RESUMEN
Granulocyte-monocyte progenitor (GMP) cells play a vital role in the immune system by maturing into a variety of white blood cells, including neutrophils and macrophages, depending on exposure to cytokines such as various types of colony stimulating factors (CSF). Granulocyte-CSF (G-CSF) induces granulopoiesis and macrophage-CSF (M-CSF) induces monopoiesis, while granulocyte/macrophage-CSF (GM-CSF) favors monocytic and granulocytic differentiation at low and high concentrations, respectively. Although these differentiation pathways are well documented, the mechanisms behind the diverse behavioral responses of GMP cells to CSFs are not well understood. In this paper, we propose a mechanism of interacting CSF-receptors and transcription factors that control GMP differentiation, convert the mechanism into a set of differential equations, and explore the properties of this mathematical model using dynamical systems theory. Our model reproduces numerous experimental observations of GMP cell differentiation in response to varying dosages of G-CSF, M-CSF, and GM-CSF. In particular, we are able to reproduce the concentration-dependent behavior of GM-CSF induced differentiation, and propose a mechanism driving this behavior. In addition, we explore the differentiation of a fourth phenotype, monocytic myeloid-derived suppressor cells (M-MDSC), showing how they might fit into the classical pathways of GMP differentiation and how progenitor cells can be primed for M-MDSC differentiation. Finally, we use the model to make novel predictions that can be explored by future experimental studies.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/metabolismo , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Factor Estimulante de Colonias de Macrófagos/metabolismo , Macrófagos/fisiología , Modelos Teóricos , Células Supresoras de Origen Mieloide/fisiología , Neutrófilos/fisiología , Animales , Diferenciación Celular , Relación Dosis-Respuesta Inmunológica , Humanos , Análisis de SistemasRESUMEN
The kinetics of stem and progenitor cell differentiation at the single-cell level provides essential clues to the complexity of the underlying decision-making circuits. In many hematopoietic progenitor cells, differentiation is accompanied by the expression of lineage-specific markers and by a transition from a non-adherent to an adherent state. Here, using the granulocyte-macrophage progenitor (GMP) as a model, we introduce a label-free approach that allows one to follow the course of this transition in hundreds of single cells in parallel. We trap single cells in patterned arrays of micro-wells and use phase-contrast time-lapse movies to distinguish non-adherent from adherent cells by an analysis of Brownian motion. This approach allowed us to observe the kinetics of induced differentiation of primary bone-marrow-derived GMPs into macrophages. The time lapse started 2 hours after addition of the cytokine M-CSF, and nearly 80% of the population had accomplished the transition within the first 20 h. The analysis of Brownian motion proved to be a sensitive and robust tool for monitoring the transition, and thus provides a high-throughput method for the study of cell differentiation at the single-cell level.
Asunto(s)
Células Progenitoras de Granulocitos y Macrófagos/citología , Dispositivos Laboratorio en un Chip , Animales , Fenómenos Biofísicos , Adhesión Celular , Diferenciación Celular , Linaje de la Célula , Diseño de Equipo , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Ratones , Microscopía de Contraste de Fase , Movimiento (Física) , Análisis de la Célula Individual/instrumentación , Imagen de Lapso de Tiempo/instrumentaciónRESUMEN
Los granulocitos son un elemento fundamental en la respuesta del organismo frente a infecciones y traumatismos. Se revisan las alteraciones cuantitativas de los neutrófilos, congénitas y adquiridas, y las terapéuticas disponibles frente a ellas (AU)
Granulocytes are a fundamental element in the body response against infections and traumas. Quantitative alterations of neutrophils congenital and acquired and therapies available against them are reviewed (AU)
Asunto(s)
Humanos , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Granulocitos , Neutropenia/fisiopatología , Neutrófilos/fisiologíaRESUMEN
The actin-binding protein filamins (FLNs) are major organizers of the actin cytoskeleton. They control the elasticity and stiffness of the actin network and provide connections with the extracellular microenvironment by anchoring transmembrane receptors to the actin filaments. Although numerous studies have revealed the importance of FLN levels, relatively little is known about the regulation of its stability in physiological relevant settings. Here, we show that the ASB2α cullin 5-ring E3 ubiquitin ligase is highly expressed in immature dendritic cells (DCs) and is down-regulated after DC maturation. We further demonstrate that FLNs are substrates of ASB2α in immature DCs and therefore are not stably expressed in these cells, whereas they exhibit high levels of expression in mature DCs. Using ASB2 conditional knockout mice, we show that ASB2α is a critical regulator of cell spreading and podosome rosette formation in immature DCs. Furthermore, we show that ASB2(-/-) immature DCs exhibit reduced matrix-degrading function leading to defective migration. Altogether, our results point to ASB2α and FLNs as newcomers in DC biology.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/fisiología , Movimiento Celular/genética , Células Dendríticas/fisiología , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Proteínas Contráctiles/genética , Proteínas Contráctiles/metabolismo , Células Dendríticas/metabolismo , Filaminas , Técnicas de Silenciamiento del Gen , Células Progenitoras de Granulocitos y Macrófagos/metabolismo , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Isoenzimas/genética , Isoenzimas/metabolismo , Isoenzimas/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Células 3T3 NIH , Proteínas Supresoras de la Señalización de Citocinas , Transfección , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitina-Proteína Ligasas/fisiologíaRESUMEN
Cyclin dependent kinase inhibitors (CDKIs) influence proliferation of hematopoietic progenitor cells (HPCs), but little is known of how they influence proliferative responsiveness of HPCs to colony stimulating factors (CSFs), alone and in combination with other hematopoietically active factors, such as the potent co-stimulating cytokine stem cell factor (SCF), or inhibition by myelosuppressive chemokines. Using mice with deletions in p18(INK4c), p21(CIP1/WAF1), or p27(KIP1) genes, and in mice with double gene deletions for either p18/p21 or p18/p27, we determined effects of absence of these CDKIs and their interactions on functional HPC numbers in vivo, and HPC proliferative responsiveness in vitro. There is a decrease in bone marrow HPC proliferation in p18(-/-) mice commensurate with decreased numbers of HPC, suggesting a positive role for p18 on HPC in vivo, similar to that for p21. These positive effects of p18 dominate negative effects of p27 gene deletion. Moreover, the CDKIs differentially regulate responsiveness of granulocyte macrophage (GM) progenitors to synergistic cell proliferation in response to GM-CSF plus SCF, which is considered important for normal hematopoiesis. Responsiveness of HPCs to inhibition by myelosuppressive chemokines is directly related to the capacity of HPCs to respond to synergistic stimulation, and their cell cycle status. P18(INK4c) gene deletion rescued the loss of chemokine suppression of synergistic proliferation due to deletion of p21(CIP1/WAF1). These findings underscore the complex interplay of cell cycle regulators in HPC, and demonstrate that loss of one can sometimes be compensated by loss of another CDKI in both, a pro- or anti-proliferative context.
Asunto(s)
Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/fisiología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/fisiología , Células Madre Hematopoyéticas/fisiología , Factor de Células Madre/fisiología , Animales , Células de la Médula Ósea/fisiología , Proliferación Celular , Células Cultivadas , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/genética , Proteínas Inhibidoras de las Quinasas Dependientes de la Ciclina/metabolismo , Citocinas/fisiología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Péptidos y Proteínas de Señalización Intercelular/fisiología , Interleucina-6/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fase S , Bazo/citología , Factor de Células Madre/farmacologíaRESUMEN
Clinical studies using bone marrow-derived proangiogenic cells (PACs) have demonstrated modest improvements of function and/or perfusion of ischemic myocardium or skeletal muscle. Because the identities of these PACs and their functional ability to promote neovascularization remain poorly understood, it is possible that a subset of robust PACs exists but is obscured by the heterogeneous nature of this cell population. Herein, we found that common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs) preferentially differentiate into PACs compared with megakaryocyte-erythrocyte progenitors, hematopoietic stem cells, and common lymphoid progenitors. In vivo hindlimb ischemia studies and Matrigel plug assays verified the enhanced neovascularization properties uniquely associated with PACs derived from CMPs and GMPs. Taken together, these observations identify CMPs and GMPs as key bone marrow progenitors for optimal PAC function in vitro and in vivo and provide a foundation for novel therapeutic approaches to modulate angiogenesis.
Asunto(s)
Células Progenitoras de Granulocitos y Macrófagos/fisiología , Isquemia/fisiopatología , Células Progenitoras Mieloides/fisiología , Neovascularización Fisiológica , Animales , Biomarcadores/metabolismo , Células de la Médula Ósea/fisiología , Células Cultivadas , Técnicas de Cocultivo , Miembro Posterior , Células Endoteliales de la Vena Umbilical Humana/fisiología , Isquemia/metabolismo , Isquemia/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Isquemia Miocárdica/fisiopatología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Factores de TiempoRESUMEN
Emerging evidence demonstrates that proangiogenic cells (PACs) originate from the BM and are capable of being recruited to sites of ischemic injury where they contribute to neovascularization. We previously determined that among hematopoietic progenitor stem cells, common myeloid progenitors (CMPs) and granulocyte-macrophage progenitor cells (GMPs) differentiate into PACs and possess robust angiogenic activity under ischemic conditions. Herein, we report that a TGF-ß1-responsive Krüppel- like factor, KLF10, is strongly expressed in PACs derived from CMPs and GMPs, â¼ 60-fold higher than in progenitors lacking PAC markers. KLF10(-/-) mice present with marked defects in PAC differentiation, function, TGF-ß responsiveness, and impaired blood flow recovery after hindlimb ischemia, an effect rescued by wild-type PACs, but not KLF10(-/-) PACs. Overexpression studies revealed that KLF10 could rescue PAC formation from TGF-ß1(+/-) CMPs and GMPs. Mechanistically, KLF10 targets the VEGFR2 promoter in PACs which may underlie the observed effects. These findings may be clinically relevant because KLF10 expression was also found to be significantly reduced in PACs from patients with peripheral artery disease. Collectively, these observations identify TGF-ß1 signaling and KLF10 as key regulators of functional PACs derived from CMPs and GMPs and may provide a therapeutic target during cardiovascular ischemic states.
Asunto(s)
Células de la Médula Ósea/citología , Células de la Médula Ósea/fisiología , Diferenciación Celular , Factores de Transcripción de la Respuesta de Crecimiento Precoz/fisiología , Factores de Transcripción de Tipo Kruppel/fisiología , Neovascularización Fisiológica , Transducción de Señal , Factor de Crecimiento Transformador beta1/fisiología , Animales , Proteínas de Unión al ADN/genética , Factores de Transcripción de la Respuesta de Crecimiento Precoz/genética , Regulación de la Expresión Génica , Células Progenitoras de Granulocitos y Macrófagos/citología , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Miembro Posterior , Isquemia/metabolismo , Isquemia/patología , Isquemia/fisiopatología , Factores de Transcripción de Tipo Kruppel/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Células Progenitoras Mieloides/citología , Células Progenitoras Mieloides/fisiología , Enfermedad Arterial Periférica/metabolismo , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Flujo Sanguíneo Regional , Factor de Crecimiento Transformador beta1/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND AIMS: Mesenchymal stromal cells (MSC) are the most popular cells used in regenerative medicine and biotechnology. The clonogenic potential of these cells is defined by colony-forming unit-fibroblasts (CFU-F). It is well known that there is an interaction between hematopoietic cells and stromal cells in disease formation pathogenesis. Therefore we hypothesized that there should be a quantitative and qualitative relationship between MSC colonies (CFU-F) and hematopoietic stem cell colonies (colony-forming unit-granulocyte-macrophages; CFU-GM) among patients with and without hematologic diseases. METHODS: Forty-two patients were included in this study. Patients were divided into three groups: group A, patients with hematologic malignancies (n =20); group B, patients with bone marrow (BM) failure (n =11); group C, patients without hematologic diseases (n =11). BM aspirates were plated in different densities for CFU-F culture. The plating density was the same for CFU-GM culture. RESULTS: CFU-GM colonies grew in 90% of group A cells and all of group B and C cells (P= 0.0001). CFU-F colonies became visible on the ninth day of plating in group A and on the eight day in groups B and C. There was no statistically significant difference between the groups for the duration of CFU-F colony formation (P= 0.12). There were differences in the morphology of the colonies among the groups. CONCLUSIONS: This is the first study that has compared the clonogenic potential of stromal cells and hematopoietic stem cells in the same subjects with and without hematologic diseases. No correlation was shown between the clonogenic potential of stromal cells and hematopoietic cells.
Asunto(s)
Trasplante de Células Madre Hematopoyéticas/métodos , Células Madre Hematopoyéticas/fisiología , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/fisiología , Células del Estroma/fisiología , Células del Estroma/trasplante , Adulto , Anciano , Enfermedades de la Médula Ósea/fisiopatología , Enfermedades de la Médula Ósea/terapia , Técnicas de Cultivo de Célula , Diferenciación Celular/fisiología , Linaje de la Célula/fisiología , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias/métodos , Femenino , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Enfermedades Hematológicas/terapia , Neoplasias Hematológicas/fisiopatología , Neoplasias Hematológicas/terapia , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Adulto JovenRESUMEN
OBJECTIVE: Emerging work has revealed an integral role of the tumor necrosis factor-alpha (TNF-alpha) nuclear factor (NF)-kappaB pathway in the regulation of hematopoiesis. TNF-alpha inhibition of hematopoietic stem/progenitor cell growth involves type I TNF-alpha receptor (TNF-RI) and type II TNF-alpha receptor (TNF-RII). However, the role of TNF-RI vs TNF-RII in mediating this response is less clear. Full induction of NF-kappaB-dependent gene expression through TNF-RI requires the transcriptional coactivator SIMPL (substrate that interacts with mouse pelle-like kinase). To address the role of SIMPL in TNF-alpha-dependent signaling in hematopoiesis, endothelial cells and hematopoietic progenitors expressing SIMPL short hairpin RNA were characterized. MATERIAL AND METHODS: In vitro gene expression and progenitor assays employing SIMPL short hairpin RNA were used to examine the requirement for SIMPL in TNF-alpha-dependent effects upon cytokine gene expression and hematopoietic progenitor cell growth. Competitive repopulation studies were used to extend these studies in vivo. RESULTS: SIMPL is required for full TNF-RI-dependent expression of NF-kappaB-controlled cytokines in endothelial cells. Hematopoietic progenitor cell expansion is not affected if progenitors lacked SIMPL or if progenitors are treated with human TNF-alpha, which signals through TNF-RI. In the absence of SIMPL, human TNF-alpha leads to a dramatic decrease in progenitor cell expansion that is not due to apoptosis. Loss of SIMPL does not affect the activity of transforming growth factor-beta1 and interferon-gamma, other known suppressors of hematopoiesis. CONCLUSIONS: Suppression of myeloid progenitor cell expansion requires signaling through TNF-RI and TNF-RII. Signals transduced through the TNF-alpha-TNF-RI-SIMPL pathway support hematopoietic progenitor cell survival, growth and differentiation.
Asunto(s)
Proteínas Portadoras/fisiología , Supervivencia Celular , Células Madre Hematopoyéticas/fisiología , Factor de Necrosis Tumoral alfa/farmacología , Animales , Apoptosis , Células de la Médula Ósea/química , Células de la Médula Ósea/metabolismo , Proteínas Portadoras/análisis , Proteínas Portadoras/genética , Diferenciación Celular , División Celular , Línea Celular , Embrión de Mamíferos , Células Endoteliales/metabolismo , Femenino , Fibroblastos/química , Fibroblastos/metabolismo , Expresión Génica/efectos de los fármacos , Células Progenitoras de Granulocitos y Macrófagos/citología , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Hematopoyesis/efectos de los fármacos , Hematopoyesis/fisiología , Células Madre Hematopoyéticas/química , Células Madre Hematopoyéticas/citología , Humanos , Péptidos y Proteínas de Señalización Intracelular , Secuencias Invertidas Repetidas , Riñón , Ratones , Ratones Endogámicos C57BL , FN-kappa B/fisiología , ARN/genética , ARN Mensajero/análisis , Receptores del Factor de Necrosis Tumoral/fisiología , Proteínas Recombinantes/farmacología , Transducción de Señal , TransfecciónRESUMEN
BACKGROUND: Umbilical cord blood (UCB) banking is a well-established activity supporting the increasing number of UCB transplantations in haematological diseases. Our aim was to analyse the UCB characteristics of UCB units from preterm deliveries and compare them to full-term deliveries. MATERIAL AND METHODS: A prospective study in 194 preterm deliveries occurring at the La Fe University Hospital in Valencia was performed. Patients between 25 and 37 weeks of gestation were included. Those cases were compared to a full-term deliveries control group. RESULTS: The cases were grouped according to the gestational age: between 25 and 33 weeks (group 1), between 34 and 37 weeks (group 2) and between 38 and 42 weeks (group 3). Among obstetric variables, only arterial pH and maternal age variables were similar for all the groups. Higher CD34(+) cell counts were observed in the group 2, while the clonogenic efficiency was higher for the most preterm deliveries. DISCUSSION: UCB from deliveries of at least 34 weeks of gestation contain sufficient hematopoietic stem cell content for unrelated banking and transplantation, even containing higher CD34(+) cell content than UCB units from full-term deliveries. However, UCB from deliveries of less than 33 weeks' gestation contain only sufficient progenitors for children under 20 kg.
Asunto(s)
Sangre Fetal/fisiología , Células Madre Hematopoyéticas/fisiología , Recien Nacido Prematuro/sangre , Adulto , Antígenos CD34/sangre , Supervivencia Celular/fisiología , Células Clonales , Células Precursoras Eritroides/fisiología , Femenino , Citometría de Flujo , Edad Gestacional , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Humanos , Recién Nacido , Células Progenitoras Mieloides/fisiología , Embarazo , Estudios Prospectivos , Estadísticas no ParamétricasRESUMEN
The constant regeneration of the blood system during hematopoiesis requires tightly controlled lineage decisions of hematopoietic progenitor cells (HPCs). Because of technical limitations, differentiation of individual HPCs could not previously be analyzed continuously. It was therefore disputed whether cell-extrinsic cytokines can instruct HPC lineage choice or only allow survival of cells that are already lineage-restricted. Here, we used bioimaging approaches that allow the continuous long-term observation of individual differentiating mouse HPCs. We demonstrate that the physiological cytokines, macrophage colony-stimulating factor and granulocyte colony-stimulating factor, can instruct hematopoietic lineage choice.
Asunto(s)
Linaje de la Célula , Factor Estimulante de Colonias de Granulocitos/fisiología , Células Progenitoras de Granulocitos y Macrófagos/citología , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Factor Estimulante de Colonias de Macrófagos/fisiología , Animales , Muerte Celular , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Ratones , Monocitos/citología , Mielopoyesis , Neutrófilos/citologíaRESUMEN
Nucleoside analogs are rationally designed anticancer agents that disrupt DNA and RNA synthesis. Fludarabine and cladribine have important roles in the treatment of hematologic malignancies. Clofarabine is a next generation nucleoside analog which is under clinical investigation. The bone marrow toxicity, tumor cell cytotoxicity and human tumor xenograft activity of fludarabine, cladribine and clofarabine were compared. Mouse and human bone marrow were subjected to colony forming (CFU-GM) assays over a 5-log concentration range in culture. NCI-60 cell line screening data were compared. In vivo, a range of clofarabine doses was compared with fludarabine for efficacy in several human tumor xenografts. The IC90 concentrations for fludarabine and cladribine for mouse CFU-GM were >30 and 0.93 microM, and for human CFU-GM were 8 and 0.11 microM, giving mouse to human differentials of >3.8- and 8.5-fold. Clofarabine produced IC90s of 1.7 microM in mouse and 0.51 microM in human CFU-GM, thus a 3.3-fold differential between species. In the NCI-60 cell line screen, fludarabine and cladribine showed selective cytotoxicity toward leukemia cell lines while for clofarabine there was no apparent selectivity based upon origin of the tumor cells. In vivo, clofarabine produced a dose-dependent increase in tumor growth delay in the RL lymphoma, the RPMI-8226 multiple myeloma, and HT-29 colon carcinoma models. The PC3 prostate carcinoma was equally responsive to clofarabine and fludarabine. Bringing together bone marrow toxicity data, tumor cell line cytotoxicity data, and human tumor xenograft efficacy provides valuable information for the translation of preclinical findings to the clinic.
Asunto(s)
Nucleótidos de Adenina/uso terapéutico , Arabinonucleósidos/uso terapéutico , Cladribina/uso terapéutico , Células Progenitoras de Granulocitos y Macrófagos/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Neoplasias/patología , Fosfato de Vidarabina/análogos & derivados , Nucleótidos de Adenina/antagonistas & inhibidores , Nucleótidos de Adenina/farmacología , Animales , Antimetabolitos Antineoplásicos/farmacología , Antimetabolitos Antineoplásicos/uso terapéutico , Arabinonucleósidos/farmacología , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/fisiología , Células Cultivadas , Cladribina/farmacología , Clofarabina , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Células HT29 , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones SCID , Modelos Biológicos , Resultado del Tratamiento , Fosfato de Vidarabina/farmacología , Fosfato de Vidarabina/uso terapéutico , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The zinc finger transcription factor GATA-2 has been implicated in the regulation of hematopoietic stem cells. Herein, we explored the role of GATA-2 as a candidate regulator of the hematopoietic progenitor cell compartment. We showed that bone marrow from GATA-2 heterozygote (GATA-2(+/-)) mice displayed attenuated granulocyte-macrophage progenitor function in colony-forming cell (CFC) and serial replating CFC assays. This defect was mapped to the Lin(-)CD117(+)Sca-1(-)CD34(+)CD16/32(high) granulocyte-macrophage progenitor (GMP) compartment of GATA-2(+/-) marrow, which was reduced in size and functionally impaired in CFC assays and competitive transplantation. Similar functional impairments were obtained using a RNA interference approach to stably knockdown GATA-2 in wild-type GMP. Although apoptosis and cell-cycle distribution remained unperturbed in GATA-2(+/-) GMP, quiescent cells from GATA-2(+/-) GMP exhibited altered functionality. Gene expression analysis showed attenuated expression of HES-1 mRNA in GATA-2-deficient GMP. Binding of GATA-2 to the HES-1 locus was detected in the myeloid progenitor cell line 32Dcl3, and enforced expression of HES-1 expression in GATA-2(+/-) GMP rectified the functional defect, suggesting that GATA-2 regulates myeloid progenitor function through HES-1. These data collectively point to GATA-2 as a novel, pivotal determinant of GMP cell fate.
Asunto(s)
Factor de Transcripción GATA2/fisiología , Células Progenitoras de Granulocitos y Macrófagos/citología , Animales , Línea Celular , Factor de Transcripción GATA2/genética , Factor de Transcripción GATA2/metabolismo , Perfilación de la Expresión Génica , Genotipo , Células Progenitoras de Granulocitos y Macrófagos/fisiología , Ratones , Ratones Mutantes , Unión Proteica , Interferencia de ARNRESUMEN
OBJECTIVE: To study the effect of weiganli ([Chinese characters: see text]) on bone marrow hemopoiesis. METHODS: The effects of weiganli on the peripheral blood picture and the number of bone marrow nucleated cells (BMCs) were observed in myelosuppressed anemic model mice, and the effects of weiganli on the growth of colony forming unit-granulocyte macrophage (CFU-GM), colony forming unit-erythroid (CFU-E), burst forming unit-erythroid (BFU-E), colony forming unit-megkaryocyte (CFU-Meg) were investigated by in vitro cell culture technique. The hemopoietic stem cells (HSCs, c-kit+) in bone marrow were double stained with fluorescent antibody PE-C-Kit and FITC-CD45, and the HSCs (c-kit+) were counted by flow cytometer with CD45/SSC (side scatter) gating. RESULTS: Peripheral blood cell counts and the number of BMCs were significantly improved after weiganli administration; and bone marrow hemopoietic stem/progenitor cells were significantly increased. CONCLUSION: Weiganli can effectively promote the recovery of hemopoietic function in the myelosuppressed anemic mice.