Search

No relevant conflicts of interest to declare.

Introduction:

To observe the proliferative effect on acute lymphocytic leukemia (ALL) cell lines caused by G-CSF, and to evaluate the synergetically anti-leukemic effect on these cell lines produced by Ara-C combined with G-CSF.With fluorescence quantitation kit and flow cytometry, G-CSFR expression density of 5 ALL cell lines were detected, and expression density changes due to G-CSF stimulation were evaluated. After treatment by G-CSF of various concentrations, changes of cell cycle and cell proliferation in ALL cell lines were examined by DNA-PREP-TM kit and Cell Counting Kit (CCK-8), respectively. After treatment by different concentration's Ara-C combined with different concentration's G-CSF, ALL cell lines’ existing cell viability were detected by CCK-8.All 5 ALL cell lines expressed G-CSFR, 2 ALL cell lines including CCRF-CEM and 697 exhibited G-CSFR expression density up-regulation tendency after G-CSF's stimulation, while 3 T-ALL cell lines including MOLT-4, Jurkat,Clone E6-1, A3 showed G-CSFR expression density down-regulation tendency after G-CSF's stimulation, which was similar in neutrophil. Cell cycle analysis showed that S phase cell proportion in MOLT-4, Jurkat,Clone E6-1, A3 cell lines had a growing tendency after G-CSF’ treatment, while no similar tendency was observed in CCRF-CEM, 697 cell lines. A relatively proliferative advantage was found in MOLT-4, Jurkat,Clone E6-1, A3 cell lines after G-CSF's treatment, but only in MOLT-4 there was a statistically significance after 10ng/ml G-CSF's stimulation. A synergetically anti-leukemic effect was observed in MOLT-4, Jurkat,Clone E6-1, A3 cell lines after Ara-C's treatment combined with G-CSF.The biological characteristics of ALL cell lines expressing G-CSFR was different from each other. Synergetically anti-leukemic effect produced by cytotoxic agent and G-CSF could occur in ALL cell lines which showed a growing S phase cell proportation after G-CSF's treatment. ALL cell's response to G-CSF, which means down-regulation of G-CSFR expression density, seems a promising indicator that a synergetically effect could be produced by Ara-C plus G-CSF. Before this indicator could be applied for guiding ALL patients’ management, clinical researches are needed to confirm it.No relevant conflicts of interest to declare.

Abstract 4330

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

Abstract 4496

Abstract 4503

Background CD4+CD25+ regulatory T cell (Treg), the most significant subset of regulatory T cells, has been found to play an important role in suppressing allogenetic immune response and controlling GVHD recently. The present single-center study monitored the ratio of CD4+CD25high T cells and the expression of FOXP3 gene in peripheral blood post-transplant to assess the reconstitution of CD4+CD25high Treg early after allogenetic stem cell transplantation and its correlations with aGVHD. Methods: 22 patients undergoing allo-HSCT were enrolled. To detect the CD4+CD25+T cells and the CD4+CD25high T cells subpopulations ratio in CD4+ T lymphocytes(CD4+CD25+/CD4+ ACD4+CD25high/CD4+) Athe CD4+CD25high T cells subpopulations ratio in CD4+CD25+ T cells(CD4+CD25high/CD4+ CD25+) Aand the expression of FOXP3 mRNA in the donor graft and periperal blood frequently after transplantation with the flow cytometry and real-time PCR assays. Results: \|[Dagger]\|@CD4 +CD25high/CD4+ and CD4+CD25high/CD4+CD25+ ratios in the donor graft were significantly lower in the aGVHD group(P<0.05)‡AEarly after transplantation, Both groups’CD4+CD25+/CD4+ ratio in peripheral blood increased significantly than that in the donor graft(P<0.05).CD4+CD25high/CD4+ and CD4+CD25high/CD4+CD25+ ratios were also significantly higher than that in donor graft in the aGVHD group, but not in the no-aGVHD group.‡BThe expression of FOXP3 in two groups after transplantation increased gradually, until six months after transplantation, its expression was still below the control level(P<0.05). The expression of FOXP3 was lower in the aGVHD group. Conclusion: \|[Dagger]\|@The early activation and proliferation of the CD4+CD25high regulatory T cells is critical for the maintainance of immune hemostasis and the control of aGVHD.‡AThe occurrence of aGVHD is related with lower ratios of CD4+CD25high regulatory T cells in CD4+T cells and activated CD4+ effector T cells.‡BaGVHD may affect the quantities of CD4+CD25high regulatory T cells and the expression of FOXP3.

BackgroundCD4+CD25+regulatory T cell (Treg), the most significant subset of regulatory T cells, has been found to play an important role in suppressing allogenetic immune response and controlling GVHD recently. The present single-center study monitored the ratio of CD4+CD25highT cells and the expression of FOXP3 gene in peripheral blood post-transplant to assess the reconstitution of CD4+CD25highTreg early after allogenetic stem cell transplantation and its correlations with aGVHD.

Idiopathic hypereosinophilic syndromes (HES) or chronic eosinophilic leukemia (CEL) comprise a spectrum of indolent to aggressive diseases characterized by persistent hypereosinophilia. Hypereosinophilia can result from the presence of a defect in the hematopoietic stem cell giving rise to eosinophilia, it can present in many myeloproliferative disorders or alternatively it may be a reactive form, secondary to many clinical conditions. The fusion gene FIP1L1-PDGFR alpha was identified in a subset of patients presenting with HES/CEL. In spite of this, the majority of HES/CEL patients do not present detectable molecular lesions and for many of them the diagnosis is based on exclusion criteria and sometimes it remains doubt. CD34-positive progenitor cells from bone marrow (BM) express BAALC and WT1. Overexpression of BAALC and WT1 were seen in patients with AML and ALL. In a subset of AML it marked poor prognosis, suggesting a role for BAALC or WT1 overexpression in acute leukemia. To explored the possibility to distinguish between HES/CEL and reactive hypereosinophilia based on the measurement of BAALC and WT1 transcript amount. Twenty-two patients with hypereosinophilia were characterized at the molecular level and analyzed for BAALC and WT1 expression. The transcription of FIP1L1-PDGFRalpha fusion gene was detected by nested RT-PCR. The relative transcript amount of BAALC and WT1 were determined by real time PCR analyses. The FIP1L1-PDGFRalpha fusion gene expressed has been identified in bone marrow mononuclear cells of 4 cases. The relative expression level of BAALC and WT1 in these 4 cases with positive FIP1L1-PDGFRalpha fusion gene expression were 2.27(0.27–6.8) and 0.39(0.002–0.90), respectively. Whereas the relative amount of transcripts of BAALC and WT1 in 18 patients with negative FIP1L1-PDGFRalpha fusion gene were 0.069(0.015–0.11) and 0.054(0–0.34) respectively. The relative amount of transcripts of BAALC and WT1 in patients with HES/CEL were 32 times and 7 times than that in those with negative FIP1L1-PDGFRalpha fusion gene, respectively. These results clearly demonstrates that BAALC and WT1 quantitative assessment allows to discriminate between HES/CEL and reactive eosinophilia and represents a useful tool for disease monitoring especially in the patients lacking a marker of clonality.

Idiopathic hypereosinophilic syndromes (HES) or chronic eosinophilic leukemia (CEL) comprise a spectrum of indolent to aggressive diseases characterized by persistent hypereosinophilia. Hypereosinophilia can result from the presence of a defect in the hematopoietic stem cell giving rise to eosinophilia, it can present in many myeloproliferative disorders or alternatively it may be a reactive form, secondary to many clinical conditions. The fusion gene FIP1L1-PDGFR alpha was identified in a subset of patients presenting with HES/CEL. In spite of this, the majority of HES/CEL patients do not present detectable molecular lesions and for many of them the diagnosis is based on exclusion criteria and sometimes it remains doubt. CD34-positive progenitor cells from bone marrow (BM) express BAALC and WT1. Overexpression of BAALC and WT1 were seen in patients with AML and ALL. In a subset of AML it marked poor prognosis, suggesting a role for BAALC or WT1 overexpression in acute leukemia. To explored the possibility to distinguish between HES/CEL and reactive hypereosinophilia based on the measurement of BAALC and WT1 transcript amount. Twenty-two patients with hypereosinophilia were characterized at the molecular level and analyzed for BAALC and WT1 expression. The transcription of FIP1L1-PDGFRalpha fusion gene was detected by nested RT-PCR. The relative transcript amount of BAALC and WT1 were determined by real time PCR analyses. The FIP1L1-PDGFRalpha fusion gene expressed has been identified in bone marrow mononuclear cells of 4 cases. The relative expression level of BAALC and WT1 in these 4 cases with positive FIP1L1-PDGFRalpha fusion gene expression were 2.27(0.27–6.8) and 0.39(0.002–0.90), respectively. Whereas the relative amount of transcripts of BAALC and WT1 in 18 patients with negative FIP1L1-PDGFRalpha fusion gene were 0.069(0.015–0.11) and 0.054(0–0.34) respectively. The relative amount of transcripts of BAALC and WT1 in patients with HES/CEL were 32 times and 7 times than that in those with negative FIP1L1-PDGFRalpha fusion gene, respectively. These results clearly demonstrates that BAALC and WT1 quantitative assessment allows to discriminate between HES/CEL and reactive eosinophilia and represents a useful tool for disease monitoring especially in the patients lacking a marker of clonality.