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1.
Cancer Med ; 2019 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-31518054

RESUMO

BACKGROUND: CXCR4 chemokine receptors play an important role in leukemia proliferation, extramedullary migration, infiltration, adhesion, and resistance to chemotherapy drugs. METHODS: The CXCR4 expression by flow cytometry in 122 acute myeloid leukemia (AML) patients between 2010 and 2014 was analyzed. RESULTS: The expression of CXCR4 in AML-M4/M5 was found to be significantly higher than that of other subtypes according to both FAB subtype and WHO classification. The FLT3-ITD mutant was significantly higher in high CXCR4 expression group (P = .0086). Our data also showed that CXCR4 expression was correlated with CD64 expression. Low CXCR4 expression on AML cells was associated with better prognosis, and the median overall survival (OS) for low CXCR4 expression patients was 318 days, compared with 206 days for patients with high CXCR4 expression (P = .045). Multivariate analysis revealed that CXCR4 expression, age, and extramedullary infiltration were independent prognostic factors. CONCLUSIONS: Our study demonstrated that CXCR4 expression in AML was an independent prognostic predictor for disease survival that could be rapidly and easily determined by flow cytometry at disease presentation.

3.
J Transl Med ; 17(1): 191, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31171000

RESUMO

BACKGROUND: Elevated protein expressions of CD markers such as IL2RA/CD25, CXCR4/CD184, CD34 and CD56 are associated with adverse prognosis in acute myeloid leukemia (AML). However, the prognostic value of mRNA expressions of these CD markers in AML remains unclear. Through our pilot evaluation, IL2RA mRNA expression appeared to be the best candidate as a prognostic biomarker. Therefore, the aim of this study is to characterize the prognostic value of IL2RA mRNA expression and evaluate its potential to refine prognostification in AML. METHODS: In a cohort of 239 newly diagnosed AML patients, IL2RA mRNA expression were measured by TaqMan realtime quantitative PCR. Morphological, cytogenetics and mutational analyses were also performed. In an intermediate-risk AML cohort with 66 patients, the mRNA expression of prognostic biomarkers (BAALC, CDKN1B, ERG, MECOM/EVI1, FLT3, ID1, IL2RA, MN1 and WT1) were quantified by NanoString technology. A TCGA cohort was analyzed to validate the prognostic value of IL2RA. For statistical analysis, Mann-Whitney U test, Fisher exact test, logistic regression, Kaplan-Meier and Cox regression analyses were used. RESULTS: In AML cohort of 239 patients, high IL2RA mRNA expression independently predicted shorter relapse free survival (RFS, p < 0.001) and overall survival (OS, p < 0.001) irrespective of age, cytogenetics, FLT3-ITD or c-KIT D816V mutational status. In core binding factor (CBF) AML, high IL2RA mRNA expression correlated with FLT3-ITD status (p = 0.023). Multivariable analyses revealed that high IL2RA expression (p = 0.002), along with c-KIT D816V status (p = 0.013) significantly predicted shorter RFS, whereas only high IL2RA mRNA expression (p = 0.014) significantly predicted shorter OS in CBF AML. In intermediate-risk AML in which multiple gene expression markers were tested by NanoString, IL2RA significantly correlated with ID1 (p = 0.006), FLT3 (p = 0.007), CDKN1B (p = 0.033) and ERG (p = 0.030) expressions. IL2RA (p < 0.001) and FLT3 (p = 0.008) expressions remained significant in predicting shorter RFS, whereas ERG (p = 0.008) and IL2RA (p = 0.044) remained significant in predicting shorter OS. Similar analyses in TCGA intermediate-risk AML showed the independent prognostic role of IL2RA in predicting event free survival (p < 0.001) and OS (p < 0.001). CONCLUSIONS: High IL2RA mRNA expression is an independent and adverse prognostic factor in AML and specifically stratifies patients to worse prognosis in both CBF and intermediate-risk AML.

4.
Ann Thorac Surg ; 2018 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-30273571

RESUMO

BACKGROUND: We investigated the frequency of c-MYC amplification in esophageal squamous cell carcinoma (ESCC), including both stage I-II and III-IVa disease, and evaluated the correlation of c-MYC amplification with clinicopathologic parameters and outcome. METHODS: In 259 ESCC resected at Zhongshan Hospital, Fudan University from January 2007 to November 2010, c-MYC amplification was analyzed using tissue microarray, with fluorescence in situ hybridization (FISH) assay. RESULTS: c-MYC gene amplification was found in 43.2% (112/259) of ESCC patients. There were significant differences between c-MYC amplification and patient age (P=0.009), and lymph node metastasis (P=0.046). The median follow-up period was 33 months (range 4-102 months). There was a survival difference between patients with different c-MYC status. Among 112 patients with c-MYC amplification, a significantly poorer prognosis was observed, with a median DFS and OS of 24.0 and 31.0 months compared to 48.0 and 48.0 months for those without c-MYC amplification (P=0.011 and 0.018). Upon univariate and multivariate analysis, site, clinical stage, lymph node metastasis, adjuvant therapy and c-MYC amplification were associated with DFS and OS. When patients were divided into stage I-II and stage III-IV subgroups, c-MYC amplification tended to associate with poorer survival, but without statistical difference (P>0.05). CONCLUSIONS: c-MYC amplification was associated with age and lymph node metastasis, and was an independent poor-prognostic factor for DFS and OS in full cohort of ESCC patients.

5.
Intern Med J ; 48(4): 439-444, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-28636094

RESUMO

BACKGROUND: Clonal Philadelphia (Ph)-negative cytogenetic abnormalities (CPCA) have been reported in chronic myeloid leukaemia (CML) patients treated with either interferon or tyrosine kinase inhibitor (TKI). However, the incidences and types of these cytogenetic abnormalities after treatment vary due to the limited populations enroled. METHODS: We analysed the frequency and types of CPCA in a cohort of 607 CML patients in the chronic phase after TKI treatment. We also followed up these CPCA with a median of 31.8 months (range from 11 to 63 months) from diagnosis and investigated their effects on disease progression. RESULTS: We found 18 out of 607 CML patients had cytogenetic abnormality in the Ph-negative cells with an incidence of 3%. In total, six types of chromosomal abnormalities have been identified in these 18 patients with the majority of them aneuploidy abnormalities, especially the trisomy 8. Four of 18 patients (22.2%) were noted to have several abnormalities in the Ph-negative cells. Furthermore, follow-up studies of these CPCA showed that they could be either persistent or transient (15 vs 3 patients), and may not affect disease progression since none of them developed transformed myelodysplasia or transformed acute myeloid leukaemia. CONCLUSION: Three percent of CML patients in the chronic phase were observed to have CPCA during TKI treatment. Our results suggest that the detection of CPCA in CML may not predict disease progression.


Assuntos
Análise Citogenética , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/diagnóstico , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Análise Citogenética/métodos , Feminino , Seguimentos , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/epidemiologia , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/epidemiologia , Masculino , Pessoa de Meia-Idade , Adulto Jovem
6.
Cell Physiol Biochem ; 42(5): 1779-1788, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28746919

RESUMO

BACKGROUND/AIMS: The transcription factor CCAAT/enhancer-binding protein α (C/EBPα) is a basic leucine zipper transcription factor that plays essential roles in tumor progression. Although decreased or absent C/EBPα expression in many cancers suggests a possible role for C/EBPα as a tumor suppressor, the functions of C/EBPα in lung adenocarcinoma remain unclear. METHODS: Here, C/EBPα expression levels in 26 lung adenocarcinoma and para-carcinoma tissue samples were detected by qRT-PCR and immunohistochemistry. Cell transwell assays, wound healing assay and three-dimensional spheroid invasion assay were performed to assess the effects of C/EBPα on migration and invasion in lung adenocarcinoma cells in vitro. Western blotting was applied to analyze the potential mechanisms. RESULTS: C/EBPα was found to be decreased in lung adenocarcinoma tissues compared to para-carcinoma tissues. Overexpression of C/EBPα significantly inhibited the migration and invasion of lung adenocarcinoma cells. In addition, C/EBPα overexpression suppressed the epithelial-mesenchymal transition (EMT) that was characterized by a gain of epithelial and loss of mesenchymal markers. Further study showed that C/EBPα suppressed the transcription of ß-catenin and downregulated the levels of its downstream targets. CONCLUSION: Our data suggest that C/EBPα inhibits lung adenocarcinoma cell invasion and migration by suppressing ß-catenin-mediated EMT in vitro. Thus, C/EBPα may be helpful as a potential target for treatment of lung adenocarcinoma.


Assuntos
Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , beta Catenina/metabolismo , Células A549 , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Proteína alfa Estimuladora de Ligação a CCAAT/genética , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Movimento Celular , Regulação para Baixo , Transição Epitelial-Mesenquimal , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Reação em Cadeia da Polimerase em Tempo Real , Transcrição Genética , Via de Sinalização Wnt
7.
Tumour Biol ; 37(3): 3817-21, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26474588

RESUMO

Interleukin-3 (IL-3) receptor α chain (CD123) plays an essential role in regulating the proliferation of hematopoietic stem cells. In the hematopoietic malignancies, CD123 expression has been found in acute myeloid leukemia (AML), B-precursor acute lymphoblastic leukemia (B-ALL), as well as dendritic cell malignancies. However, whether CD123 is also expressed in T-acute lymphoblastic leukemia (T-ALL) remains unknown. Using multi-parameter flow cytometry, we analyzed CD123 expression in 160 consecutive diagnostic T-ALL patients, including 88 pediatric T-ALL cases and 72 adult T-ALL cases. The minimal residual disease (MRD) was detected after one course of induction therapy to evaluate the treatment effects. CD123 expression was detected in 24 out of 88 (27 %) pediatric T-ALLs and 30 out of 72 (42 %) adult T-ALLs. Further analysis revealed that CD123 expression is associated with the maturation stage of T-ALLs. The frequencies of CD123-positive cases decreased from 83 to 40 % and 21 % in early T-precursor ALLs, T-precursor ALLs, and mature T-ALLs, respectively. Interestingly, we detected the CD4+CD8+ double-positive leukemic cells in 22 immature and 34 mature T-ALL patients. Of note, only 4 % of these patients expressed CD123. In addition, we found that 79 % of CD33+ and 64 % of CD117+ immature T-ALL patients also expressed CD123. However, CD123 expression did not predict the outcomes of the first course of induction therapy in T-ALL patients. In conclusion, we found that CD123 is preferentially expressed in immature T-ALL. Moreover, CD123 expression is strongly associated with cross-lineage expression of myeloid markers in early T-precursor ALL.


Assuntos
Subunidade alfa de Receptor de Interleucina-3/metabolismo , Neoplasia Residual/metabolismo , Leucemia-Linfoma Linfoblástico de Células T Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células T Precursoras/metabolismo , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Neoplasia Residual/diagnóstico , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patologia , Proteínas Proto-Oncogênicas c-kit/metabolismo , Lectina 3 Semelhante a Ig de Ligação ao Ácido Siálico/metabolismo , Adulto Jovem
8.
Mol Clin Oncol ; 3(4): 833-838, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26171191

RESUMO

Resistance to chemotherapy is a major challenge for leukemia treatment. It has been suggested that leukemia stem cells (LSCs), a small pool of self-renewing leukemic cells, play important roles in development of chemotherapy resistance. The expression of cluster of differentiation 96 (CD96), a potential marker for LSCs, was investigated in CD34+CD38- cells of 105 acute leukemia (AL) patients by flow cytometry. The data showed that all the CD34+, CD34+CD38- and CD34+CD38-CD96+ proportions were much higher in AL compared to the normal control (P<0.01), while a clear difference was identified in the CD34+CD38- and CD34+CD38-CD96+ proportions between acute lymphoid leukemia and acute myeloid leukemia (AML). However, all the AML patients with >15% CD34+CD38- cells achieved complete remission (CR), suggesting that as an LSC-rich population, the amount of CD34+CD38- cells may not be positively associated with the proportion of refractory LSCs. The mean percentage of the co-presence of CD96 expression itself was similar in AML patients with CR and non-CR (P>0.05). However, the CR rate was significantly higher in the AML population with <10% CD96 expressed, which indicated that a distinct sub-group of CD34+CD38-CD96+ cells may still contribute to the drug resistance or poor prognosis.

9.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 23(3): 637-41, 2015 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-26117008

RESUMO

OBJECTIVE: To construct a lentivirus vector carrying SARI gene and to investigate its biological effects on K562 cells. METHODS: SARI was amplified from the plasmid containing SARI cDNA and subcloned into pLOV.CMV.eGFP virus vector. After sequencing, lentivirus packaging, titering, the viruses of SARI-pLOV.CMV.eGFP were harvested and tansfected into the K562 cells. Real-time quantitive PCR and Western blot were performed to validate the SARI expression at the level of mRNA and protein respectively. Simultaneously, the proliferation, apoptosis and cell cycle of K562 cells were detected by CCK-8 and flow cytometry respectively. RESULTS: The SARI overexpressed lentivirus vector was successfully constructed. The mRNA and protein levels of SARI increased significantly in the pLOV.CMV.eGFP-SARI group, which was confirmed by Q-PCR and Western blot; as compared with blank and mock groups, SARI over-expression leaded to significant proliferation inhibition and increased apoptosis of K562 cells, without visible effects on cell cycle. CONCLUSION: the over-expression of SARI gene obviously suppresses the cell proliferation of the K562 cells as well as promotes the apoptosis. The results implied that the induction of the SARI gene expression may be an important candidate therapeutic method for the CML.


Assuntos
Expressão Gênica , Apoptose , Fatores de Transcrição de Zíper de Leucina Básica , Ciclo Celular , Linhagem Celular , Proliferação de Células , DNA Complementar , Vetores Genéticos , Humanos , Células K562 , Lentivirus , Plasmídeos , Transfecção , Proteínas Supressoras de Tumor
10.
Mol Med Rep ; 10(1): 39-44, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24788122

RESUMO

The aim of the present study was to examine the effects of epithelial-mesenchymal transition (EMT) and apoptosis of renal tubular epithelial cells on the prognosis of immunoglobulin A (IgA) nephropathy. Renal biopsy tissues from 74 cases of IgA nephropathy were divided into a mild mesangial proliferation group (27 cases), a focal hyperplasia group (28 cases) and a proliferative sclerosis group (19 cases). The blood pressure, serum creatinine and 24 h urinary protein excretion of all patients were detected. To define EMT, α-smooth muscle actin (α-SMA), vimentin and collagen fibers were assessed. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The blood pressure, serum creatinine and 24 h urinary protein excretion of patients with IgA nephropathy altered with increasing pathological grade. All clinical indices of patients in the proliferative sclerosis group were higher than those of the other two groups, and the 24 h urinary protein excretion of the focal hyperplasia group was statistically higher than that of the mild mesangial proliferation group. The expression of tubular interstitial α-SMA, vimentin and collagen fibers increased with the pathological grade and was closely correlated with clinical indices, including collagen fibers and 24 h urinary protein excretion. TUNEL-positive cells increased with the exacerbation of pathological changes. The EMT and apoptosis of renal tubular epithelial cells reflected the clinical severity of IgA nephropathy. α-SMA, vimentin and the apoptotic index may be used as important markers for evaluating the prognosis of IgA nephropathy.


Assuntos
Apoptose , Glomerulonefrite por IGA/patologia , Túbulos Renais/metabolismo , Actinas/metabolismo , Adolescente , Adulto , Colágeno/metabolismo , Progressão da Doença , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Transição Epitelial-Mesenquimal , Feminino , Glomerulonefrite por IGA/metabolismo , Humanos , Hiperplasia/metabolismo , Túbulos Renais/patologia , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Proteínas/análise , Esclerose/metabolismo , Vimentina/metabolismo , Adulto Jovem
11.
Eur J Clin Invest ; 43(11): 1140-6, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23992300

RESUMO

BACKGROUND: Increased bone marrow (BM) hematogones (HGs) are often observed in patients with marrow regenerating status. Many studies have focused on the role of HGs in acute lymphoblastic leukaemia (ALL), but very little has been done to understand their effects on acute myeloid leukaemia (AML). MATERIALS AND METHODS: Through immunophenotyping, HGs were quantified in 471 BM samples from 292 postchemotherapy AML cases. These samples were analysed to determine whether there is any relationship between HGs percentages and French-American-British (FAB) subtypes or risk stratification of AML. RESULTS: HGs were identified in 57.75% of 471 patient samples (271) with a mean percentage of 3.87 ± 0.25%. No significant differences were found amongst different FAB subtypes of AML (P > 0.05). However, significant differences (P < 0.05) in HG numbers were noted between AML patients experiencing haematological complete remission (HCR) and those who have relapsed. HGs were identified in 59.9% of samples under HCR with a mean per cent of 3.98 ± 0.31%, and 36.7% of individuals who have relapsed have detectable HGs with a mean per cent of 1.75 ± 0.47. In addition, HGs in patients groups with low risk or intermediate risk were elevated when compared with high-risk groups (P < 0.05), whilst no significant difference was found between low-risk patients and intermediate-risk patients (P > 0.05). Patients with >0.1% of HGs had a significantly better median leukaemia-free survival (LFS) and overall survival (OS) than those with <0.1% of HGs (P < 0.01). CONCLUSIONS: Therefore, our data indicate that HGs in bone marrow may be used as a favourable prognostic factor that predict for a better outcome of AML patients.


Assuntos
Células da Medula Óssea/patologia , Leucemia Mieloide Aguda/patologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Leucemia Mieloide Aguda/mortalidade , Leucemia Mieloide Aguda/terapia , Masculino , Pessoa de Meia-Idade , Prognóstico , Recidiva , Indução de Remissão , Medição de Risco , Adulto Jovem
12.
Chin Med J (Engl) ; 125(9): 1669-71, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22800843

RESUMO

A female patient diagnosed with acute myelocytic leukemia M5a (AML-M5a) relapsed 986 days after her allogeneic peripheral blood stem cell transplantation (allo-PBSCT) from an unrelated male donor with matched human leukocyte antigen (HLA). Three re-induction chemotherapies were administered, and partial remission was achieved. The patient was given repetitive infusion of cytokine-induced killer (CIK) cells expanded from recipient peripheral mononuclear cells of full donor chimerism due to loss of contact of quondam donor for donor lymphocyte infusion (DLI) and rejection of second transplantation. The patient achieved complete cytogenetical remission. This strategy might overcome the obstacle of donor unavailability and present an appealing new therapeutic alternative to donor-recruited adoptive immunotherapy for relapsed disease at post-transplantation.


Assuntos
Células Matadoras Induzidas por Citocinas/transplante , Transplante de Células-Tronco Hematopoéticas , Leucemia/terapia , Adulto , Feminino , Humanos
13.
Cancer Immunol Immunother ; 59(2): 195-202, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19847424

RESUMO

Identification and purification of antigen-specific T cells without altering their functional status are of high scientific and clinical interest. Staining with major histocompatibility complex (MHC)-peptide multimers constitutes a very powerful method to study antigen-specific T-cell subpopulations, allowing their direct visualization and quantification. MHC-peptide multimers, such as dimers, tetramers, pentamers, streptamers, dextramers and octamers have been used to evaluate the frequency of CD8(+) T cells, specific for tumor/leukemia-associated antigens as well as for viral antigens, e.g., CMVpp65 and EBV-EBNA. Moreover, MHC-peptide multimers have been used for rapid and efficient ex vivo isolation and expansion of T cells. A recent development in the field of MHC-peptide multimers led to the purification of CD8(+) T cells specific for leukemia antigens. This might help to select leukemia-specific donor lymphocyte infusions (DLIs), thus allowing dissection of the noxious graft-versus-host disease (GvHD) from beneficial anti-viral and even anti-leukemic effects. This review covers different types of MHC-peptide multimers and their applications, as well as the impact that multimers might have on further development of DLIs.


Assuntos
Transferência Adotiva/métodos , Antígenos de Neoplasias/imunologia , Separação Celular/métodos , Peptídeos/imunologia , Linfócitos T/imunologia , Linfócitos T/transplante , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/transplante , Citometria de Fluxo , Doença Enxerto-Hospedeiro/imunologia , Humanos , Leucemia/imunologia , Leucemia/terapia , Complexo Principal de Histocompatibilidade/imunologia , Peptídeos/química , Multimerização Proteica
14.
J Immunol Methods ; 343(2): 140-7, 2009 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-19248785

RESUMO

Peptide vaccination constitutes a novel immunotherapeutical approach for the treatment of patients with solid tumors, lymphoma and leukemia. Moreover it might be of use in hematooncological patients for the prevention and therapy of infections like cytomegalovirus (CMV) reactivation due to immunosuppression. To meet good manufacturing practice (GMP) criteria, we introduce here a bio-assay to validate peptide vaccines for peptide content and bio-activity. As a paradigm for peptide vaccine preparation the immunogenic CMV peptide 495-503 NLVPMVATV lyophilisate was resolubilized in dimethyl sulfoxide, phosphate buffered saline and admixed with Montanide. Addition of different amounts of peptide (10-80 microg) to a mixed lymphocyte peptide culture (MLPC) resulted in the generation of interferon (IFN) gamma and granzyme B releasing CD8(+) CMV tetramer(+) T cells in a dose dependent manner. The combination of FACS and ELISPOT results allowed the definition of the peptide amount in a vaccine preparation. Storage at +/-4 degrees C over 24 h did not result in a significant change of the immunogenicity of the vaccine. In contrast, cryopreservation of the vaccine at -20 degrees C resulted in a loss of immunogenicity. Quantitation of tumor/viral antigen peptides admixed with adjuvants, such as incomplete Freund's adjuvant (IFA), is feasible through bio-assays as the modified ELISPOT/FACS assay described here, meeting GMP criteria for multi-center trials.


Assuntos
Antígenos Virais/análise , Vacinas contra Citomegalovirus/imunologia , Leucemia/imunologia , Linfoma/imunologia , Transplante de Células-Tronco , Antígenos Virais/imunologia , Linfócitos T CD8-Positivos/imunologia , Criopreservação , Vacinas contra Citomegalovirus/normas , Ensaio de Imunoadsorção Enzimática , Granzimas/imunologia , Humanos , Interferon gama/imunologia , Vacinas de Subunidades/imunologia , Vacinas de Subunidades/normas , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/normas
15.
J Huazhong Univ Sci Technolog Med Sci ; 28(5): 520-4, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18846330

RESUMO

The homocysteine (Hcy)-induced tissue factor (TF) expression in human vascular smooth muscle cells (VSMCs) and the effect of Hcy on the activity of nuclear factor-kappaB (NF-kappaB) and the expression of inducible nitric oxide synthase (iNOS) were investigated. Human umbilical artery VSMCs were cultured by tissue explanting method, identified by alpha-actin immunohistochemistry, and incubated with different concentrations of Hcy/PTDC (NF-kappaB inhibitor). Semi-quantitative RT-PCR was performed to detect the expression of TF mRNA in VSMCs. Flow cytometry was used to assay the expression of TF protein on the surface of VSMCs and the expression of iNOS in VSMCs. Western blot was carried out to detect the expression of NF-kappaB protein in nuclei. The results showed that Hcy could induce VSMCs expressing TF mRNA significantly after the VSMCs were incubated with Hcy at concentrations of 10, 100, 500 micromol/L respectively. There was low expression level of TF protein on the surface of the resting VSMCs and Hcy could also induce VSMCs expressing TF protein on the cell surface in different concentrations. Additionally, Hcy could rapidly induce the activation of NF-kappaB and this effect could be significantly inhibited by PDTC. Hcy alone could not induce the expression of iNOS in VSMCs. It was concluded that Hcy could significantly induce the expression of TF in VSMCs and enhance the activation of NF-kappaB, subsequently mediate TF gene expression and protein synthesis. NF-kappaB-mediated expression of TF in VSMCs might be the important mechanism of atherosclerosis and thrombosis induced by Hcy.


Assuntos
Homocisteína/farmacologia , Músculo Liso Vascular/citologia , Músculo Liso Vascular/metabolismo , Tromboplastina/metabolismo , Células Cultivadas , Humanos , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tromboplastina/genética , Veias Umbilicais/citologia
16.
Clin Infect Dis ; 46(10): e96-105, 2008 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-18419478

RESUMO

BACKGROUND: Cytomegalovirus (CMV) disease represents a serious complication after allogeneic peripheral blood stem cell (PBSC) transplantation. If possible, stem cell donors for transplantation are selected on the basis of their CMV serostatus. However, the cytomegalovirus-specific immune status can be further characterized by measuring CMV phosphoprotein 65-specific CD8(+) T cell frequencies using tetramers, pentamers, and streptamers. We therefore investigated the specificity and sensitivity of all 3 methods and compared the results to patient serostatus. METHODS: Twenty-three samples from CMV-seropositive healthy volunteers and 15 samples from CMV-seropositive patients before and after allogeneic PBSC transplantation were stained with tetramers, pentamers, or streptamers and analyzed by flow cytometry. RESULTS: Similar frequencies of CD8(+) and multimer(+) T cells could be measured by all 3 multimer technologies. The lowest background signals (< or =0.02%) were obtained using tetramer technology. Frequencies of 0.19%-2.48% of CMV phosphoprotein 65 495-503-specific CD8(+) T cells were detected in healthy volunteers. Antigen-specific T cells were detected in only 11 (48%) of 23 seropositive healthy volunteers. CMV antigenemia before day 100 after allogeneic PBSC transplantation occurred in 2 of 3 patients without any specific T cells. CONCLUSION: These findings demonstrate the power of multimer staining and a certain limitation of serologic testing to define appropriate donors for transplantation. Therefore, whenever possible, CMV-seropositive donors of transplants to seropositive recipients should be screened for their CD8(+) T cell frequency. All 3 multimer technologies can be used, yielding similar results. The streptamer technology additionally offers the advantage of selecting CMV phosphoprotein 65-specific CD8(+) T cells at the good manufacturing practice level for adoptive T cell transfer.


Assuntos
Linfócitos T CD8-Positivos/química , Linfócitos T CD8-Positivos/imunologia , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/imunologia , Fosfoproteínas/análise , Coloração e Rotulagem/métodos , Proteínas da Matriz Viral/análise , Adulto , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Infecções por Citomegalovirus/terapia , Feminino , Citometria de Fluxo , Humanos , Imunoterapia Adotiva , Subpopulações de Linfócitos/química , Subpopulações de Linfócitos/imunologia , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade
17.
Cytometry B Clin Cytom ; 74(1): 25-9, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18061959

RESUMO

New WHO classification has been widely applied in the diagnosis of leukemia. To elucidate the immunophenotype of acute myeloid leukemia (AML) and characterize the correlation among morphological, immunological, cytogenetic, and clinical features, we studied the bone marrow immunophenotypes of 180 AML patients in China by flow cytometry. The results showed that CD34, CD2, CD14, CD19, CD56, and HLA-DR were correlated with FAB subtypes. Amongst the 180 patients enrolled in this study, 122 cases were also subjected to karyotype analysis by G-banding technology and abnormal karyotypes were detected in 69 out of 122 patients. Correlation assay showed that t(8;21) was only present in 16 AML-M2 patients, and strongly associated with the individual or combinational expressions of CD15/CD19/CD34/CD56. As to M3, although lymphoid lineage antigens were observed in a considerable number of patients, they were never detected in t(15;17) positive patients. The expressions of CD22, CD56, and TdT showed significant correlation with the overall presence of abnormal karyotype. Additionally, the expressions of CD4, CD7, CD14, CD56, and TdT were positively correlated with clinical features such as white blood cell count, platelet count, and patient's age. In conclusion, immunophenotype analysis was useful for AML diagnosis and classification. At the same time, the data also suggested that the karyotype abnormalities and clinical features were tightly linked with abnormal antigen expression characteristics in AML patients. As one of the largest correlative study performed in China, the results highlighted the importance of a morphological, immunological, and cytogenetic classification of AML that might constitute a working basis for future studies aimed at a better definition of clinicopathological features and optimal treatment strategy for these leukemias.


Assuntos
Medula Óssea/patologia , Citometria de Fluxo/métodos , Imunofenotipagem , Leucemia Mieloide Aguda/patologia , Adolescente , Adulto , Idoso , Medula Óssea/imunologia , Criança , Pré-Escolar , Aberrações Cromossômicas , Bandeamento Cromossômico , Citogenética , Feminino , Humanos , Lactente , Cariotipagem , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/imunologia , Masculino , Pessoa de Meia-Idade
18.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 14(3): 455-9, 2006 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-16800919

RESUMO

To study the effects of supernatant derived from acute myeloid leukemia (AML) cell lines on proliferation and apoptosis of CD4(+) and CD8(+) T cell subsets and to investigate the mechanism by which AML escapes from immune recognition, lymphocytes were labeled with CFSE and were stimulated with anti-CD3 and anti-CD28 in presence or absence of supernatants from three AML cell lines (HL-60, NB4, U937). After culture, cell suspensions were labeled with 7AAD and CD4 PE (or CD8 PE). Cells were then detected by flow cytometry and their proliferation and apoptosis were analyzed. The results showed that supernatants from two of three cell lines (HL-60 and NB4) inhibited the proliferation of CD4(+) and CD8(+) T cells, and the degree of inhibition showed a dose-dependent way. Similarly, the apoptosis of stimulated CD4(+) T cells was inhibited, but stimulated CD8(+) T cells remained unaffected by supernatant from HL-60 and NB4. In contrary, the apoptosis of proliferative CD8(+) T cells were increased significantly by HL-60 and NB4 supernatant. It is concluded that soluble factors derived from AML cell lines inhibit the proliferation of CD4(+) and CD8(+) T cells and induce the apoptosis of proliferative CD8(+) T cells, that may be one of the mechanisms by which the immunity was suppressed.


Assuntos
Apoptose/fisiologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD8-Positivos/citologia , Leucemia Mieloide Aguda/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Proliferação de Células , Células Cultivadas , Meios de Cultura , Células HL-60 , Humanos , Leucemia Mieloide Aguda/patologia , Linfócitos T/citologia , Células Tumorais Cultivadas , Células U937
19.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 14(1): 119-22, 2006 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-16584606

RESUMO

This study was aimed to analyze the proportion of T cell subsets, CD4(+) CD25(high) regulating T cells (Tr) in peripheral blood of B-NHL patients and their change regularity, and to investigate the immunosuppression mechanism and influence of chemotherapy on immunosuppression function of B-NHL patients. The peripheral blood was collected from 42 patients with B-NHL, 36 patients with B-NHL who achieved partial remission (PR) or complete remission (CR) after 4 - 6 cycles of chemotherapy and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets and Tr cells. The results showed that the proportion of CD3(+) and CD4(+) T cells, and the ratio of CD4/CD8 in patients with B-NHL group was significantly less than those in the healthy controls, i.e. (68.33 +/- 15.27)% versus (72.06 +/- 9.26)%; (34.47 +/- 12.84)% versus (42.45 +/- 9.2)%; 1.36 +/- 0.26 versus 1.92 +/- 0.20, but the prevalence of the CD4(+) CD25(high) Tr cells was significantly higher than those in the healthy group [(4.10 +/- 1.21)% versus (2.04 +/- 1.03)%, P < 0.001]. The ratio of CD4/CD8 in chemotherapy group was lower than that in control, but the proportion of CD4(+) CD25(high) Treg cells in chemotherapy group was higher than those before chemo-/radio-therapy and the control. It is concluded that the relative increase of CD4(+) CD25(high) Tr cells in peripheral blood of B-NHL patients may be related to immunosuppression and tumor progression.


Assuntos
Linfoma de Células B/sangue , Linfoma de Células B/imunologia , Linfócitos T Reguladores/imunologia , Adolescente , Adulto , Antígenos CD4/análise , Feminino , Humanos , Tolerância Imunológica/imunologia , Subunidade alfa de Receptor de Interleucina-2/análise , Masculino , Subpopulações de Linfócitos T/imunologia
20.
Artigo em Inglês | MEDLINE | ID: mdl-17219964

RESUMO

The proportion and changes of CD4+CD25high regulatory T cells (Trs) in peripheral blood of non-small cell lung cancer (NSCLC) patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets (CD3+, CD4+ and CD8+) and CD4+CD25high Tr cells. The results showed that the proportion of CD4+CD25high Tr cells in NSCLC group was significantly higher than in control group [(4.36 +/-2.07) % vs (2.04+/-1.03) %, P<0.01]. The proportion of CD4+CD25 high Tr cells in late stage was higher than that in early stage [stages I +II (2.26+/-0.6) %; stage III (3.28+/-1.38) %; stage IV (6.06 +/-4.08) %] (P<0.05). Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4+CD25high Tr cells in peripheral blood was worse (P=0.0026). In conclusion, the relative increase in CD4+CD25high Tr cells in peripheral blood may be related to immunosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4+CD25+high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4+CD25high Tr cell therapy to treat NSCLC patients may be an effective strategy.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Carcinoma Pulmonar de Células não Pequenas/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Idoso , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Subunidade alfa de Receptor de Interleucina-2/sangue , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Linfócitos T Reguladores/patologia
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