RESUMO
Vascular calcification (VC) is a common complication of chronic kidney disease (CKD) that has a detrimental effect on patients' survival and prognosis. The aim of this study was to develop and validate a practical and reliable prediction model for VC in CKD5 patients. The medical records of 544 CKD5 patients were reviewed retrospectively. Multivariate logistic regression analysis was used to identify the independent risk factors for vascular calcification in patients with CKD5 and then created a nomogram prediction model. The area under the receiver operating characteristic curve (AUC), Hosmer-Lemeshow test, and decision curve analysis (DCA) were used to assess model performance. The patients were split into groups with normal and high serum uric acid levels, and the factors influencing these levels were investigated. Age, BUN, SUA, P and TG were independent risk factors for vascular calcification in CKD5 patients in the modeling group (P < 0.05). In the internal validation, the results of model showed that the AUC was 0.917. No significant divergence between the predicted probability of the nomogram and the actual incidence rate (x2 = 5.406, P = 0.753) was revealed by the calibration plot and HL test, thus confirming that the calibration was satisfactory. The external validation also showed good discrimination (AUC = 0.973). The calibration chart and HL test also demonstrated good consistency. Besides, the correlation analysis of serum uric acid levels in all CKD5 patients revealed that elevated uric acid levels may be related to gender, BUN, P, and TG.
Assuntos
Falência Renal Crônica , Calcificação Vascular , Humanos , Nomogramas , Ácido Úrico , Estudos Retrospectivos , Calcificação Vascular/etiologiaRESUMO
This study was purposed to evaluate the long-term outcome and the safety of autologous peripheral blood mononuclear cells (PBMNC) treated by interleukin 2 (IL-2) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the therapy of patients with aplastic anemia (AA). The therapy of 49 patients admitted BG in hospital from April 2001 to December 2007 were analyzed retrospectively. PBMNC were isolated and cultured for 48 hours in presence of IL-2 and GM-CSF. Cells were collected, and 6 × 10(6) - 1 × 10(8) PBMNC were intravenously injected weekly for 4 - 22 months. Hematopoietic recovery was evaluated by examinations of peripheral blood, bone marrow aspirates and bone marrow biopsy. Flow cytometry was used to assess the peripheral T cell subsets before and after treatment. Polymerase chain reaction was performed to observe the clonal diversity of T cell receptor variable ß-chain (TCR-Vß) recombination. The results showed that 37 cases were cured and none of them relapsed during the follow-up, 5 cases were in partial remission, 3 cases got improvement, and 4 cases showed no response. The total efficiency reached up to 91.8%. The ratios of CD4(+)/CD8(+) subsets were abnormal in 39 patients prior to the treatment, and 31 cases restored to the normal range after cell transfusions. Analysis on the clonal diversity of TCR-Vß recombination in 11 patients showed the transition from monoclonal or biclonal spectratype to polyclonal one. No long-term side effects were documented. It is concluded that the treatment with PBMNC treated by IL-2 and GM-CSF is generally safe and effective. The underlying mechanisms may be in relation to the restoration of cell immunity.
Assuntos
Anemia Aplástica/terapia , Monócitos/transplante , Transplante de Células-Tronco de Sangue Periférico/métodos , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Seguimentos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/uso terapêutico , Humanos , Interleucina-2/uso terapêutico , Masculino , Pessoa de Meia-Idade , Transplante Autólogo , Adulto JovemRESUMO
BACKGROUND AIMS: Aplastic anemia (AA) is a rare but potentially life-threatening disease. There is a need for the development of new, more effective and less toxic therapies for treating AA. The safety and efficacy of an immune cell-based therapy for AA was examined. METHODS: Thirty-one patients with idiopathic AA received intravenous infusions of ex vivo-activated autologous and allogeneic immune cells at least once a week. Response to therapy was assessed by symptoms, transfusion dependency, blood counts, bone marrow biopsy and survival. RESULTS: Of the 31 patients, 25 (81%) had either complete (11, 35%) or partial (14, 45%) responses, while six (19%) showed no response to the therapy. The overall survival rates at 3 years were 90%. CONCLUSIONS: The therapy described appears to be safe and effective. The data from this pilot study suggest that a larger, controlled study is warranted.
Assuntos
Anemia Aplástica/imunologia , Anemia Aplástica/terapia , Hematopoese/imunologia , Imunoterapia Adotiva , Leucócitos Mononucleares/metabolismo , Adolescente , Adulto , Anemia Aplástica/induzido quimicamente , Anemia Aplástica/patologia , Benzeno/efeitos adversos , Calcimicina/farmacologia , Contagem de Células , Criança , Progressão da Doença , Feminino , Seguimentos , Humanos , Interleucina-2/imunologia , Interleucina-2/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Resultado do TratamentoRESUMO
OBJECTIVE: To assess the reaction of cytokines induced killer (CIK) cells treatment in hematopoietic injury at different levels on patients with benzene poisoning and seek a novel, safe and effective immunotherapy for benzene poisoning. METHODS: CIK cells were in vitro activated by interleukin-2 (IL-2) and granulocyte-macrophage-colony-stimulating factor (GM-CSF) from the peripheral blood mononuclear cells (PBMC). Thirty-two patients with benzene poisoning were treated with CIK cells. Nineteen patients with mild or moderate benzene poisoning in the control group were treated with VitB4, batilol, leucogen, inosine and stanozolol. The results for treatment of 12 patients with aplastic anemia induced by severe benzene poisoning (the efficacy rate and the case fatality rate) were analyzed. The change of T-lymphocyte subset analyzed by flow cytometry was also observed before and after treatment. RESULTS: For mild or moderate benzene poisoning, the increase of WBC and RLT in CIK group was higher than that in the control group (P < 0.05). The CD(4)/CD(8) levels were significantly increased after CIK treatment. And for severe benzene poisoning, the effective rate of the CIK group was 91.7% and the mortality rate was 0%. CONCLUSION: CIK treatment is safe and effective for hematopoietic injury caused by benzene poisoning. The mechanism may be related with the immune modulation of CIK treatment on immunodeficiency of patients with benzene poisoning.
Assuntos
Benzeno/intoxicação , Células Matadoras Induzidas por Citocinas/imunologia , Imunoterapia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto JovemRESUMO
We have previously shown that a novel immunotherapy using ex vivo activated immune cells is capable of promoting survival and hematopoietic recovery in mice after combined chemotherapy and radiotherapy. In this study, we investigated whether the immunotherapy with ex vivo activated immune cells had the same beneficial effects after syngeneic and semiallogeneic bone marrow transplantation (BMT) in BALB/c mice subjected to a lethal dose of total body irradiation (TBI). Immune cells were cultured in vitro with a combination of cytokines and a calcium ionophore for 2 days and subsequently injected to mice daily for 4 days starting 1 day after BMT. The immunotherapy enhanced survival and multilineage peripheral blood recovery in BMT mice with limited numbers of transplanted bone marrow cells when a low dose of ex vivo activated immune cells were used. However, the beneficial effects were completely lost when a higher dose of the same therapeutic immune cells were tested, and instead the immunotherapy significantly exacerbated complications associated with the lethal radiation and BMT. This detrimental effect appeared to be the result of strong in vivo nonspecific immune responses induced by either activated therapeutic immune cells or interaction between therapeutic immune cells and MHC-mismatched bone marrow cells transplanted or both. Our data suggest that the immunotherapy with appropriately selected dosages may be beneficial to BMT but vigorous in vivo immune responses soon after BMT may exacerbate post-transplant complications.
Assuntos
Transplante de Medula Óssea/imunologia , Rejeição de Enxerto/prevenção & controle , Imunoterapia/métodos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/transplante , Animais , Linfócitos B/imunologia , Técnicas de Cultura de Células , Feminino , Hematopoese/imunologia , Células Matadoras Naturais/imunologia , Ativação Linfocitária/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Monócitos/imunologia , Linfócitos T/imunologia , Transplante Homólogo , Transplante IsogênicoRESUMO
Recovery of multilineage hematopoiesis from severe myelosuppression due to chemotherapy and radiotherapy remains a clinical problem. The authors have developed a simple immunotherapy to treat this disease in a mouse model. Syngeneic spleen cells or xenogeneic human peripheral mononuclear cells were cultured ex vivo with a combination of IL-2 at 500 IU/mL, GM-CSF at 200 U/mL, and calcium ionophore A23187 at 100 ng/mL for 2 days and injected intravenously into mice that had previously received a lethal dose of carboplatin and radiation. The therapy was highly effective: a single injection of activated cells enhanced survival and simulated multilineage recovery of hematopoiesis. Ex vivo activated immune cells produced multiple cytokines, including several hematopoietic growth factors. Adherent cells were found to be more potent than nonadherent cells in promoting survival, and the therapy alone was capable of mobilizing peripheral blood stem cells in normal mice.
Assuntos
Anemia Aplástica/terapia , Transplante de Células/métodos , Imunização Passiva/métodos , Mielopoese/efeitos dos fármacos , Anemia Aplástica/etiologia , Animais , Antígenos CD34/análise , Medula Óssea/efeitos dos fármacos , Medula Óssea/patologia , Medula Óssea/efeitos da radiação , Carboplatina/toxicidade , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Células Cultivadas , Citocinas/metabolismo , Contagem de Eritrócitos , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Imunossupressores/toxicidade , Interleucina-2/farmacologia , Contagem de Leucócitos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/transplante , Camundongos , Camundongos Endogâmicos BALB C , Mielopoese/efeitos da radiação , Contagem de Plaquetas , Baço/citologia , Células-Tronco/química , Células-Tronco/citologia , Taxa de Sobrevida , Transplante Heterólogo , Transplante Isogênico , Irradiação Corporal Total/efeitos adversosRESUMO
OBJECTIVE: To explore the methods for rapid in vitro culture of the dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) under serum-free conditions and ascertain whether intracellular signal transduction pathway differs between calcium ionophore (CI) and tumor necrosis factor (TNF)- alpha during their induction of dendritic cell differentiation. METHODS: PBMCs isolated from healthy donors were plated in serum-free medium supplemented with 50 ng/ml rhGM-CSF. Cells cultured overnight were induced to differentiate with 100 ng/ml A23187 or 50 ng/ml TNF-alpha, given before or 30 min after pre-treatment with 0.5 mug/ml cyclosporine A (CsA). After culture for 40 h, the cell morphology was observed under phase-contrast microscope, and the surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was employed to assess the proliferation of the allogeneic T cells. RESULTS: PBMCs of healthy donors treated with 50 ng/ml rhGM-CSF in combination with 100 ng/ml CI or 50 ng/ml TNF-alpha for 40 h exhibited typical morphology of DCs with rapidly decreased CD14 expression and increased expressions of CD83 and co-stimulatory molecules (CD80 and CD86), showing also enhanced ability of stimulating allogeneic T cell proliferation. Calcineurin antagonist CsA inhibited the differentiation induced by CI, but not that induced by TNF-alpha. CONCLUSIONS: Under serum-free conditions, both CI and TNF-alpha are capable of inducing rapid DC differentiation from human PBMCs, but the intracellular signal transduction of CI-induced differentiation is different from that induced by TNF-alpha.
Assuntos
Diferenciação Celular/fisiologia , Células Dendríticas/citologia , Monócitos/citologia , Transdução de Sinais , Calcimicina/farmacologia , Células Cultivadas , Meios de Cultura Livres de Soro , Ciclosporina/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Proteínas Recombinantes , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
AIM: To explore the intracellular signal transduction pathway in the differentiation of human peripheral blood mononucleocytes (PBMCs) towards dendritic cells (DCs) induced by calcium ionophore (CI). METHODS: PBMCs isolated from a healthy donor were cultured with A23187 plus rhGM-CSF, 100 microg/L each. In some experiments, PBMCs were cultured for 30 minutes with W-7 (10 micromol/L), CsA(0.5 mg/L) or KT5926(1 micromol/L) before addition of rhGM-CSF and A23187. After culture for 40 hours, morphological change of the cells were observed under phase contrast microscope; surface markers on treated PBMCs were analyzed by flow cytometry; the proliferation of allogeneic human T cells stimulated by the treated PBMCs was detected by MTT colorimetry. RESULTS: PBMCs of the healthy donor cocultured with rhGM-CSF plus CI for 40 hours had the typical morphology of DCs, with decreased CD14 expression, and increased CD83, CD80 and CD86 expressions. The proliferation of allogeneic T cells stimulated by PBMCs treated with A23187 plus rhGM-CSF was strengthened. But the morphological changes, surface marker expressions and the ability to enhancing proliferation of allogeneic T cells were inhibited to different degrees by W-7, CsA or KT5926. CONCLUSION: The differentiation of PBMCs towards DCs by CI may be modulated by Ca (2+)/calmodulin and multiple signal transduction pathways downstream.
Assuntos
Calcimicina/farmacologia , Células Dendríticas/citologia , Ionóforos/farmacologia , Leucócitos Mononucleares/fisiologia , Transdução de Sinais , Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Canais de Cálcio/farmacologia , Carbazóis/farmacologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Dendríticas/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Imunoglobulinas/metabolismo , Indóis/farmacologia , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/imunologia , Receptores de Lipopolissacarídeos/metabolismo , Glicoproteínas de Membrana/metabolismo , Sulfonamidas/farmacologia , Antígeno CD83RESUMO
Aplastic anemia is a bone marrow failure disorder characterized by pancytopenia and a hypocellular marrow. Benzene is one of the etiologic agents capable of inducing the disease. With modest to severe aplastic anemia, one previously untreated patient and 13 patients who had failed immunosuppressive therapy were studied. Peripheral blood mononuclear cells from patients were expanded in vitro with a combination of cytokines and a calcium-mobilizing agents for 2 days, and the activated cells were infused intravenously once a week. In some cases, we used allogenic leukocytes instead of autologous cultured lymphocytes. After 6-35 weeks of the treatment, all patients had multilineage responses to this therapy and achieved complete disease remission, defined as normal blood count, independence from transfusion, and normal bone marrow histology. The therapy was safe and well tolerated with minimal side effects. The cultured cells produced interleukin-1 and induced immune responses in vivo. Serum interleukin-2 and interferon- gamma were detected following cell infusion. Finally, patients had sustained responses to the therapy and no relapse was found up to 18 months after cellular therapy.
Assuntos
Anemia Aplástica/terapia , Imunoterapia/métodos , Transplante de Células-Tronco/métodos , Adulto , Anemia Aplástica/etiologia , Benzeno/intoxicação , Contagem de Células Sanguíneas , Transfusão de Sangue , Medula Óssea/anatomia & histologia , Medula Óssea/patologia , Relação CD4-CD8 , Calcimicina/farmacologia , Ensaio de Imunoadsorção Enzimática , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Hemoglobinas/análise , Humanos , Interferon gama/sangue , Interleucina-1/sangue , Interleucina-2/análise , Interleucina-2/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/transplante , Masculino , Exposição Ocupacional/efeitos adversos , Recidiva , Indução de Remissão , Transplante de Células-Tronco/efeitos adversosRESUMO
OBJECTIVE: To explore the effect of treatment with immunocyte therapy on benzene-induced haemopoietic dysfunction. METHODS: Mono-nuclear cells (MNC) were separated from 40 - 50 ml peripheral blood in patients and mixed with interleukin-2 and granulocyte macrophage colony stimulating factor (GM-CSF) for six day cultivation. The new formed immunocytes were collected and transfused into the patients. Bone marrow aspiration and biopsy were taken before and after therapy for all patients with severe benzene poisoning. Blood samples were stained by flow cytometry for detecting CD(4) and CD(8) positive cells. RESULTS: Of 20 patients with chronic benzene poisoning, 9 were severe benzene poisoning. All examination including blood count, bone marrow biopsy and T cell subpopulation restored to normal after immunocyte therapy. Laboratory tests (liver and kidney function, and myocardial enzymes) were observed periodically and showed normal during therapy. Follow-up study (the longest time was more than 15 months) showed that bone marrow haemopietic function of all treated patients were in normal range. CONCLUSION: Bone marrow haemopoietic dysfunction caused by benzene poisoning may be closely related to disorder of immune function. Immunocyte therapy may significantly improve bone marrow haemopoietic dysfunction induced by benzene poisoning.