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OBJECTIVE: To establish a visual reporting system for evaluating the activity of collagen â α 1 chain (COL1A1) gene promoter in immortalized human hepatic stellate cells, so as to estimate the activation status of the cells and provide a new cell model for the screening and study of anti-hepatic fibrosis drugs. METHODS: The promoter sequence of human COL1A1 was amplified from the genomic DNA of human hepatocarcinoma cell line HepG2. Based on the pLVX-AcGFP1-N1 plasmid, the recombinant plasmid pLVX-COL1A1-enhanced green fluorescent protein (EGFP) was constructed, in which the enhanced green fluorescent protein gene expression was regulated by the COL1A1 promoter. The monoclonal cell line was acquired by stably transfecting pLVX-COL1A1-EGFP into the immortalized human hepatic stellate cell line LX-2 by the lentivirus packaging system and screening. The cell line was treated with transforming growth factor-ß1 (TGF-ß1) or co-treated with TGF-ß1 and drugs with potential anti-hepatic fibrosis effects. The EGFP fluorescence intensity in cells was analyzed by the fluorescence microscope and ImageJ 1.49 software using a semi-quantitative method. The COL1A1 and EGFP mRNA were detected by reverse transcription real-time quantitative PCR (RT-qPCR), and corresponding proteins were detected by Western blot. RESULTS: The recombinant plasmid pLVX-COL1A1-EGFP with the expression of EGFP regulated by COL1A1 promoter was successfully constructed. Kozak sequence was added to enhance the expression of EGFP, which was identified by double digestion and sequencing. The LX-2 monoclonal cell line LX-2-CE stably transfected with pLVX-COL1A1-EGFP was obtained. After co-treatment with TGF-ß1 and 5 µmol/L dihydrotanshinone â with potential anti-hepatic fibrosis effect for 24 h, the total fluorescence intensity and the average fluorescence intensity of LX-2-CE were lower than those in TGF-ß1 single treatment group (P < 0.05), the intracellular mRNA and protein levels of COL1A1 and EGFP were also lower than those in the TGF-ß1 single treatment group (P < 0.05). CONCLUSION: A reporter system for estimating activation of hepatic stellate cells based on COL1A1 promoter regulated EGFP expression is successfully constructed, which could visually report the changes in COL1A1 expression, one of the activation-related markers of hepatic stellate cells, in vitro. It provides a new cell model for the screening and study of anti-hepatic fibrosis drugs.
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Células Estreladas do Fígado , Fator de Crescimento Transformador beta1 , Humanos , Fator de Crescimento Transformador beta1/farmacologia , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Cirrose Hepática/genética , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo I/farmacologia , RNA Mensageiro/metabolismoRESUMO
OBJECTIVE: To investigate the effect of CD8+ CD28- T cells on acute graft-versus-host disease(aGVHD) after haploidentical hematopoietic stem cell transplantation(haplo-HSCT). METHODS: The relationship between absolute count of CD8+ CD28- T cells and aGVHD in 60 patients with malignant hematological diseases was retrospectively analyzed after haplo-HSCT, and the differences in the incidence rate of chronic graft-versus host disease(cGVHD), infection and prognosis between different CD8+ CD28- T absolute cells count groups were compared. RESULTS: aGVHD occurred in 40 of 60 patients after haplo-HSCT, with an incidence rate of 66.67%. The median occurrence time of aGVHD was 32.5(20-100) days. At 30 days after the transplantation, the absolute count of CD8+ CD28- T cells of aGVHD group was significantly lower than that of non-aGVHD group (P =0.03). Thus the absolute count of CD8+ CD28- T cells at 30 days after transplantation can be used to predict the occurrence of aGVHD to some extent. At 30 days after transplantation, the incidence rate of aGVHD in the low cell count group (CD8+ CD28- T cells absolute count < 0.06/µl) was significantly higher than that in the high cell count group (CD8+ CD28- T cells absolute count ≥0.06/µl,P =0.011). Multivariate Cox regression analysis further confirmed that the absolute count of CD8+ CD28-T cells at 30 days after transplantation was an independent risk factor for aGVHD, and the risk of aGVHD in the low cell count group was 2.222 times higher than that in the high cell count group (P =0.015). The incidence of cGVHD, fungal infection, EBV infection and CMV infection were not significantly different between the two groups with different CD8+ CD28- T cells absolute count. The overall survival, non-recurrent mortality and relapse rates were not significantly different between different CD8+ CD28- T cells absolute count groups. CONCLUSION: Patients with delayed CD8+ CD28- T cells reconstitution after haplo-HSCT are more likely to develop aGVHD, and the absolute count of CD8+ CD28- T cells can be used to predict the incidence of aGVHD to some extent. The absolute count of CD8+ CD28- T cells after haplo-HSCT was not associated with cGVHD, fungal infection, EBV infection, and CMV infection, and was also not significantly associated with the prognosis after transplantation.
Assuntos
Antígenos CD28 , Linfócitos T CD8-Positivos , Doença Enxerto-Hospedeiro , Transplante de Células-Tronco Hematopoéticas , Humanos , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Estudos Retrospectivos , Prognóstico , Transplante Haploidêntico , Doença Aguda , Masculino , Feminino , AdultoRESUMO
OBJECTIVE: To evaluate the efficacy and safety of unmanipulated haploidentical allogeneic peripheral blood stem cells transplantation (PBSCT) on hematologic diseases. METHODS: Patients who underwent unmanipulated HLA-mismatched/haploidentical PBSCT from July 2007 to December 2011 were investigated retrospectively. RESULT: Forty-nine patients with hematologic diseases underwent unmanipulated human leukocyte antigen (HLA)-mismatched/haploidentical PBSCT with myeloablative conditioning. All patients were mismatched at the allele level for HLA-A, HLA-B, HLA-Cw, HLA-DRB1 and HLA-DQ1. Fifteen patients were mismatched in 5 loci, 11 patients in 4 loci, 7 patients in 3 loci, 5 patients in 2 loci, and 11 patients in 1 locus. The median numbers of mononuclear cells and CD34⺠cells infused at transplantation were 10.01(7.05-25.34) × 108/kg and 4.51 (2.01-11.47) × 106/kg, respectively. Patients achieved myeloid and platelet engraftment at a median of 14 (10-25) days and 22 (10-135) days, respectively. The cumulative incidence of acute graft versus host disease (aGVHD) on day 100 was (61.6 ± 7.3)%, and the 2-year cumulative incidence of chronic graft versus host disease (cGVHD) was (42.6 ± 8.5)%. One hundred-day transplantation related mortality (TRM) rate and 2-year cumulative TRM rate were (14.7 ± 5.1)% and (30.9 ± 8.8)%, respectively. The 2-year cumulative incidence estimate of relapse was (25.4 ± 7.0)%. The 2-year cumulative overall survival rate was (58.1 ± 8.8)% and 2-year disease-free survival rate was (53.9 ± 8.4)% with an 11.5-months median follow-up. CONCLUSION: Unmanipulated PBSCT is a promising protocol for patients with hematologic diseases in HLA-mismatched/haploidentical transplant settings.
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Doenças Hematológicas/cirurgia , Histocompatibilidade , Transplante de Células-Tronco de Sangue Periférico/métodos , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Antígenos HLA/imunologia , Haploidia , Doenças Hematológicas/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Condicionamento Pré-Transplante , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: The impacts of previous cardio-cerebrovascular disease (pre-CCVD) on the outcomes of hematopoietic cell transplantation (HCT) are not well described. Patients with pre-CCVD may often be poor candidates for HCT. This study aimed to investigate the impact of pre-CCVD on transplant outcomes. METHODS: A retrospective study was conducted between patients with and without pre-CCVD who consecutively received allogeneic or autologous HCT between November 2013 and January 2020 with a matching of age and disease status. The cardiovascular complications and HCT outcomes of the two groups were evaluated and compared. The primary endpoints were post-transplant cardio-cerebrovascular disease (post-CCVD) and non-relapse mortality (NRM). We used a multivariable Cox proportional hazard model and the Fine-Gray competing risk regressions for analyses to estimate the hazard ratios (HRs). RESULTS: The outcomes of 23 HCT recipients with pre-CCVD were compared with those of 107 patients in the control group. No significant differences were noted in terms of engraftment, overall survival (OS) (67.00% vs. 67.90%, Pâ=â0.983), or relapse (29.78% vs. 28.26%, Pâ=â0.561) between the pre-CCVD group and the control group. The cumulative incidences of 2-year NRM were similar between patients with pre-CCVD and the controls (14.68% vs. 17.08%, Pâ=â0.670). However, pre-CCVD was associated with an increased incidence of post-CCVD (HR: 12.50, 95% confidence interval [CI]: 3.88-40.30, Pâ<â0.001), which was an independent risk factor for increased NRM (HR: 10.29, 95% CI: 3.84-27.62, Pâ<â0.001) and inferior OS (HR: 10.29, 95% CI: 3.84-27.62, Pâ<â0.001). CONCLUSIONS: These findings suggest that the existence of pre-CCVD before transplantation might not result in increased mortality directly but superpose the toxicity of the transplantation procedure, leading to a risk of post-CCVD. Post-CCVD was a powerful predictor for high NRM and inferior OS. Further risk stratification of pre-CCVD is needed to reduce NRM in various transplantation settings.
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Transtornos Cerebrovasculares , Transplante de Células-Tronco Hematopoéticas , Transtornos Cerebrovasculares/etiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Condicionamento Pré-Transplante , Transplante AutólogoRESUMO
OBJECTIVE: To study the molecular characteristics and clinical significance of elderly patients with acute myeloid leukemia (AML). METHODS: Dideoxy sequencing was used to analyze the mutation spectrum and clinical significance of 51 hematopathy-related genes in 52 patients with newly diagnosed elderly AML. The efficacy of 39 patients receiving DCAG chemotherapy was also analyzed. RESULTS: The mutational frequency was high in elderly AML patients (98.1%, 51/52), and there were some coexistence or mutual exclusion between different mutations. Both the number of mutations and the incidence of epigenetic mutations DNMT3A, TET2 (P<0.01), as well as FLT3-ITD (P<0.05) increased with age. c-KIT mutations were most common in favorable-risk AML (P<0.01), while NPM1 and DNMT3A were common in intermediate-risk AML (P<0.05), especially in AML with normal karyotype. The complete remission rate of elderly AML patients receiving DCAG chemotherapy was 71.8% (28/39). CONCLUSION: Elderly AML patients have specific molecular characteristics, and the incidence of methylation-related gene mutations is very high, showing a certain significance for clinical diagnosis and treatment.
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Leucemia Mieloide Aguda , Proteínas Nucleares , Idoso , Humanos , Leucemia Mieloide Aguda/genética , Mutação , Taxa de Mutação , Proteínas Nucleares/genética , Nucleofosmina , PrognósticoRESUMO
Many oncogenes are involved in the progression from low-grade squamous intraepithelial lesions (LSILs) to high-grade squamous intraepithelial lesions (HSILs); which greatly increases the risk of cervical cancer (CC). Thus, a reliable biomarker for risk classification of LSILs is urgently needed. The prolyl isomerase Pin1 is overexpressed in many cancers and contributes significantly to tumour initiation and progression. Therefore, it is important to assess the effects of cancer therapies that target Pin1. In our study, we demonstrated that Pin1 may serve as a biomarker for LSIL disease progression and may constitute a novel therapeutic target for CC. We used a the novel Pin1 inhibitor KPT-6566, which is able to covalently bind to Pin1 and selectively target it for degradation. The results of our investigation revealed that the downregulation of Pin1 by shRNA or KPT-6566 inhibited the growth of human cervical cancer cells (CCCs). We also discovered that the use of KPT-6566 is a novel approach to enhance the therapeutic efficacy of cisplatin (DDP) against CCCs in vitro and in vivo. We showed that KPT-6566-mediated inhibition of Pin1 blocked multiple cancer-driving pathways simultaneously in CCCs. Furthermore, targeted Pin1 treatment suppressed the metastasis and invasion of human CCCs, and downregulation of Pin1 reversed the epithelial-mesenchymal transition (EMT) of CCCs via the c-Jun/slug pathway. Collectively, we showed that Pin1 may be a marker for the risk of progression to HSIL and that inhibition of Pin1 has anticancer effects against CC.
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OBJECTIVE: To investigate the clinical features,therapy and prognosis of patients with peripheral T cell lymphoma(PTCL), and to find out the prognostic factors of the disease. METHODS: The clinical data of 73 patients with PTCL were reviewed.The median pre-treatment disease course was 3 months.Fifty-five patients were males, and 18 were females, with the median age of 42 years.Five patients received the combined chemo-radio therapy, 65 received chemotherapy alone, and the other 3 patients were treated with auto hematopoietic stem cell transplantation (HSCT). RESULTS: Of all the patients, the overall 3 -year and 5-year survival rates were 38% (28 /73) and 22% ( 16 /73) respectively.The survival rates decreased with the progression of the Ann Arbor stages.The survival rate of the patients with B symptom (fever, night sweat, and weight loss) or the international prognostic factors index ( IPI)>2 was lower than those of the patients without B symptom or IPI<2.The patients with the increased CA125 or D-dimer lever had the worst curative effect. CONCLUSION: Peripheral T cell lymphoma is highly aggressive with poor prognosis.The clinical features,Ann Arbor staging, IPI and B symptom are important prognostic factors.CA125 and D-dimer may be also important prognostic factors.
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Protocolos de Quimioterapia Combinada Antineoplásica , Linfoma de Células T Periférico/diagnóstico , Linfoma de Células T Periférico/terapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Antígeno Ca-125/sangue , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , Humanos , Linfoma de Células T Periférico/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
OBJECTIVE: To evaluate the efficacy and safety of low-dose amphotericin B (AmB) in different antifungal strategies for treatment of invasive fungal disease(IFD) in patients with hematologic malignancies. Metheds: The clinical dada of the patients were collected and analyzed retrospectively and the levels of creatinine (Cr), urea nitrogen (BUN) and potassium (K+) before and after using low-dose AmB were compared and statistically analyzed. RESULTS: Among 97 cases, 2 cases were diagnosed as invasive fungal disease (IFD), 11 cases were diagnosed as clinical probable IFD, 15 cases were diagnosed as possible IFD, 69 cases were undefined IFD. The response rate of all patients treated with low-dose AmB was 69.4%, the response rate for targed therapy was 72.7%, the response rate for diagnosis-driven therapy was 63.6%, the response rate of empirical therapy was 75%, the efficacy of the combination with other antibiotics was 50%, 66.7% and 75%. According to all the patients received AmB, only 7 cases was detected with higher level of Cr (7.2) than normal and this level come back to normal with in 7 days after drug withdrew. Although the Cr level in serum after 1 day of drug withdrew was higher than that before administration of drug(64.86±3.00 vs 58.76±1.67 µmol/L) and was with statistical difference(P<0.05), but did not show significant difference in comparison with the level after drug withdrew 7 days (58.43±1.68 µmol/L,P>0.05). CONCLUSION: AmB injection is an effective and safe method in empirical therapy and diagnosis-driven antifungal therapy for neutropenic, febrile patients with hematological malignancies.
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Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Micoses/tratamento farmacológico , Creatinina , Neoplasias Hematológicas/complicações , Humanos , Micoses/etiologia , Estudos RetrospectivosRESUMO
Tyrosine kinase inhibitor (TKI) therapy significantly improved the prognosis and outcome of patients with chronic myeloid leukemia(CML). Long-term therapy of TKI drugs was often accompanied with financial burden and the rise of chronic adverse effects. At present, the treatment-free remission (TFR) has been gradually regarded as the new ultimate aim to the patients with long-term CML. In clinical trials, the patients with the therapy of imatinib stopping TKI treatment after acquired deep molecular reaction still maintained remission. Here, the research progress on discontinuation of TKI therapy and how to better grasp the safety of drug withdrawal strategy are reviewed. However, the radical cure of CML needs more further research.
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Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Objetivos , Humanos , Mesilato de Imatinib , Prognóstico , Proteínas Tirosina QuinasesRESUMO
OBJECTIVE: To investigate the clinical characteristics, therapeutic efficacy survival and prognosis of patients with adult acute lymphoblastic leukemia (ALL) accompanied by central nervous system leukemia (CNSL). METHODS: The clinical and cerebrospinal fluid (CSF) features, diagnosis and treatment, therapeutic efficacy and survival rate of 21 cases of acute lymphoblastic leukemia (ALL) with central nervous system involvement (CNSL) were analyzed retrospectively. RESULTS: Out of 21 cases, 10 cases were B cell acute lymphoblastic leukemia(B-ALL), 6 cases were T cells acute lymphoblastic(T-ALL), 4 cases were determined as no clear typing, 1 case was Burkitt lymphoma/leukemia, 7 patients had CNSL at the time of diagnosis, and 14 patients were showed CNS relapse. Clinical manifestations included headache, facial paralysis, limb weakness and blurred vision, etc. Their median follow-up time was 19(6-40) months,from them 10 cases died, 7 cases survived, 4 cases were lost to follow up. Patients had CNSL at the time of diagnosis, their peripheral blood LDH≥600 U/L or not achieving complete remission (CR) after 1 course of treatment with poor prognosis, and the difference is significant (P< 0.05). Radiotherapy and allogeneic stem cell transplantation (allo-HSCT) could improve the patient's survival. Multivariate analysis showed that the LDH and allo-HSCT was significantly correlated with survival time (P=0.048, P=0.013). CONCLUSION: There are no specific clinical manifestations, CSF features and imaging manifestations of ALL accompanied by CNSL, and the diagnosis of CSF is needed to find the leukemia cells in CSF. The factors for poor prognosis include LDH≥600 U/L, no CR of patients after 1 course of treatment, existence of CNSL at the diagnosis. ALL patients with CNSL have a poor prognosis. Intrathecal injection combined with systemic chemotherapy, radiation therapy and allo-HSCT after CR is the feasible and effective treatment regimen.
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Neoplasias do Sistema Nervoso Central , Leucemia-Linfoma Linfoblástico de Células Precursoras , Adulto , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas , Humanos , Prognóstico , Recidiva , Indução de Remissão , Estudos Retrospectivos , Taxa de Sobrevida , Transplante Homólogo , Resultado do TratamentoRESUMO
OBJECTIVE: To investigate the inductive therapeutic effects of imatinib combined with VP low dose regiment on adult patients with Ph-positive acute lymphoblastic leukemia (Ph(+) ALL). METHODS: Fourteen newly diagnosed adult patients with Ph(+) ALL were treated with VP regimen, and imatinib (400 mg/d) was added at the 8(th) day. VP regimen would be stopped when neutropenia lasted for 1 week or complicated with infection, fever, etc. Therapeutic effects were assessed by bone marrow morphology and quantitative analysis of BCR/ABL:ABL at the 28(th) - 33(rd) day. Patients could be treated with imatinib combined with chemotherapy for consolidation and maintenance therapy or were treated with allogeneic hematopoietic stem cell transplantation after complete remission. RESULTS: Fourteen cases obtained CR1 after first course of treatment, the median decline of BCR/ABA:ABL was 55.89 (10.25 -180.97) %; during the induction chemotherapy, pulmonary infection occurred in 3 patients, diarrhea in 1 patients, facial edema in 3 patients, however, all these patients were cured after symptomatic treatment, only 1 patient died of relapse after transplantation. CONCLUSION: In the period of tyrosine kinase inhibitor (TKI), inductive chemotherapy combined with imatinib and low dose VP can obtaine satisfactory CR rate and decrease the toxicity of the traditional drugs. It is suggested that TKI combined with VP regimen chemotherapy can achieve CR1 and make possible for allo-HSCT, from which patients can achieve the long-term survival.
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Leucemia-Linfoma Linfoblástico de Células Precursoras , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica , Medula Óssea , Cisplatino , Proteínas de Fusão bcr-abl , Transplante de Células-Tronco Hematopoéticas , Humanos , Mesilato de Imatinib , Quimioterapia de Indução , Neutropenia , Inibidores de Proteínas Quinases , Recidiva , Indução de Remissão , Transplante Homólogo , VindesinaRESUMO
The petroleum ether, chloroform, and methyl alcohol extracts of wild and cultivated Rhodiola saccharinensis A. Bor and Rhodila angusta Nakai taken from Changbai Mountains, and obtained by the same isolation procedure, were for the first time studied with Fourier transform infrared spectrometry(FTIR), and the spectra obtained were compared with those of a sort of goods named Rhodiola Cake (Hong Jing Tian Kuai). It was shown that the infrared spectra of the wild and cultivated Rhodiola saccharinensis A. Bor and Rhodila angusta Nakai are different, which can be used to determine and evaluate their respective species.
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Identificação Biométrica/métodos , Rhodiola/química , Especificidade da Espécie , Espectrofotometria Infravermelho/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Algoritmos , China , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/normas , Extratos Vegetais/análise , Extratos Vegetais/química , Padrões de Referência , Pesquisa , SolventesRESUMO
This study was aimed to explore the efficacy and associated complications of haploidentical peripheral blood stem cell transplantation (hi-PBSCT) without ex vivo T-cell depletion in treatment of hematological malignancies. 15 high-risk patients received HLA 1-3 loci (A, B, or DRB1) mismatched hi-PBSCT. The modified Bu/Cy or TBI/Cy regimen was used for preconditioning of patients. The anti-thymocyte globulin, cyclosporin A, methotrexate and mycophenolate mofetil were used for GVHD prophylaxis. 4 cases were administrated with anti-CD25 monoclonal antibody. G-CSF-mobilized peripheral blood stem cells were infused, with the median number of infused nucleated cells was 8.16 (3.92-10.86)x10(8)/kg and that of CD34+ cells was 4.51 (1.27-5.95)x10(6)/kg. The results showed that the rapid engraftment was observed in all cases. The median times of neutrophil recovery>or=0.5x10(9)/L and platelet recovery>or=20x10(9)/L were 14 (11-19) and 22 (11-52) days after transplantation respectively. 6 cases developed acute GVHD of grade I-II, and 2 cases experienced chronic extensive GVHD. Infection within 100 days after hi-PBSCT was documented in all cases. 8 cases were subjected to bacterial infection, and six got cytomegalovirus infection. Relapse occurred in five cases. Overall survival of patients was 46.7% (7/15), with a median follow-up of 213 (42-589) days. In conclusion, hi-PBSCT provides an effective alternative treatment for high-risk patients in lack of matched donors, and to reduce the high transplantation-related mortality.
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Neoplasias Hematológicas/cirurgia , Transplante de Células-Tronco de Sangue Periférico/métodos , Condicionamento Pré-Transplante/métodos , Adolescente , Adulto , Feminino , Sobrevivência de Enxerto , Doença Enxerto-Hospedeiro , Haploidia , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Depletion of T and B cells from the graft is prerequisite for haploidentical transplantation to decrease the risk of GVHD and EBV-associated lymphoproliferative disease. This study was aimed to investigate the performance of T-cell and B-cell simultaneous depletion from mobilized peripheral blood stem cells (PBSCs) for the first time in China, using anti-CD3 and anti-CD19 antibodies conjugated to magnetic microbeads by the CliniMACS device. The depletion efficiency of T-cell and B-cells was analyzed by flow cytometry; the function of the stem cells after depletion was evaluated using colony assays. The results indicated that the mononuclear cell count prior to T- and B-cell depletion was 4.88 x 10(10). After depletion, the percentage of T cells was 0.02% with a log (10) depletion of 4.4. The percentage of B cells was less than 0.01% with a log (10) depletion of at least 3.3. The product contained not only CD34(+) stem cells, but also NK cells, monocytes and granulocytes. After T- and B-cell depletion the purity of CD34(+) cells was 0.98%, the number of CD34 cells was 1.84 x 10(8) and their recovery rate was 69.7%. The number of NK cells was 2.54 x 10(9) and the recovery rate of NK cells was 71.7%. In vitro colony assays showed no negative impact on function of the hematopoietic stem cells. In conclusion, the CliniMACS system can be used to efficiently deplete T and B cells from PBSCs simultaneously, without adverse effect on biological function of hematopoietic stem cells. This study provides technical platform for haploidentical hematopoietic stem cell transplantation.