骨髓移植诱导特异性免疫耐受同种皮肤移植的动物实验研究

目的　证实通过动物实验模型的骨髓移植可以诱导同种皮肤移植的免疫耐受。方法　将 114只日本白色家兔和Dutch家兔分为对照组和实验组 ,日本白色家兔作为供体 ,Dutch家兔作为受体。对照组 ,在不使用免疫抑制剂的情况下 ,将 12只日本白色家兔与 12只Dutch家兔行相同面积的背部全厚皮肤互换移植 ,观察其成活时间。实验组 ,将 4 5只日本白色家兔和 4 5只Dutch家兔行全厚皮肤移植的同时行骨髓移植 ,然后将作为受体的Dutch家兔分为A ,B ,C ,D四组 ,分别行非致死量的γ射线全身照射的骨髓细胞移植及同种皮肤移植 ,观察移植皮肤的成活时间。结果　对照组 ,供体与受体移植皮肤的平均成活时间分别为 (12 .0± 1.7)天和 (10 .3± 1.3)天。实验组 ,A ,B ,C ,D四组移植皮肤的平均成活时间分别为 (6 1.0± 7.2 )、(80 .7± 10 .4 )、(78.8± 12 .7)、(88.0± 6 .0 )天。结论　通过骨髓移植导特异性免疫耐受同种皮肤移植的动物实验 ,旨在为临床应用提供了理论基础及可靠依据 ,为同种组织重建提供一个新方法     

骨髓移植诱导免疫耐受家兔全耳同种移植

目的为了减轻免疫诱导过程中对受体的侵袭,进一步探讨以家兔为动物模型,通过异体骨髓移植诱导同种皮肤移植免疫耐受.方法通过骨髓腔内直接注射异体骨髓细胞,进行骨髓移植.结果皮片的平均存活时间是(88.0±6.0)天(P<0.001).为了探讨其普遍性,我们用此诱导方法,还进行了同种全耳移植.平均存活时间是(146.0±15.1)天,并有1只移植耳存活时间已超过1年,未见任何排斥反应.结论本研究证明了以家兔为动物模型,在不使用免疫抑制剂的情况下,通过骨髓腔内直接注射异体骨髓细胞,进行骨髓移植的方法,不仅在皮肤移植上可以诱导长期稳定的特异性免疫耐受,而且在实质性器官耳的同种移植上取得了良好的效果.     

骨髓移植诱导免疫耐受家兔全耳同种移植

目的　为了减轻免疫诱导过程中对受体的侵袭 ,进一步探讨以家兔为动物模型 ,通过异体骨髓移植诱导同种皮肤移植免疫耐受。方法　通过骨髓腔内直接注射异体骨髓细胞 ,进行骨髓移植。结果　皮片的平均存活时间是 (88.0± 6 .0 )天 (P <0 .0 0 1 )。为了探讨其普遍性 ,我们用此诱导方法 ,还进行了同种全耳移植。平均存活时间是 (1 4 6 .0± 1 5 .1 )天 ,并有 1只移植耳存活时间已超过 1年 ,未见任何排斥反应。结论　本研究证明了以家兔为动物模型 ,在不使用免疫抑制剂的情况下 ,通过骨髓腔内直接注射异体骨髓细胞 ,进行骨髓移植的方法 ,不仅在皮肤移植上可以诱导长期稳定的特异性免疫耐受 ,而且在实质性器官耳的同种移植上取得了良好的效果     

供者基因转染受者细胞诱导特异性免疫耐受的实验研究

目的　探讨供体特异性基因片段MHCClassI类抗原分子RT1.AacDNA在诱导免疫耐受中的作用和可能机制。方法　采用大鼠同种异体心脏异位移植模型,通过供体MHCClassI类抗原的RT1.AacDNA基因片段转染受体成肌细胞（MB）并接种自体胸腺,观察移植物存活时间,判断受体免疫耐受产生和维持的状态。结果　经胸腺接种转染供体基因的自体成肌细胞并同时服用CsA,移植物平均存活时间高达(96.13±12.91)d,明显高于其它实验组（P＜0.05）;动态混合淋巴细胞反应（MLR）,无论外周输注或胸腺接种其对照组cpm值均高于各自实验组;CD4     

供体脾细胞胸腺注射对大鼠肢体移植存活的研究

目的 :通过大鼠肢体移植模型 ,旨在分析供体脾细胞注射对大鼠肢体移植中免疫耐受的诱导作用。方法 :选择雄性Wistar和SD大鼠为供、受体 ,对照组为胸腺注射脾细胞培养液 ,实验组为供体脾细胞注射 ,进行了 1 6例异体肢体移植动物实验。观察大鼠移植肢体排斥反应时间及存活时间。结果 :对照组肢体平均存活时间为 ( 9.38± 1 .92 )d ;实验组移植肢体存活时间为 ( 1 5.38± 2 .97)d。结论 :供体脾细胞胸腺注射大鼠肢体移植术后能够明显延长移植肢体的存活时间。     

转染细胞毒性T淋巴细胞相关抗原4免疫球蛋白基因的同供体大鼠树突状细胞对胰岛移植物存活的影响

目的　探讨转染细胞毒性T淋巴细胞相关抗原4(转染CTLA4Ig基因)的同供体大鼠DC细胞对同种大鼠胰岛移植的影响。方法　链尿菌素(STZ) 60mg/kg体重腹腔内注射制作SD大鼠糖尿病模型,胶原酶法分离、Ficoll 40 0密度梯度离心法纯化胰岛,GM CSF +IL 4诱生培育的方法,获得高纯度的DC ,含目的基因CTLA4Ig重组腺病毒AdvCTLA4Ig ,转染同供体DC细胞,与胰岛细胞同时移植于糖尿病受体大鼠肾包膜下,免疫组织化学、Dot ELISA、逆转录 聚合酶链反应(RT PCR)检测CTLA4Ig基因在实验组和对照组DC细胞中的表达,并检测受体大鼠的血糖浓度变化,同时观察受体存活情况。结果　实验组DC细胞有CTLA4Ig表达,而对照组DC细胞没有CTLA4Ig表达,实验组正常血糖维持时间(17.3±2 .4)d较对照组(10 .1±1.5 )d、空白对照组(8.3±1.2 )d显著延长,实验组、对照组和空白对照组受体大鼠存活天数分别为(3 4.5±3 .4)d、(14 .7±2 .3 )d和(11.2±1.4)d ,实验组高于对照组(P <0 .0 1)。结论　表达CTLA4Ig基因的DC细胞可能诱异胰岛移植免疫耐受,延长胰岛移植物的存活时间     

胸腺注射供体脾细胞诱导移植心脏免疫耐受的实验研究

目的探讨诱导心脏移植免疫耐受的方法及其产生的可能机制. 方法采用大鼠腹部心脏移植模型,随机分成未处理(Ⅰ)组,胸腺注射供体脾细胞(Ⅱ)组,腹腔注射兔抗鼠淋巴细胞血清(Ⅲ)组,胸腺注射供体脾细胞联合应用兔抗鼠淋巴细胞血清(Ⅳ)组,每组6只大鼠.Ⅱ、Ⅳ组在移植前2 1 d将供体脾细胞2.5×107个注射到受体胸腺,Ⅲ、Ⅳ组受体腹腔注射兔抗鼠淋巴细胞血清(ALS)1 ml,然后行异位心脏移植.观察移植心脏存活时间,供心病理学改变及供、受体间的混合淋巴细胞反应(MLR). 结果Ⅳ组供心平均存活时间(MST)为(81.8±7.6)d,较Ⅰ组(7.3±1.0)d、Ⅱ组( 7.8±1.0)d、Ⅲ组(8.2±1.2)d显著延长,差异有显著性意义(P＜ 0.01 );供心仅见少量炎性细胞浸润;供、受体间MLR较正常对照组显著降低,差异有显著性意义(P＜0.01). 结论胸腺注射供体脾细胞联合应用ALS能成功诱导心脏移植的免疫耐受;胸腺内特异性T细胞克隆消除可能与免疫耐受的形成有关.     

胸腺修饰诱导大鼠心脏移植耐受与白细胞介素-2,10的关系

目的在手术当天进行胸腺修饰,诱导大鼠同种心脏移植免疫耐受,并对其可能机制作初步分析.方法通过胸腺注射和围手术期短程使用FK506来诱导心脏移植耐受,观察供心存活天数、混合淋巴细胞反应及受体鼠血清中白细胞介素(IL)-2、IL-10水平的变化.结果无处理组、对照组、经典诱导组和实验组供心存活时间分别为(6.8±1.9)、(17.4±5.1)、(73.8±8.6)、(55.0±24.7)d,实验组与经典诱导组比较差异无统计学意义(P＞0.05).无处理组、经典诱导组和实验组的供受体脾细胞混合淋巴细胞培养刺激效应分别为198.72%、95.80%、67.94%,实验组和经典诱导组较无处理组增殖反应均明显降低(P＜0.05),而两者间增殖反应差异无统计学意义(P＞0.05).IL-10水平在无处理组移植心脏被排斥时为(48.10±5.14)ng/L较移植前(52.60±10.14)ng/L差异无统计学意义(P＞0.05);而在实验组早期呈低水平表达,为(36.10±2.30)ng/L,术后中期(281.80±65.44)ng/L晚期(80.90±12.39)ng/L较移植前水平高得多,差异有统计学意义(P＜0.05).受体IL-2水平在无处理组发生排斥时为(159.80±59.19)ng/L较移植前(54.80±8.42)ng/L明显升高,差异有统计学意义(P＜0.05).结论心脏移植手术当日胸腺内注射供体同种抗原,与术前21 d胸腺注射的经典诱导组同样能诱导宿主对移植物的低反应状态;IL-2的水平与排斥反应的发生有关,而IL-10可能是免疫耐受的特异性指标,IL-10更可能与免疫耐受的维持有关.     

FK506预处理下骨髓移植诱导同种异体小鼠皮肤移植耐受的实验研究

目的探讨短期大剂量FK506作为宏嵌合诱导供体特异性耐受中,骨髓移植前对受体预处理方法的可行性及临床实用性. 方法 100只雄性C57BL/6和60只雌性BALB/C小鼠分别作为皮肤移植的供体和受体,雄性ICR小鼠15只作为无关第三品系用以检测移植耐受状态的特异性.将60只受体小鼠随机分为5组,即无处理对照组、单纯FK506组、单纯骨髓细胞移植(BMT)组、实验组(FK506 BMT)和无关供体对照组,每组12只.FK506诱导及维持方案是皮肤移植前对受体小鼠先给予大剂量FK506腹腔注射(3 mg/kg×2 d),移植当天尾静脉输注2×107个骨髓细胞,再以小剂量FK506(0.5 mg/kg×7 d)短期维持治疗.观察皮肤移植存活时间、对第三方皮肤的排斥反应及对供体鼠的单向混合淋巴细胞反应,并用多聚酶链式反应(PCR)检测嵌合体的形成. 结果常规剂量的骨髓输注或短期FK506治疗并不能延长移植物的存活时间,也没有宏嵌合形成.实验组皮肤移植物存活时间(24.0±1.5)d比无处理对照组(9.6±1.1)d、单纯FK506组(10.5±1.6)d、单纯骨髓细胞移植组(10.3±1.5)d、无关供体对照组(9.8±1.1)d明显延长(P<0.05).混合淋巴细胞反应实验组供者特异性抑制率80.55%±14.10%明显高于单纯FK506组38.65%±12.43%及单纯骨髓细胞移植组35.41%±8.99%(P<0.05),实验组宏嵌合呈阳性. 结论采用短期大剂量FK506这一温和的非照射预处理方法,可获得一定程度的免疫耐受,延长移植物存活.移植前输注供体骨髓细胞能够促进宏嵌合的形成及移植物的存活.     

大鼠T细胞疫苗的制备及其抗移植皮肤排斥作用的实验研究

目的 探讨T细胞疫苗(TCV)的制备方法及其抗移植皮肤排斥反应的作用.方法 制备针对特定SD大鼠的供体特异性T细胞疫苗,将其免疫受体Wistar大鼠;然后取免疫前和每次免疫后第五天的受体Wistar的淋巴细胞(反应细胞)与供体SD的淋巴细胞(刺激)进行体外单向混合淋巴细胞反应(MLR),以MTT法检测细胞免疫增值反应情况,比较疫苗接种后诱导淋巴细胞反应受抑制的情况:再将SD大鼠皮肤移植到TCV免疫后的Wistar大鼠,观察皮肤移植反应并统计移植物存活的时间.排斥反应的移植物行病理检查.结果 TCV组受体淋巴细胞反应程度比接种前显著减弱(P＜0.05);特异性TCV组皮肤移植物存活时间较非特异性TCV组及对照组延长(P＜0.05).移植皮肤排斥反应病理表现更轻.结论 TCV经腹腔接种可以诱导出针对同种抗原特异性免疫耐受,TCV能够延长同种异体移植皮肤的存活时间,有一定抗移植排斥反应作用.     

Facial restoration by transplantation

Hundred years ago, Sir Harold Gillies laid a foundation to the modern plastic surgery trying to reconstruct facial defects of severely disfigured soldiers of World War I. Some years later, Joseph Murray experimented with rejection of skin grafts aimed for treatment of burned patients who sustained their injuries on battlefields of World War II. In 1954, the acquired expertise and intensive research allowed him to perform the first successful kidney transplantation in the world at Peter Bent Brigham Hospital in Boston. For his achievements in organ transplantation he was awarded Nobel Prize in 1990. The face transplantation appears to be a natural evolution of the work of these two extraordinary plastic surgeons. The first case of partial face transplant from 2005 in France revealed the world that facial restoration by transplantation is superior to conventional reconstruction methods. Since 2009, our team has performed 7 cases of face transplantation at Brigham and Women's Hospital, which is to our best knowledge the largest living single center face transplant cohort in the world. In this article, we want to reflect on the experience with face transplantation at our institution from the past years. We aim to briefly review the key points of the know-how which was given to us from the care of these unique patients.     

Angiogenesis by endothelial cell transplantation.

PURPOSE: Myocardial angiogenesis may improve regional perfusion and perhaps function after cardiac injury. We evaluated the effect of endothelial cell transplantation into a myocardial scar on angiogenesis and ventricular function, as an alternative to angiogenic gene or protein therapy.Methods and Results: A transmural myocardial scar was created in the left ventricular free wall of rat hearts by cryoinjury. Allogeneic aortic endothelial cells were injected into the scar 2 weeks after cryoinjury. A cluster of transplanted cells was identified at the site of injection 1 day and 1 week after transplantation, but not after 2 weeks. The size of this cluster of transplanted cells decreased as vascular density in the transplanted scar tissue increased with time. Six weeks after transplantation, vascular density was significantly greater in transplanted hearts than in control hearts. Regional blood flow, by microsphere analysis, was greater in the transplanted rats. Systolic and diastolic ventricular function was similar between groups. In a second series of experiments, syngeneic aortic endothelial cells labeled with bromodeoxyuridine were transplanted 2 weeks after cryoinjury. Vascular density in the transplanted scar was greater than in controls. Labeled transplanted endothelial cells were identified forming part of the newly developed blood vessels. No difference in vascular density was found between allogeneic and syngeneic cell transplantation. Vascular endothelial growth factor was not expressed at greater levels in the transplanted cells or the myocardial scar. CONCLUSION: Transplanted endothelial cells stimulated angiogenesis, were incorporated into the new vessels, and increased regional perfusion in myocardial scar tissue, but did not improve global function in this cryoinjury rat model.     

Transmission of tuberculosis by kidney transplantation

Tuberculosis occurred in two patients, each of whom received a kidney from the same cadaver donor whose cerebrospinal fluid cultures grew Mycobacteria following organ donation. Although the degree of immunosuppression and graft function were similar in the recipients, one died of disseminated tuberculosis. Kidneys contaminated with certain pathogens, including Mycobacteria, should not be transplanted. Transplant recipients with tuberculosis require prompt antituberculous therapy, and may require transplant nephrectomy for persistent evidence of urinary tract tuberculosis.     

Cost–utility analysis of living-donor kidney transplantation followed by pancreas transplantation versus simultaneous pancreas–kidney transplantation

For a type I diabetic with end-stage renal disease, the choice between a kidney-alone transplant from a living-donor (KA–LD) and a simultaneous pancreas–kidney (SPK) transplant remains a difficult one. The prevailing practice seems to favor KA–LD over SPK, presumably due to the superior long-term renal graft survival in KA–LD and the elimination of the lengthy waiting time on the cadaver transplant list. In this study, two treatment options, KA–LD followed by pancreas-after-kidney (PAK) and SPK transplant, are compared using a cost–utility decision analysis model. The decision tree consisted of a choice between KA–LD+PAK and SPK. The analysis was based on a 5-yr model and the measures of outcome used in the model were cost, utility and cost–utility. The expected 5-yr cost was $277 638 for KA–LD+PAK and $288 466 for SPK. When adjusted for utilities, KA–LD+PAK at a cost of $153 911 was less cost-effective than SPK at a cost of $110 828 per quality-adjusted year. One-way sensitivity analyses were performed by varying patient and graft survival probabilities, utilities and cost. SPK remained the optimal strategy over KA–LD+PAK across all variations. Two-way sensitivity analysis showed that in order for KA–LD+PAK to be at least as cost-effective as SPK, 5-yr pancreas and patient survival rates following PAK would need to surpass 86 and 80%. In conclusion, according to the 5-yr cost–utility model presented in this study, KA–LD followed by PAK is less cost-effective than SPK as a treatment strategy for a type I diabetic with end-stage renal disease. For patients interested in the benefits of a pancreas transplant, it would be reasonable to offer SPK as the optimal treatment, even if a living kidney donor is available.