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Objective To investigate the therapeutic effects of haploidentical hematopoietic stem-cell transplantation (Haplo-PBSCT) for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy. Methods Eighty-nine cases of AML in first relapse after complete remission by standard DA/Hi-Ara-C regimens induction chemotherapy were evaluated retrospectively. Fiftythree cases were grafted by haplo-PBSCT and 26 cases were treated with iDA/Mid-Ara-C or MA/ Mid- Ara-C agents. Results The second remission rate in haplo-PBSCT group and continuous chemotherapy group was 86. 7 % (46/53 cases) and 38. 1% (9/23 cases) respectively (P＜0. 01). Survival postprogression (SPP) at 36th month was 43. 4 % (23/53 cases) in haplo-PBSCT group and 11.5 % (3/26 cases) in continuous chemotherapy group (P ＜ 0. 05). Conclusion Haplo-PBSCT could significantly increase the second remission rate and prolong the survival time of patients with acute myeloid leukernia in first relapse after complete remission by standard induction chemotherapy.
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BACKGROUND: It is easy to established human solid tumor nude mouse model, but for leukemia which is difficult. We inhibited immune system further by radioactive ray or CTX, to decrease cost and increase the stability.OBJECTIVE: To establish a human acute myeloblastic leukemia M2 Kasumi-1 models containing AML/ETO positive genes in BALB/c nude mouse. METHODS: Nude mice were randomly divided into three groups: CTX group was injected CTX 2 mg/day in abdominal cavity for two days, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein next day; irradiation group was exposed to total body irradiation, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein that day; untreated group was inoculated with 8×10~5/mouse Kasumi-1 cells by caudal vein injection. Three additional mice were considered as the normal control group. The blood smearing and bone morrow slides were detected, immunity type of BMC was detected using flow cytometry, loading of leukemic cellular tumor was detected using RT-PCR, and positive ratio of AML/ETO fusion gene was detected using FISH method. RESULTS AND CONCLUSION: After inoculated into untreated nude mice by caudal vein injection for 14 days, the ratio of leukemia cell in blood smearing was 3.5%, and over 40% in bone marrow slides, which was equal to the results of FISH and FCM. The increasing of tumor loading was time-dependent. For irradiation group and CTX treated group, the tumor loading was higher that untreated group, and the cells also survived more than 60 days. AML/ETO band was observed by RT-PCR in all experimental groups, for normal mice it was negative. The results indicated that the systemic disseminated leukemia model was established successfully by caudal vein injection 8×10~5/mouse Kasumi-1 cells in the three experimental groups.
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BACKGROUND: The effect of Chinese traditional medicine 3'-meisoindigo as well as indimbin derivatives on normal immunocytes is less reported while it is used for antitumor.OBJECTIVE: To investigate the effect of 3'-meisoindigo on the proliferation of the splenocytes and thymocytes of C56BL/6 mouse.DESIGN, TIME AND SETTING: The randomized cytopathology observation was performed between August 2007 and January 2008 at Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong Province, China.MATERIALS: C57BL/6 mice, clean, male, 6-8 weeks old, weighing (20±2) g.METHODS: The thymus gland and the spleen of C57BL/6 mice were ground to get the single-cell suspension and cells were treated by 5, 10, 15, 20 and 25 μ mol/L 3'-meisoindigo,respectively. Cells without any drug treatment were used as blank control and cells treated by concanavalin A were used as positive control.MAIN OUTCOME MEASURES: Proliferation of splenocytes and thymocytes detected using MTT method; IL-12 activity in culture supematant detected using ELISA method; the cell cycle, apoptosis rate, cell death rate and intracellular reactive oxygen species level detected using flow cytometry; the mRNA level of bcl-2 and cdk2 detected using reverse transcription-polymerase chain reaction; the expression rate of Bcl-2, CDK2 and Bax detected using fluorescence microscope.RESULTS: After treating for 24 hours, 15, 20 and 25 μ mol/L 3'-meisoindigo can significantly inhibit the proliferation of thymocytes and splenocytes (P < 0.05) and the inhibition was dose-dependent and time-dependent. Cells resumed proliferation after removing the 3'-meisoindigo, although they had been treated by 25 μ mol/L 3'-meisoindigo for 72 hours. The secretion of IL-12 was markedly reduced in all 3'-meisoindigo groups versus control groups at each time point (P < 0.05). 15 μ mol/L 3'-meisoindigo could decrease the mRNA expression level of apoptosis-related protein bcl-2 and cyclin cdk2 gene, decrease the expression level of BCL-2 protein and CDK protein, increase Bax expression level, decrease Bcl-2/Bax ratio markedly and started apoptosis.15 μ mol/L 3'-meisoindigo arrested cells in the G2/M stage of the thymocytes and splenocytes, and intracellular reactive oxygen species level elevated dose-dependently and time-dependently.CONCLUSION: In certain concentration range, 3'-meisoindigo can reversibly inhibit the proliferation of thymocytes and splenocytes of C57BL/6 mouse and can inhibit IL-12 secretion in parallel, and start apoptosis.
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<p><b>OBJECTIVE</b>To observe the influence of decreasing conditioning regimen intensity on the engraftment of HLA haplotype peripheral blood stem cell transplantation.</p><p><b>METHOD</b>Twelve patients with leukemia, including 4 in complete remission, whose HLAs were full matched with donors, and 8 with refractory leukemia, whose HLAs were mismatched, were transplanted with G-CSF mobilized allogeneic peripheral blood stem cells after conditioned with a regimen consisting of fludarabine (30 mg/m(2) x 6 days), busulfan (4 mg/kg x 2 days) and cyclophosphamide (30 approximately 60 mg/kg x 2 days) (FBC). Donor lymphocytes were infused at day + 30, + 60 and + 90 after transplantation, respectively. Hematopoietic reconstitution was observed. Engraftment was documented by the analysis of short tandem repeats with polymerase chain reaction (STR-PCR).</p><p><b>RESULT</b>Patients in HLA haplotype group received a mean number of 4.87 x 10(8)/kg donor mononuclear cells (MNC), with CD(34)(+) cells of 4.58 x 10(6)/kg and patients in HLA identical group a mean number of 4.85 x 10(8)/kg MNC with CD(34)(+) cells of 4.47 x 10(6)/kg. The mean time of white blood cell count more than 1.0 x 10(9)/L was 14 (10 approximately 18) days in HLA matched patients and 29 (11 approximately 90) days in HLA haplotype group. One three locus mismatched patient failed to engraft, but auto-hematopoiesis was recovered on day + 50. Full donor chimerism was observed in all patients except one with mixed chimera. The mixed chimera was converted into full donor chimera after three times donor lymphocyte infusion. One each died from severe acute GVHD, severe VOD and severe chronic GVHD in HLA haplotype group, and one from chronic GVHD in HLA identical group.</p><p><b>CONCLUSION</b>Patients survived engraftment was not influenced by decreasing conditioning intensity as in this regimen. Haplotype stem cells could be engrafted durable in recipients by this regimen combined with donor lymphocyte infusion.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Busulfan , Therapeutic Uses , Cyclophosphamide , Therapeutic Uses , Graft Survival , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Histocompatibility Testing , Immunosuppressive Agents , Therapeutic Uses , Leukemia , Therapeutics , Transplantation Conditioning , Transplantation Tolerance , Vidarabine , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Ly49A transfected mouse spleen cells on graft versus host disease (GVHD) and graft versus leukemia (GVL) effect after haploidentical allogeneic bone marrow transplantation in mice.</p><p><b>METHODS</b>Ly49A gene was transfected into spleen cells of C57BL/6 mice by retrovirus and the expression rate of Ly49A receptor was evaluated by flow cytometry. The murine model of haploidentical allogeneic acute GVHD was established by using C57BL/6(H - 2b) mouse as donor, and (BALB/c x C57BL/6) F1(H - 2d/b) (CB(6)F(1)) mouse as the recipient which was injected EL9611 cells before transplantation. After irradiation (TBI, (60)Co 10.5 Gy), the recipient received mixed graft of spleen cells and bone marrow cells to establish a GVHD model. The effects of Ly49A transfected spleen cells on GVHD and GVL post haploidentical allogeneic bone marrow transplantation were detected with this model.</p><p><b>RESULTS</b>The expression rate of Ly49A receptor was (42.20 +/- 4.87)%, (18.67 +/- 2.48)% and (18.73 +/- 3.82)% for pLXSN-Ly49A, pLXSN transfected and untransfected spleen cells respectively. Among haploidentical allo-BMT (C57BL/6(H - 2b)-->CB6F1(H - 2d/b)) groups, the survival time was (7.80 +/- 3.36) days for irradiation group; (21.70 +/- 2.87) days for cyclophosphomide therapy group; (29.40 +/- 6.43) days for mixed bone marrow cells and spleen cells transplantation group; (29.10 +/- 7.39) days for mixed bone marrow cells and pLXSN transfected spleen cells transplantation group and (45.00 +/- 12.38) days for mixed bone marrow cells and Ly49A transfected spleen cells transplantation group, which was much longer than that of any other groups (P = 0.000).</p><p><b>CONCLUSION</b>The Ly49A transfected spleen cell transplantation could alleviate GVHD and retain GVL effect in the acute GVHD model post haploidentical allo-BMT.</p>