抗胸腺细胞球蛋白在同胞全相合异基因外周血造血干细胞移植中的应用

 目的　探讨抗胸腺细胞球蛋白（ATG）在同胞全相合异基因外周血造血干细胞移植（allo-PBSCT）中应用的可行性及价值。方法　57例血液系统恶性肿瘤患者行同胞全相合PBSCT,采用改良的Bu／Cy方案或TBI／Cy方案预处理,予ATG+环胞菌素A（CsA）+霉酚酸酯（MMF）+短程甲氨蝶呤（MTX）预防移植物抗宿主病（GVHD）。结果　57例患者均达到稳定植活,急性GVHD（aGVHD）发生率为36.8 %,其中I～Ⅱ度aGVHD 31.6 %,Ⅲ度aGVHD 5.2 %,无Ⅳ度aGVHD发生。慢性GVHD（cGVHD）发生率为33.3 %,其中2例表现为广泛型。38.6 %患者在移植早期出现感染,50.9 %患者出现巨细胞病毒（CMV）血症。5年总生存（OS）率及无病生存率（DFS）分别为71.35 %和68.61 %。结论　同胞全相合allo-PBSCT预处理阶段加用低剂量的ATG可能降低GVHD发生率,患者移植后淋巴增殖性疾病及机会感染发生率无明显增加,但可能增加高危患者的复发率。     

HLA半相合外周血活化干细胞治疗晚期实体瘤的疗效

目的：观察HLA半相合外周血活化干细胞（HLA haploidentical peripheral blood stem cells,haploPBSCs）对晚期难治性实体瘤患者的临床抗肿瘤疗效以及不良反应。方法：入组42例全部为2004年10月至2007年10月天津医科大学肿瘤医院生物治疗科收治的晚期难治性恶性肿瘤患者（所有入组患者均知情同意,试验工作经医院伦理委员会批准）,其中卵巢癌12例,肾癌9例,肺癌8例,乳腺癌8例,结肠癌2例,胃癌2例,恶性黑素瘤1例。供者为患者健康直系亲属,进行haploPBSCs的动员、采集和体外rhIL2活化。经HLA半相合外周血干细胞治疗后,分别通过CT/PETCT检查、RESIST标准、KPS评分、临床症状缓解率等指标来评估HLA半相合外周血干细胞的临床疗效及不良反应情况。结果：42例患者接受1个疗程治疗后,全体患者中位无进展生存期（PFS）为6个月,临床获益率（CR+PR+SD）为73.8%;患者生活质量总获益率为76.2%,生活质量评分（KPS）较治疗前平均提高20分（0～30分）。其中,KIR不相合方向为GVH组的临床获益率、无进展生存期、生活质量总获益率均显著优于HVG(或相合)组\[94.1% vs 60.0%,（13.4±1.3）vs（8.0±0.9）个月,89.5% vs 65.2%,均P<005\];供受者关系为母子/女组的治疗有效率、患者生存期和生活质量均显著优于父子/女组（均P<0.05）;肾癌和卵巢癌的临床获益率分别为90.0%和81.8%,相对于其他类肿瘤类型高,在治疗反应性和敏感性上可能占优势。结论：HLA半相合外周血活化干细胞治疗后,机体产生非特异性的抗肿瘤作用,对改善患者症状和提高生活质量有显著效果。     

IL-2短期活化的HLA半相合的外周血单个核细胞治疗晚期恶性实体瘤初步观察

目的:初步探讨IL-2短期活化的HLA半相合的异体外周血单个核细胞(allo-mPBMCs)治疗晚期转移性/化疗抗性实体瘤的可行性。方法:11例晚期转移性/化疗抗性恶性实体瘤患者,供者为与患者HLA半相合直系亲属。供者经10μg/kg·dG-CSF动员3天后采集PBMC,体外用大剂量rhIL-2培养后回输患者。观察治疗后临床反应,采用流式细胞仪、LDH和ELISA试剂盒检测动员、活化前后表型,IL-2短期活化前后的非特异性杀瘤活性以及患者回输前后血清中各种细胞因子的含量变化。结果:11例患者回输的HLA半相合allo-mPBMCs总数达(2.5～4.3)×1010。体外经G-CSF动员后T细胞,尤其是CD4+T细胞显著下降,而NK细胞比例显著上升;经大剂量IL-2短期活化后,各种细胞亚群比例未见改变,但活化细胞(包括CD69+和CD25+细胞)亚群比例显著升高。体外实验中可观察到IL-2短期活化的allo-mPBMCs具有类似LAK样的较高的非特异性杀瘤活性。经过治疗,11例患者血清中Th1类细胞因子,如IL-2、IL-12、IFN-γ、TNF-α含量明显升高,而Th2类细胞因子,如TGF-β含量明显下降。治疗后8例观察到临床症状的缓解,或肿瘤标志物的降低,或影像学资料的改善,其中1例治疗后4个月达到部分缓解,而3例出现进展,其中2例死亡。结论:IL-2短期活化的HLA半相合的allo-mPBMCs具有明显的抗肿瘤效应,为治疗晚期转移性/化疗抗性实体瘤提供了新的希望。     

化疗联合亲缘HLA不全相合造血干细胞输注治疗血液系统恶性疾病

 目的　明确化疗联合亲缘HLA不全相合造血干细胞输注治疗血液系统恶性疾病的疗效及安全性。 方法　9例血液系统恶性疾病患者化疗后36 h回输亲缘HLA不全相合造血干细胞,评价其疗效、造血恢复时间及并发症。结果　9例患者包括4例急性髓系白血病、1例急性B淋巴细胞白血病、2例多发性骨髓瘤、1例霍奇金淋巴瘤、1例弥漫大B细胞淋巴瘤;年龄29～67岁;共接受治疗19例（次）,每个疗程平均回输单个核细胞计数（3.12±1.29）×108／kg,CD+34 细胞计数（1.71±1.00）×106／kg;CD+3 细胞计数（2.13±0.99）×108／kg。完全缓解4例,部分缓解1例,疾病进展4例。随访2～14个月,生存4例,死亡5例。9例患者输注亲缘HLA不全相合造血干细胞过程顺利,无病例检测到供者嵌合状态,未出现移植物抗宿主病。结论　化疗联合亲缘HLA不全相合造血干细胞输注治疗血液系统恶性疾病操作简单且经济,供者来源丰富且易获得,具有良好的安全性和耐受性。     

活化半相合混合骨髓移植GVHD反应的动物实验研究

目的:观察活化半相合混合骨髓移植的GVHD反应.方法:以急性放射病615小鼠模型为受鼠,615×C57BL/6杂交F1代小鼠为半相合供鼠.半相合鼠骨髓和脾细胞中混合一定比例的同基因脾细胞,经白细胞介素-2体外活化后,进行活化半相合混合骨髓移植.观察移植后小鼠死亡率、白细胞系造血重建、脾结节、体内混合淋巴细胞培养、嵌合体及病理学改变,比较不同移植方式的GVHD反应.结果:活化半相合骨髓移植组与未活化半相合骨髓移植组具有明显的GVHD反应.供受鼠3:1活化半相合混合骨髓移植不能明显降低GVHD反应,供受鼠脾细胞比例1:1和2:1移植组可以明显减轻GVHD反应.表现为外周血白细胞及骨髓造血恢复快,体内混合淋巴细胞反应降低及嵌合体百分率升高.结论:活化半相合混合骨髓梃移植方式可以降低GVHD反应,并与同基因和异基因脾细胞比例有关.     

家族相关的HLA部分不合的异基因骨髓移植

目的 :为了扩大异体造血干细胞移植供者造血干细胞的来源 ,探索 HL A部分不完全相合的家属供者的异体外周血干细胞移植治疗白血病的方法和效果 ,以克服家庭缩小 ,家族性 HL A相合供者不足的困难。方法 :对两例无 HL A相合家族供者的白血病患者进行 HL A一个位点不合的家族供者的异体外周血造血干细胞移植 ,给予抗淋巴细胞球蛋白和 FK5 0 6,以防治 GVHD。结果 :2例病人均移植成功 ,1例 HL A- A位点不合的病人顺利植入 ,未发生 GVHD。 1例 HL A- B位点不合的病人发生了 度 GVHD,经治疗后缓解。随访 >6个月均显示供者在染色体改变 ,无白血病表现。结论 :以 HL A不完全相合的家族供者进行异体造血干细胞移植治疗白血病是可行的有效方法 ,抗淋巴细胞球蛋白及 FK 5 0 6是防治 GVHD的有效方法     

非清髓性异基因造血干细胞移植治疗恶性肿瘤

非清髓性异基因造血干细胞移植,主要是通过移植物抗肿瘤效应来根除恶性肿瘤细胞,与经典的清髓性异基因造血干细胞移植相比,其移植相关并发症和病死率降低,对年龄较大、体质差的患者是一种较为安全可靠的治疗手段.     

非清髓性异基因造血干细胞移植治疗恶性肿瘤

非清髓性异基因造血干细胞移植 ,主要是通过移植物抗肿瘤效应来根除恶性肿瘤细胞 ,与经典的清髓性异基因造血干细胞移植相比 ,其移植相关并发症和病死率降低 ,对年龄较大、体质差的患者是一种较为安全可靠的治疗手段     

HLA半相合造血干细胞移植治疗恶性血液病的临床观察

背景与目的:异基因造血干细胞移植(allogeneichematopoieticstemcelltransplantation,allo-HSCT)是目前治疗恶性血液系统疾病的最有效手段。但仅有25%～30%的患者能找到人类白细胞抗原(humanleukocyteantigen,HLA)相合的亲缘供者;在无关人群中找到相合供者的概率是1/5万～1/10万,甚至更低。若进行HLA半相合造血干细胞移植(hematopoieticstemcelltransplantation,HSCT),则有90%的患者能找到供者,本文旨在探索HLA半相合HSCT治疗恶性血液病的可行性。方法:25例恶性血液病患者进行HLA半相合(其中单倍体20例)的亲缘供者HSCT。采用延长、强化联合免疫抑制促进植入及使用抗胸腺细胞球蛋白(antithymocyteglobulin,ATG)和/或加舒莱(抗CD25单抗)加强预防移植物抗宿主病(graft-versus-hostdisease,GVHD),粒细胞集落刺激因子(granulocytecolonystimulatinfactor,G-CSF)动员的骨髓(bonemarrow,BM)或加外周血干细胞(peripheralbloodstemcell,PBSC)混合移植方案。结果:所有患者均获得造血重建及达完全供者植入。21例(21/25)发生急性GVHD(aGVHD),其中Ⅰ度8例,Ⅱ度6例,Ⅲ度2例,Ⅳ度5例(其中3例为同胞部分相合),Ⅱ～Ⅳ度和Ⅲ～Ⅳ度aGVHD累积发生率分别为48.0%和28.6%。12例(12/25)发生慢性(c)GVHD,均为局限性。16例患者无病生存,1年预计无病生存率(disease-freesurvival,DFS)为(64.00±2.98)%,1年预计总生存率(overallsurvival,OS)为(64.0±3.08)%。9例死亡,其中1例死于复发,8例死于移植相关合并症,其中肺部感染4例,Ⅳ度GVHD3例,白质脑病1例。结论:HLA配型半相合的HSCT是治疗无亲缘和无关供体全相合的高危恶性血液病的有效方法,但风险较大,需在严密监测下慎重使用。     

异基因造血干细胞移植治疗白血病的临床研究

目的探讨异基因造血干细胞移植治疗白血病。方法脐血移植治疗儿童白血病3例,异基因外周血造血干细胞移植治疗白血病3例。结果6例患者完全植入,+15～+25天白细胞(WBC)>1.0×109/L,+35天～+51天血小板(Plt)>20×109/L。例1出现急性GVHDⅠ度,例5出现急性GVHDⅣ度,例6出现肝静脉闭塞综合征(VOD)、急性GVHDⅠ度、出血性膀胱炎(HC),例2移植后6个月复发,放弃治疗死亡。余5例正常生活或工作。结论异基因造血干细胞移植治疗白血病是安全有效的方法。     

Antineoplastic effects of partially HLA-matched irradiated blood mononuclear cells in patients with renal cell carcinoma.

PURPOSE: Vaccines, cytokines, and other biologic-based therapies are being developed as antineoplastic agents. Many of these agents are designed to induce an autologous immune response directed against the malignancy. In contrast, hematopoietic stem-cell transplantation is being developed as a form of allogeneic immunotherapy. This study tests the tolerance and antineoplastic activity of sequential infusions of partially HLA-matched allogeneic blood mononuclear cells (obtained from relatives) when administered outside of the context of a hematopoietic stem-cell transplantation. The cells are irradiated to prevent graft-versus-host disease. PATIENTS AND METHODS: Fifteen patients with relapsed or refractory malignancies for which no standard therapy was available were enrolled onto a clinical trial designed to assess the tolerability and antineoplastic effects of irradiated partially HLA-matched blood mononuclear cells obtained from relatives. RESULTS: There was disease regression in three patients with metastatic renal cell carcinoma during treatment. There was disease progression in six patients with metastatic renal cell carcinoma and two patients with metastatic melanoma during treatment. There was no change in disease state in several other patients. CONCLUSION: Irradiated allogeneic blood mononuclear cells administered outside the context of hematopoietic stem-cell transplantation may induce disease responses in patients with relapsed or refractory malignancies. Transfusion of irradiated allogeneic blood mononuclear cells should be developed further as a novel therapeutic antineoplastic approach.     

The combined administration of partially HLA-matched irradiated allogeneic lymphocytes and thalidomide in advanced renal-cell carcinoma: a case report

Renal-cell carcinoma (RCC) is susceptible to immune therapy including the use of the nonmyeloablative allogeneic transplantation (NST). However, NST can produce severe toxicity, might not be appropriate for many patients with metastatic RCC. Other novel allogeneic immunotherapies are designed to induce an autologous immune response directed against the malignancy. In single-arm phase II trials, thalidomide has demonstrated a modest activity in the treatment of advanced RCC. Here we present a case report in which a patient with advanced RCC in the absence of transplant conditioning, that was receiving thalidomide, was infused with partially HLA-matched irradiated allogeneic lymphocytes. In this patient a complete response with weak acute graft-versus-host disease (GVHD) was observed. No evidence of the disease was present over the subsequent 36 months survival of the patient, suggesting that the infusions may have played a major role in the antineoplastic effect. A potential mechanism of this protocol may involve a host-versus-graft reactions-mediated antitumor effect against the malignancy. In addition, the present results suggest that a combination protocol with alternate treatment (e.g., chemotherapy) schedules merit further investigation in the management of various malignancies.     

晚期原发性肝癌治疗的研究进展

原发性肝癌位居全球最常见恶性肿瘤的第五位,在癌症相关死亡原因中排名第三位.原发性肝癌治疗方法的选择很大程度上取决于肝脏功能,肿瘤大小,是否存在转移灶和血管侵犯.目前,原发性肝癌的根治方法包括肝切除术和肝移植手术,但是很多患者确诊时已进入晚期,失去了治愈的机会.全身系统性治疗对于晚期原发性肝癌的患者变得尤为重要,其中包括:分子靶向治疗、全身系统性化疗和免疫治疗.本文就近年来晚期原发性肝癌治疗的研究进展作一综述.     

Nonmyeloablative transplantation for solid tumors: a new frontier for allogeneic immunotherapy

The failure of conventional chemotherapy to improve survival in a large percentage of patients with advanced solid tumors has prompted the development of alternative anticancer approaches. Conventional allogeneic hematopoietic stem cell transplantation (HSCT) relies on myeloablative conditioning to eradicate the underlying disease, as well as suppress the patient’s immune response, allowing engraftment of the donor’s lymphohematopoietic system. Such preparative regimens are frequently associated with serious hematologic and nonhematologic toxicities, resulting in substantial morbidity and mortality. A significant curative component of allogeneic HSCT is the immune-mediated graft-versus-tumor (GVT) effect. Nonmyeloablative preparative regimens were designed to suppress host immunity to allow for sufficient engraftment of the donor immune system for the subsequent generation of GVT effects. These relatively low-dose preparative regimens are generally well tolerated and are associated with a reduction in the risk of transplant-related mortality. Nonmyeloablative HSCT provides a safer platform to explore the efficacy of allogeneic HSCT in patients with solid tumors. Initial reports have demonstrated that GVT may occur against several different solid tumors, including renal cell carcinoma, ovarian cancer, breast cancer and others. Based on these preliminary encouraging results, further exploration of nonmyeloablative HSCT for solid tumors is clearly warranted. The development of strategies to decrease graft-versus-host disease while enhancing post-transplant antitumor immunity will hopefully be forthcoming in the near future.     

DC-CIK联合化疗治疗进展期胃癌的临床研究

目的:观察DC-CIK过继免疫治疗联合化疗对进展期胃癌的临床疗效。方法:将60例患者分成实验组及对照组,前组采取DC-CIK、过继免疫治疗,并给予XELOX方案化疗。后组单独给予XELOX方案化疗,比较两组患者的近期疗效、免疫功能影响及安全性,对结果进行统计学分析。结果:实验组RR及DCR率分别为66.7%、76.7%,对照组发生PR 12例、SD 3例,对应RR及DCR率分别为40%、50%,两者相比均有统计学意义（P＜0.05）;治疗结束后实验组CD3+ 、CD4+ 、CD8+ 、CD3+CD56+双阳性细胞的比例均较对照组明显升高,两组对比有统计学意义,其它不良反应可耐受。结论:DC-CIK联合化疗治疗进展期胃癌优于常规化疗,安全性良好,但尚需进一步扩大样本量研究,值得临床推广应用。     

Changes of peripheral blood lymphocyte subsets in a long-term survivor with advanced renal cell carcinoma

We present the case of an 11-year survivor with advanced renal cell carcinoma (T4N2M1). This patient received long-term interferon-alpha (IFN-α)-based immunotherapy. Measurement of peripheral blood lymphocyte subsets suggested that reduction of the IFN-α dose might be associated with a marked increase of cytotoxic T cells, suppressor T cells, activated suppressor/cytotoxic T cells, and natural killer cells among the peripheral blood lymphocytes.     

Adjuvant therapy for advanced renal cell carcinoma

Introduction: Locally advanced, non-metastatic renal cell carcinoma (RCC) is conventionally managed with surgery. However, patients are at a high risk of RCC recurrence and have poor survival outcomes. An effective adjuvant systemic treatment is needed to improve on these outcomes. Targeted molecular and immune-based therapies have been investigated, or are under investigation, but their role in this setting remains unclear.

Areas covered: A comprehensive search of PubMed and ClinicalTrials.gov was performed for relevant literature. The following topics pertinent to adjuvant therapy in RCC were evaluated: strategies for patient selection, cytokine-based immunotherapy, vaccine therapy, VEGF and non-VEGF targeted molecular agents, and immune checkpoint inhibitors.

Expert commentary: Strong evidence for the incorporation of adjuvant therapy in high-risk RCC is lacking. Multiple targeted molecular therapies have been examined with only one approved for use. Genetic and molecular-based prognostic models are needed to determine who may benefit from adjuvant therapy. Developing adjuvant therapy strategies in the future depends on the results of important ongoing trials with immunotherapy and targeted agents.     

DC-CIK细胞治疗晚期转移性鼻咽癌的疗效

目的：评价DC-CIK细胞治疗晚期转移性鼻咽癌患者的疗效。方法：回顾性分析2011年8月至2015年6月解放军第八一医院肿瘤生物治疗科行DC-CIK细胞治疗的29例晚期转移性鼻咽癌患者。观察DC-CIK细胞治疗前后患者的疗效、安全性及EB病毒VCA-IgA抗体水平和淋巴细胞亚群的变化。结果：29例鼻咽癌患者经DC-CIK细胞治疗后客观缓解率为0,疾病控制率为65.5%,中位生存时间为42个月,1~4年生存率都为68.0%,无严重不良反应。治疗后29例患者的外周血EB病毒VCA-IgA抗体显著低于治疗前\[（30.88±3.91）U/ml vs (49.86±5.02) U/ml, P<0.05）\]。经细胞治疗后CD8+ T细胞显著上升（P<0.05）、CD4+/ CD8+ T细胞比值显著下降（P<0.05）。结论：DC-CIK细胞治疗转移性鼻咽癌安全、可行,能产生一定的临床获益。     

细胞因子在移植相关移植物抗宿主病诊断中的作用

目的探讨异基因造血干细胞移植（allo—HSCT）后外周血细胞因子（CK）表达水平的变化及其与移植物抗宿主病（GVHD）的关系。方法21例行allo—HSCT的血液病和实体瘤患者,根据预处理方案分为非清髓性（A组）和清髓性allo—HSCT组（B组）;根据发生GVHD情况分为1组（A组发生aGVHD）,2组（A组发生cGVHD）,3组（B组发生aGVHD）,4组（B组未发生GVHD）,5组（A组未发生GVHD）。采用半定量反转录聚合酶链反应（RT—PCR）和双抗体夹心酶联免疫吸附法（ELISA）对21例allo—HSCT患者动态监测外周血可溶性白细胞介素-2受体（sIL-2R）、干扰素-γ（IFN-γ）、转化生长因子β1（TGF—β1）表达变化情况。结果21例患者均获得造血重建,A、B两组发生GVHD情况无差别（χ^2=3．711,P=0．144）;RT—PCR方法及ELISA法检测CK,术后患者IL-2R、IFN-γ的表达均逐渐增高,IL-2R在＋7天时已明显高于术前（P〈0．05）,发生GVHD时达高峰,TGF-β1的表达随时间延长而逐渐降低,发生GVHD时最低;经治疗后,各指标逐渐接近术前水平;三指标在A组和B组表达不同（P〈0．01）;发生GVHD组与无GVHD组相比,差异有统计学意义（P〈0．001）。结论slL-2R、IFN-γ、TGF—β1可以作为allo—HSCT后预测aGVHD早期发生的指标;slL-2R、TGF—β1独立于其他参数,更能影响GVHD发生;RT—PCR和ELISA两种方法检测CK的敏感性无差别。