FasL基因修饰树突状细胞的抗白血病作用及其机制

目的: 选择性去除骨髓移植物中异基因反应性淋巴细胞，特异性抑制移植物抗宿主病（graft versus host disease, GVHD）并保留移植物的抗白血病(graft versus leukemia, GVL)作用。方法：分别把未经处理的、溶T淋巴细胞处理的和以FasL基因修饰树突状细胞（FasLDC）处理的     

二甲基肼诱导昆明小鼠大肠癌的研究

目的: 选择性去除骨髓移植物中异基因反应性淋巴细胞,特异性抑制移植物抗宿主病(GVHD).方法: 用携带有FasL基因的重组腺病毒转染Balb/c小鼠来源的树突状细胞(dendritic cells, DC),并与C57BL/6小鼠骨髓细胞移植物共培养,把经过这种处理的骨髓细胞移植物移植给Balb/c受体小鼠(C57BL/6→Balb/c小鼠GVHD 模型 ,H-2b→H-2d),然后观察、比较各组GVHD表现.结果: 致死剂量照射的受体鼠在接受经FasL-DC处理的供体骨髓细胞移植后,没有出现明显的GVHD表现,生存期显著延长,3个月时生存率80%以上.但对照组2周后均出现了明显的GVHD症状,腹泻、脱毛和靶组织淋巴细胞浸润等,生存期没有超过30 d.结论: 转染FasL基因的DC可有效去除骨髓移植物中异基因反应性T淋巴细胞,移植用这种方法处理过的骨髓,能够有效抑制GVHD的发生.     

选择性去除骨髓移植物中异基因反应性淋巴细胞

目的:选择性去除骨髓移植物中异基因反应性淋巴细胞,诱导异基因特异性低反应性.方法:用携带有FasL基因的重组腺病毒转染BALB/c小鼠来源的DC,并与C57BL/6小鼠脾淋巴细胞共培养,通过混合淋巴细胞培养等方法检测异基因特异性低反应性.结果:转染FasL基因的DC诱导了对BALB/c异基因反应性T淋巴细胞凋亡,2次混合淋巴细胞反应显著降低,但并没有抑制针对第三方的反应.结论:转染FasL基因的DC体外可有效去除骨髓移植物中异基因反应性T淋巴细胞,移植用这种方法处理过的骨髓,有希望在抑制GVHD的同时,不影响移植物抗肿瘤复发和抗感染的功能.     

吲哚胺2，3-二氧化酶基因修饰的DCs对小鼠移植物抗宿主病的抑制作用

目的:探讨小鼠树突状细胞(dendritic cells,DCs)转染吲哚胺2,3-二氧化酶(indolamine 2,3-dioxygenase,IDO)基因后对移植物抗宿主病(graft versus host disease,GVHD)的抑制作用。方法:用携带IDO基因的重组腺病毒感染BALB/c小鼠来源的树突状细胞,后者与C57BL/6小鼠骨髓移植物共培养,将经过处理的骨髓移植给BALB/c小鼠(C57BL/6→BALB/c小鼠GVHD模型,H-2~h→H-2~d),观察、比较各组GVHD表现(包括GVHD评分、生存期、病理学改变),并行嵌合体检测及观察混合淋巴细胞反应(mixed lymphocyte reaction,MLR)。结果:致死剂量照射的受体小鼠接受经IDO处理的骨髓移植(bone marrow transplantation,BMT)后,未出现明显的GVHD反应,生存期显著延长,3个月时生存率大于80%;对照组均在移植后2周左右出现明显的GVHD表现,生存期未超过1个月。IDO-DC治疗组未出现明显的组织病理学损害;3个月时IDO-DC治疗组仍然保持比较高的嵌合;IDO-DC组的T细胞对C57BL/6、BALB/c淋巴细胞的反应性与对C3H淋巴细胞反应性相比显著降低(P<0.05)。结论:经IDO基因修饰的DCs可以选择性去除骨髓移植物中异基因反应性T细胞,从而特异性地抑制GVHD并能及早地免疫重建。     

非清髓性异基因骨髓移植治疗白血病的动物实验研究

[目的]探讨非清髓性异基因骨髓移植及加供者淋巴细胞输注治疗小鼠白血病的疗效.[方法]荷L7212白血病的615(H-2K)小鼠,于接种白血病细胞后第2天接受60Co-γ射线全身照射(TBI 8.5Gy或5Gy)分为若干组,照射当天移植供鼠BALB/C(H-2d)小鼠的骨髓细胞(5×106)和脾细胞(1.5×107),移植后第2天腹腔注射环磷酰胺(200mg/kg);供者淋巴细胞输注组分别于移植后第7天、14天、21天再次输注供鼠脾细胞5×106、1×107、2×107,观察受鼠的移植物植入、移植物抗宿主病(GVHD)、受鼠生存时间及移植相关并发症等.[结果]非清髓性预处理能保证移植物的稳定植入,非清髓性异基因骨髓移植组小鼠生存时间为22.3±4.8天,与非清髓空白组14.7±3.4天和传统移植组18.3±3.2天比较均有显著性差异(P＜0.05),供者淋巴细胞输注(DLI)组小鼠平均生存时间明显延长,为34.3±2.5天,与非清髓移植组比较均有显著性差异(P＜0.05),且无明显GVHD表现和病理学改变,移植相关并发症减少.[结论]非清髓性异基因骨髓移植能在减轻GVHD的同时保留一定的移植物抗白血病(GVL)效应,移植后行DLI可在减轻移植相关并发症的基础上进一步增强GVL效应.     

CD自杀基因系统对T淋巴细胞作用的实验研究

异基因骨髓移植是治愈白血病及某些实体肿瘤的有效手段,但其并发症——移植物抗宿主病(GVHD)目前仍具有相当高的发病率和死亡率,是影响移植成功的主要障碍.在移植前体外去除供者骨髓中的T淋巴细胞可有效减轻GVHD)的发生,但T细胞的去除可使移植物失去移植物抗白     

供受体的不同T细胞比例对GVHD影响的研究

目的 :建立大鼠异基因骨髓移植模型 ,探讨供受体的不同T细胞比例对GVHD严重程度的影响。方法 :受体SD大鼠接受TBI+CTX预处理 ,分别按 1∶1、2∶1、4∶1的移植物与受体T细胞比例回输供体WISTAR大鼠骨髓细胞 ,观察受体生存期、临床及病理GVHD评分。结果 :实验组大鼠按移植物与受体T细胞比例为 1∶1和 2∶1移植时生存期、临床及病理GVHD评分与 4 :1时差异有显著性 (P <0 0 5 )。结论 :当移植时移植物内T细胞和经预处理后受体残存T细胞比例为 1∶1和2∶1时GVHD程度较轻 ,而 4∶1时GVHD明显加重。     

非清髓性异基因造血干细胞移植在实体瘤治疗中的应用

异基因干细胞移植用于实体瘤的治疗有很大的潜力。传统的清髓性造血干细胞移植采用大剂量化疗药物对受者进行预处理,虽然起到一定的杀伤肿瘤细胞作用,但增加了移植相关死亡率（transplant-related mortality,TRM）。随着对非清髓性预处理的深入研究,人们将非清髓性造血干细胞移植用于实体瘤的治疗。与清髓性干细胞移植比较,非清髓性移植减轻了TRM,且拓宽了移植适应证。移植物抗宿主病（graft versus host disease,GVHD）是异基因移植后出现的毒性反应,甚至成为致命性的并发症。移植物抗肿瘤（graft versus tumor,GVT）效应是异基因移植治疗实体瘤的关键,而GVT效应常伴随GVHD的出现。因此,如何在保留GVT效应的同时降低GVHD是我们所面临挑战。目前,通过改变预处理方案、加强对移植物的处理、改变免疫抑制疗法等3种策略使用于GVHD的防治,取得了一定效果,为异基因造血干细胞移植治疗实体瘤带来了广阔前景。     

次要组织相容性抗原在异基因造血干细胞移植中应用的研究进展

异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation, alloHSCT)是目前治愈血液系统恶性疾病的有效手段。由于次要组织相容性抗原（minor histocompatibility antigens, mHags）本身的特性及其免疫学效应,在alloHSCT后的移植物抗宿主病（graft versus host disease, GVHD）和移植物抗白血病效应（graft versus leukemia effect, GVL）中都发挥了重要的作用。研究发现,一些mHags分布广泛,在各种细胞中均有表达,如HA3、HA4等;另外一些则限制性表达在造血细胞起源的细胞表面,如HA1、HA2等。目前正在研究利用mHags分布的差异应用于alloHSCT中,以达到加强GVL和减少GVHD的目的。     

自杀基因在异基因骨髓移植中的应用

异基因骨髓移植(allo-BMT)是治愈恶性血液系统疾病的重要方法之一,但其并发症移植物抗宿主病(GVHD)限制了其在临床上的发展应用.国内外不少研究表明自杀基因转染T淋巴细胞可控制移植物抗宿主病.     

MHC半相合脾加骨髓细胞诱发H22荷瘤鼠的抗肿瘤效应

目的：观察MHC半相合脾加骨髓细胞移植抗小鼠H22实体瘤的效果。方法：以皮下接种H22肝癌细胞的BALB/c×C57BL/6杂交F1代雌性小鼠为受鼠,以健康雌性F1、雄性C57BL/6、雄性C3H小鼠为MHC全相合、半相合、不相合供鼠,观察移植后的抑瘤情况；观察供鼠细胞经~(60)Co照射的MHC半相合移植对WBC、生化和嵌合体的影响；比较供鼠细胞经与不经~(60)Co照射的MHC半相合移植的GVHD情况。结果：供鼠细胞经/不经~(60)Co照射的MHC半相合移植小鼠的肿瘤明显较小,与单纯化疗未进行移植者比较,差异具有统计学意义(P＜0．05)；但受鼠未经化疗预处理的MHC半相合细胞输注没有出现抗肿瘤效应；供鼠细胞经7．5 Gy ~(60)Co照射的MHC半相合移植能明显降低GVHD反应,且对外周血白细胞、生化无不良影响。结论：经7．5 Gy~(60)Co照射的MHC半相合脾加骨髓细胞移植能对H22肝癌细胞产生移植物抗肿瘤效应并降低GVHD反应。     

Immunomodulation following chemotherapy

Summary In the last decade, immunomodulation has emerged as a mode of therapy capable of mediating the regression of cancer in some patients. This article reviews our experience with immunomodulation following transplant and non-transplant chemotherapy. We used interferon and cyclosporine A following conventional chemotherapy in a non-transplant setting for a B 16 melanoma in a murine model. This combination generated cells with MHC-unrestricted cytotoxicity. We have also used immunotherapy in the transplant setting with IL-2 activated PBSC in patients with breast cancer. Of the 28 patients treated, 20 developed GVHD and the average time to reconstitution was 12 days (comparable to a control group). This article also raises the possibility of extending immunomodulation to breast cancer patients in the nontransplant setting to induce an antitumor immune response following cytoreductive chemotherapy.Presented at the symposium "New Approaches in the Therapy of Breast Cancer", Georgetown University Medical Center, Washington DC, October 1994, generously supported by an education grant from Bristol-Myers Squibb.     

肝癌肝移植受者急性移植物抗宿主病存在移植物抗肿瘤效应吗?(英文)

We aimed to access if acute graft-versus-host disease （aGVHD） in liver transplantation recipients of hepatocellular carcinoma （HCC） might develop a graft-versus-tumor effect （GVT） other than immunological damage which would benefit prophylaxis of tumor recurrence. Methods： Dynamic observation of 3 cases of liver transplantation recipients of HCC and cirrhosis, which developed manifestations of fever, skin rash, watery diarrhea, pancytopenia and were finally diagnosed as aGVHD. Two of which got recovered from intravenously pulse methylprednisolone, high-dose intravenous immunoglobulin, antibiotics administration simultaneously and promptly withdrawal of oral immunosuppressants. Two survivors were follow-up regularly with biological monitoring and imaging surveillance for tumor recurrence thereafter. Results： Two recipients survived healthily with stable liver graft function and normal serum AFP level and blood routine test. No sign of tumor recurrence was found in repeat imaging examinations for liver graft, lung, brain and other tissue or organs within a period of 96 months and 17 months to date, respectively. Conclusien： Despite of the fatal damage to according organs and tissue, it suggest that aGVHD in liver recipients of HCC may also develop a GVT effect and benefit prophylaxis of tumor recurrence and result in a long-term healthy recipients survival.     

Might liver transplantation recipients with primary hepatocellular carcinoma benefit from GVT effect of aGVHD?

Objective:We aimed to access if acute graft-versus-host disease (aGVHD) in liver transplantation recipients of hepatocel ular carcinoma (HCC) might develop a graft-versus-tumor ef ect (GVT) other than ...     

Autograft mediated adoptive immunotherapy of cancer in the context of autologous stem cell transplantation

The infused stem cell autograft in autologous stem cell transplantation (ASCT) has been viewed mainly as hematologic rescue from the myelosuppressive side effect of conditioning regimens. However, recent reports have shown that the immune effector cells collected at the same time as the stem cells can produce an autologous graft-versus-tumor effect, similar to the graft-versus-tumor effect seen in allogeneic stem cell transplantation without the detrimental effects of graft-versus-host disease. In this article, we review the different immune effector cells collected and infused from the stem cell autograft and their association with clinical outcome post-ASCT, suggesting that ASCT can be viewed not only as a therapeutic maneuver to recover bone marrow function after deliver high-dose chemotherapy, but also as an adoptive immunotherapeutic intervention capable of eradicating residual tumor cells in patients with cancer.     

Combination of anti‐CD4 antibody treatment and donor lymphocyte infusion ameliorates graft‐versus‐host disease while preserving graft‐versus‐tumor effects in murine allogeneic hematopoietic stem cell transplantation

Allogeneic hematopoietic stem cell transplantation (allo‐HSCT) is not only a well‐established immunotherapy for hematologic malignancies, but is potentially useful for treating solid tumors refractory to available therapies. However, application of allo‐HSCT to solid tumors is limited, despite the beneficial antitumor effects, by the risk of graft‐versus‐host disease (GVHD). CD4⁺ T cells have been implicated in several aspects of GVHD, and suppress antitumor CD8⁺ T‐cell responses. In the present study, we investigated clinically applicable allo‐HSCT protocols designed to maximize antitumor effects while reducing the risk of GVHD. We used a mouse model of allo‐HSCT with s.c. tumors. We found that myeloablative conditioning was associated with better inhibition of tumor growth but with severe acute GVHD. Early treatment with anti‐CD4 mAb substantially ameliorated GVHD while preserving antitumor effects, leading to improved survival in myeloablative allo‐HSCT. Late treatment with anti‐CD4 mAb also ameliorated GVHD to some extent. Donor lymphocyte infusion in GVHD mice treated with anti‐CD4 mAb further suppressed tumor growth without exacerbating GVHD. Collectively, our results suggest that myeloablative allo‐HSCT followed by anti‐CD4 mAb treatment and donor lymphocyte infusion could be a potent and safe immunotherapy for patients with cancers refractory to available therapies.     

HLA Homozygosity and Shared HLA Haplotypes in the Development of Transfusion-Associated Graft-Versus-Host Disease

We review the pathogenesis, epidemiology and patient cases of transfusion-associated graft-versus-host disease (TA-GVHD) in the context of host inability to eliminate viable donor T-lymphocytes. This review is based on the published English-language literature pertaining to TA-GVHD, including case reports with HLA data on transfusion recipients as well as blood donors. The role of shared histocompatibility antigens between the donor and the recipient in promoting TA-GVHD is discussed critically. Since TA-GVHD is usually a fatal disease for which effective therapy is lacking, prevention is of utmost importance and guidelines for gamma-irradiation of cellular blood products are presented.

TA-GVHD has been described in immunodeficient as well as in immunocompetent hosts, and following blood product transfusions from related as well as from unrelated donors. This review includes analysis of patient data in each of these settings and probability estimates based on principles of population genetics. We emphasize that transfusion of blood products from individuals who are homozygous at the HLA loci to heterozygous recipients who share that HLA haplotype occurs at a frequency proportional to the genetic homogeneity of the population and that the process mediating TA-GVHD in such instances appears to be independent of the host's immune status.     

HLA-CW基因分型在造血干细胞移植中的意义探讨

目的探讨HLA-CW位点基因分型在造血干细胞移植(hematopoicetic stem cell transplantation-HSCT)中的作用,对HLA-CW基因位点相合与不相合造血干细胞移植的效果进行分析.方法采用聚合酶链反应-序列特异性引物扩增(polymerase chain reaction-sequence specific primers,PCR-SSP)方法对HSCT供、受体共42个样本进行HLA-CW位点等位基因分析.结果HLA-CW基因全相合者9例造血重建,存活200 d以上7例,1例存活303d复发,2例存活44 d(死于感染),无病生存率(66.7±3.4)%.1例(11.1%)发生急性移植物抗宿主病(aGVHD),3例(33.3%)发生慢性移植物抗宿主病(cGVHD).HLA-CW基因不相合者12例造血重建,存活200 d以上6例,1例存活105 d复发,6例存活75 d死亡,无病生存率(41.7±5.4)%(P＜0.05).6例(50%)发生aGVHD;5例(41.7%)发生cGVHD(P＞0.05).结论HLA-CW基因不相合导致的aGVHD及相关死亡率明显高于CW基因相合者,显示CW基因不相合也是造成aGVHD发生率增高和影响移植效果的重要因素.     

不同淋巴细胞输注对非清髓小鼠异基因骨髓干细胞移植的影响

目的:建立小鼠非清髓异基因骨髓干细胞移植模型,探讨不同的供者淋巴细胞输注(donor lympho-cytes infusion DLI)对移植效果的影响及其机制。方法:受者鼠为Balb/c小鼠,供者鼠为C57b l/6小鼠,依据不同DLI将受者鼠随机分为A、B、C、D和E 5组,每组5只。5组均给予预处理,A组预处理后于移植d1给予供者小鼠全脾细胞1.0×107个;B组分别于移植后d1、7、14给予供者小鼠全脾细胞0.3×107个;C组于移植后d1、7给予去除CD8 细胞供者鼠脾细胞0.5×107个;D组于移植后d1、7给予供者鼠全脾细胞0.5×107个,E组为对照组,只作预处理,不作干细胞移植及DLI处理。观察各组嵌合状态、造血恢复情况、移植物抗宿主反应(graft versus host d isease GVHD)。结果:未移植组嵌合率为22.33%,为非特异性粘着,移植组小鼠嵌合率均在80%以上,与未移植组相比差别显著;除C组一只小鼠死亡外其余均存活,并于移植后20天自行恢复造血功能,但C组白细胞计数小于2.0×109/L持续到d16,恢复明显比其余组慢,GVHD评分A、B组间无明显统计学差异(P=0.173),C、D组间相比有统计学差异(P=0.033)。结论:建立非清髓小鼠异基因骨髓干细胞移植模型条件适宜,且去除CD8 细胞的DLI移植后GVHD比未去除组要严重,移植效果相对较差。