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1.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 19(3): 781-6, 2011 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-21729571

RESUMO

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 Jovem
2.
Artigo em Chinês | MEDLINE | ID: mdl-17997890

RESUMO

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 Jovem
3.
Di Yi Jun Yi Da Xue Xue Bao ; 24(11): 1263-6, 2004 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-15567774

RESUMO

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/farmacologia
4.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 20(5): 540-3, 2004 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-15367342

RESUMO

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 CD83
5.
Artigo em Chinês | MEDLINE | ID: mdl-14761426

RESUMO

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.


Assuntos
Anemia Aplástica/terapia , Benzeno/intoxicação , Transplante de Células-Tronco de Sangue Periférico/métodos , Adulto , Anemia Aplástica/induzido quimicamente , Anemia Aplástica/imunologia , Medula Óssea/imunologia , Medula Óssea/patologia , Feminino , Citometria de Fluxo , Seguimentos , Humanos , Masculino , Doenças Profissionais/induzido quimicamente , Doenças Profissionais/imunologia , Doenças Profissionais/terapia , Resultado do Tratamento
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