细胞因子活化的供体淋巴细胞输注的疗效观察以及作用机制的初步研究
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摘要
一、细胞因子活化的供体淋巴细胞输注的疗效观察
目的观察细胞因子活化的供体淋巴细胞输注(新型DLI)治疗异基因造血干细胞移植(allo-HSCT)术后急性白血病复发患者的疗效。
方法回顾性分析16例allo-HSCT术后复发并接受新型DLI或常规DLI治疗的急性白血病患者的临床资料,用多参数微小残留病检测系统,包括骨髓形态学、流式细胞术(FCM)、实时定量PCR(RQ-PCR)、复合扩增荧光标记短串联重复序列(STR)-PCR结合毛细管电泳、R显带或荧光原位杂交(FISH)综合评价比较新型DLI和常规DLI疗效差异。新型DLI采用干扰素-α联合供体淋巴细胞输注,常规DLI为直接输注单采的供体淋巴细胞。
结果新型DLI组8例,治疗时疾病状态包括血液学复发(HR)7例,复发倾向1例,常规DLI组8例,治疗时疾病状态包括HR7例,分子学复发1例。新型DLI组与常规DLI组的诱导缓解率分别为62.5% vs 25%,对HR的诱导缓解率分别为57.14% vs 14.28%,诱导缓解中位时间分别为7天vs 23天。GVHD发生率分别为62.5% vs 25%,全血细胞减少发生率分别为62.5% vs 75%。1年总生存率分别为50% vs 12.5%。
结论单中心小样本临床分析结果显示,新型DLI治疗移植后复发急性白血病的诱导缓解率为62.5%,诱导缓解中位时间7天,持续缓解时间长,是治疗移植后复发的有效手段,未来有必要进一步探索个体化治疗策略,针对患者疾病状态、复发时间及移植类型设计新型DLI的治疗剂量,以最大程度产生GVL效应,降低重症GVHD的发生,延长患者的无病生存。
二、细胞因子活化的供体淋巴细胞输注作用机制的初步研究
目的初步探索细胞因子活化的供体淋巴细胞输注(新型DLI)可能的作用机制。
方法1.血细胞单采法收集经G-CSF动员后的外周血干细胞, Ficoll密度梯度离心法分离得到单个核细胞(MNC),用不同浓度的IFN-α(500U/ml、1000 U/ml、2000U/ml、4000U/ml)活化的MNC与SHI-1细胞系共孵育不同时间(2、3和7天),MTT法检测IFN-α活化的MNC对SHI-1细胞系的杀伤效应;2.利用SHI-1细胞系分组实验,摸索达到最佳细胞毒效应的时间和细胞因子浓度节点,应用FCM检测IFN-α对淋巴细胞和树突状细胞(DC)免疫表型的影响;3.收集4例移植后复发患者接受新型DLI治疗前后的外周血,FCM检测治疗前后的外周血中淋巴细胞亚群及淋巴细胞表面活化标志的变化。
结果1. IFN-α能增强MNC对SHI-1的杀伤效率,高浓度IFN-α(4000U/ml)在作用时间为2天和3天时,即能显著增强MNC对SHI-1的杀伤效应,当作用时间延长至第7天时,常规浓度(1000U/ml)亦能显著增强MNC对SHI-1的杀伤效率;2.达到最佳细胞毒效应的IFN-α浓度为4000U/ml,作用时间为3天,经过IFN-α培养3d的MNC,CD3~+CD4~+细胞比例升高,淋巴细胞活化标志CD69表达升高,DC表面成熟标志不同程度升高;3.治疗有效的2例患者在治疗前后CD3~+细胞、CD3~+CD4~+细胞、CD3~+CD8~+细胞均维持在稳定水平,而治疗无效的2例患者均出现不同程度的下降。治疗有效的患者均出现淋巴细胞活化标志HLA-DR表达升高。4例患者治疗前后NK细胞、CD4~+CD25~+细胞、Tγδ细胞均未见明显变化。
结论1.体外细胞系实验证实,经过7天的共孵育,高浓度和低浓度IFN-α均能增强供体淋巴细胞(G-CSF动员的PBSC)的抗白血病效应;2. IFN-α可能通过促进DC成熟活化,增加对肿瘤抗原的提呈,从而刺激T淋巴细胞增殖和活化,增强其杀伤效应;3.新型DLI可能通过活化传统T细胞发挥GVL效应。
PartⅠClinical Research of Cytokine-Activated Donor Lymphocyte Infusion
Objective:To evaluate the effect and efficacy of cytokine-actived donor lymphocyte infusion (the novel DLI) in patients who relapsed after allogeneic hematopoietic stem cell transplantations (allo-HSCT).
Methods:Sixteen patients with acute leukemia were retrospectively analyzed, who relapsed after allo-HSCT and were treated with the novel DLI or conventional DLI. Through multi-parameter detection of minimal residual disease, including morphology examination, flow cytometry (FCM), real time quantitive PCR (RQ-PCR), multiplex PCR amplification of short tandem repeats markers (STR-PCR), karyotype analysis with R-banding and fluorescence in situ hybridization, the therapeutic response of the novel DLI and conventional DLI were campared. Treatment regimen of two kinds of DLI were: interferon-αfollowed by DLI or infused donor lymphocyte by leukapheresis, respectively.
Result:Eight patients received the novel DLI, in which 7 patients had hematological relapse and 1 patient had recurrence tendency. Other 8 patients received conventional DLI, in which 7 patients had hematological relapse and 1 patient had molecular relapse. Complete remission (CR) rate induced by the novel DLI and conventional DLI was 62.5% vs 25%, respectively. Complete remission rate for patients with hematological relapse was 57.14% vs 14.28%, respectively. The median time required for a response to CR was 7d vs 23d, respectively. In the group of receiving the novel DLI, the occurrence of GVHD and pancytopenia were 62.5% and 62.5%, whereas in the group of receiving conventional DLI, 2 of 8 patients developed GVHD and 6 patients (75%) developed pancytopenia. The overall survival at 1 year were 50% and 12.5% respectively.
Conclusions:This pilot study in our institution here shows that a CR rate of 62.5% could be achieved used by the novel DLI treatment in relapsed patiens after allo-HSCT. The median remission-induction time is 7 days, and the remission state is durable. The novel DLI may be an effective therapy for patients with acute leukemia relapsed after allo-HSCT. In future, the optimal regimen of the novel DLI should be explored according to patients’disease stage, relapsing time after HSCT and transplantation modality, in order to enhance GVL and minimize GVHD as well as prolong disease-free survival.
PartⅡPreliminary Study of Mechanisms for Cytokine-Activated Donor Lymphocyte Infusion
Objective:To investigate the mechanism of cytokine-actived donor lymphocyte infusion (the novel DLI).
Methods:1. G-CSF-primed peripheral blood stem cell (PBSC) was collected by leukapheresis. PBSC was separated by ficoll density gradient centrifugation to obtain mononuclear cells (MNC). The SHI-1 cell strains were incubated with MNC activated by various concentrations of interferon-α(500, 1000, 2000 and 4000U/ml) and different times (2, 3 and 7 days). The tumoricidal activity of interferon-αactived MNC against SHI-1 was measured by MTT; 2. The key point of time and cytokine concentration was explored while the cytotoxicity of MNC against SHI-1 was maximal, the chage of surface molecules expression of lymphocytes and dendritic cells (DC) incubated with interferon-αwas analyzed by flow cytometry (FCM); 3. Peripheral blood of 4 patients who were treated with the novel DLI was collected. The immunophenotype of lymphocytes after treatment was analyzed by FCM.
Result:1. Interferon-α(IFN-α) enhanced the cytotoxicity of MNC against SHI-1 cell strains. After incubation for 2 and 3 days, high concentration of IFN-α(4000U/ml) could significantly enhance the cytotoxicity of MNC. While incubation for 7 days, conventional concentration of IFN-α(1000U/ml) could enhance the cytotoxicity of MNC against SHI-1 cell strains significantly; 2.The optimal key point of cytokine concentration and time are 4000U/ml and 3 days. The percentage of CD3~+CD4~+ cell increased significantly and the activated marker CD69 of T lymphocyte also increased markedly. The expression level of CD83, CD86 and HLA-DR of DC also increased; 3. CD3~+、CD3~+CD4~+ and CD3~+CD8~+ cell maintained at the stable level among the patients who had a response to the novel DLI, but decreased at the different level in the patients who had no response to the novel DLI. HLA-DR, the activated marker of lymphocyte, increased in the patients with response of the novel DLI. While the percentages of NK cell、CD4~+CD25~+ cell and Tγδcell did not changed during the novel DLI for all the 4 patients.
Conclusions:1. In vitro, both high and low concentration of IFN-αcan enhance MNC activity to kill the target cell after incubation for 7 days; 2. IFN-αmay promote the maturation of DC and enhance the capability of presenting tumor antigen, which can stimulate T cell to active and proliferate and mediate the tumoricidal activity; 3. The GVL effect of the novel DLI may be mediated by traditional T lymphocyte.
目的观察细胞因子活化的供体淋巴细胞输注(新型DLI)治疗异基因造血干细胞移植(allo-HSCT)术后急性白血病复发患者的疗效。
方法回顾性分析16例allo-HSCT术后复发并接受新型DLI或常规DLI治疗的急性白血病患者的临床资料,用多参数微小残留病检测系统,包括骨髓形态学、流式细胞术(FCM)、实时定量PCR(RQ-PCR)、复合扩增荧光标记短串联重复序列(STR)-PCR结合毛细管电泳、R显带或荧光原位杂交(FISH)综合评价比较新型DLI和常规DLI疗效差异。新型DLI采用干扰素-α联合供体淋巴细胞输注,常规DLI为直接输注单采的供体淋巴细胞。
结果新型DLI组8例,治疗时疾病状态包括血液学复发(HR)7例,复发倾向1例,常规DLI组8例,治疗时疾病状态包括HR7例,分子学复发1例。新型DLI组与常规DLI组的诱导缓解率分别为62.5% vs 25%,对HR的诱导缓解率分别为57.14% vs 14.28%,诱导缓解中位时间分别为7天vs 23天。GVHD发生率分别为62.5% vs 25%,全血细胞减少发生率分别为62.5% vs 75%。1年总生存率分别为50% vs 12.5%。
结论单中心小样本临床分析结果显示,新型DLI治疗移植后复发急性白血病的诱导缓解率为62.5%,诱导缓解中位时间7天,持续缓解时间长,是治疗移植后复发的有效手段,未来有必要进一步探索个体化治疗策略,针对患者疾病状态、复发时间及移植类型设计新型DLI的治疗剂量,以最大程度产生GVL效应,降低重症GVHD的发生,延长患者的无病生存。
二、细胞因子活化的供体淋巴细胞输注作用机制的初步研究
目的初步探索细胞因子活化的供体淋巴细胞输注(新型DLI)可能的作用机制。
方法1.血细胞单采法收集经G-CSF动员后的外周血干细胞, Ficoll密度梯度离心法分离得到单个核细胞(MNC),用不同浓度的IFN-α(500U/ml、1000 U/ml、2000U/ml、4000U/ml)活化的MNC与SHI-1细胞系共孵育不同时间(2、3和7天),MTT法检测IFN-α活化的MNC对SHI-1细胞系的杀伤效应;2.利用SHI-1细胞系分组实验,摸索达到最佳细胞毒效应的时间和细胞因子浓度节点,应用FCM检测IFN-α对淋巴细胞和树突状细胞(DC)免疫表型的影响;3.收集4例移植后复发患者接受新型DLI治疗前后的外周血,FCM检测治疗前后的外周血中淋巴细胞亚群及淋巴细胞表面活化标志的变化。
结果1. IFN-α能增强MNC对SHI-1的杀伤效率,高浓度IFN-α(4000U/ml)在作用时间为2天和3天时,即能显著增强MNC对SHI-1的杀伤效应,当作用时间延长至第7天时,常规浓度(1000U/ml)亦能显著增强MNC对SHI-1的杀伤效率;2.达到最佳细胞毒效应的IFN-α浓度为4000U/ml,作用时间为3天,经过IFN-α培养3d的MNC,CD3~+CD4~+细胞比例升高,淋巴细胞活化标志CD69表达升高,DC表面成熟标志不同程度升高;3.治疗有效的2例患者在治疗前后CD3~+细胞、CD3~+CD4~+细胞、CD3~+CD8~+细胞均维持在稳定水平,而治疗无效的2例患者均出现不同程度的下降。治疗有效的患者均出现淋巴细胞活化标志HLA-DR表达升高。4例患者治疗前后NK细胞、CD4~+CD25~+细胞、Tγδ细胞均未见明显变化。
结论1.体外细胞系实验证实,经过7天的共孵育,高浓度和低浓度IFN-α均能增强供体淋巴细胞(G-CSF动员的PBSC)的抗白血病效应;2. IFN-α可能通过促进DC成熟活化,增加对肿瘤抗原的提呈,从而刺激T淋巴细胞增殖和活化,增强其杀伤效应;3.新型DLI可能通过活化传统T细胞发挥GVL效应。
PartⅠClinical Research of Cytokine-Activated Donor Lymphocyte Infusion
Objective:To evaluate the effect and efficacy of cytokine-actived donor lymphocyte infusion (the novel DLI) in patients who relapsed after allogeneic hematopoietic stem cell transplantations (allo-HSCT).
Methods:Sixteen patients with acute leukemia were retrospectively analyzed, who relapsed after allo-HSCT and were treated with the novel DLI or conventional DLI. Through multi-parameter detection of minimal residual disease, including morphology examination, flow cytometry (FCM), real time quantitive PCR (RQ-PCR), multiplex PCR amplification of short tandem repeats markers (STR-PCR), karyotype analysis with R-banding and fluorescence in situ hybridization, the therapeutic response of the novel DLI and conventional DLI were campared. Treatment regimen of two kinds of DLI were: interferon-αfollowed by DLI or infused donor lymphocyte by leukapheresis, respectively.
Result:Eight patients received the novel DLI, in which 7 patients had hematological relapse and 1 patient had recurrence tendency. Other 8 patients received conventional DLI, in which 7 patients had hematological relapse and 1 patient had molecular relapse. Complete remission (CR) rate induced by the novel DLI and conventional DLI was 62.5% vs 25%, respectively. Complete remission rate for patients with hematological relapse was 57.14% vs 14.28%, respectively. The median time required for a response to CR was 7d vs 23d, respectively. In the group of receiving the novel DLI, the occurrence of GVHD and pancytopenia were 62.5% and 62.5%, whereas in the group of receiving conventional DLI, 2 of 8 patients developed GVHD and 6 patients (75%) developed pancytopenia. The overall survival at 1 year were 50% and 12.5% respectively.
Conclusions:This pilot study in our institution here shows that a CR rate of 62.5% could be achieved used by the novel DLI treatment in relapsed patiens after allo-HSCT. The median remission-induction time is 7 days, and the remission state is durable. The novel DLI may be an effective therapy for patients with acute leukemia relapsed after allo-HSCT. In future, the optimal regimen of the novel DLI should be explored according to patients’disease stage, relapsing time after HSCT and transplantation modality, in order to enhance GVL and minimize GVHD as well as prolong disease-free survival.
PartⅡPreliminary Study of Mechanisms for Cytokine-Activated Donor Lymphocyte Infusion
Objective:To investigate the mechanism of cytokine-actived donor lymphocyte infusion (the novel DLI).
Methods:1. G-CSF-primed peripheral blood stem cell (PBSC) was collected by leukapheresis. PBSC was separated by ficoll density gradient centrifugation to obtain mononuclear cells (MNC). The SHI-1 cell strains were incubated with MNC activated by various concentrations of interferon-α(500, 1000, 2000 and 4000U/ml) and different times (2, 3 and 7 days). The tumoricidal activity of interferon-αactived MNC against SHI-1 was measured by MTT; 2. The key point of time and cytokine concentration was explored while the cytotoxicity of MNC against SHI-1 was maximal, the chage of surface molecules expression of lymphocytes and dendritic cells (DC) incubated with interferon-αwas analyzed by flow cytometry (FCM); 3. Peripheral blood of 4 patients who were treated with the novel DLI was collected. The immunophenotype of lymphocytes after treatment was analyzed by FCM.
Result:1. Interferon-α(IFN-α) enhanced the cytotoxicity of MNC against SHI-1 cell strains. After incubation for 2 and 3 days, high concentration of IFN-α(4000U/ml) could significantly enhance the cytotoxicity of MNC. While incubation for 7 days, conventional concentration of IFN-α(1000U/ml) could enhance the cytotoxicity of MNC against SHI-1 cell strains significantly; 2.The optimal key point of cytokine concentration and time are 4000U/ml and 3 days. The percentage of CD3~+CD4~+ cell increased significantly and the activated marker CD69 of T lymphocyte also increased markedly. The expression level of CD83, CD86 and HLA-DR of DC also increased; 3. CD3~+、CD3~+CD4~+ and CD3~+CD8~+ cell maintained at the stable level among the patients who had a response to the novel DLI, but decreased at the different level in the patients who had no response to the novel DLI. HLA-DR, the activated marker of lymphocyte, increased in the patients with response of the novel DLI. While the percentages of NK cell、CD4~+CD25~+ cell and Tγδcell did not changed during the novel DLI for all the 4 patients.
Conclusions:1. In vitro, both high and low concentration of IFN-αcan enhance MNC activity to kill the target cell after incubation for 7 days; 2. IFN-αmay promote the maturation of DC and enhance the capability of presenting tumor antigen, which can stimulate T cell to active and proliferate and mediate the tumoricidal activity; 3. The GVL effect of the novel DLI may be mediated by traditional T lymphocyte.
引文
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[6]Dazzi F, Szydlo RM, Cross NC, et al. Durability of responses following donor lymphocyte infusions for patients who relapse after allogeneic stem cell transplant- tation for chronic myeloid leukemia. Blood. 2000; 96: 2712-2716.
[7]Porter DL, Antin JH. Donor leukocyte infusions in myeloid malignancies: new strategies. Best Pract Res Clin Haematol. 2006;19:737-755.
[8]Dazzi F, Fozza C. Disease relapse after haematopoietic stem cell transplantation: Risk factors and treatment. Best Pract Res Clin Haematol. 2007;20:311-327. [9]Falkenburg JH, van de Corput L, Marijt EW, et al. Minor histocompatibility antigens in human stem cell transplantation. Exp Hematol 2003;31:743-751.
[10]Marijt WA, Heemskerk MH, Kloosterboer FM, et al. Hematopoiesis-restricted minor histocompatibility antigens HA-1- or HA-2-specific T cells can induce com- plete remissions of relapsed leukemia. Proc Natl Acad Sci USA. 2003;100: 2742 -2747.
[11]Abbas AK, Murphy KM, Sher A. Functional diversity of helper T lymphocytes. Nature. 1996;383: 787-793.
[12]K?gi D, Vignaux F, Ledermann B, et al. Fas and perforin pathways as major mechanisms of T cell- mediated cytotoxicity. Science. 1994;265:528-530.
[13]Collins RH, Shpilberg O, Drobyski WR, et al. Donor leukocyte infusions in 140 patients with relapsed malignancy after allogeneic bone marrow transplantation. J Clin Oncol 1997;15:433-444.
[14]Radich JP, Gehly G, Gooley T, et al. Polymerase chain reaction detection of the BCR-ABL fusion transcript after allogeneic marrow transplantation for chronic myeloid leukemia: results and implications in 346 patients.Blood.1995;85:2632 -2638.
[15]Guerrasio A, Martinelli G, Saglio G, et al. Minimal residual disease status in transplanted chronic myelogenous leukemia patients: low incidence of polymerase chain reaction positive cases among 48 long disease-free subjects who received unmanipulated allogeneic bone marrow transplants. Leukemia .1992;6:507-512.
[16]Miyamura K, Tahara T, Tanimoto M, et al. Long persistent BCR-ABL positive transcript detected by polymerase chain reaction after marrow transplant for chronic myelogenous leukemia without clinical relapse: a study of 64 patients. Blood.1993; 81:1089-1093.
[17]Zaccaria A, Rosti G, Sessarego M, et al. Relapse after allogeneic bone marrow transplantation for Philadelphia chromosome positive chronic myeloid leukemia: cytogenetic analysis of 24 patients. Bone Marrow Transplant.1988;3:413-423.
[18]Arthur CK, Apperley JF, Guo AP, et al. Cytogenetic events after bone marrow transplantation for chronic myeloid leukemia in chronic phase. Blood.1988;71:1179- 1186.
[19]Michael S. Adoptive allogeneic immunotherapy - history and future perspectives. Transfusion Science. 2000;23:133-150.
[20]Rezvani K, Rahemtulla A, Cummins M, et al. Response of CML to donor lymphocyte infusions (DLI): factors in?uencing the effective cell dose and implica- tions for the definition of refractoriness. Blood. 2000; 96:195.
[21]Mackinnon S, Papadopoulos EB, Carabasi MH, et al. Adoptive immunotherapy evaluating escalating doses of donor leukocytes for relapse of chronic myeloidleukemia after bone marrow transplantation: separation of graft-versus-leukemia responses from graft-versus-host disease. Blood. 1995; 86:1261-1268.
[22]Dazzi F, Szydlo RM, Craddock C, et al. Comparison of single-dose and escalating -dose regimens of donor lymphocyte infusion for relapse after allografting for chronic myeloid leukemia. Blood. 2000; 95:67-71.
[23]Guglielmi C, Arcese W, Dazzi F, et al. Donor lymphocyte infusion for relapsed chronic myelogenous leukemia: prognostic relevance of the initial cell dose. Blood. 2002; 100:397- 405.
[24]Giralt S, Hester J, Huh Y et al. CD8-depleted donor lymphocyte infusion as treatment for relapsed chronic myelogenous leukemia after allogeneic bone marrow transplantation. Blood. 1995; 86:4337-4343.
[25]徐文鑫,翁开枝。免疫细胞在白血病免疫治疗应用中的研究进展。医学综述。2006;12:739-742。
[26]Loren AW, Porter DL. Donor leukocyte infusions for the treatment of relapsed acute leukemia after allogeneic stem cell transplantation. Bone Marrow Transplant. 2008;41:483-493.
[27]Liebowitz D, Lee K, CH J. Costimulatory approaches to adoptive immunotherapy. Curr Opin Oncol. 1998; 10:533–541.
[28]Porter DL, Levine BL, Bunin N, et al. A phase I trial of donor lymphocyte infusions expanded and activated ex vivo via CD3/CD28 co-stimulation. Blood. 2006; 107: 1325–1331.
[29]Kerr BM, Hsu AKW, Jones KL, et al. Adoptive immunotherapy to treat leukemic relapse following allogeneic hematopoietic stem cell transplantation. Clin Applied Immunol Rev. 2005;5:77–93.
[30]Fontaine P, Roy-Proulx G, Knafo L, et a1. Adoptive transfer of minor histocom- mpatibility antigen-specific T lymhpocytes eradicates leukemia cells without causing graft-versus-host disease. Nat Med. 2001;7:789-794.
[31]Molldrem JJ, Lee PP, Wang C, et al. A PR1-human leukocyte antigen-A2 tetramer canbe used to isolate low-frequency cytotoxic T lymphocytes from healthy donors that selectively lyse chronic myelogenous leukemia. Cancer Res.1999;59: 2675-2681.
[32]Gao L, Bellantuono I, Elsasser A, et al. Selective elimination of leukemic CD34(+) progenitor cells by cytotoxic T lymphocytes specific for WT1. Blood. 2000;95: 2198–2203.
[33]Zeis M, Siegel S, Wagner A, et al. Generation of cytotoxic responses in mice and human individuals against hematological malignancies using survivin-RNA- transfected dendritic cells. J Immunol. 2003;170:5391–5397.
[34]Biagi E, Marin V, Giordano-Attianese GM, et al. Chimeric T-cell receptors-new challenges for targeted immunotherapy in hematologic malignancies. Haemato logica. 2007;92:381- 388 .
[35]Greiner J, Dohner H, Schmitt M. Cancer vaccines for patients with acute myeloid leukemia-definition of leukemia-associated antigens and current clinical protocols targeting these antigens. Haemato logica. 2006;91:1653-1661.
[36]Wang J, Shaw JI, Mullen CA. Down-regulation of antihost alloractivity after bone marrow transplant permits relapse of hematological malignancy. Cancer Res. 2002;6: 208-212.
[37]Goulmy E, Voogt P, van Els C, et al. The role of minor histocompatibility anti- gens in GVHD and rejection: a mini-review. Bone Marrow Transplant.1991;7: 49-51.
[38]Borgham JA, Bredius RG, Hazenberg MD, et a1. Early determinants of long-term T-cell reconstitution after hematopoietic stem cell transplantation for severe combined Immunodeficiency. Blood. 2006;108:763-769.
[39]刘芯,谭获。特异诱导移植物抗白血病效应的研究进展。肿瘤防治杂志。2004;11:992-995。
[40]Drobyski WR, Gendelman M, Vodanovic-Jankovic S, et a1. Elimination of leuke- mia in the absence of lethal graft-versus-host disease after allogenic bone marrow tra nsplantation.1mmunol. 2003;170:3046-3053.
[41]Litvinova E, Maury S, Boyer O, et a1. Graft-versus-leukemia effect after suicide-gene-mediated control of graft-versus-host disease. Blood. 2002;100:2020-2025.
[42]Bordignon C, Bonini C, Verzeletti S, et al. Transfer of the HSV-tk gene into donor peripheral blood lymphocytes for in vivo modulation of donor anti-tumor immunity after allogeneic bone marrow transplantation. Hum Gene Ther. 1995;6: 813 -819.
[43]Bonini C, Ferrari G, Verzeletti S, et al. HSV-TK gene transfer into donor lymphocytes for control of allogeneic graft-versus-leukemia. Science. 1997;276: 1719 -1724.
[44]Koh CY, Blazar BR, George T, et al. Augmentation of antitumor effects by NK cell inhibitory receptor blockade in vitro and in vivo. Blood. 2001;97:3132-3137.
[45]Passweg JR, Tichelli A, Meyer-Monard S, et al. Purified donor NK-lymphocyte infusion to consolidate engraftment after haploidentical stem cell transplantation. Leukemia. 2004;18:1835-1838.
[46]Koehl U, S?rensen J, Esser R, et al. IL-2 activated NK cell immunotherapy of three children after haploidentical stem cell transplantation. Blood Cells Mol Dis. 2004;33:261-266.
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[7]Dazzi F, Fozza C. Disease relapse after haematopoietic stem cell transplantation: Risk factors and treatment. Best Pract Res Clin Haematol. 2007; 20:311-327.
[8]Porter DL, Antin JH. Donor leukocyte infusions in myeloid malignancies: new strategies. Best Pract Res Clin Haematol. 2006;19:737-755.
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[1]Wang Y, Xue MX, Wu XH, et al. Successful treatment of relapsed Philadelphia chromosome positive acute lymphoblastic leukemia with T315I mutation after haplo-identical hematopoietic stem cell transplantation with donor lymphocyte transfusion and interferonα-2b. Leuk Res. 2009;33:e111-113.
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[1]Weiden PL, Flournoy N, Thomas ED, et al. Antileukemic effects of graft- versus- host disease in human recipients of allogeneic marrow grafts. N Engl J Med. 1979; 300:1068–1073.
[2]Horowitz MM, Gale RP, Sondel PM, et al. Graft-versus-leukemia reactions after bone marrow transplantation. Blood. 1990; 75:555–562.
[3]Kolb HJ, Mittermuller J, Clemm C, et al. Donor leukocyte transfusions for treatment of recurrent chronic myelogenous leukemia in marrow transplant patients. Blood.1990; 76:2462–2465.
[4]Kolb HJ. Graft-versus-leukemia effects of transplantation and donor lymphocytes. Blood. 2008;112: 4371-4383.
[5]Ljunggren HG, Karre K. In search of the missing self : MHC molecules and NK cell recogition. Immunol Today.1990;11:237-244.
[6]Dazzi F, Szydlo RM, Cross NC, et al. Durability of responses following donor lymphocyte infusions for patients who relapse after allogeneic stem cell transplant- tation for chronic myeloid leukemia. Blood. 2000; 96: 2712-2716.
[7]Porter DL, Antin JH. Donor leukocyte infusions in myeloid malignancies: new strategies. Best Pract Res Clin Haematol. 2006;19:737-755.
[8]Dazzi F, Fozza C. Disease relapse after haematopoietic stem cell transplantation: Risk factors and treatment. Best Pract Res Clin Haematol. 2007;20:311-327. [9]Falkenburg JH, van de Corput L, Marijt EW, et al. Minor histocompatibility antigens in human stem cell transplantation. Exp Hematol 2003;31:743-751.
[10]Marijt WA, Heemskerk MH, Kloosterboer FM, et al. Hematopoiesis-restricted minor histocompatibility antigens HA-1- or HA-2-specific T cells can induce com- plete remissions of relapsed leukemia. Proc Natl Acad Sci USA. 2003;100: 2742 -2747.
[11]Abbas AK, Murphy KM, Sher A. Functional diversity of helper T lymphocytes. Nature. 1996;383: 787-793.
[12]K?gi D, Vignaux F, Ledermann B, et al. Fas and perforin pathways as major mechanisms of T cell- mediated cytotoxicity. Science. 1994;265:528-530.
[13]Collins RH, Shpilberg O, Drobyski WR, et al. Donor leukocyte infusions in 140 patients with relapsed malignancy after allogeneic bone marrow transplantation. J Clin Oncol 1997;15:433-444.
[14]Radich JP, Gehly G, Gooley T, et al. Polymerase chain reaction detection of the BCR-ABL fusion transcript after allogeneic marrow transplantation for chronic myeloid leukemia: results and implications in 346 patients.Blood.1995;85:2632 -2638.
[15]Guerrasio A, Martinelli G, Saglio G, et al. Minimal residual disease status in transplanted chronic myelogenous leukemia patients: low incidence of polymerase chain reaction positive cases among 48 long disease-free subjects who received unmanipulated allogeneic bone marrow transplants. Leukemia .1992;6:507-512.
[16]Miyamura K, Tahara T, Tanimoto M, et al. Long persistent BCR-ABL positive transcript detected by polymerase chain reaction after marrow transplant for chronic myelogenous leukemia without clinical relapse: a study of 64 patients. Blood.1993; 81:1089-1093.
[17]Zaccaria A, Rosti G, Sessarego M, et al. Relapse after allogeneic bone marrow transplantation for Philadelphia chromosome positive chronic myeloid leukemia: cytogenetic analysis of 24 patients. Bone Marrow Transplant.1988;3:413-423.
[18]Arthur CK, Apperley JF, Guo AP, et al. Cytogenetic events after bone marrow transplantation for chronic myeloid leukemia in chronic phase. Blood.1988;71:1179- 1186.
[19]Michael S. Adoptive allogeneic immunotherapy - history and future perspectives. Transfusion Science. 2000;23:133-150.
[20]Rezvani K, Rahemtulla A, Cummins M, et al. Response of CML to donor lymphocyte infusions (DLI): factors in?uencing the effective cell dose and implica- tions for the definition of refractoriness. Blood. 2000; 96:195.
[21]Mackinnon S, Papadopoulos EB, Carabasi MH, et al. Adoptive immunotherapy evaluating escalating doses of donor leukocytes for relapse of chronic myeloidleukemia after bone marrow transplantation: separation of graft-versus-leukemia responses from graft-versus-host disease. Blood. 1995; 86:1261-1268.
[22]Dazzi F, Szydlo RM, Craddock C, et al. Comparison of single-dose and escalating -dose regimens of donor lymphocyte infusion for relapse after allografting for chronic myeloid leukemia. Blood. 2000; 95:67-71.
[23]Guglielmi C, Arcese W, Dazzi F, et al. Donor lymphocyte infusion for relapsed chronic myelogenous leukemia: prognostic relevance of the initial cell dose. Blood. 2002; 100:397- 405.
[24]Giralt S, Hester J, Huh Y et al. CD8-depleted donor lymphocyte infusion as treatment for relapsed chronic myelogenous leukemia after allogeneic bone marrow transplantation. Blood. 1995; 86:4337-4343.
[25]徐文鑫,翁开枝。免疫细胞在白血病免疫治疗应用中的研究进展。医学综述。2006;12:739-742。
[26]Loren AW, Porter DL. Donor leukocyte infusions for the treatment of relapsed acute leukemia after allogeneic stem cell transplantation. Bone Marrow Transplant. 2008;41:483-493.
[27]Liebowitz D, Lee K, CH J. Costimulatory approaches to adoptive immunotherapy. Curr Opin Oncol. 1998; 10:533–541.
[28]Porter DL, Levine BL, Bunin N, et al. A phase I trial of donor lymphocyte infusions expanded and activated ex vivo via CD3/CD28 co-stimulation. Blood. 2006; 107: 1325–1331.
[29]Kerr BM, Hsu AKW, Jones KL, et al. Adoptive immunotherapy to treat leukemic relapse following allogeneic hematopoietic stem cell transplantation. Clin Applied Immunol Rev. 2005;5:77–93.
[30]Fontaine P, Roy-Proulx G, Knafo L, et a1. Adoptive transfer of minor histocom- mpatibility antigen-specific T lymhpocytes eradicates leukemia cells without causing graft-versus-host disease. Nat Med. 2001;7:789-794.
[31]Molldrem JJ, Lee PP, Wang C, et al. A PR1-human leukocyte antigen-A2 tetramer canbe used to isolate low-frequency cytotoxic T lymphocytes from healthy donors that selectively lyse chronic myelogenous leukemia. Cancer Res.1999;59: 2675-2681.
[32]Gao L, Bellantuono I, Elsasser A, et al. Selective elimination of leukemic CD34(+) progenitor cells by cytotoxic T lymphocytes specific for WT1. Blood. 2000;95: 2198–2203.
[33]Zeis M, Siegel S, Wagner A, et al. Generation of cytotoxic responses in mice and human individuals against hematological malignancies using survivin-RNA- transfected dendritic cells. J Immunol. 2003;170:5391–5397.
[34]Biagi E, Marin V, Giordano-Attianese GM, et al. Chimeric T-cell receptors-new challenges for targeted immunotherapy in hematologic malignancies. Haemato logica. 2007;92:381- 388 .
[35]Greiner J, Dohner H, Schmitt M. Cancer vaccines for patients with acute myeloid leukemia-definition of leukemia-associated antigens and current clinical protocols targeting these antigens. Haemato logica. 2006;91:1653-1661.
[36]Wang J, Shaw JI, Mullen CA. Down-regulation of antihost alloractivity after bone marrow transplant permits relapse of hematological malignancy. Cancer Res. 2002;6: 208-212.
[37]Goulmy E, Voogt P, van Els C, et al. The role of minor histocompatibility anti- gens in GVHD and rejection: a mini-review. Bone Marrow Transplant.1991;7: 49-51.
[38]Borgham JA, Bredius RG, Hazenberg MD, et a1. Early determinants of long-term T-cell reconstitution after hematopoietic stem cell transplantation for severe combined Immunodeficiency. Blood. 2006;108:763-769.
[39]刘芯,谭获。特异诱导移植物抗白血病效应的研究进展。肿瘤防治杂志。2004;11:992-995。
[40]Drobyski WR, Gendelman M, Vodanovic-Jankovic S, et a1. Elimination of leuke- mia in the absence of lethal graft-versus-host disease after allogenic bone marrow tra nsplantation.1mmunol. 2003;170:3046-3053.
[41]Litvinova E, Maury S, Boyer O, et a1. Graft-versus-leukemia effect after suicide-gene-mediated control of graft-versus-host disease. Blood. 2002;100:2020-2025.
[42]Bordignon C, Bonini C, Verzeletti S, et al. Transfer of the HSV-tk gene into donor peripheral blood lymphocytes for in vivo modulation of donor anti-tumor immunity after allogeneic bone marrow transplantation. Hum Gene Ther. 1995;6: 813 -819.
[43]Bonini C, Ferrari G, Verzeletti S, et al. HSV-TK gene transfer into donor lymphocytes for control of allogeneic graft-versus-leukemia. Science. 1997;276: 1719 -1724.
[44]Koh CY, Blazar BR, George T, et al. Augmentation of antitumor effects by NK cell inhibitory receptor blockade in vitro and in vivo. Blood. 2001;97:3132-3137.
[45]Passweg JR, Tichelli A, Meyer-Monard S, et al. Purified donor NK-lymphocyte infusion to consolidate engraftment after haploidentical stem cell transplantation. Leukemia. 2004;18:1835-1838.
[46]Koehl U, S?rensen J, Esser R, et al. IL-2 activated NK cell immunotherapy of three children after haploidentical stem cell transplantation. Blood Cells Mol Dis. 2004;33:261-266.