异基因免疫治疗促进移植物抗肿瘤效应的探讨

 造血干细胞移植后细胞免疫缺陷是疾病复发的一个重要原因,同时异基因造血干细胞移植也可以认为是一种过继性免疫治疗,能克服受者细胞免疫缺陷和促进移植物抗肿瘤（GVT）效应,清除微小残留病和预防疾病复发。如何提高GVT效应仍需要进一步明确,重点介绍2011年美国血液学会（ASH）年会上相关的新进展。     

胃肠道移植物抗宿主病治疗的进展

0 引言 利用异基因造血干细胞移植( allogeneichematopoietic stem cell-transplantation,allo-HSCT)中的移植物抗肿瘤反应( graft-versus-tumor effect,GVT)治疗恶性血液病和实体瘤如肾癌、结直肠癌、胃癌等取得了良好的效果.但伴随其严重的并发症-移植物抗宿主病( graft-versus-host disease,GVHD)的发病率和死亡率仍非常高,阻碍了alloHSCT发展,影响了allo-HSCT治疗效果.GVHD 的靶器官主要有皮肤、肝脏和胃肠道等,由于胃肠道移植物抗宿主病( gastrointestinal GVHD,GI GVHD)发生率高,临床症状重,对患者全身影响大,是增加患者痛苦的重要因素之一,故针对GI GVHD的治疗就显得尤为重要.本文根据国内外GI GVHD的最新进展作一综述,希望对GI GVHD的治疗提供一些新的参考.     

减轻异基因造血干细胞移植GVHD增强GVT的研究

异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation,allo-HSCT)是一种有效治疗恶性肿瘤及非恶性疾病的方法,近年来成为治愈某些恶性血液性肿瘤疾病的主要手段之一,明显提高了患者的长期生存率[1];但移植物抗宿主病(graft-versus-host disease,GVHD)仍然是allo-HSCT的主要并发症,是移植相关死亡及影响患者生存质量的重要原因。GVHD是由移植物中所含供者T细胞识别宿主抗原而攻击宿主组织器官所产生的病理损伤,并导致宿主多脏器损伤的一个疾病过程。以T细胞为靶点的免疫抑制剂及T细胞去除术的应用是目前GVHD…     

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.     

急性移植物抗宿主病的诊疗进展

移植物抗宿主病（GVHD）是异基因造血干细胞移植最常见的并发症,分为急性（aGVHD）和慢性（cGVHD）2种类型,其中aGVHD是供者T细胞识别不匹配的宿主多态性组织相容性抗原（MHC）而发生的免疫应答,具有独特发病机制,病理改变及临床表现复杂,可累及多个器官.文章就目前国内外对aGVHD的诊疗进展进行综述.     

协同刺激因子在移植物抗宿主病发生中作用的初步研究

 目的　探讨协同刺激因子（CM）在异基因造血干细胞移植（allo-HSCT）相关移植物抗宿主病（GVHD）中的作用。方法　根据预处理方案不同,将21例行allo-HSCT的血液病和实体瘤患者分为A组（NST组）和B组（清髓性HSCT组）;所有患者在移植前后不同时间,应用流式细胞术（FCM）检测外周血CD+4 T细胞表面CM（CD28、CD80、CD152）的表达;用短串联重复序列聚合酶链反应（STR-PCR）方法检测微卫星嵌合体形成。结果　21例患者移植后均获得造血功能重建,经STR-PCR检测均转为混合嵌合体（MC）或完全嵌合体（CC）;不同预处理方案GVHD发生率差异无统计学意义（χ2＝3.711,P＝0.144）;COX回归分析结果显示：CD+4 CD+152能明显影响GVHD发生（χ2＝13.128,P＜0.0001）;术后,外周血T细胞表面CD+4 CD+28、CD+4 CD+80表达逐渐增高,GVHD时达高峰,经过治疗和控制GVHD后,逐渐下降;而T细胞表面CD+4 CD+152表达则逐渐降低,GVHD时最低,经治疗和控制GVHD后,又逐渐上升。结论　清髓性HSCT和NST的GVHD发生率无差别;T细胞表面CM的表达与GVHD发生有关,并且在清髓性HSCT和NST的表达不同;B7-CD28／CD152共刺激途径在GVHD发生中起重要作用。     

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.     

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。     

IL-4及IL-18水平变化与急性移植物抗宿主病的系

目的 探讨异基因造血干细胞移植(allo- HSCT)前后受者血清IL-4及IL-18的变化及与急性移植物抗宿主病(aGVHD)的关系,为临床早期诊断aGVHD提供可靠细胞因子.方法 20例血液病患者行allo-HSCT,于移植前后不同时间点采集外周血,采用ELISA法检测其血清IL-4及IL -18水平(pg/ml).16例自体造血干细胞移植(auto-HSCT)患者作为对照组.结果 实验组和对照组在预处理及干细胞回输前后血清IL-4及IL -18水平变化差异无统计学意义(P＞0.05).实验组有6例患者发生aGVHD,其中Ⅰ～Ⅱ度3例,Ⅲ～Ⅳ度3例,血清IL-4及IL -18水平变化早于临床症状出现.aGVHD诊断时血清IL-4及IL -18水平分别显著低于和高于移植后7天(P＜0.05),抗aGVHD治疗后其水平又明显升高和下降(P＜0.05).结论 血清IL-4及IL-18水平和aGVHD的发生存在相关性,其水平检测有助于aGVHD的预测和早期诊断.     

移植物抗宿主病的治疗及移植相关肝损伤的研究进展

异基因造血干细胞移植使治愈血液系统恶性疾病成为可能.但在移植后出现的移植物抗宿主病及重要脏器损伤越来越成为移植后的主要并发症及非复发死亡的重要原因.目前移植物抗宿主病的治疗以免疫抑制为主,但部分患者疗效欠佳,并可能减弱移植物抗白血病效应、增加感染机会.对于如何预防及早期识别移植物抗宿主病,不同药物方案对比以及对于肝脏相...     

Clinical Application of Serum Tumor Abnormal Protein (TAP) in Colorectal Cancer Patients

Objective: To explore the association of serum tumor abnormal protein (TAP) with other serologicalbiomarkers e.g. carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), carbohydrate antigen19-9 (CA19-9) and its clinical application in colorectal cancer (CRC) patients. Methods: Patients (N=98) wereenrolled into this study with histologically or cytologically confirmed CRC. Using a test kit, the level of TAPwas determined, while chemiluminescence was used to measure the levels of some other common serologicalbiomarkers e.g. CEA, CA125 and CA19-9. Results: The area of TAP condensed particulate matter decreased afterchemotherapy compared with before chemotherapy when CT or MRI scans showed disease control. In contrast,it increased with disease progression (P<0.05). Furthermore, a statistically significant difference was confirmed inmonitoring of TAP and common serological biomarkers e.g. CEA and CA19-9 (p<0.05). Conclusions: DetectingTAP in CRC patients has high sensitivity and specificity and can be used as a new independent indicator forclinically monitoring CRC patients in the course of chemotherapy.     

新辅助化疗、术中动脉灌注化疗对结直肠癌患者VEGF的影响

目的 探讨新辅助化疗、术中动脉灌注化疗对结直肠癌患者VEGF的影响及临床意义.方法 收集井冈山大学附属医院、南昌大学第二附属医院结直肠癌病例117例,随机分为新辅助化疗组61例、术中动脉灌注化疗组56例,健康人群36例作为空白对照组.比较各组治疗前、手术1周后VEGF水平差异.结果 健康人群组VEGF浓度明显低于结直肠癌组(P＜0.05).新辅助化疗前后VEGF无明显差别(P＞0.05),术中动脉灌注化疗后VEGF水平明显低于术前(P＜0.05).结论 新辅助化疗不能显著降低VEGF水平,术中动脉灌注化疗能显著降低VEGF水平.术中动脉灌注化疗有望降低结直肠癌术后转移、复发率,是安全有效,简单易行的.     

结直肠癌患者组织与粪便中K—ras癌基因12位点突变的分析

目的：探讨直肠癌（CRC）患者的组织与粪便中K-ras癌基因12位点突变检测结果的联系及检出率差异,为建立结直肠癌早期筛检方法提供科学依据。方法：应用聚合酶链反应－限制性片段长度多态性分析（PCR－RFLP）方法对45例原发性结直肠癌患者手术切除的组织及术前粪便标本中K-ras癌基因12位点的突变情况进行检测,结果采用卡方检验及卡伯值（Kappa value)进行统计分析。结果：癌组织、远端手术切缘粘膜、粪便标本中的K-ras癌基因12位点突变检出率分别为35．56％（16／45）、4．44％（2／45）、37．78％（17／45）。癌组织与粪便标本检测结果的关联检验为：X^2=33．09,P＜0．01;差别检验为：X^2=0,P＞0．05;观察一致率为93．33％,卡柏值为0．86。结论：粪便中K-ras癌基因12位点突变检测结果与癌组织的检测结果存在联系,其检出率无显著性差异,可靠性极好;且粪便取材方便,无创伤性,可能作为结直肠癌高危人群的筛检方法。     

Parallel Evaluation of Circulating Tumor DNA and Circulating Tumor Cells in Metastatic Colorectal Cancer

Background

Tissue biopsy is the gold standard for tumor genotyping, but it is an invasive procedure providing a single snapshot into tumor heterogeneity. Liquid biopsy approaches, encompassing the analysis of circulating tumor DNA (ctDNA) or circulating tumor cells (CTCs), have been proposed as an alternative, with the potential of providing a comprehensive portrait of the tumor molecular landscape. In metastatic colorectal cancer (mCRC), both CTCs and ctDNA analysis have been investigated, but comparative analyses are limited.

Clinical Trial of Continuous Infusion of 5-Fluorouracil Using an Ambulatory Pump for Metastatic Colorectal Cancer

A clinical trial was conducted in order to evaluate the anti-tumoreffect and toxicity of a continuous ifusion of 5-fluorouracil(5-FU) for metastati colorectal cancer. Two-hundred and fiftymg/m²/day 5-FU was administered as a continuous infusion throughan indwelling central venous catheter with ambulatory pump.Twenty patients with metastatic colorectal cancer which couldbe measured or evaluated were enrolled in the trial. The objectiveresponse rate was 35% (95% confidence interval, 14–56%).The response rates by site were 33% in liver, 17% in lung, 60%in lymph nodes, 50% in adrenal gland and 50% in primary lesion.The major toxicity was stomatitis (50%; grades 2 and 3 on EasternCooperative Oncology Group (ECOG) criteria) and hand-foot syndrome(40%; grades 2 and 3 on ECOG criteria). The sequence of toxicitywas stomatitis first, followed by hand-foot syndrome. The mediancumulative dose of 5-FU from the initiation of therapy to theonset of toxicity was 7125 mg in stomatitis and 17,875 mg inhand-foot syndrome. These toxicities were mild and reversibleafter a short interruption to the 5-FU infusion. Neither hematologicaltoxicity nor serious catheter-related complications were observed.We concluded that continuous infusion of 5-FU was a feasibletreatment for the patient with metastatic colorectal cancer,and manageable on an out-patient basis     

血清多种肿瘤标志物联合检测对结直肠癌的诊断价值

目的：探讨血清多种肿瘤标志物联合检测对结直肠癌的诊断价值。方法选择82例结直肠癌患者作为结直肠癌组,70例结直肠良性病变患者作为良性病变组,以及70例同期于我院进行健康体检的患者作为对照组。采用电化学发光免疫分析法对3组血清样本进行CEA、CA199、CA125、CA242及CA50等肿瘤标志物检测。结果结直肠癌组各项肿瘤标志物表达水平显著高于良性病变组及对照组,差异具有统计学意义（P＜0．05）。随着结直肠癌分期的进展及淋巴结转移,各项肿瘤标志物表达水平也逐渐升高,差异具有统计学意义（ P＜0．05）。此外,采用5种肿瘤标志物联合检测与肿瘤标志物单项检测比较,具有更高的敏感度及准确性（ P＜0．05）。结论血清多种肿瘤标志物联合检测对于结直肠疾病的鉴别诊断以及结直肠癌TNM分期判断都具有积极的意义,可作为结直肠癌早期诊断的重要辅助手段。     

载脂蛋白E对结直肠癌细胞侵袭迁移能力的影响

目的 观察载脂蛋白E（apolipoprotein E, ApoE）对结直肠癌细胞侵袭迁移能力的影响。方法 在结直肠癌细胞系SW48中转染ApoE过表达质粒及其特异性siRNA,实时荧光定量PCR（Real timeqPCR）检测转染ApoE的表达,利用Transwell小室观察ApoE对细胞侵袭迁移能力的影响,通过免疫印迹检测ApoE对基质金属蛋白酶（matrix metalloproteinase, MMP）家族蛋白表达的影响。结果 RTPCR检测结果显示：过表达ApoE组细胞中ApoE mRNA水平显著高于空白对照白组和阴性对照组,差异有统计学意义（P=0.008）,ApoE沉默组中ApoE mRNA水平与阴性对照组比,差异有统计学意义（P=0.013）;Transwell侵袭实验结果显示,穿膜细胞数：空白对照组、阴性对照组、过表达ApoE组、si-ApoE组依次为：（209±17）、（228±11）、（67±9）、（428±15）个/视野,过表达ApoE组和si-ApoE组与阴性对照组相比,差异有统计学意义（P<0.05）;Transwell迁移实验结果显示,空白对照组（326±18）个/视野,阴性对照组（338±21）个/视野,过表达ApoE组（133±11）个/视野,与阴性对照组相比,差异有统计学意义（P<0.05）,si-ApoE组（518±13）个/视野与阴性对照组相比,差异有统计学意义（P<0.05）。同时免疫印迹实验显示,过表达ApoE组细胞中MMP-2及MMP-9的表达下降,组织基质金属蛋白酶抑制剂2（tissue inhibitors of metalloproteinase-2, TIMP-2）的表达升高,而si-ApoE组MMP-2及MMP-9表达升高,TIMP-2的表达降低。结论 ApoE可影响结直肠癌细胞的侵袭、迁移能力,并且可能通过影响TIMP-2、MMP-2及MMP-9的表达发挥上述功能。     

miR-455 Functions as a Tumor Suppressor Through Targeting GATA6 in Colorectal Cancer

Emerging evidence indicates that microRNAs (miRNAs) are often aberrantly expressed in human cancers. Meanwhile, the importance of miRNAs in regulating multiple cellular biological processes has been appreciated. The aim of this study was to investigate the significance of miR-455 and identify its possible mechanism in regulating colorectal cancer (CRC) progression. We found that the expression of miR-455 was sharply reduced in CRC tissues and cell lines. Importantly, the low expression of miR-455 was associated with poor overall survival of CRC patients. Overexpression of miR-455 in CRC cell lines significantly inhibited cell proliferation and migration in vitro. Moreover, GATA-binding protein 6 (GATA6), whose expression can be inversely regulated by miR-455 in CRC cell lines, was validated as a direct target of miR-455. Overall, our results revealed that miR-455 functions as a tumor suppressor, and its downregulation may contribute to CRC progression. Our study may provide a novel therapeutic target for CRC in the future.