RESUMO
We investigated the roles of CD3McAb and rhIL-2 activated bone marrow in the killing and purging of leukemia cells. Cytotoxicity of activated bone marrow was detected with MTT assay. CFU-GM level in activated bone marrow and the destruction of leukemia cells were measured using the semi-solid cell culture. Immune activation markers in activated bone marrow were detected by indirect immunofluorescence assay. Bone marrow activated by CD3McAb and rhIL-2 displayed significantly upregulated the killing and purging abilities on the leukemia cell line K562 and HL-60. Such effects were superior to that of bone marrow activated by rhIL-2 or CD3McAb alone (P < 0.05, P < 0.01). Activation by rhIL-2 and (or) CD3McAb exerted no obvious influence on CFU-GM level in bone marrow. Compared with bone marrow activated by rhIL-2 or CD3McAb alone, the synergistic effect of both CD3McAb+ and hIL-2 caused significant increase of CD3(+), CD8(+), CD19(+), CD25(+), CD38(+), and CD56(+) levels. Our study indicates that CD3McAb enhanced the killing and purging effects of rhIL-2 activated bone marrow on leukemia cells.
Assuntos
Anticorpos Monoclonais/farmacologia , Células da Medula Óssea/efeitos dos fármacos , Purging da Medula Óssea , Interleucina-2/farmacologia , Adolescente , Adulto , Anticorpos Monoclonais/imunologia , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Purging da Medula Óssea/métodos , Transplante de Medula Óssea , Complexo CD3/imunologia , Complexo CD3/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Feminino , Células Progenitoras de Granulócitos e Macrófagos , Células HL-60 , Humanos , Imunofenotipagem , Células K562 , Leucemia/metabolismo , Leucemia/patologia , Leucemia/terapia , Masculino , Pessoa de Meia-Idade , Fenótipo , Adulto JovemAssuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Pré-Escolar , Criança , Adolescente , Pessoa de Meia-Idade , Adulto Jovem , Atenção Primária à Saúde , Células Progenitoras de Granulócitos e Macrófagos/imunologia , Deformidades Congênitas dos Membros/reabilitação , Doenças Negligenciadas/complicações , Doenças do Sistema Nervoso Periférico/complicações , Hanseníase Virchowiana/epidemiologia , Mycobacterium leprae , Brasil , Doenças Negligenciadas/epidemiologia , Doenças Negligenciadas/prevenção & controle , Esquema de Medicação , Hanseníase/prevenção & controle , Monitoramento Epidemiológico , Prevalência , Resistência a MedicamentosRESUMO
The effects of the traditional Chinese drug Jianpi Bushen Prescription (JBP) were investigated on expressions of Wnt3a and Cyclin D1 genes in radiation-damaged mice. The radiation damage model was induced in Kumming mice by single total body irradiation treatment for 9 days. Mice were divided into the radiation group, low-dose (100%) JBP group, high-dose (200%) JBP group, or batyl alcohol group (positive control), which were administered twice a day for 9 days. mRNA and protein expressions of Wnt3a were detected in bone marrow mononuclear cells by real-time polymerase chain reaction and Western blot, whereas Cyclin D1 mRNA was detected by in situ hybridization. Wnt3a expressions were significantly downregulated in the radiation damage model group compared with all other groups (P < 0.05). The positive cell rate of Cyclin D1 mRNA expression and the number of granulocyte macrophage colonies were significantly decreased in the radiation damage model group relative to all other groups (P < 0.05). Furthermore, mRNA and protein expressions of Wnt3a, the positive cell rate of Cyclin D1 mRNA expression in bone marrow cells, and the number of granulocyte macrophage colonies were all significantly higher in the low-dose JBP group than in the high-dose JBP group (P < 0.05). In summary, JBP plays a protective role on radiation-induced bone marrow through the activation of the Wnt3a signaling pathway, and promotes the transcription and expression of Cyclin D1.
Assuntos
Ciclina D1/metabolismo , Expressão Gênica/efeitos dos fármacos , Lesões Experimentais por Radiação/tratamento farmacológico , Protetores contra Radiação/farmacologia , Proteína Wnt3A/metabolismo , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Células da Medula Óssea/efeitos da radiação , Células Cultivadas , Ciclina D1/genética , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/efeitos da radiação , Células Progenitoras de Granulócitos e Macrófagos/metabolismo , Masculino , Medicina Tradicional Chinesa , Camundongos , Lesões Experimentais por Radiação/genética , Lesões Experimentais por Radiação/metabolismo , Transdução de Sinais , Irradiação Corporal Total , Proteína Wnt3A/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Synadenium umbellatum Pax. is widely found in South America and empirically used in Brazil for the treatment of several diseases, mainly cancer. The aim of the study was to investigate cell death mechanisms induced by Synadenium umbellatum Pax. using Ehrlich ascites tumor (EAT) cells, as well as the myelotoxicity potential of this plant. MATERIALS AND METHODS: S. umbellatum cytotoxicity was evaluated in EAT cells by trypan blue exclusion and MTT reduction test and the mechanisms involved in EAT cell death were investigated by light and fluorescence microscopy, flow cytometry and immunocytochemistry. Investigation of S. umbellatum myelotoxicity was performed by clonogenic assay of colony forming unit- granulocyte macrophage (CFU-GM). RESULTS AND CONCLUSION: Our results demonstrated that S. umbellatum decreased the viability of EAT cells using both methods. Morphological analyses revealed that S. umbellatum-treatment induced EAT cell death by apoptotic pathway. We demonstrated the occurrence of reactive oxygen species (ROS) overgeneration, increased intracellular Ca(2+) concentration, alteration in mitochondrial membrane potential, phosphatydylserine externalization, and activation of caspases 3, 8, and 9. However, S. umbellatum produced myelotoxicity in bone marrow cells in a concentration-dependent manner. In comparison to EAT cells, the effects of S. umbellatum in bone marrow cells were 8-fold lower. Taken together, our results showed that S. umbellatum induced apoptosis in EAT cells at several levels and seems more toxic to tumor cells than to normal bone marrow cells.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma de Ehrlich/patologia , Euphorbiaceae , Animais , Antineoplásicos Fitogênicos/toxicidade , Cálcio/metabolismo , Carcinoma de Ehrlich/metabolismo , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Ciclo Celular/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Citometria de Fluxo , Células Progenitoras de Granulócitos e Macrófagos/efeitos dos fármacos , Células Progenitoras de Granulócitos e Macrófagos/patologia , Imuno-Histoquímica , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Microscopia de Fluorescência , Estresse Oxidativo/efeitos dos fármacos , Fosfatidilserinas/metabolismo , Componentes Aéreos da Planta , Plantas Medicinais , Espécies Reativas de Oxigênio/metabolismo , Células Tumorais CultivadasRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The Asháninkas, indigenous people of Peru, use cat's claw (Uncaria tomentosa) to restore health. Uncaria tomentosa has antioxidant activity and works as an agent to repair DNA damage. It causes different effects on cell proliferation depending on the cell type involved; specifically, it can stimulate the proliferation of myeloid progenitors and cause apoptosis of neoplastic cells. Neutropenia is the most common collateral effect of chemotherapy. For patients undergoing cancer treatment, the administration of a drug that stimulates the proliferation of healthy hematopoietic tissue cells is very desirable. It is important to assess the acute effects of Uncaria tomentosa on granulocyte-macrophage colony-forming cells (CFU-GM) and in the recovery of neutrophils after chemotherapy-induced neutropenia, by establishing the correlation with filgrastim (rhG-CSF) treatment to evaluate its possible use in clinical oncology. MATERIALS AND METHODS: The in vivo assay was performed in ifosfamide-treated mice receiving oral doses of 5 and 15 mg of Uncaria tomentosa and intraperitoneal doses of 3 and 9 µg of filgrastim, respectively, for four days. Colony-forming cell (CFC) assays were performed with human hematopoietic stem/precursor cells (hHSPCs) obtained from umbilical cord blood (UCB). RESULTS: Bioassays showed that treatment with Uncaria tomentosa significantly increased the neutrophil count, and a potency of 85.2% was calculated in relation to filgrastim at the corresponding doses tested. An in vitro CFC assay showed an increase in CFU-GM size and mixed colonies (CFU-GEMM) size at the final concentrations of 100 and 200 µg extract/mL. CONCLUSIONS: At the tested doses, Uncaria tomentosa had a positive effect on myeloid progenitor number and is promising for use with chemotherapy to minimize the adverse effects of this treatment. These results support the belief of the Asháninkas, who have classified Uncaria tomentosa as a 'powerful plant'.
Assuntos
Unha-de-Gato , Proliferação de Células/efeitos dos fármacos , Células Progenitoras Mieloides/efeitos dos fármacos , Neutropenia/tratamento farmacológico , Neutrófilos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Administração Oral , Animais , Unha-de-Gato/química , Células Cultivadas , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Sangue Fetal/citologia , Filgrastim , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Células Progenitoras de Granulócitos e Macrófagos/efeitos dos fármacos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Humanos , Ifosfamida , Injeções Intraperitoneais , Masculino , Medicina Tradicional , Camundongos , Camundongos Endogâmicos BALB C , Células Progenitoras Mieloides/metabolismo , Células Progenitoras Mieloides/patologia , Neutropenia/induzido quimicamente , Neutropenia/metabolismo , Neutropenia/patologia , Neutrófilos/metabolismo , Neutrófilos/patologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/administração & dosagemRESUMO
We studied the effects of Chlorella vulgaris (CV) on the interaction between stromal and hematopoietic stem cells in normal and Ehrlich ascites tumor (EAT)-bearing mice. Long-term bone marrow culture (LTBMC), cytokine production, spleen mononuclear cells (SMC) proliferation (SCP), colony stimulating activity (CSA), and NK cells activity were evaluated. In tumor bearers, reduced capacity of stromal cell layer to support the growth and differentiation of granulocyte-macrophage progenitor cells (CFU-GM), concomitantly to decreased numbers of total nonadherent cells in LTBMC and reduced local production of IL-6 and IL-1α, were observed. Presence of the tumor has not altered the number of stromal adherent cells. CV treatment restored the ability of stromal cells from EAT-bearing mice to produce IL-6 and IL-1α, which was consistent with increased number of nonadherent cells and higher ability to display CFU-GM in vitro. EAT growth increased SCP, serum CSA, and IL-10 production and concurrently depressed NK cell activity and the secretion of IL-2, IFN-γ, and TNF-α. Treatment of tumor-bearing mice with CV augmented CSA, SMC proliferation, NK cell activity, and the production of IL-2, IFN-γ, and TNF-α, whereas IL-10 levels where reduced. Our results suggest that CV modulates immunehematopoietic cell activity and disengages tumor-induced suppression of these responses.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Carcinoma de Ehrlich/tratamento farmacológico , Carcinoma de Ehrlich/imunologia , Chlorella vulgaris , Fatores Imunológicos/uso terapêutico , Mielopoese , Animais , Células da Medula Óssea/imunologia , Células da Medula Óssea/metabolismo , Carcinoma de Ehrlich/metabolismo , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Fatores Estimuladores de Colônias/sangue , Fatores Estimuladores de Colônias/metabolismo , Citocinas/metabolismo , Suplementos Nutricionais , Células Progenitoras de Granulócitos e Macrófagos/citologia , Células Progenitoras de Granulócitos e Macrófagos/imunologia , Células Progenitoras de Granulócitos e Macrófagos/metabolismo , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fitoterapia , Organismos Livres de Patógenos Específicos , Baço/citologia , Baço/imunologia , Baço/metabolismo , Células Estromais/imunologia , Células Estromais/metabolismoRESUMO
In this study we demonstrated that the oral administration of ß-1,3-glucan (Imunoglucan®) protects mice from a lethal dose of Listeria monocytogenes (LM) when administered prophylactically for 10 days at the doses of 150 and 300 mg/kg, with survival rates up to 40%. These doses also prevented the myelosuppression and the splenomegaly caused by a sublethal infection with LM, due to increased numbers of granulocyte-macrophage progenitors (CFU-GM) in the bone marrow. Investigation of the production of colony-stimulating factors revealed an increased colony-stimulating activity (CSA) in the serum of infected mice pre-treated with Imunoglucan®. The treatment also restored the reduced ability of stromal cells to display myeloid progenitors in long-term bone marrow cultures (LTBMC) and up-regulated IL-6 and IL-1α production by these cells in the infected mice, which was consistent with higher number of non-adherent cells. Additional studies to investigate the levels of interferon-gamma (INF-γ) in the supernatant of splenocyte cultures demonstrated a further increase in the level of this cytokine in infected-treated mice, compared to infected controls. In all cases, no differences were observed between the responses of the two optimal biologically effective doses. In contrast, no significant changes were produced by the treatment with the 50mg/kg dose. In addition, no changes were observed in normal mice treated with the three doses used. All together our results suggest that orally given Imunoglucan® indirectly modulates immune activity and probably disengages Listeria induced suppression of these responses by inducing a higher reserve of myeloid progenitors in the bone marrow in consequence of biologically active cytokine release (CSFs, IL-1α, IL-6, and INF-γ).
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Hematopoese/efeitos dos fármacos , Hematopoese/imunologia , Células-Tronco Hematopoéticas/imunologia , Listeriose/prevenção & controle , beta-Glucanas/uso terapêutico , Adjuvantes Imunológicos/administração & dosagem , Administração Oral , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/imunologia , Técnicas de Cultura de Células , Células Cultivadas , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Células Progenitoras de Granulócitos e Macrófagos/citologia , Células Progenitoras de Granulócitos e Macrófagos/efeitos dos fármacos , Células Progenitoras de Granulócitos e Macrófagos/imunologia , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Interferon gama/imunologia , Interferon gama/metabolismo , Interleucina-1alfa/biossíntese , Interleucina-1alfa/imunologia , Interleucina-6/biossíntese , Interleucina-6/imunologia , Listeria monocytogenes/efeitos dos fármacos , Listeriose/complicações , Listeriose/imunologia , Listeriose/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Esplenomegalia/etiologia , Esplenomegalia/imunologia , Esplenomegalia/prevenção & controle , beta-Glucanas/administração & dosagemRESUMO
The effects of a dry extract of the roots of Angelica sinensis (Oliv.) Diels (ASE) on the growth and differentiation of granulocyte-macrophage progenitor cells (CFU-GM) in normal and Listeria monocytogenes-infected mice were studied. Myelosuppression concomitant with increased numbers of spleen CFU-GM was observed in infected mice. Prophylactic administration of ASE (10, 25, and 50 mg/kg) stimulated marrow myelopoiesis in a dose-dependent manner and reduced spleen colony formation to control values. The dose of 50 mg/kg ASE was the optimal biologically active dose in infected mice, and this dose schedule significantly increased survival of mice infected with a lethal dose of L. monocytogenes, with survival rate up to 30%. Investigation of the production of colony-stimulating factors revealed a dose-dependent increased colony-stimulating activity in the serum of infected mice, with higher response produced by the 50 mg/kg dose. Notably, no effects were observed with the 100 mg/kg dose, compared with infected nontreated controls. Further studies to investigate the production of factors such as inteferon-γ and tumor necrosis factor-α demonstrated increased levels of both cytokines in mice infected with L. monocytogenes and treated with 50 mg/kg ASE. We propose that ASE indirectly modulates immune activity and probably disengages Listeria-induced suppression of these responses by inducing a higher reserve of myeloid progenitors in the bone marrow in consequence of biologically active cytokine release (colony-stimulating factors, interferon-γ, and tumor necrosis factor-α).