免疫调节细胞的临床应用研究进展

1．1 器官移植面临的难题 器官移植是近半个世纪以来医学发展的前沿。对于心、肝、肺等脏器功能衰竭者，移植是目前惟一有效的治疗手段。移植医学的发展面临的主要障碍：一是随着器官移植近期效果的不断提高，对移植的需求不断增长，导致供器官的严重短缺；二是经历了50多年的发展，移植的长期效果并没有得到明显的提高。所有器官移植后，10年存活率均低于50％。各种原因致受者需要二次移植，也使器官供体需求增加。据文献报道，美国等待肾移植的受者名单中有20％属再次或多次移植患者。免疫抑制剂的应用极大地提高了移植物的存活率，但是排斥反应一直是困扰器官移植的难题，移植受者长期服用免疫抑制剂所带来的毒副作用，也影响着移植物及受者的存活。     

树突状细胞:诱导移植免疫耐受的工具细胞和靶细胞

目前，树突状细胞（dendritic cells，DC）生物学研究的热点已由诱导免疫应答转向诱导免疫耐受。经典的髓细胞样DC（mDC）及最近新发现的类浆细胞样DC（pDC）亚群的致耐受作用已经明确；DC诱导耐受的特性可能与调节性T细胞（Treg）的作用密切相关；DC细胞具有独特的获取并交叉呈递外来抗原的能力。因此，在非炎性环境中DC可以诱导抗原特异性免疫耐受。本文综述了这三方面的研究现状，并探讨了DC诱导器官移植免疫耐受的应用前景。     

浅谈高致敏肾移植受者术前处理策略

由于移植水平的不断提高和新型高效免疫抑制剂的问世，肾脏移植的适应证也在不断扩大，高危肾移植受者接受移植也越来越多。所谓高危顾名思义应该是同时存在有可能危及肾移植受者生命安全或影响移植肾近期或远期存活的诸多因素。如合并糖尿病、动脉硬化、冠心病、心功能不全、传染性肝病，或二次移植、多次输血、长期血液透析及妊娠等原因，使受者致敏，有广泛的多价抗HLA抗体，移植时易发生超急性排斥反应（超排）及各种难以逆转的急性排斥反应（AR），导致移植失败，严重者甚至可能危及受者生命。笔者主要针对后者即高致敏肾移植受者的术前处理作一粗浅阐述。     

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目的 报告国内首例同种异体肝小肠联合移植，并就肝肠联合移植适应证、排斥诊断与治疗、感染防治、肠功能恢复与营养支持进行讨论。方法 供肠长380cm，供肠肠系膜上动脉、门静脉分别与受体腹主动脉、下腔静脉端侧吻合；供肝行改良背驮式肝移植。术后免疫抑制剂采用FK506 激素 骁悉 赛尼哌方案。排斥监测采用内镜指导下肠粘膜活检病理学检查及临床观察。并采取措施防治感染和促进移植肠功能的恢复。结果病人恢复顺利，移植肠和移植肝功能良好，未发生排斥反应，术后3个月时已摆脱全肠外营养，依靠肠内营养维持营养状态。结论 肝小肠联合移植对肠衰竭合并全肠外营养所致肝功能损害病人，是可行、有效的治疗方法。     

肝移植术后胆道并发症33例分析*

目的：胆道并发症是肝移植术后常见的并发症,严重影响肝移植患者的生存率和生活质量。课题拟进一步明晰认识肝移植术后胆道并发症的病因并确立早期防治措施。 方法：①对象：解放军总医院第二附属医院全军器官移植中心于2002-04/2007-09完成的254例原位肝移植患者中,术后发生胆道并发症者33例,回顾分析患者的临床资料。②方法：胆管重建方式为胆总管端端吻合术。并发胆漏者,予置管充分引流;吻合口狭窄者经皮肝穿刺胆道造影或经内镜逆行胰胆管造影球囊扩张术,必要时放置胆道支架;非吻合口狭窄者行经皮肝穿刺胆道造影联合胆道镜治疗,严重者行二次肝移植术。 结果：18例胆漏患者均通过腹腔引流、鼻胆管引流治愈;9例吻合口狭窄患者经内镜逆行胰胆管造影或经皮肝穿刺胆道造影球囊扩张术治愈,其中3例放置了胆道支架;6例非吻合口狭窄患者中,2例经皮肝穿刺胆道造影联合胆道镜治疗后痊愈,2例接受了二次肝移植,2例恢复不佳。 结论：改进手术方法、缩短移植物冷热缺血时间及保护胆管血供可降低肝移植术后胆道并发症的发生,治疗的关键在于早期诊断、合理选择治疗方法,处理得当患者可长期存活。     

肝移植106例术后急性排斥反应的诊断和治疗

目的:总结肝移植急性排斥反应的新表现及诊治经验. 方法:回顾我中心106例肝移植资料,对其中发生急性排斥反应病例的临床表现、病理组织学改变、诊治方案加以分析. 结果:106例肝移植中,术后发生急性排斥反应17例(16.0%);其中临界型1例,轻度6例,中度7例,重度3例. 这些病例在具备病理组织学改变的同时缺乏典型的临床特征和肝功能生化指标的改变. 结论:由于强效免疫抑制剂的应用,肝移植术后急性排斥反应发生的临床症状和体征不典型,我们应该重视急性排斥反应的诊断并积极治疗.     

肝癌患者肝移植术前外周血AFP mRNA的表达与肿瘤复发及患者生存的关系

目的:探讨原发性肝细胞肝癌患者在肝移植术前外周血AFP mRNA的表达及其与术后肿瘤复发、患者预后的关系.方法:以2003年2月～2004年2月收入天津市第一中心医院的31例肝细胞肝癌行原位肝移植患者为研究对象,采用实时定量RT-PCR检测患者术前外周血AFP mRNA表达水平,均采用尸体供肝经典原位肝移植.以SPSS 12.0软件分析AFP mRNA的表达与患者术后肿瘤复发及生存的相关性.结果:31例患者中有10例AFP mRNA表达阳性,表达量为57～10 400 copy/μg,中位数为360 copy/μg.患者Child-pugh分级,肿瘤的大小、个数,病理Edmond分级与术前AFP mRNA的表达水平无关.肿瘤TNM分期为Ⅲ～Ⅳ期的患者AFP mRNA的阳性表达率显著高于Ⅰ～Ⅱ期患者(P=0.001);脉管浸润的患者AFP mRNA的表达水平显著高于无脉管浸润者(P=0.029).肝移植术前AFP mRNA阴性组的无瘤生存及总体生存显著高于AFP mRNA阳性组.结论:术前AFP mRNA表达的水平是肿瘤复发的危险因素,是术前可能存在肝外微转移的指标,可作为预后的判断因子.     

老年活体供肾移植术后供者安全性及受者移植效果的分析

Objective To investigate the safety for donors and the effectiveness for recipients of living-related donor (LRD) kidney transplantation from elder donors. Methods 251 cases of LRD kidney transplantation were reviewed. According to the age of LRDs, the patients were divided into 2 groups:≥55 years group (group A) and <55 years (group B). The parameters studied included serum creatinine (Cr), glomerular filtration rate (GFR), creatinine clearance (Ccr), perioperative complications, average hospital stay, and acute rejection rate of LRDs and recipients were compared. Results (1)There was no significant difference in serum Cr between groups A and B at different time points (P>0.05). (2)There was no significant difference in Ccr between two groups pre-donation (P = 0.45). But at the 10th day after the donor nephrectomy, Ccr level in group A was significant lower than in group B (P<0.01). (3)Total GFR pre-donation, remaining renal GFR, and remaining renal GFR on the day 10 after donation had no significant difference in both groups A and B (P>0.05). Remaining renal GFR on the day 10 before and after donation had no significant difference in group A (P>0.05), but on the day 10 after donation that was significantly increased in group B as compared with that pre-donation (P<0.01). (4) The serum Cr of recipients at different time points after transplantation had no significant difference between two groups (P>0.05). (5) The mean hospital stay after donation of LRDs and recipients had no significant difference between two groups. (6) The incidence of recipients" acute rejection was 6.50 % (5/77) in group A, and 5.75%(10/174) in group B within 6 months after operation (P = 0.95). Conclusion Transplantations performed from the elderly donors will yield similar results from younger donors if the eider donors are evaluated or assessed as the standards.     

老年活体供肾移植术后供者安全性及受者移植效果的分析

Objective To investigate the safety for donors and the effectiveness for recipients of living-related donor (LRD) kidney transplantation from elder donors. Methods 251 cases of LRD kidney transplantation were reviewed. According to the age of LRDs, the patients were divided into 2 groups:≥55 years group (group A) and <55 years (group B). The parameters studied included serum creatinine (Cr), glomerular filtration rate (GFR), creatinine clearance (Ccr), perioperative complications, average hospital stay, and acute rejection rate of LRDs and recipients were compared. Results (1)There was no significant difference in serum Cr between groups A and B at different time points (P>0.05). (2)There was no significant difference in Ccr between two groups pre-donation (P = 0.45). But at the 10th day after the donor nephrectomy, Ccr level in group A was significant lower than in group B (P<0.01). (3)Total GFR pre-donation, remaining renal GFR, and remaining renal GFR on the day 10 after donation had no significant difference in both groups A and B (P>0.05). Remaining renal GFR on the day 10 before and after donation had no significant difference in group A (P>0.05), but on the day 10 after donation that was significantly increased in group B as compared with that pre-donation (P<0.01). (4) The serum Cr of recipients at different time points after transplantation had no significant difference between two groups (P>0.05). (5) The mean hospital stay after donation of LRDs and recipients had no significant difference between two groups. (6) The incidence of recipients" acute rejection was 6.50 % (5/77) in group A, and 5.75%(10/174) in group B within 6 months after operation (P = 0.95). Conclusion Transplantations performed from the elderly donors will yield similar results from younger donors if the eider donors are evaluated or assessed as the standards.     

负向调节树突状细胞对心脏移植大鼠调节性T细胞的影响

目的:探讨受者树突状细胞(DC)与体外光化学法(PUVA)处理的供者脾淋巴细胞共培养,对心脏移植受者CD4+CD25+调节性T细胞(Treg)及移植心存活时间的影响。方法:以DA大鼠为供者,Lewis大鼠为受者,SD大鼠为无关供者,建立大鼠腹部异位心脏移植模型。分离正常的供者脾淋巴细胞(SP),制备经PUVA处理的供者脾淋巴细胞(PU-VA-SP)。在体外将DA大鼠PUVA-SP或SP与受者骨髓来源的DC共同培养,收集经上述处理后受者DC,流式细胞术(FCM)检测其表面分子CD80、CD86以及OX6的表达状况。根据受者术前静脉输注的成分将实验动物随机分为3组:①Control组:单纯输注PBS,n=7;②SPDC组:输注加载供者SP的受者大鼠DC,n=8;③PUVA-SPDC组:输注加载PUVA处理的供者SP的受者大鼠DC,n=8。移植术前7d,经外周静脉给受者输注与PUVA-SP共培养后的DC(PUVA-SPDC组)、正常受者DC(DC组)或只输注PBS(Control组),移植术后观察受者移植物的存活时间,检测受者外周血中CD4+CD25+T细胞、CD4+CD25highT细胞的比例及其Foxp3表达状况。过继转移PUVA-SPDC组受者大鼠的T细胞后,检测正常LEW大鼠对供者抗原或无关抗原的DTH反应。结果:受者DC与供者未经处理的脾淋巴细胞(SP)混合培养后,其表面分子CD80、CD86以及OX6的表达分别为16.6%±0.72%、36.5%±0.87%及65.6%±1.45%,明显高于未处理DC组(3.53%±0.27%、13.0%±0.57%及27.7%±1.23%)(P0.01);而受者DC与PUVA处理过的供者脾淋巴细胞(PUVA-SP)混合培养后,其表面分子的表达仍保持较低的水平,CD80、CD86以及OX6的表达分别为3.9%±0.12%、13.4%±0.59%及28.0%±1.73%,与未经任何处理的受者DC无统计学差异(P0.05)。移植术后,PUVA-SPDC组受者,外周血CD4+CD25+T、CD4+CD25highT细胞占CD4+T的比例分别是18.97%和3.81%,明显高于输注DC组(4.40%和0.81%)和Control组(3.11%和0.09%)的大鼠(P0.01)。PUVA-SPDC组大鼠外周血CD4+CD25+T细胞中Foxp3表达率为29%±1.73%,CD4+CD25highT细胞中Foxp3阳性率高达95%±1.67%,均明显高于对照组(12%±0.58%和19.3%±2.03%)及DC(16.3%±0.88%和52.0%±1.73%)组(P0.01)。PUVA-SPDC组大鼠移植心存活时间为(27.3±0.98)d,与Control组(6.7±0.29)d及DC组(11.0±0.32)d相比,明显延长(P0.01)。过继转移实验显示,接受PUVA-SPDC组受者T细胞的正常LEW大鼠对DA大鼠抗原的刺激呈特异性免疫低应答状态。结论:PUVA-SPDC能够在移植受者体内诱生CD4+CD25+Foxp3+Treg,同时诱导抗原特异性免疫低反应,进而延长异基因移植物的存活时间。