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  • Geng, Suxia  (7)
  • 2005-2009  (7)
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  • 2005-2009  (7)
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  • 1
    Online Resource
    Online Resource
    Analysis T Cell Receptor Excision Circles (TRECs) in Patients with Graft-Versus-Host Disease (GVHD) after Allogeneic Stem Cell Transplantation.
    American Society of Hematology ; 2006
    In:  Blood Vol. 108, No. 11 ( 2006-11-16), p. 5272-5272
    Details
    In: Blood, American Society of Hematology, Vol. 108, No. 11 ( 2006-11-16), p. 5272-5272
    Abstract: The thymic-dependent pathway that involves generation of new naive T cells from donor-derived precursor cells accounts for the more durable reconstitution of the T-cell compartment and generates a more diverse TCR repertoire. Thymic function and production of recent thymic emigrants (RTEs) may be directly evaluated through the quantification, by real-time polymerase chain reaction (PCR), of the T-cell receptor excision circles (TRECs). Following hematopoietic stem cell transplant (HSCT), there is a prolonged period of profound immune deficiency, which continues for years after HSCT. The factors that inhibit thymic function may include age, graft-versus-host disease (GVHD), and direct thymic damage from chemoradiotherapy. GVHD after HSCT also leads to thymic insufficiency, possibly by direct attack on the thymic stroma by allogeneic effector cells. The aim of our study is to analysis T cell receptor excision circles (TRECs) in patients with GVHD after allogeneic stem cell transplantation. We used real-time polymerase chain reaction (PCR) to quantify SjTRECs in 12 patients with GVHD(9 males, 3 females; median age 32 years old), who underwent HLA-matching sibling BMT and/or peripheral blood stem cell transplantation (PBSCT) at our department. Quantitative detection of sjTRECs in DNA of peripheral blood mononuclear cells from 13 normal individualals. The median value of sjTRECs copies P1 000 PBMCs was 4.37±3.64 in normal indiviuals. However, the decreased levels of TRECs were most profound in the group of patients with active chronic GVHD at the time of study. it was 0.26±0.22 copies P1 000 PBMCs in 12 patients with GVHD (P 〈 0. 00007). We conclude that measurement of sjTREC may provide an important tool for predicting thymus-dependent T-cell reconstitution in GVHD patients after transplantation.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V108.11.5272.5272
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2006
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 2
    Online Resource
    Online Resource
    Using Multiparametric Flow Cytometry to Analysis the Immunophenotypic Abnormalities of Bone Marrow Cells in Patients with MDS-RAEB.
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4856-4856
    Details
    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
    URL: Article
    DOI: 10.1182/blood.V114.22.4856.4856
    RVK:
    XA 33000
    RVK:
    XA 10000
    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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  • 3
    Online Resource
    Online Resource
    Monitoring JAK3 mRNA Levels in Patients with GVHD after Allogeneic Hematopoietic Stem Cell Transplantation by Real-Time Quantitative PCR.
    American Society of Hematology ; 2006
    In:  Blood Vol. 108, No. 11 ( 2006-11-16), p. 5273-5273
    Details
    In: Blood, American Society of Hematology, Vol. 108, No. 11 ( 2006-11-16), p. 5273-5273
    Abstract: Background: Graft-versus-host disease (GVHD) represents a major hurdle impeding the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT). JAK3 plays pivotal roles in the initiation of cytokine-triggered signaling transduction. Recent studies about the murine GVHD model show that targeting JAK3 in donor lymphocytes with a chemical inhibitor such as WHI-P131 may be useful in the prevention of severe GVHD. However, the level of JAK3 expression in patients with GVHD has not been reported. The current study is to develop a reliable and rapid real-time quantitative PCR (Q-PCR) method using Taqman Probe for detecting JAK3 in allo-HSCT recipients. Methods: 20 patients who received myeloablative-conditioning regimen for hematological malignancies and cyclosporine for GVHD prophylaxis have been investigated in this still ongoing study clinically. One of these patients developed acute GVHD (aGVHD), 9 patients developed chronic GVHD (cGVHD), and 10 patients showed no evidence of GVHD during the first 365 days post-transplant. RNA was extracted from white blood cells and cDNA templates were synthesized using 1μg RNA. The standard curve method was used for relative quantitation of JAK3. Results: There is a clear trend for a increasing expression of JAK3 in 9 patients with GVHD. Interestingly, a significant decrease of JAK3 gene expression when the severity of GVHD was remission (P=0.04). One of the patient with cGVHD, the level of JAK3 gene expression has no changed before and after treatment. However, these patients post HSCT without GVHD have a comparable level of JAK3 expression before myeloablative conditioning regimen(P 〉 0.05). Conclusion: Based on our preliminary results, JAK3 mRNA level was obviously correlated with the development of GVHD. Analysis JAK3 mRNA expression may be useful in evaluating GVHD patients with allogeneic stem cell transplantation.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V108.11.5273.5273
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2006
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 4
    Online Resource
    Online Resource
    Mesenchymal Stem Cells Relieve Chronic GVHD Via Modulation the Ratio of CD8+CD28-/CD8+CD28+T Cells.
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4501-4501
    Details
    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
    URL: Article
    DOI: 10.1182/blood.V114.22.4501.4501
    RVK:
    XA 33000
    RVK:
    XA 10000
    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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  • 5
    Online Resource
    Online Resource
    Differential Protein Expression of Serum in Patients with RAEB Myelodysplastic Syndromes.
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4845-4845
    Details
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4845-4845
    Abstract: Abstract 4845 To screen molecular markers for refractory anemia with excess blasts in transformation (RAEB) in myelodysplastic syndromes (MDS) by serum proteome profiling. Methods Serum protein were isolated respectively from patients with RAEB, acute myeloid leukemia or normal person by 2-dimensional electrophoresis(2-DE). The imaging films were obtained and the differential reacting protein spots were recognized. Then the differential reacting proteins were obtained in the replica gel by matching analysis,and identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) and database searching. Results Seven differentially expressed proteins in RAEB were found by 2-DE. Of the 7 proteins, 4 were identified with significantly statistical difference, including dipeptidyl peptidase (DPP/CD26), polymerase (DNA directed) kappa, PRO2044 and albumin-like. Conclusion 2-DE-based serum proteome profile can help to identify serum proteomic biomarkers related to MDS. DDP/CD26 has increased serum levels in RAEB subtype MDS, suggesting significant action in advanced MDS. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V114.22.4845.4845
    RVK:
    XA 33000
    RVK:
    XA 10000
    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
    Different Dosage of Bortezomib Impacts the Prevention of Acute Graft-Versus-Host Disease.
    American Society of Hematology ; 2009
    In:  Blood Vol. 114, No. 22 ( 2009-11-20), p. 4499-4499
    Details
    In: Blood, American Society of Hematology, Vol. 114, No. 22 ( 2009-11-20), p. 4499-4499
    Abstract: Abstract 4499 The proteasome inhibitor bortezomib which prevents nuclear factor kappaB (NF-kappaB) activation, blocks T-cell activation, induces T cell and dendritic cells apoptosis, and inhibits dendritic cells mature. Bortezomib has been used successfully in the treatment of graft-versus-host disease (GVHD) and autoimmune diseases in animal models. As recent reports, bortezomib not only could prevent GVHD but also induce severe GVHD which depends on the time of administration. We explored whether the dosage of bortezomib impacts the prevention of aGVHD. For this purpose, different dosage of bortezomib (10ug/kg, 100ug/kg, 800ug/kg) were administrated to recipient C57BL/6 (H2b) mice which were lethally irradiated and given transplants of bone marrow cells and splenocytes from major histocompatibility complex (MHC)-disparate BALB/c (H2d) donors. The median survival time of control group was 31 days (19 to 37), low dosage group (10ug/kg) was 27.5 days (range: 18 to 42), intermediate dosage group(100ug/kg) was 45 days (range: 32 to 67), and high dosage group(800ug/kg) was 33 days (range:22 to 67). The survival time of intermediate dosage group compared with control group and other dosage group increased significantly (P 〈 0.05). On day +30,all the inter-dose group mice were alive(100%), and the control group, the low-dose group and the high-dose group remained 40%(2/5), 30% (3/10) and 60% (6/10) mice, respectively. On day +60, the inter-dsoe group and the high-dose group remained three and one alive, respectively. Pathologic assessment of skin and liver during aGVHD on day +30 showed less lesions in inter-dose group and high-dose group than low-group and control group. Interestingly, pathology also revealed that significant increases in gastrointestinal lesions occurred following high dose (800ug/kg) bortezomib administration in aGVHD. The results show that intermediate dosage bortezomib could effectively improve the survival time and protect injury from aGVHD. In this study, we detected T cell apoptosis by Annexin-V/PI. The percentages of the apoptosis cells in control group, low-, inter- and high dosage group were 4.00±0.14, 4.65±0.2, 12.13±0.14 and 12.37±0.75, respectively. The result indicated T cell apoptosis induced by bortezomib were more evident in the inter-dose group and high-dose group than low-dose group and control group (P 〈 0.05). Also, the proportion of CD4+CD25+ regulatory T cells were significantly increase in the inter-dose group and high-dose group than other two groups (inter-, high-dose group, and contral group, low- dose group were 30.25±6.32%, 35.25±2.14%, and 21.71±0.52%, 23.15±4.12%, respectively; P 〈 0.05). Otherwise, the level of serum IL-2 decreased and IL-4 increased significantly in intermediate and high dosage group, the low dosage bortezomib didn't affect the leverl of serum IL-2 and IL-4. Intermediate dosage of bortezomib prevents aGVHD without presenting obvious adverse effects. The effect of bortezomib on preventing aGVHD are relevant to T cell apoptosis, regulating IL-2 and IL-4 levels and increasing regulatory T cell level. These results indicated that the prevention effects of proteasome inhibition with bortezomib on aGVHD are critically dependent on the dosage of bortezomib administration. And these results establish the basis for the clinical use of bortezomib in the management of graft-versus-host disease (GVHD). 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
    URL: Article
    DOI: 10.1182/blood.V114.22.4499.4499
    RVK:
    XA 33000
    RVK:
    XA 10000
    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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  • 7
    Online Resource
    Online Resource
    The Studies on the T-Cell Receptor Repertoire Diversity and Flow Cytometric Analysis in a Patient with MAS/HPS after Unrelated Peripheral Blood Stem Cell Transplantation.
    American Society of Hematology ; 2007
    In:  Blood Vol. 110, No. 11 ( 2007-11-16), p. 4970-4970
    Details
    In: Blood, American Society of Hematology, Vol. 110, No. 11 ( 2007-11-16), p. 4970-4970
    Abstract: Macrophage activation syndrome (MAS) /Hemophagocytic syndrome (HPS) is characterized by proliferation of activated macrophages under conditions such as infection(C Clin Infect Dis 2004)lymphoma(Aouba A Am J Hematol 2004), autoimmune disease(Kaneko K Clin Rheumatol 2005), solid organ transplantation(Akamatsu N,Transplant Proc 2006;). There have been several reports of MAS /HPS after hematopoietic stem cell transplantation, involving not only allogeneic,but also autologous transplantation(Sreedharan A Bone Marrow Transplantation,2006). Generally, MAS /HPS is a cytokine-related disorder.But at present, its clinical characteristics remain unknown. We firstly study here the T-cell receptor repertoire diversity and flow cytometric analysis in MAS /HPS after unrelated peripheral blood stem cell transplantation. The CDR3 of TCR Vα and Vβ subfamily genes were amplified in peripheral blood mononuclear cells from the patient with MAS/HPS after unrelated peripheral blood stem cell using RT-PCR for detection of the distribution of TCR Vα and Vβ repertoire, the PCR products were further analyzed by genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR Vα and VβT cells. Lymphocyte subsets in the peripheral blood were detected by monoclone antibody and flow cytometry including T lymphocyte subsets and NK cells. Flow-cytometric analysis showed CD56+ CD16+ cell 68.65% and CD3+ cell 11.79% in the lymphocyte population;CD16+CD69+ cell 68.51% and CD25+CD16+ cell 31.59% in NK cell. In the T lymphocytic subsets, CD25 + CD3+ cell 62%; CD69+CD3+ cell 75.81%; CD25CD4+ cell 0.81%,CD25CD8+ cell 3.48%; CD69CD4+ cell 0.31%, CD69+CD8+ cell 16.86%.The results show that the main activated lymphocytes is NK cell in patient at diagnosed with MAS/HPS. Of interest, it was only after the addition of high-dose IVIG 1g/kg/d for two days (Ostronoff et al BMT2006) to the treatment that MAS remitted. There are 23 Vα and 15Vβ subfamily T cells could be identified in this time, and the clonal expansion T cells could be found in TCR Vα5, 13, 20; TCR Vβ4, 11, 15 and 21subfamilies. Billiau et al (Blood 2005)describes the immunohistochemical findings on liver tissues from 5 children with MAS in the context of a different type of hemophagocytic syndrome (HPS) in liver transplantation. This study is the first directly to substantiates the presumed immunopathogenesis of MAS by documenting in situ expression of IFN-γ+ by activated CD8+ lymphocytes, and of IL-6 and TNF-α+ by hemophagocytosing macrophages, on liver tissues of patients with MAS. We found no evidence of potential infectious, autoimmune or malignant triggers of R-HPS in our patient, despite extensive investigations. We conclued that the skew distribution and clonal expansion of TCR Vα and Vβ subfamily T cells underscore the primary role of T cells in the pathogenesis of MAS/HPS.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V110.11.4970.4970
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2007
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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