RESUMO
Colony stimulating factors (CSF) have been considered to modulate liver regeneration (LR) after partial hepatectomy (PH) at the tissue level. However, it remains unclear about precise mechanism of action of CSF in regeneration at the cellular level. Therefore, eight rat liver cell types were isolated by Percoll gradient centrifugation and magnetic beads. CSF-mediated signaling pathway genes were obtained by searching the related pathway databases and their expression profiles in 8 hepatic cell types were measured using rat Genome 230 2.0 Microarray. RT-PCR was performed to assess the reliability of chip results. The result showed a large difference in expression profiles of CSF-mediated signaling pathway genes between different cell types; most genes involved in CSF-mediated signaling pathways were mainly unregulated across liver cell samples. The implication of these genes in LR was analyzed by the bioinformatics and systems biology method. According to chip results and gene synergy, a significant enhancement of the CSF3-mediated Pi3k/Akt pathway at 30-36 h in hepatocytes and at 24 h in biliary epithelial cells post-PH could be associated with active proliferation in these two cell types; the striking decrease in Jak/Stat cascade activity in hepatic stellate cells at 2 and 12 h post- PH or even inactive in dendritric cells during the whole LR implied that proliferation of these two cell types is possibly regulated by other signaling pathways. These data suggest the potential relevance of CSF in liver regeneration at the cellular level.
Assuntos
Fatores Estimuladores de Colônias/farmacologia , Regulação da Expressão Gênica , Hepatócitos/metabolismo , Regeneração Hepática/genética , Fígado/metabolismo , Animais , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Perfilação da Expressão Gênica , Hepatectomia , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Janus Quinases/genética , Janus Quinases/metabolismo , Células de Kupffer/citologia , Células de Kupffer/efeitos dos fármacos , Células de Kupffer/metabolismo , Fígado/cirurgia , Análise em Microsséries , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição STAT/genética , Fatores de Transcrição STAT/metabolismo , Transdução de SinaisAssuntos
Humanos , Citocinas/classificação , Pulmão , Fatores Estimuladores de Colônias/classificação , Fatores Estimuladores de Colônias/farmacologia , Citocinas/farmacologia , Interferons/classificação , Interferons/farmacologia , Interleucinas/classificação , Interleucinas/farmacologia , Fator de Necrose Tumoral alfa/classificação , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Hematopoietic cells can be transformed through the acquisition of autocrine growth factor production. Because of their ability to inhibit autocrine growth, antibodies directed against the growth factor or its receptor may have therapeutic potential. However, these agents may also inhibit normal cell development. We have developed two monoclonal antibodies, 4G8 and 2F2, directed against a protein of 110 to 150 Kd that interacts with the interleukin-3 (IL-3) receptor (R) complex. These antibodies inhibit IL-3-induced proliferation of nonleukemic and leukemic IL-3-dependent cell lines, as well as the autonomous growth of WEHI-3B in vitro and in vivo. These results suggest the possibility that anti-IL-3R antibodies may be useful in the treatment of some leukemias. However, the effect of anti-IL-3R antibodies on normal myeloid development in vitro has not been examined. We examined the effect of 4G8 and 2F2 on the growth in vitro of colony-forming unit granulocyte-macrophage (CFU-GM) colonies induced by IL-3, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), and macrophage-CSF (M-CSF). Our results show that while 4G8 and 2F2 inhibited CFU-GM colony formation induced by IL-3, they augmented colony formation induced by the other hematopoietins. 4G8 and 2F2 also enhanced G-CSF-induced proliferation of 32Dc13 and GM-CSF-induced proliferation of PT18, confirming that the effect on CFU-GM was a direct effect. Finally, 4G8 and 2F2 inhibited G-CSF-induced differentiation of 32Dc13, similar to low levels of IL-3; yet, neither 4G8 nor 2F2 blocked binding of G-CSF to its receptor. These results indicate that, in the absence of IL-3 and in the presence of other hematopoietins, 4G8 and 2F2 can function as weak IL-3 agonists. These studies suggest that antibodies such as 4G8 and 2F2, directed against components of the IL-3R, could potentially augment myeloid growth in vivo, rather than inhibit myeloid growth.
Assuntos
Anticorpos Monoclonais , Diferenciação Celular/fisiologia , Fatores Estimuladores de Colônias/farmacologia , Proteínas de Neoplasias/imunologia , Proteínas de Neoplasias/fisiologia , Neutrófilos/citologia , Receptores de Interleucina-3/fisiologia , Animais , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Linhagem Celular , Ensaio de Unidades Formadoras de Colônias , Fator Estimulador de Colônias de Granulócitos/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Interleucina-3/metabolismo , Interleucina-3/farmacologia , Cinética , Leucemia Experimental , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , Proteínas de Neoplasias/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Proteínas Recombinantes/farmacologiaAssuntos
Fatores Estimuladores de Colônias/uso terapêutico , Leucopenia/tratamento farmacológico , Síndrome da Imunodeficiência Adquirida/complicações , Anemia Aplástica/tratamento farmacológico , Antineoplásicos/efeitos adversos , Transplante de Medula Óssea , Fatores Estimuladores de Colônias/efeitos adversos , Fatores Estimuladores de Colônias/farmacologia , Avaliação de Medicamentos , Hematopoese/efeitos dos fármacos , Humanos , Leucopenia/etiologia , Síndromes Mielodisplásicas/tratamento farmacológico , Neoplasias/complicações , Proteínas Recombinantes/uso terapêuticoRESUMO
Evidence is provided that conditioned medium from a macrophage-like cell line contains molecules of approximately 45 kd molecular weight with granulocyte colony-stimulating factor (G-CSF)-like activity as well as with the property of inducing granulocytes to phagocytose latex particles and to mature morphologically. This type of differentiation was found to be induced on either bone marrow or induced granulocytes, but not on resident or induced macrophages. On the other hand, resident but not induced macrophages are shown to induce these types of activities when challenged by bacterial lipopolysaccharides. Evidence that macrophages produce a factor that is mitogenic for fibroblasts is also provided. This activity was measured by the induction of increased proliferation by either low-density or saturated cultures of fibroblasts. Human recombinant G-CSF was employed and found also to possess these dual capabilities of inducing both the proliferation and differentiation of granulocytes as well as the proliferation of fibroblasts. Finally, a mechanism for the regulation of myeloid cell production and differentiation is described in which G-CSF produced by macrophages not only induces granulocytes to differentiate but induces fibroblasts to proliferate and secrete macrophage colony-stimulating factor (M-CSF), which in turn makes myeloid monocyte precursors proliferate and secrete more G-CSF.
Assuntos
Fatores Estimuladores de Colônias/farmacologia , Fibroblastos/citologia , Granulócitos/fisiologia , Macrófagos/metabolismo , Fagocitose , Animais , Células da Medula Óssea , Diferenciação Celular , Divisão Celular , Linhagem Celular , Fatores Estimuladores de Colônias/biossíntese , Fatores Estimuladores de Colônias/metabolismo , Feminino , Fibroblastos/metabolismo , Fator Estimulador de Colônias de Granulócitos , Granulócitos/citologia , Fator Estimulador de Colônias de Macrófagos , Masculino , Camundongos , Peso Molecular , Proteínas Recombinantes/farmacologiaRESUMO
Leishmania organisms are important pathogens, causing diseases worldwide. Standard therapies are often toxic and are not always effective. The effect of recombinant human granulocyte-macrophage and macrophage colony-stimulating factors (GM-CSF and M-CSF) on intramacrophage survival of Leishmania mexicana amazonensis (Lma) were compared with those of interferon-gamma (IFN-gamma). Macrophages previously infected with Lma were treated with or without GM-CSF and M-CSF. Compared with no cytokine treatment, treatment with GM-CSF (0.1-100 ng/ml) or M-CSF (1:3.5 X 10(6) - 1:3.5 X 10(3) dilutions) caused a significant dose-dependent reduction in intracellular parasites, 427 +/- 20 (mean +/- SE) Lma/100 macrophages. GM-CSF or M-CSF in combination with IFN-gamma resulted in more effective inhibition of intracellular parasites. Thus, the cytostatic activity appears to require interaction between cytokines, macrophages, and amastigotes. These cytokines are potential therapeutic agents for visceral leishmaniasis.
Assuntos
Fatores Estimuladores de Colônias/farmacologia , Substâncias de Crescimento/farmacologia , Leishmania mexicana/imunologia , Macrófagos/parasitologia , Monócitos/parasitologia , Animais , Células Cultivadas , Meios de Cultura , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Humanos , Interferon gama/farmacologia , Fator Estimulador de Colônias de Macrófagos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Proteínas Recombinantes/farmacologiaRESUMO
The recently described MB-02 human cell line requires Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) for continuous growth and terminally differentiates into enucleate, hemoglobinized cells in response to erythropoietin. Here, analysis of globin production now demonstrates that uninduced MB-02 cells produce alpha globin and the fetal globin chains G gamma and A gamma in a ratio of 1:3. Addition of erythropoietin results in de novo synthesis of beta globin chains and a marked increase in total Hb/cell. Thus, the MB-02 cell line partially recapitulates the fetal to adult globin switch that occurs during erythroid and human fetal development and provides a new clonal human erythroid progenitor system for investigating the biochemical and molecular events involved in globin gene switching.
Assuntos
Fatores Estimuladores de Colônias/farmacologia , Eritropoetina/farmacologia , Hemoglobina Fetal/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Genes/efeitos dos fármacos , Substâncias de Crescimento/farmacologia , Hemoglobina A/genética , Linhagem Celular , Hemoglobina Fetal/biossíntese , Globinas/biossíntese , Globinas/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Hemoglobina A/biossíntese , HumanosRESUMO
Patients with refractory carcinoma were treated with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) by intravenous (IV) infusion. During the period of treatment, studies of polymorphonuclear leukocyte superoxide (O2-) release in response to formylmethionylleucylphenylalanine (fMLP) and phorbol myristate acetate (PMA) and studies of chemotaxis in response to fMLP and C5a were performed. We observed that patients receiving rhGM-CSF in vivo exhibited primed O2- release after stimulation both with fMLP and PMA. Chemotaxis, however, was not enhanced by the treatment. These data suggest that host defenses may be enhanced by this treatment and that rhGM-CSF may be a useful therapeutic adjunct in compromised patients.
Assuntos
Fatores Estimuladores de Colônias/farmacologia , Substâncias de Crescimento/farmacologia , Neutrófilos/fisiologia , Carcinoma/fisiopatologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Humanos , Neutrófilos/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Superóxidos/metabolismoAssuntos
Fatores Biológicos/uso terapêutico , Imunoterapia , Neoplasias/terapia , Fatores Biológicos/administração & dosagem , Fatores Biológicos/farmacologia , Ensaios Clínicos como Assunto , Fatores Estimuladores de Colônias/farmacologia , Fatores Estimuladores de Colônias/uso terapêutico , Avaliação Pré-Clínica de Medicamentos , Humanos , Infusões Intravenosas , Interferon gama/administração & dosagem , Interferon gama/farmacologia , Interferon gama/uso terapêutico , Interleucina-2/administração & dosagem , Interleucina-2/farmacocinética , Interleucina-2/uso terapêutico , Fator de Necrose Tumoral alfa/administração & dosagem , Fator de Necrose Tumoral alfa/uso terapêuticoRESUMO
In this paper we describe a crude pregnant mouse uterus and embryo extract (PMUE) prepared from CFW/ep mice which was able to stimulate the proliferation of high-proliferative-potential colony-forming cells (HPP-CFC) of bone marrow of normal mice, in vitro, in semisolid agar culture system. The development of that primitive murine progenitor cells requires the presence of a macrophage-stimulating factor (CSF-1) plus a synergistic factor (SF). The biological activity of both factors was present in our extracts. The higher SF activity was found in uterine plus placental tissues extracts. The SF was precipitated over 45 per cent ammonium sulfate saturation, and behaved as a nondialyzable substance, remained unaffected by trypsin digestion, and was heat-stable (70 degrees C for 15 min).