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  • Zhou, Maohua  (7)
  • 2005-2009  (7)
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  • 2005-2009  (7)
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  • 1
    In: Blood, American Society of Hematology, Vol. 110, No. 11 ( 2007-11-16), p. 4967-4967
    Abstract: Chronic graft-versus-host disease (cGVHD) continue to cause significant morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). It remains the most frequent complication following allogeneic hematopoietic stem cell transplantation, occurring in 30% to 70% of long-term survivors.[Lee SJ, Blood 2002 ]. An experienced clinician may be able to make a diagnosis of chronic GVHD based on classic manifestations such as lichenoid or sclerodermatous skin changes. If the findings are not typical, a tissue biopsy is warranted to aid the diagnosis and to determine if infection is present (A.L. Gilman Gilman and J. Serody seminhematol.2005). Recently, there are many research in lymphocyte subsets on cGVHD by using flowcytometry.It helps us to understand the mechsim about it, and could be help for diagnosis, staging, and response criteria in this disorder. However, the change of lymphocytes subsets is still unknown in the different stage of cGVHD. We are here to study the lymphocyte subsets in the peripheral blood with different stage of cGVHD after allo-HSCT. Peripheral blood mononuclear cells were isolated from anticoagulated venous blood using lymphocyte separation medium according to manufacturer’s nstructions.Freshly isolated peripheral blood mononuclear cells were analyzed. Fluorescein sothiocyanate conjugated monoclonal antibodies (Becton Dickinson, mAb) By using FACScalibur, (Becton Dickinson).The data of the two-group comparison was done on the analysis of variance and Students t test.According to the National Institutes of Health Consensus Development Project on Criteria for Clinical Trials in chronic Graft-versus-Host Disease in 2005, we analysis the diffierent groups with non-cGVHD, mild, moderate and severe cGVHD. The proportions of the lymphocyte subsets are different with the different stage of cGVHD. The more serious of cGVHD, the more proportions of the total lymphoctyes. B lymphocyte has significant difference with different stage of cGVHD, although the percentage of the total lymphocytes is 2.14%∼16.14% (P=0.0023). There is significant difference on the percent of CD8+ CD45RO+ cell among the non-cGVHD, mild cGVHD, moderate cGVHD or severe cGVHD(44.56%,46.59%,56.77% and 61.68%, respectively. P=0.0475). But the percent of CD4+CD25+ cell is higher than that of CD8+CD25+ cell (31.86%∼43.75%/4.50%∼7.08%)The percent of CD69+CD4+,CD69+CD8+ (CD95+CD4+, CD95+CD8+, CD28+ CD4+, CD28+CD8+ cell are not significant different within the different statue cGVHD. We can draw a conclusion as below: It may help for clinician to early diagnosis,stage using routine monoclone antibodies by flowcytometry. the percents of the lymphocyte subsets are different with the different status of cGVHD. With the more serious of cGVHD, the more proportions of the total lymphocytes; CD4+CD25+ cell ; CD3+CD8+ cell and B cell increase significantly, the percent of CD45RO+CD8+ cell and CD3+CD4+ cell drease at the same time. It may help for early diagnoses and treatment for the cGVHD.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2007
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  • 2
    Online Resource
    Online Resource
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4853-4853
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4853-4853
    Abstract: Abstract 4853 Chronic myelomonocytic leukemia(CMML) was incorporated into the FBA classification of myelodysplastic syndromes(MDS) because dysplastic changes are commonly found in blood and bone marrow cells of patients with this order. However, sometimes doctors misdiagnose it because it may be distinguishable by only the presence of monocytosis( 〉 1.0×109/L) in the blood.CMML is always a rather controversial type of MDS, some of patients present charaterication of myeloproliferation. In a further step, the WHO classification now includes CMML in a category of mixed myeloproliferative/myelodysplastic disorders, together with atypical CML and JMML,and proposed to separate CMML into CMML I and CMML II [Harris et al. Journal of Clinical Oncology,1999; Bennett International Journal of Hematology,2000]. Prognosis is also extremely variable in CMML.The median survival was about 19 months in 288 CMML patients included in Düsseldorf MDS Registry. There was no significant difference in median survival of MPD-CMML and MDS-CMML[Nosstinger et al. Leukemia Researsh.2001]. Germing reported the median of CMML patients who developed AML(18%) was 14 months,as compared to 20 months for patients who did not develop AML. [Germing et al. Leukemia and Lymphoma. 2004] .In 2002, the M.D.Anderson Prognostic Score(MDAPS) using lymphocyte counts, hemoglobin level, medullary blast count, and presence of immature myeloid precursors in blood was developed by Onida[Onida et al. Blood. 2002]. Here we analyzed the clinical characterization of CMML in our hospital. 16 cases of CMML diagnosed according to the criteria of WHO classification were retrospectively analyzed. The median age was 51 years,and female/male was 11/5.Most of patients were median or older men with splenomegaly(37%), figure(30%), bone pain(25%), hepatomegaly(13%). White blood counts varyied from hypoleukocytosis to normal to hyperleukocytosis, median count was 27×109/L (1.3-74×109/L), median platelet count was 97×109/L(12-576×109/L),median hemoglobin count was 91g/l(62-138 g/l), median monocyte count was 4.6×109/L(2-14.5×109/L), median lymphocyte count was 4.4×109/L(1.0-33.8×109/L), LDH was elevated. Bone marrow was obviously active with dyshaematopoiesis, median blast cells of granulocyte were 8.5%(1%-18%), immature monocyte was 0-2.5%. 44% of patients had eosinophilic cells and/or basophil cells increase in bone marrow, and decrease in NAP, median score was 13(0-62). Immunophenotypic feature was CD13+CD14+CD56+,and FISH: BCR-ABL(-).The clinical course of CMML could remain stable for many years only low dose chemotherapy or without any treatment. On the other hand, there were patients with rapidly progressive disease. The Median survival was 20 months. CMML-I cases average survival was 50+ months, CMML-II was 18+ months≤ MDAPS showed low risk group had a average survival of 60+ months, intermediate group 1 and 2 was 30+ months and 17+ months,high risk patients with a average survival of only 1+ months. Only one case transformed into M4 as late as 3+ years after diagnosis of CMML(6%). In conclusion, the new criteria of WHO classification presents clinical feature of CMML better, and it can provide more effective prognostic parameters for risk assessment with MDAPS. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2009
    detail.hit.zdb_id: 1468538-3
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  • 3
    In: Blood, American Society of Hematology, Vol. 110, No. 11 ( 2007-11-16), p. 4358-4358
    Abstract: Patients with relapsed or refractory leukemia have less chances of obtaining remission than patients with newly diagnosed.It is reported that more than 80% of acute myeloid leukemia(AML) patients have myeloid blast cells that express the CD33 surface antigen. This antigen also is present on the leukemic stem cells at least some patients with chronic myeloid leukemia(CML)and acute lymphoblastic leukemia(ALL). It is absent from normal hematopoietic stem cells and nonhematopoietic cells and tissues. Gemtuzumab is a humanized anti-CD33 antibody conjugated to calicheamicin, a potent anti tumor antibiotic derived from a bacterium. It is conditionally approved in the US for treatment of CD33+ AML in first relapse in patients over 60 years[Sievers et al.Journal of Clinical Oncology,2001]. Here we evaluate the efficacy and safety of Gemtuzumab -based regimens. The study population comprise 11 patients with CD33-positive refractory leukemia (determined as CD33-antigen expression in over 50% of leukemic blasts by bone marrow aspirates and immunophenotyping), including two with myeloid blast phase of CML, one with refractory ALL, two relapsed after Auto-stem cell transplantation(Auto-SCT). The median age was 47 years. Four cases who were over 60 years or after Auto-SCT treated with single agent. The other seven cases treated with mylotarg and cytotoxic agents, including mylotarg plus idarubine (MI); Gemtuzumab plus fludarabine, cytarabine, CsA(MFAC); or plus mitoxantrone, Ara-C (MMA).The overall response rate was 54% (6/11), with 36%(4/11) patients obtaining complete remission (CR) and 18% (2/11) achieving CRp.The median overall survival time after treatment was 4.8 months, and the median overall survival time after CR was 8.2 months.Two patients with myeloid blast phase of CML achieved CR with BCR/ABL(−), the survival time after CR was 5+months and 20+months respectively, but failed in second relapse. One patient received Allo-SCT after CR with refractory ALL is still alive at present (21months) with disease free. The median time to ANC recovery of 0.5×109/L was 15 days.The common adverse events was myelosuppression (100% Grade 4 neutropenia and thrombocytopenia).Significant non-hematologic toxicitics included infection(96%), infusion-related chills and fever (55%).Although hepatic dysfunction and mucositis were observed,they were generally infrequent and not severe(Grade 1–2).Six patients (55%) developed Hepatic veno-occlusive disease(VOD), four of them were either over 60 years old or received Auto-SCT before although they received only single agent mylotarg therapy, but it was transient and no one died from it. In conclusion, patients with CD33-positive refractory leukemia, Gemtuzumab -based regimens have a comparable response rate and offer a more favorable toxicity profile, expecially for the patients with myeloid blast phase of CML. It is also effective for the patient with refractory ALL. In the treatment, we should pay attention to the hepatic condition of ones who are over 60 years old or after stem cell transplantation, attemps to avoid and treat VOD are warranted. Above all, When patients with refractory leukemia got remission, allogenic-SCT should be done as soon as possible
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2007
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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  • 4
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4856-4856
    Abstract: Abstract 4856 Objective The multiparametric flow cytometry is becoming a very useful tool of the diagnosis and prognostication for patients with Myelodysplastic Syndrome(MDS). This study was aimed at using multiparametric flow cytometry to explore the immunophenotypic abnormalities of bone marrow cells from patients with RAEB. Methods We collected BM samples from 12 MDS-RAEB patients (6 male, 6 female, Median Age 67.5) and 20 non-MDS patients (11 male, 9 female, median age 32.5, 7 AA, 5 PNH, 3 IDA, 1 ALL, 2 CML, 2 MM). The multiparametric flow cytometric analysis was performed using an extensive panel of monoclonal antibodies. We used the conventional and secondary gating strategies to analysis the BM cells compartments such as the percents of blast cells and the expression of lineage and maturation-associated antigens of BM hemopoietic cells quantified. Results Compared with the non-MDS group, the proportion of blast cells increased significantly in the MDS-RAEB group (P=0.001), but the percentages of nucleated erythrocyte, lymphocyte, monocyte and granulocyte were no significant difference (P=0.954, P=0.893, P=0.730 and P=0.182). As the percentage of blasts cells increasing, the survival time became shorter (13 ± 6 vs 35 ± 15 months; P=0.02). The expressions of haemopoietic stem/progenitor cell surface marker CD34+ and T lymphocyte surface marker CD7+ on blast cells were much higher by secondary gating method than non-MDS group (P=0.009, P=0.002, respectively), while no significant difference of the expression of CD56 (P=0.375). The expressions of CD7+ and CD56+ on lymphocyte were no significant difference between the two groups (P=0.195, P=0.369, respectively), however the expression of CD19+ may be different (P=0.039). The expressions of CD33+ and CD13+ on granulocyte were no significant difference between the two groups (P=0.289, P=0.744, respectively). However, the expression levels of CD15+CD11b+, CD15+CD11b-, CD10+, HLA-DR, CD56+ in the MDS-RAEB group were significantly higher than those in the non-MDS group, specially, the levels of CD10+, HLA-DR and CD56+ were much higher (P=0.016, P=0.011, P=0.005, P=0.005 and P=0.005, respectively) and these patients showed a shorter median overall survival (15 ± 5 vs 36 ± 10 months; p = 0.03). Conclusions The percentage of blast cells increased in MDS-RAEB patients and the expressions of CD34+, CD7+ on blast cells and the expressions of CD10+, HLA-DR and CD56+ on granulocyte is higher than non-MDS group. It will be necessary to increase the number of cases (MDS-RAEB) to confirm our findings. Using multiparametric flow cytometry can find the immunophenotypic abnormalities more sensitively, accurately and objectively, and this new approach could provide much more useful information in the diagnosis and prognosis of MDS-RAEB patients. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2009
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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  • 5
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4843-4843
    Abstract: Abstract 4843 Background The myelodysplastic syndromes (MDS) are a group of clonal heterogeneous bone marrow disorders characterized by peripheral cytopenias, ineffective hematopoiesis, and unilineage or multilineage dysplasia. Multiparametric flow cytometry is increasingly being used as an adjunct to the establishing of MDS. While many antigens have been described to be aberrantly expressed in MDS the findings are generally heterogeneous and there is no consistent finding that would be present in all cases with MDS. Aim To investigate the immunophenotypic features of MDS and non-MDS patients and the characteristic of subtypes of MDS. Methods BM samples were collected from 22 MDS patients including 3 RA (2 male, 1 female, median age 57), 3 RAS (2 male, 1 female, median age 72), 12 RAEB (6 male, 6 female, median age 67.5), 4 MDS-AML (2 male, 2 female, median age 69.5) and 20 non-MDS (11 male, 9 female, median age 32.5, 7 AA, 5 PNH, 3 IDA, 1 ALL, 2 CML, 2 MM). The multiparametric flow cytometric analysis was performed using an extensive panel of monoclonal antibodies and using the conventional and secondary gating strategies to analysis the immunophenotypic features of BM cells. Results This study showed that the proportion of blast cells increased significantly than non-MDS group (P=0.002). As the disease progressing, the percentage of blast cells became higher and significantly difference compared to the non-MDS group (P=0.226, P=0.464, P=0.001 and P=0.000, respectively). The expressions of CD34+ and CD7+ on blast cells were significantly difference between MDS and non-MDS groups (P=0.005, and P=0.002, respectively). Compared with the subtypes of MDS and non-MDS group, the expressions of CD34+ and CD7+ on blast cells became high gradually (P=0.534, P=0.487, P=0.009, P=0.004 and P=0.294, P=0.166, P=0.002, P=0.001) and the high percentage of blast cells and high expression levels of CD34+ and CD7+ might indicate poor prognosis. The expression of CD7+ on lymphocytes was similar with CD34+ and CD7+ on blast cells, but the expressions of CD19+ and CD56+ on lymphocytes were no significantly difference (P=0.076, P=0.252, respectively). The expressions of antigens on granulocytes showed that the expressions of CD15+CD11b+, CD10+ and HLA-DR were significantly difference between MDS and non-MDS groups(P=0.000, P=0.009 and P=0.007, respectively), meanwhile, as the disease progressing, the expression rates of CD15+CD11b+, CD10+ and HLA-DR in subtypes of MDS increased gradually and the survival time of these patients who had over-expression of these antigens was shorter than control group(P=0.002). However, the expressions of CD33+, CD13+, CD56+ and CD15+CD11b- were significantly difference between subtypes of MDS and non-MDS group (P=0.059, P=0.588, P=0.063 and P=0.207, respectively). Conclusions Our results showed that using multiparametric flow cytometry to analyze the immunophenotypic features of BM cells could provide clinically useful information for the diagnosis, classification and prognosis of MDS patients. Particularly, the percentage of blast cells, the expression of CD34+ and CD7+ on blast cells, the expression of CD7+ on lymphocytes and the expression of CD15+CD11b+, CD10+ and HLA-DR on granulocytes may provide the much more useful information. However, further studies including larger number of patients with a longer follow-up are necessary to confirm these results. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2009
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 6
    Online Resource
    Online Resource
    American Society of Hematology ; 2005
    In:  Blood Vol. 106, No. 11 ( 2005-11-16), p. 3922-3922
    In: Blood, American Society of Hematology, Vol. 106, No. 11 ( 2005-11-16), p. 3922-3922
    Abstract: Objective Allogeneic transplantation of recombinant human granulocyte colony-stimulating factor (rhG-CSF) -mobilized peripheral blood stem cells (PBSCs) is now being increasingly performed, but safety considerations for hematologically normal PBSC donors have not been fully addressed. The effection of G-CSF on donors’ lymphocytes have not been defined clearly. Our study was to detect and analyze the phenotypical and functional properties of lymphocytes from allo-PBSC donors treated with recombinant human G-CSF by flow cytometry. Methods Thirty-four HLA-identical sibling donors (13male, 21female; median age, 35 years) were treated by subcutaneous injection with rhG-CSF at a dose of 5 μg/kg twice daily for 4–6 consecutive days to mobilize HSCs to the peripheral blood. Leukapheresis was performed using a continuous flow blood cell separator (COBE Spectra, Lakewood, CO) on 1 to 2 consecutive days beginning on day 4 of rhG-CSF administration. Donor blood samples, which were obtained before the first administration of G-CSF (pre-G), on day 4 of G-CSF administration (post-G), and one week after the last rhG-CSF were analyzed by 3-color flow cytometry. Monoclonal antibodies included: CD3, CD4, CD8, CD20, CD16, CD56, CD25, CD69, CD45RA, CD45RO, CD28, CD95. Results Absolute counts of lymphocytes, B cells, T cells, CD4+ and CD8+cells post-G were significantly elevated two more times than pre-G (P & lt;0.05), but these changes recovered when the last G-CSF administrated one week later. The percentage of CD3+, CD4+, CD8+ and the ratio of CD4 and CD8 T cells had no significantly difference between pre-G and post-G. The percentage of CD4+CD25+, CD4+CD45RO+ and CD4+CD28+ T cells were significantly decreased post-G (45.26±9.68% to 37.34±11.12%; 65.53±13.74% to 52.32±13.47%; 94.75±4.02% to 91.74±8.72%, P=0.00329, 0.0003 and 0.0947, respectively). We found that CD4+CD28+ T cells(91.77 ± 6.15, P=0.00218) were still lower than pre-G when stopped administrate G-CSF one week later, while others changes have recovered to the Pre-G level. Conclusion: The study shows that CD4+CD28+ T lymphoctye subsets were still lower than pre-G, although most of other change recover one week later after mobilization
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2005
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 7
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4501-4501
    Abstract: Abstract 4501 Chronic graft-versus-host disease (GVHD) is a major complication of allogeneous hematopoietic stem cell transplantation, and it is regarded as a type of Th2 disease and seems to be a T-cell-mediated disorder in which immune tolerance to self-antigens is broken. Mesenchymal stem cells (MSCs) are multipotent cells which can extenuate refractory (GVHD) have been recently reported; however, the clinical data on MSC relieves chronic GVHD are generally lacking, and the machanisms remain to be investigated. One area of recent interest and controversy is the role of regulatory T cells (CD4+/CD25high T cells) in chronic GVHD. We examined the lymphocyte subsets and regulatory T cells of patients with refractory chronic GVHD were treated with basic immunosuppression plus MSCs derived from volunteer donors. Between April 2005 and October 2008, 19 patients were treated with their basic immunosuppression plus in vitro expanded bone-marrow-derived MSC as a compassionate treatment for resistant chronic GVHD. The median MSC dose given was 0.6×106/kg per body weight. Flow cytometric analysis was perfromed to analysis lymphocyte composition and activation at the time before MSC infusion(pre-MSC) and 3 months after MSC infusion (post-MSC). The study was approved by the Ethics Committee of the Guangdong General Hospital. All patients and the MSC donors provided written informed consent. In the present study, there were no significant changes in the proportion of T cells, B cells, NK cells and activation in no-response patients between pre-MSC and post-MSC. In the patients with response, the total lymphocyte was greater post-MSC transfusion than pre-MSC transfusion, and the proportion of CD4+ T cells were increased CD+8 T cells were decreased (the P value was 0.001 and 0.018, respectively). However, the proportion of CD4+CD25+ T cell and CD8+CD25+ T cells hadn't significantly changed post-MSC transfusion compared to pre-MSC infusion in patients with response. The T lymphocyte subsets of CD8+CD28+ cells, which are active CTLs that participate transplantation immunity and rejection reaction, were down-regulated when the chronic GVHD improved post-MSC infusion (P=0.023). By contrast, CD8+CD28- cells, which regulate cells to induce immune tolerance and inhibit antibody production and imbalance in the production of Th1 and Th2 cytokines, were increased in patients responsed to MSC infusion (P=0.025), and the ratio of CD8+CD28- cells to CD8+CD28+ was increased. Importantly, the increase of CD8+CD28- T cells and decrease of CD8+CD28+ T cells with improvement of cGVHD indicate that these factors may be involved in the development of chronic GVHD and that MSC suppresses chronic GVHD via modulation of the ratios of CD8+CD28- to CD8+CD28+ T cells. However, further immunological studies specifically addressing this issue are needed to confirm this possibility. This work was supported by Science and Technology Planning Project of Guangdong Province (2006B36005003,2007A032100003) and Science and Technology Planning Project of Guangzhou (2006Z2-E4071,2008A1-E4011-4,2008A1-E4011-5). Disclosures: No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2009
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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