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ABSTRACT: This study was purposed to investigate the expression of B7-H1 gene in leukemia cells and its clinical significance. The expression of B7-H1 mRNA was detected by SYBR Green I real-time quantitative PCR in a panel of 9 leukemia cell lines, 4 leukemia cell lines induced with IFN-γ, the bone marrow mononuclear cells (BMMNC) from 59 initial leukemia patients and 10 dendritic cells (DC) derived from BMMNC of initial leukemia patients, 2 solid tumour cell lines and BMMNC from 10 normal persons. The correlation between the clinical features of 59 acute leukemia patients and the expression level of B7-H1 mRNA in leukemia cells was analyzed. The results showed that the lower level of B7-H1 mRNA expression was found in leukemia cell lines and primary acute leukemia cells, but the expression level of B7-H1 mRNA was up-regulated significantly in the leukemia cell lines induced by IFN-γ and DC derived from BMMNC of leukemia patients. The expression level of B7-H1 mRNA in non complete remission (CR) patients after therapy was significantly higher than that in CR patients. It is concluded that the expression level of B7-H1 mRNA in leukemia cells is lower, but is up-regulated when affected by some factors. A correlation exists between the expression level of B7-H1 gene in leukemia cells and response of patients to therapy.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 06/2012; 20(3):541-4.
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ABSTRACT: Dendritic-like leukemia cells (DLLC) originating from leukemic cells could potentially induce a T cell-mediated anti-leukemia immune response. It has been demonstrated that B7-H1, a newly identified homologue of CD80/CD86, is abundant in human carcinomas and dendritic cells (DC), can exert co-stimulatory and immune regulatory functions. We demonstrated that B7-H1 was significantly expressed on AML cells and was strongly enhanced after differentiation to DLLC. Blockade of B7-H1 expressed on DLLC results in increased T cell proliferation and Th1 cytokine production, and decreased Th2 cytokine production. Importantly, autologous CTLs induced by DLLC treated with B7-H1 mAb showed significantly increased specific cytotoxcity against AML blasts. We further demonstrated that a significant decrease in IL-12 production, increase in IL-10 production by DLLC, and an increased CD4(+)CD25(+)Foxp3(+) T regulatory population lead to the defective T cell immune response that is induced by B7-H1 up-regulation on DLLC. Our data suggest that up-regulated B7-H1 on DLLC acts as a strong inhibitor of anti-leukemia T cell response, and that blockade of B7-H1 can improve DLLC-mediated anti-leukemia immunity.
Leukemia research 03/2009; 33(7):948-57. · 2.36 Impact Factor
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ABSTRACT: To inhibit the expression of beta-catenin and investigate the effect of the beta-catenin gene on Jurkat and K562 cells.
siRNA specifically knocking down the expression of beta-catenin was used to testify the function of beta-catenin in Jurkat and K562 cells. Real time polymerase chain reaction and Western blot were performed respectively to testify the mRNA level and protein level of beta-catenin. Growth curve was determined by counting viable cells using trypan blue refusal-dyed method. The proliferation of cells was assayed by clonogenic counting and MTT method. The apoptotic cells were measured by Annexin V/PI staining. The cell cycle analysis was performed based on propidium iodide staining.
Compared with the control group (transfected with siRNA directed against scramble gene), the survival, colonogenicity, and proliferation of the Jurkat and K562 cells were significantly decreased in experimental group transfected with beta-catenin siRNA. The colonogenicity was decreased from 31.9 +/- 5.55 (siRNA) to 25.0 +/- 5.13 (control) in Jurkat cells, and from 47.33 +/- 8.52 (siRNA) to 39.33 +/- 6.26 (control) in K562 cells (both P <0.05). The inhibition rate was (49.3 +/- 9.86)% (siRNA) and (15.1 +/- 6.55)% (control) respectively in Jurkat cells, and (39.4 +/- 7.56)% (siRNA) and (10.1 +/- 6.89)% (control) in K562 cells (both P <0.05). In addition, the apoptotic rate increased from (23.5 +/- 2.82)% (control group) to (55.9 +/- 2.22)% (experiment group) in Jurkat cells and from (14.9 +/- 8.54)% (control group) to (27.9 +/- 15.3)% (experiment group) in K562 cells. However, cell cycle analysis revealed no obvious phases change both in Jurkat and in K562 cells.
Knock-down of beta-catenin gene may decrease the proliferation, survival, and clonogenicity in Jurkat cells and K562 cells.
Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae 06/2008; 30(3):290-5.
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ABSTRACT: To detect the expression of B7-H1 gene in bone marrow mononuclear cells (BMMNCs) from leukemia patients and explore its clinical implications.
The B7-H1 mRNA expression levels of BMMNCs from 74 newly diagnosed leukemia patients and 10 normal volunteers were detected by real-time quantitative PCR. At the same time, BMMNCs from 12 patients in complete remission (CR) after chemotherapy and 5 in relapse were followed up. The correlation between the clinical features of 74 de novo leukemia patients and the expression level of B7-H1 gene was analyzed.
The mRNA expression level of B7-H1 gene in BMMNCs from de novo leukemia patients (RQ = 0.125) was lower than that from normal control (RQ=1). When patients achieved CR the gene expression level (RQ = 69.07) was significantly higher than that before CR (P = 0.001). After relapsed, its level (RQ=4) was still higher than that before CR (P > 0.05). No clinical parameters such as gender, age, peripheral white blood count, blast cells ratio in BM, CD34 positive cells were significantly correlated with the expression level of B7-H1 except the response to therapy. The initial expression level of B7-H1 gene in non CR patients after therapy was significantly higher than that in CR patients (RQ = 26. 91, P = 0.005).
The mRNA expression level of B7-H1 gene in newly diagnosed leukemia patients is lower than that in normal controls, and is higher in CR patients than in newly diagnosed patients. There is a correlation between the gene expression level and responsiveness to therapy.
Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 12/2007; 28(12):837-40.
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ABSTRACT: This study was aimed to investigate the expression of beta-catenin in leukemic cell lines and its relationship with pathogenesis of leukemia, semi-quantitative RT-PCR and Western blot were performed to detect the expression of beta-catenin in a panel of 15 human hematopoietic cell lines (U937, KG1a, Jurkat, K562, Namalwa, HEL, HUT78, Raji, Daudi, CEM, LCL-H, HL-60, NB4, J6-1, Ramos). Immunocytochemistry was performed in some of these cell lines to detect the location of beta-catenin. The results showed that the beta-catenin gene was widely expressed in most leukemic cell lines in various degree, the high expression of beta-catenin was found is U937, KG1a, Jurkat, K562 and Namalwa cells, middle expression of beta-catenin was observed in HEL, HUT78, Raji, Daudi and CEM cells, lower expression of beta-catenin was observed in LCL-H, HL-60, NB4, J6-1 and Ramos cells. The expression level of beta-catenin protein was identical to the expression level of beta-catenin mRNA. The expression of beta-catenin could be found in nuclei of all cells mentioned above, but their levels were different between them. Abundant beta-catenin also could be observed in nuclei of some leukemic cells by immunocytochemistry. It is concluded that overexpression of beta-catenin in leukemia cells, as a key mediator of Wnt signaling transduction pathway, indicates that the Wnt signaling transduction pathway may be aberrantly activated in leukemia.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 11/2007; 15(5):919-22.
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ABSTRACT: This study was aimed to quantitatively detect the expression level of beta-catenin and bcr/abl in different phases of chronic myeloid leukemia (CML) and to analyze their potential relationship and significance in the progression of CML. First, the total RNA isolated from BMMNC of patients with CML and donors was reversely transcribed into cDNA. The real-time quantitative PCR method was used to analyze the expression level of beta-catenin and bcr/abl. The expression level of beta-catenin and bcr/abl in different phases of CML was compared and the correlation was analyzed between the two genes. The results showed that the beta-catenin gene in BMMNC of blast crisis of CML patients was expressed significantly higher than that in chronic phase (p < 0.001) and accelerated phase (p = 0.016) of CML patients and in normal donors (p = 0.004). The expression of bcr/abl in blast crisis of CML was statistically higher than that in chronic phase of CML (p = 0.001). The expression levels of beta-catenin and bcr/abl were correlated with each other in CML patients (r = 0.620, p < 0.001). It is concluded that the beta-catenin gene in blast crisis of CML patients express higher than that in chronic phase and accelerated phase of CML, and its expression level is correlated with the level of bcr/abl expression. The increased expression of beta-catenin may be account partly for the blast crisis of CML.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 10/2007; 15(5):931-5.
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ABSTRACT: To assess the prognostic value of biological features and therapy-related factors in multiple myeloma (MM).
123 patients with newly diagnosed MM between January 1998 and May 2005 were enrolled in this retrospective study. Biological features at presentation and therapy-related factors were analysed. The overall survival (OS) and time to progression (TTP) were estimated by Kaplan-Meier analysis and the distribution of OS and TTP were compared using log-rank test. Cox regression was used to identify the independent prognostic factors.
(1) The univariate analysis indicated that more immature plasma cells in bone marrow biopsy, C-reactive protein >8. Omg/L, CD117 expression, serum beta2-microglobulin (beta2-MG) (3.5 approximately 5.5 mg/L), abnormal cytogenetics aberration of chromosome 13 (Delta13), hypodiploid, poor response to chemotherapy, interferon(IFN) therapy less than 6 months were associated with shorter OS(P <0.05). Lytic bone lesions at presentation, more immature plasma cells in bone marrow biopsy, serum beta2-MG (3.5 approximately 5.5 mg/L), poor response to chemotherapy, and IFN therapy less than 6 months as well as abnormal cytogenetics, hypodiploid and Delta13 were associated with shorter TTP (P <0.05). (2) Multivariable COX analysis indicated IFN therapy more than 6 months was a protective factor for OS and TTP, and more immature plasma cells in bone marrow biopsy was an independent poor prognostic factor for TTP.
The morphology of myeloma cells is useful for assessing the prognosis. And IFN therapy more than 6 months could lengthen OS and TTP.
Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 05/2007; 28(5):330-4.
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Ya-Fei Wang,
Heng-Xing Meng,
Wei Ge,
Zhen Yu,
Yun-Tao Li,
Qiao-Chuan Li,
Chang-Chun Wan,
Yan Xu, Xin Li,
Zheng-Jun Li,
Guo-Rong Wang,
Sheng-Guo You,
Lu-Gui Qiu
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ABSTRACT: To compare the expansion efficiency and function of dendritic cells derived from CB-CD34+ cells and MPB-CD34+ cells by using two-step culture method, enriched CB-CD34+ cells or MPB-CD34+ cells with immunoadsorption were primarily cultured in the presence of FL, SCF, TPO, GM-CSF for 10 days, and then further cultured with a combination of GM-CSF, IL-4, TNF-alpha, CD40Ab and PGE2 to induce DC. The DC phenotypes were detected by flow cytometry, the expansion efficiency and cell function were evaluated by mix-lymphocyte reaction (MLR), IL-12 level was detected by using ELISA and the chemotactic function mediated by secondary lymphoid tissue chemokine (SLC) was determined with Transwell plate. The results indicated that after 10 days of expansion, there were no significant difference in the percentage of CD14+CD1a- cells between CB and MPB [(40.48 +/- 16.85)% vs (28.07 +/- 23.19)%, P > 0.05], but the expansion of total cells in CB was higher than that in MPB (388.88 +/- 84.63-fold vs 79.67 +/- 10.32-fold, P < 0.01), so the yield of CD14+CD1a- cells from CB was significantly higher than that from MPB too (189.42 +/- 25.02-fold vs 28.74 +/- 23.27-fold, P < 0.01). The percentage of CD83+ DCs cultured with CD40Ab/PGE2 derived from CB were higher than those cultured with TNF-alpha derived from MPB respectively [(34.52 +/- 11.22)% vs (3.70 +/- 2.27)% and (36.69 +/- 13.36)% vs (7.34 +/- 3.364)% respectively, P < 0.01]. In the same circumstance, the yield of CD83+ DCs derived from CB was much more than that from MPB (198.72 +/- 117.53 times vs 33.95 +/- 6.19 times, P < 0.01). There were no difference in stimulating capacity, IL-12 secretion and migration capacity between DCs derived from CB and MPB. It is concluded that DCs induced from CB-CD34+ cells by two-step culture possess similar functions with that from MPB-CD34+ cells, but the yield of DCs from CB CD34+ cells is much more than that from MPB CD34+ cells.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 01/2007; 14(6):1163-7.
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ABSTRACT: To Summarize and compare the effects of the different post-remission treatment on long-term survival in acute promyelocytic leukemia (APL) patients.
The long-term survival and relapse of 111 APL patients with different post-remission treatment were retrospectively analyzed. The patients were divided into four groups. Group 1: All-trans-retinoic acid (ATRA) + As(2)O(3) + chemotherapy; Group 2: ATRA + chemotherapy; Group 3: As(2)O(3) + chemotherapy; Group 4: chemotherapy alone. The median follow-up time was 32 (6 - 185) months.
A total of 85 patients survived without diseases, 18 patients died and 8 patients dropped out. In the four groups, the patients survived without disease were (36/40, 23/30, 21/26, 5/15), respectively. The cases of patients survived over 3 years were (16/40, 12/30, 11/26, 1/15), respectively. The patients died were (3/40, 5/30, 5/26, 5/15), respectively. The estimated 5-year overall survival (OS) and relapse-free survival (RFS) were (78.3 +/- 4.9)%, (76.9 +/- 5.1)%, respectively. There were 21 patients relapsed.
The APL patients receiving combined post-remission therapy had better OS and RFS those receiving chemotherapy alone. Sequential therapy combining ATRA, As(2)O(3) and chemotherapy is the best post-remission therapy for long-term survival of APL patients.
Zhonghua nei ke za zhi [Chinese journal of internal medicine] 10/2006; 45(9):741-3.
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Xin Li,
Yao-Zhong Zhao,
Zeng-Jun Li,
Yun-Tao Li,
Yan Li,
Chang-Chun Wan,
Qiao-Chuan Li,
Shu-Hui Deng,
Ren-Chi Yang,
Ming-Zhe Han,
Lu-Gui Qiu
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ABSTRACT: This study was aimed to investigate various factors influencing long-term survival in patients with acute promyelocytic leukemia. A single institutional retrospective study with long-term follow-up was performed to better define the prognostic factors and a rationale for the use of ATRA, chemotherapy, and As(2)O(3) in the treatment of newly diagnosed APL patients. Newly diagnosed patients with APL entering complete remission (CR) were followed up for 6 to 185 months (n = 170) from January 1990 to December 2004. Univariate and multivariate analysis of 8 potential factors influencing survival and prognosis were carried out with Log-Rank and Cox regression method, including sex, age, initial WBC count, the level of lactic hydrogenase (LDH), first induction regimen, time from induction therapy to CR, post-remission therapy, negative or positive rate of PML-RAR alpha and follow-up of reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the estimated 5-year overall survival (OS) and relapse-free survival (RFS) were 80.9% +/- 4.0% and 71.0% +/- 4.0% respectively. The 23 patients relapsed at the median time of 15 months (6 - 70) after CR. Univariate analysis revealed that initial WBC count, first induction regimen, time from induction therapy to CR, type of post-remission therapy and persistent negative RT-PCR in remission were important prognostic factors for long-term survival. Multivariate study demonstrated that only type of post-remission therapy was associated with RFS and OS. It is concluded that the post-remission treatment combining ATRA, As(2)O(3) and chemotherapy would significantly improve the long-term survival of APL patients entering CR(1).
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 07/2006; 14(3):437-41.
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ABSTRACT: To investigate the effects of stromal cell-derived factor 1 (SDF-1) and platelet factor 4 (PF4) on the homing-related function of expanded ex vivo umbilical cord blood CD34(+) cells, purified cord blood CD34(+) cells were cultured in serum-free medium containing a HGF combination of FL + SCF + TPO (FST) with either 100 ng/ml SDF-1 alone, 100 ng/ml PF4 alone, or both of these 2 cytokines. The expansion rate of CD34(+) cells, colony formation, homing-related functions including expression of homing-related adhesion molecules of expanded CD34(+) cell, adhesion activity and chemotactic function of the re-selected expanded CD34(+) cells were evaluated at different time points. The results showed that expansion rate of CD34(+) cells and expansion multiple of CFU in SDF-1 groups were higher than those in control. The expression of CD49e on the expanded CD34(+) cells was remarkable up-regulated, in contrast, expression of CXCR-4 on the expanded CD34(+) cells was remarkable down-regulated in SDF-1 groups. The expression of CD49e, CD54 and CXCR-4 on the expanded CD34(+) cells were remarkably up-regulated in the PF4 groups. In all the SDF-1 group, PF4 group and SDF-1 plus PF4 group, the ability of expanded CD34(+) cells adhering to fibronectin layer were higher than those in the control on day 10. Spontaneous migration rate of expanded CD34(+) cells in SDF-1 groups were higher than those in control, while SDF-1-induced migration rate were lower than those in control on day 10. SDF-1-induced migration rate in PF4 groups were higher than those in control on day 10. Spontaneous and SDF-1-induced migration rate of expanded CD34(+) cells in the SDF-1 plus PF4 groups were higher than those in control on day 10. It is concluded that, SDF-1 and PF4 can up-regulate expression of adhesion molecules on expanded CD34(+) cells, and retain the adherent and migration ability of expanded CD34(+) cells, which is helpful for the homing of expanded CD34(+) cells. In short, SDF-1 and PF4 are helpful for the homing-related function of the expanded UCB HSPC.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 03/2006; 14(1):83-8.
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ABSTRACT: To compare the expression of cell adhesion molecules (CAMs) among VLA-4 (CD49 d), VLA-5 (CD49e), LFA-1 (CD11a), L-selectin (CD62L), and PECAM-1 (CD31) which are more related to the homing of hematopoietic stem and progenitor cells (HSPC) on the ex vivo expanded CD34+ subset with that of fresh isolated AC133+ cells.
AC133+ cells selected from fresh cord blood (CB) samples were cultured in QBSF-60 serum-free media in the presence of Flt-3 ligand + SCF + TPO (FST), with initial addition of IL-3 for up to 2 week. Expansion potential and the expression of above CAMs were evaluated at day 0, day 7, day 10 and day 14.
(1) Simultaneously numerical expansion of various HSPC was constantly observed during the culture, and the fold expansion of AC133+ cells and CD34+ cells on day 14 were 33.50 and 64.56 respectively; (2) The number of CD34+ subsets expressing the above adhesions were all increased at different degrees (from 20 fold to 160 fold). (3) The expressions of CD11a, CD49d, and CD49e on ex vivo expanded CD34+ cells were increased as compared to their baseline levels, but the percentage of CD62L+ and CD31+ subpopulations in CD34+ cells were decreased.
Our short-term culture system can not merely support the simultaneous expansion of CB derived AC133+ cells, but the expanded hematopoietic progenitors may well sustained the expression of homing-related adhesion molecules.
Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae 03/2002; 24(1):7-10.
