose Lateral Amiotrófica/terapia , Células Sanguíneas/metabolismo , Células Sanguíneas/transplante , Cisteína Endopeptidases/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas Virais/metabolismo , Adenoviridae/genética , Esclerose Lateral Amiotrófica/genética , Animais , Cisteína Endopeptidases/genética , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Sangue Fetal/citologia , Fator 2 de Crescimento de Fibroblastos/genética , Vetores Genéticos/fisiologia , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Mutação/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Superóxido Dismutase-1/genética , Transfecção , Fator A de Crescimento do Endotélio Vascular/genética , Proteínas Virais/genéticaRESUMO
We previously reported that blood clot combined with biphasic calcium phosphate microparticles constitute a biomaterial (BRB) that can repair a bone critical defect in rat and induces subcutaneous bone formation in mice. The granulocyte colony-stimulating factor (G-CSF) is the agent most commonly used in human to enrich blood with hematopoietic stem and progenitor cells (HSPCs) as well as granulocytes (GCs). Moreover, recent data also suggest that it can mobilize mesenchymal stem cells (MSCs). Here, we asked whether the osteoinductive properties of the BRB could be further enhanced by G-CSF, either by replacing normal blood by G-CSF-mobilized blood (BRBe) or by treating the recipient animals with G-CSF. The experiments performed in C57BL/6 mice showed that G-CSF induces a marked increase of circulating HPCs and GCs, but not of MSCs. BRBe prepared with G-CSF-enriched blood induced a slight but significant decrease of subcutaneous bone formation compared to BRB prepared with normal blood. Additional injection of G-CSF to the recipient mice had no significant effect on the bone formation induced by BRB or BRBe. Altogether these results indicate that, in this model of ectopic implantation, cell mobilization induced by G-CSF has a negative effect on the osteoinductive property of this blood/BCP composite.
Assuntos
Materiais Biocompatíveis/química , Regeneração Óssea/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos/farmacologia , Hidroxiapatitas/química , Animais , Células Sanguíneas/citologia , Células Sanguíneas/efeitos dos fármacos , Células Sanguíneas/transplante , Substitutos Ósseos/química , Granulócitos/citologia , Granulócitos/efeitos dos fármacos , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Humanos , Teste de Materiais , Camundongos , Camundongos Endogâmicos C57BL , Próteses e Implantes , RatosRESUMO
BACKGROUND: We investigated the effects of transendocardial CD34(+) cell transplantation in patients with ischemic cardiomyopathy. METHODS AND RESULTS: In a prospective crossover study, we enrolled 33 patients with ischemic cardiomyopathy with New York Heart Association class III and left ventricular ejection fraction <40%. In phase 1, patients were treated with medical therapy for 6 months. Thereafter, all patients underwent transendocardial CD34(+) cell transplantation. Peripheral blood CD34(+) cells were mobilized by granulocyte colony stimulating factor, collected via apheresis, and injected transendocardially in the areas of hibernating myocardium. Patients were followed up for 6 months after the procedure (phase 2). Two patients died during phase 1 and none during phase 2. The remaining 31 patients were 85% men, aged 57±6 years. In phase 1, we found no change in left ventricular ejection fraction (from 25.2±6.2% to 27.1±6.6%; P=0.23), N-terminal pro B-type natriuretic peptide (from 3322±3411 to 3672±5165 pg/mL; P=0.75) or 6-minute walk distance (from 373±68 to 411±116 m; P=0.17). In contrast, in phase 2 there was an improvement in left ventricular ejection fraction (from 27.1±6.6% to 34.9±10.9%; P=0.001), increase in 6-minute walk distance (from 411±116 to 496±113 m; P=0.001), and a decrease in N-terminal pro B-type natriuretic peptide (from 3672±5165 to 1488±1847 pg/mL; P=0.04). The average number of injected CD34(+) cells was 90.6±7.5×10(6). Higher doses of CD34(+) cells and a more diffuse distribution of transendocardial cell injections were associated with better clinical response. CONCLUSIONS: Transendocardial CD34(+) cell transplantation may be associated with improved left ventricular function, decreased N-terminal pro B-type natriuretic peptide levels, and better exercise capacity in patients with ischemic cardiomyopathy. These effects seem to be particularly pronounced in patients receiving diffusely distributed cell injections and high-dose cell therapy. CLINICAL TRIAL REGISTRATION URL: http://www.clinicaltrials.gov. Unique identifier: NCT01350310.
Assuntos
Células Sanguíneas/patologia , Cardiomiopatias/terapia , Isquemia/terapia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/patologia , Adulto , Antígenos CD34/metabolismo , Células Sanguíneas/transplante , Estudos Cross-Over , Feminino , Seguimentos , Fator Estimulador de Colônias de Granulócitos/metabolismo , Mobilização de Células-Tronco Hematopoéticas , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Estudos Prospectivos , Recuperação de Função Fisiológica , Resultado do TratamentoRESUMO
The use of a cell-based vaccine composed of autologous whole blood cells loaded with mRNA is described. Mice immunized with whole blood cells loaded with mRNA encoding antigen develop anti-tumor immunity comparable to DC-RNA immunization. This approach offers a simple and affordable alternative to RNA-based cellular therapy by circumventing complex, laborious and expensive ex vivo manipulations required for DC-based immunizations.
Assuntos
Células Sanguíneas/metabolismo , Vacinas Anticâncer/imunologia , RNA Mensageiro/metabolismo , Animais , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Antígenos de Neoplasias/metabolismo , Células Sanguíneas/citologia , Células Sanguíneas/transplante , Vacinas Anticâncer/genética , Vacinas Anticâncer/metabolismo , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Células Dendríticas/transplante , Modelos Animais de Doenças , Eletroporação , Feminino , Imunoterapia , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/metabolismo , Melanoma Experimental/mortalidade , Melanoma Experimental/patologia , Melanoma Experimental/terapia , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/química , Taxa de SobrevidaRESUMO
Diabetes can impair wound closure, which can give rise to major clinical problems. Most treatments for wound repair in diabetes remain ineffective. This study aimed to investigate the influence on wound closure of treatments using expanded human cord blood CD34(+) cells (CB-CD34(+) cells), freshly isolated CB-CD34(+) cells and a cytokine cocktail. The test subjects were mice with streptozotocin-induced diabetes. Wounds treated with fresh CB-CD34(+) cells showed more rapid repair than mice given the PBS control. Injection of expanded CB-CD34(+) cells improved wound closure significantly, whereas the injection of the cytokine cocktail alone did not improve wound repair. The results also demonstrated a significant decrease in epithelial gaps and advanced re-epithelialization over the wound bed area after treatment with either expanded CB-CD34(+) cells or freshly isolated cells compared with the control. In addition, treatments with both CB-CD34(+) cells and the cytokine cocktail were shown to promote recruitment of CD31(+)-endothelial cells in the wounds. Both the CB-CD34(+) cell population and the cytokine treatments also enhanced the recruitment of CD68-positive cells in the early stages (day 3) of treatment compared with PBS control, although the degree of this enhancement was found to decline in the later stages (day 9). These results demonstrated that expanded CB-CD34(+) cells or freshly isolated CB-CD34(+) cells could accelerate wound repair by increasing the recruitment of macrophages and capillaries and the reepithelialization over the wound bed area. Our data suggest an effective role in wound closure for both ex vivo expanded CB-CD34(+) cells and freshly isolated cells, and these may serve as therapeutic options for wound treatment for diabetic patients. Wound closure acceleration by expanded CB-CD34(+) cells also breaks the insufficient quantity obstacle of stem cells per unit of cord blood and other stem cell sources, which indicates a broader potential for autologous transplantation.
Assuntos
Antígenos CD34/metabolismo , Células Sanguíneas/transplante , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/terapia , Sangue Fetal/citologia , Transplante de Células-Tronco Hematopoéticas , Cicatrização , Animais , Movimento Celular , Proliferação de Células , Separação Celular , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Humanos , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICRRESUMO
AIM: To explore how families with children at home managed 4-12 months after an adult family member was discharged from the hospital to home following bone marrow or blood cell transplantation. BACKGROUND: Studies of these families have been limited in number. The Family Management Style Framework offers a conceptual framework relevant to families with children who have an adult member who has undergone a bone marrow or blood cell transplant. DESIGN: This was a descriptive, qualitative, one-point-in-time study. METHOD: Content analysis in relation to the three conceptual components of the Family Management Style Framework was employed; inductive thematic analysis determined the emergent conceptual dimensions present in the data. The sample was composed of 29 participants: 15 transplant recipients and 14 significant others who met the study criteria in 2008 and 2009. Each patient and his or her significant other were interviewed; the audio-recorded interviews were analysed after transcription. FINDINGS: The three conceptual components of the Family Management Style Framework: (1) definition of the situation; (2) management behaviours; and (3) perceived consequences, were validated as relevant for this sample across all participants. New conceptual dimensions were also present, inclusive of recovery view, support base, financial picture, management domains, perceived consequences to the whole family, and perceived consequences to the dyad relationships. CONCLUSION: There are several overarching findings in this study, notably that recovery at home is a family experience. Implications for nursing practice include the need for effective transitional care for the patient and family.
Assuntos
Células Sanguíneas/transplante , Transplante de Medula Óssea/psicologia , Saúde da Família , Adolescente , Adulto , Aloenxertos , Atitude Frente a Saúde , Criança , Relações Familiares , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Percepção , Adulto JovemRESUMO
Cell therapy is a major discipline of regenerative medicine that has been continually growing over the last two decades. The aging of the population necessitates discovery of therapeutic innovations to combat debilitating disorders, such as stroke. Menstrual blood and Sertoli cells are two gender-specific sources of viable transplantable cells for stroke therapy. The use of autologous cells for the subacute phase of stroke offers practical clinical application. Menstrual blood cells are readily available, display proliferative capacity, pluripotency and angiogenic features, and, following transplantation in stroke models, have the ability to migrate to the infarct site, regulate the inflammatory response, secrete neurotrophic factors, and have the possibility to differentiate into neural lineage. Similarly, the testis-derived Sertoli cells secrete many growth and trophic factors, are highly immunosuppressive, and exert neuroprotective effects in animal models of neurological disorders. We highlight the practicality of experimental and clinical application of menstrual blood cells and Sertoli cells to treat stroke, from cell isolation and cryopreservation to administration.
Assuntos
Células Sanguíneas/transplante , Menstruação/sangue , Células de Sertoli/transplante , Transplante de Células-Tronco , Acidente Vascular Cerebral/terapia , Animais , Separação Celular , Criopreservação , Feminino , Humanos , Masculino , Acidente Vascular Cerebral/patologia , Transplante AutólogoRESUMO
Minor histocompatibility antigens (mHags) encoded by the Y-chromosome (H-Y-mHags) are known to play a pivotal role in allogeneic haematopoietic cell transplantation (HCT) involving female donors and male recipients. We present a new H-Y-mHag, YYNAFHWAI (UTY(139-147)), encoded by the UTY gene and presented by HLA-A*24:02. Briefly, short peptide stretches encompassing multiple putative H-Y-mHags were designed using a bioinformatics predictor of peptide-HLA binding, NetMHCpan. These peptides were used to screen for peptide-specific HLA-restricted T cell responses in peripheral blood mononuclear cells obtained post-HCT from male recipients of female donor grafts. In one of these recipients, a CD8+ T cell response was observed against a peptide stretch encoded by the UTY gene. Another bioinformatics tool, HLArestrictor, was used to identify the optimal peptide and HLA-restriction element. Using peptide/HLA tetramers, the specificity of the CD8+ T cell response was successfully validated as being HLA-A*24:02-restricted and directed against the male UTY(139-147) peptide. Functional analysis of these T cells demonstrated male UTY(139-147) peptide-specific cytokine secretion (IFNγ, TNFα and MIP-1ß) and cytotoxic degranulation (CD107a). In contrast, no responses were seen when the T cells were stimulated with patient tumour cells alone. CD8+ T cells specific for this new H-Y-mHag were found in three of five HLA-A*24:02-positive male recipients of female donor HCT grafts available for this study.