10-羟基喜树碱对异基因大鼠心脏移植物急性排斥反应的抑制作用

目的　探讨 10 羟基喜树碱 (HCPT)对移植物急性排斥反应的免疫抑制作用。方法　以SD大鼠为供者 ,Wistar大鼠为受者 ,行大鼠同种异体异位心脏移植。用中药制剂 10 羟基喜树碱(HCPT)预防和治疗移植物急性排斥反应。结果　HCPT 1.0mg·kg-1·d-1组与对照组比较 ,移植物存活时间明显延长 (6.0 5d/ 11.45d ,P <0 .0 0 1) ;HCPT 2 .0mg·kg-1·d-1组作用更为显著 (>2 41.6d) ,其中 3只大鼠于术后 60d停用HCPT ,移植心脏长期存活 ,超过 73 0d ,并经证实已形成免疫耐受。大鼠心脏移植物发生急性排斥反应时 ,HCPT 10 .0mg·kg-1·d-1能使排斥反应完全逆转 ,移植心脏功能完全恢复正常。较大剂量HCPT可导致轻微的胃肠道反应 ,但未观察到明显的肝、肾、心、脾、睾丸和造血系统损害。结论　HCPT对大鼠移植心脏急性排斥反应具有显著的抑制作用 ,且具有剂量相关性     

羟喜树碱和环孢素A对异基因大鼠心脏移植物急性排斥反应的协同抑制作用

目的 探讨羟喜树碱（OPT）和环孢素A（CsA）对异基因心脏移植物急性排斥反应的协同抑制作用。方法 以纯系SD大鼠为供者,纯系Wistar大鼠为受者,行异体颈部异位心脏移植。40只受者大鼠心脏移植后随机分成4组。A组接受安慰剂。B组接受 OPT1.0mg·kg-1·d-1,腹腔注射。E组接受CsA　10mg·kg-1·d-1,导管灌胃。F组联合应用OPT和CsA,方法剂量同B、E组。结果A组平均存活期为（6.05±0.76）d,B组为（11.45±1.99）d,E组为（14.45±4.85）d,F组有5只大鼠平均存活期为（45.00±19.43）d,另5只大鼠在移植60d后停用OPT和 CsA,存活期超过730 d,并经试验证明形成特异性免疫耐受。结论OPT和CsA的联合应用显著降低异基因大鼠心脏移植物急性排斥反应,明显延长心脏移植存活期。     

小鼠心脏移植急性排斥反应体外模型的建立

我们在成功建立大鼠腹部、颈部及小鼠颈部心脏移植模型的基础上,建立了小鼠心脏移植急性排斥反应体外模型,现将结果报道如下。 一、材料与方法 1.实验动物:健康雄性BALB/c和C57小鼠,体重20～25 g,由华中科技大学同济医学院器官移植研究所提供。 2.心肌细胞的分离:采用离体心脏Langendorff灌注法分离小鼠心肌细胞,取BALB/c小鼠心脏,用37℃氧含的含有胰蛋白酶和胶原酶的消化液灌注,30min左右心脏变软,颜色呈粉红半透明状,取下心脏,剪取心室肌,将之剪成1     

茯苓酸抗大鼠心脏移植急性排斥反应的实验研究

目的 探讨化合物茯苓酸对心脏移植急性排斥反应的抑制作用,为从中药中筛选出具有明确免疫抑制作用和低毒副作用的免疫抑制剂打下了良好的基础. 方法 以32只Wistar大鼠为供体,32只SD大鼠为受体,建立大鼠腹腔异位心脏移植模型,移植后的大鼠随机分为4组,每组8对.茯苓酸小剂量组:茯苓酸1 mg/(kg·d)橄榄油溶液灌胃;茯苓酸大剂量组:茯苓酸10 mg/(kg·d)橄榄油溶液灌胃;环孢菌素A(CsA)组: CsA 5 mg/(kg·d)橄榄油溶液灌胃;对照组:橄榄油溶液0.5 ml/(kg·d)灌胃.灌胃至观察终点.术后观察移植心存活时间,心脏跳动情况和心肌组织学改变,用酶联免疫吸附测定法(ELISA)检测血清白细胞介素2(IL-2)和γ干扰素(IFN-γ) 的含量.结果 茯苓酸大剂量组、茯苓酸小剂量组和CsA组移植心存活时间分别为24.90±0.99 d、15.50±1.60 d和26.80±0.88 d,均较对照组(6.10±1.10 d)显著延长(q=22.363, P=0.000; q=44.793, P=0.000; q=49.272, P=0.000).茯苓酸大剂量组、茯苓酸小剂量组和CsA组血清IL-2水平均低于对照组(q=14.483, P=0.000; q=3.705, P=0.000; q=21.418, P=0.000); 茯苓酸大剂量组低于茯苓酸小剂量组(q=10.778, P=0.000).茯苓酸大剂量组、茯苓酸小剂量组和CsA组血清IFN-γ水平均低于对照组(q=16.508, P=0.000; q=4.281, P=0.000; q=19.621, P=0.000);茯苓酸大剂量组低于小剂量组(q=14.975, P=0.000).术后第7 d移植心病理检查结果显示:茯苓酸小剂量组、茯苓酸大剂量组和CsA组心肌病理损害较对照组明显减轻. 结论 茯苓酸对心脏移植急性排斥反应具有明显的抑制作用.     

补体抑制治疗与心脏移植超急性排斥反应

心脏移植超急性排斥反应(hyperacute rejection,HAR)与补体系统激活有关,迅速导致移植心脏功能丧失.动物实验中使用眼睛蛇毒因子、可溶性补体受体1、补体调节蛋白以及C5单克隆抗体等补体抑制剂,可以从补体级联反应的不同水平抑制同种或异种心脏移植HAR的发生.抑制补体激活将在临床心脏移植HAR的预防及治疗中显示出重要的作用.     

茯苓醇提取物抗心脏移植急性排斥反应的实验研究

目的　研究茯苓醇提取物对心脏移植急性排斥反应的抑制作用。方法　以Wistar大鼠为供者、SD大鼠为受者 ,建立异位 (腹腔 )心脏移植模型 ,移植前按组分别以橄榄油、茯苓醇提取物及环孢素A灌胃 ,并设SD大鼠空白对照组。术后观察移植心的存活时间 ,测定各组术后第 7d外周血中白细胞介素 2 (IL 2 )和γ干扰素 (IFN γ)含量以及CD3+、CD4 +、CD8+细胞和CD4 +细胞与CD8+细胞的比值 (CD4 +/CD8+) ,并观察移植心的病理变化。结果　接受茯苓醇提取物 2 5mg·kg-1·d-1或 5 0mg·kg-1·d-1灌胃的大鼠 ,移植心存活时间显著延长 ,病理损害程度减轻 ,外周血IL 2及IFN γ的含量以及CD3+、CD4 +、CD8+细胞百分比和CD4 +/CD8+比值降低 ,与环孢素A 5mg·kg-1·d-1灌胃的结果相当。结论　茯苓醇提取物对心脏移植急性排斥反应具有明显的抑制作用。     

塞来昔布对大鼠心脏移植急性排斥反应的抑制作用

目的观察塞来昔布对大鼠心脏移植急性排斥反应的抑制作用。方法以SD大鼠为供者,Wistar大鼠为受者,进行40次腹部异位心脏移植。采用HE染色和原位末端标记(TUN EL)技术检测移植心切片,进行排斥反应的病理分级并计算移植心肌细胞的凋亡指数(AI)。结果移植心的细胞凋亡主要发生于心肌细胞;移植后第3、5d,塞来昔布治疗组心肌细胞凋亡指数分别为:1.03±0.42和3.28±2.42;对照组分别为2.35±1.51和11.35±3.46;两组比较,差异有统计学意义(P<0.001)。结论细胞凋亡是心脏移植急性排斥中组织损伤的重要机制;塞来昔布能明显抑制心肌细胞凋亡。     

细辛脂素对心脏移植急性排斥反应的作用研究

心脏移植是治疗终末期心脏病的有效方法，而现有的免疫抑制剂具有较强的毒副作用，并且价格昂贵，所以国内外众多学者把研究开发新型免疫抑制剂的重点转向了资源丰富、价格低廉的中草药。细辛是一种比较常用的中药，具有免疫抑制作用，但目前其抗排斥作用还鲜有报道，本研究证明其具有抗排斥反应的作用，对临床有重要的理论和实际意义。     

供体特异性输注对心脏移植急性排斥反应中细胞因子表达的影响

我们采用全血输注的方法 ,观察同种大鼠心脏移植术后移植心局部细胞因子网络在基因水平的变化 ,探讨细胞因子网络参与供体特异性输注 (ＤＳＴ)的机制。一、材料与方法1.材料 :健康雄性Ｗｉｓｔａｒ及ＳＤ大白鼠 ,近交系 ,受体为Ｗｉｓｔａｒ大白鼠 ,体重80～ 10 0 ｇ ,供体为ＳＤ大白鼠 ,体重 2 5 0～ 30 0 ｇ。分为Ａ组 :对照组 ;Ｂ组 :ＤＳＴ组。每组 12例 ,其中 7例用于动态取标本 ,其余 5例用于观察移植物存活时间 ,共计 2 4例。药品 :0 .3%戊巴比妥钠 (美国Ｓｉｇｍａ公司 ) ,ＤＳＴ组所用全血按无菌原则在移植 7ｄ前从供体舌下静脉抽…     

蛋白质谱技术预测大鼠心脏移植后急性排斥反应

目的 探讨蛋白质谱技术对大鼠心脏移植后急性排斥反应的预测.方法 建立颈部异位异种大鼠心脏移植模型.(1)急性排斥A和B组,供受体不作预处理.(2)治疗A和B组,受体术前1d腹腔注射环孢素A(CsA) 20 mg/kg,术后每天10 mg/kg.A和B组分别在术后5d和7d取血液标本及移植物(供心)标本.(3)空白组,取大鼠移植前血液及心肌组织作标本.应用蛋白质谱技术对各组的血清及心肌组织蛋白质样本进行质谱测定,统计分析各组之间的差异蛋白质,并比较分析差异蛋白质的峰强度.结果 与空白组血清比较,急性排斥A组与之存在7个差异蛋白质峰,急性排斥B组与之存在11个差异蛋白质峰.急性排斥A组与治疗A组比较,有4个差异蛋白质峰,急性排斥B组与治疗B组比较,有6个差异蛋白质峰.与空白组心肌组织蛋白质比较,急性排斥A和B组与之分别存在7个和13个差异蛋白质峰.以上比较差异均有统计学意义(P＜0.01).结论 正常与急性排斥反应后不同时相点血清及心肌组织中的蛋白存在明显差异,这种差异蛋白可能与急性排斥反应相关.     

Acute Antibody-Mediated Rejection Following Heart Transplantation

Acute antibody-mediated rejection (AMR) in heart transplantation is often associated with hemodynamic compromise, and is associated with increased mortality and development of accelerated transplant coronary artery disease (TCAD). The diagnosis of AMR has historically been controversial and outcomes with aggressive immunosuppressive therapy including plasmapheresis and cyclophosphamide are poor. Advances in diagnostic techniques like the demonstration of immunopathologic evidence for antibody-mediated rejection by deposition of the complement split product C4d in tissue and detection of anti-HLA antibodies by flow cytometry will assist in further characterizing AMR. Immunosuppression targeting B-lymphocytes and use of m-TOR inhibitors to alter the predilection to develop TCAD and improve survival in AMR remains to be proven.     

肝移植术后一过性肝内胆汁淤积症与急性排斥反应的鉴别

目的探讨肝移植术后一过性肝内胆汁淤积症（transient intrahepatic cholestasis，TIHC）与急性排斥反应（acute rejection，AR）的鉴别方法。方法按诊断标准将30例肝移植患者分为TIHC组与AR组，观察比较2组的谷丙转氨酶（ALT）及直接胆红素（DBIL）在术前1d，术后早期（术后3d、7d及21d）的变化特点。结果术后第7d对比术后第3d，TIHC组DBIL明显升高而ALT无明显变化；AR组DBIL明显升高同时ALT呈升高趋势。经过针对性治疗，至术后第21d，2组ALT及DBIL均明显下降。结论肝移植术后DBIL及ALT的变化特点对于TIHC与AR的鉴别有一定意义。     

Increased β-Myosin Heavy Chain in Acute Cellular Rejection Following Human Heart Transplantation

BACKGROUND: Increased expression of smooth muscle and nonmuscle myosin heavy chains has been previously reported in animal models of cardiac allograft rejection. However, altered expression of beta-myosin heavy chain in human cardiac rejection has not been determined. METHODS: Two-dimensional (2D)-gel electrophoresis of endomyocardial biopsies taken from patients with (Grade 3A, n = 6) and without (Grade 0, n = 6) acute rejection were analyzed. Increased expression of two protein spots (MW approximately 12 kDa) were identified in the presence of acute rejection and were further characterized by mass spectrometry analysis. In patients who had acute rejection, protein expression was subsequently analyzed by immunoblotting on biopsies preceding, during, and following treatment of rejection. RESULTS: Mass spectrometric analysis of the protein spots detected 6 and 22 tryptic peptides, respectively. Protein sequence database search analysis identified the first protein as beta-myosin heavy chain and the second spot consisted of proteins of unidentified nature that may represent novel proteins. Immunoblotting analysis showed 1.4 x fold increase (p < 0.01) of protein expression of beta-myosin heavy chain expression in the presence of acute rejection. CONCLUSIONS: To our knowledge, this is the first 2D-gel study to describe increased expression of beta-myosin heavy chain and other proteins of unidentified nature in association with human cardiac allograft rejection.     

A Peripheral Blood Diagnostic Test for Acute Rejection in Renal Transplantation

Monitoring of renal graft status through peripheral blood (PB) rather than invasive biopsy is important as it will lessen the risk of infection and other stresses, while reducing the costs of rejection diagnosis. Blood gene biomarker panels were discovered by microarrays at a single center and subsequently validated and cross‐validated by QPCR in the NIH SNSO1 randomized study from 12 US pediatric transplant programs. A total of 367 unique human PB samples, each paired with a graft biopsy for centralized, blinded phenotype classification, were analyzed (115 acute rejection (AR), 180 stable and 72 other causes of graft injury). Of the differentially expressed genes by microarray, Q‐PCR analysis of a five gene‐set (DUSP1, PBEF1, PSEN1, MAPK9 and NKTR) classified AR with high accuracy. A logistic regression model was built on independent training‐set (n = 47) and validated on independent test‐set (n = 198)samples, discriminating AR from STA with 91% sensitivity and 94% specificity and AR from all other non‐AR phenotypes with 91% sensitivity and 90% specificity. The 5‐gene set can diagnose AR potentially avoiding the need for invasive renal biopsy. These data support the conduct of a prospective study to validate the clinical predictive utility of this diagnostic tool.     

Perioperative Infliximab Application Ameliorates Acute Rejection Associated Inflammation After Intestinal Transplantation

As we have shown in the past, acute rejection‐related TNF‐α upregulation in resident macrophages in the tunica muscularis after small bowel transplantation (SBTx) results in local amplification of inflammation, decisively contributing to graft dysmotility. Therefore, the aim of this study is to investigate the effectiveness of the chimeric‐monoclonal‐anti‐TNF‐α antibody infliximab as perioperative single shot treatment addressing inflammatory processes during acute rejection early after transplantation. Orthotopic, isogenic and allogenic SBTx was performed in rats (BN‐Lewis/BN‐BN) with infliximab treatment. Vehicle and IV‐immunoglobulin‐treated animals served as controls. Animals were sacrificed after 24 and 168 h. Leukocyte infiltration was investigated in muscularis whole mounts by immunohistochemistry, mediator mRNA expression by Real‐Time‐RT‐PCR, apoptosis by TUNEL and smooth muscle contractility in a standard organ bath. Both, infliximab and Sandoglobulin® revealed antiinflammatory effects. Infliximab resulted in significantly less leukocyte infiltration compared to allogenic controls and IV‐immunoglobulin, which was accompanied by lower gene expression of MCP‐1 (24 h), IFN‐γ (168 h) and infiltration of CD8‐positive cells. Smooth muscle contractility improved significantly after 24 h compared to all controls in infliximab treated animals accompanied by lower iNOS expression. Perioperative treatment with infliximab is a possible pharmaceutical approach to overcome graft dysmotility early after SBTx.     

Impact of Acute Rejection Episodes on Long-Term Graft Survival Following Simultaneous Kidney-Pancreas Transplantation

Although it is well established that acute rejection is one of the major risk factors for chronic graft loss following kidney transplantation, its effect on long-term graft survival following simultaneous kidney-pancreas transplants (SKPTs) is less well known. We analyzed a large cohort of SKPTs and cadaver kidney transplants reported to the United Network for Organ Sharing database during 1988-97, to determine the impact of acute rejection episodes on long-term kidney and pancreas graft survival. Only patients whose kidney and pancreas grafts had survived for at least 1 year were included. Other potential risk factors influencing long-term graft survival were included in the analysis. Of the 4251 SKPTs, 45% had no acute rejection, 36% had kidney only rejection, 3% had pancreas only rejection, and 16% had both kidney and pancreas rejection within the 1st year post transplant. The 5-year kidney and pancreas graft survival rates adjusted for other risk factors were 91% and 85%, respectively; for those with no acute rejection episodes, 88% and 84%, respectively; for those with kidney only rejection, 94% and 83%, respectively; for those with pancreas only rejection; and 86% and 78%, respectively, for those with both kidney and pancreas rejection. The relative risk (RR) of kidney graft failure was 1.32 when acute rejection involved the kidney graft only, while the RR was 1.53 when the rejection involved both organs. We conclude that acute rejection episodes have a negative impact on the long-term kidney graft survival in the SKPT population similar to that in the cadaver kidney transplant population. Patients who had acute rejection episodes of both kidney and pancreas have the worst long-term graft survival.     

肝细胞移植治疗急性肝功能衰竭的免疫排斥研究

目的研究肝细胞腹腔移植治疗急性肝功能衰竭的免疫排斥反应。方法用胶原酶灌注法分离幼猪及BALB/c和C57BL/6小鼠肝细胞;用腹腔注射CCl4的方法建立C57BL/6急性肝功能衰竭模型;将实验动物分为同基因移植组、同种异基因移植组和异种移植组。各组动物肝细胞移植入急性肝功能衰竭小鼠腹腔内,观察受体小鼠存活情况,比较各组动物T细胞亚群、免疫球蛋白水平和细胞因子的变化。结果①受体小鼠存活情况:同基因移植组为8/10,同种异基因移植组为6/10,异种移植组为3/10,同基因移植组和同种异基因移植组受体的生存情况明显好于异种移植组(P<0.05)。②T细胞亚群变化:移植后7 d内,同基因移植组外周血中CD4+和CD8+T细胞均无明显变化,同种异基因移植组CD4+T细胞于移植后3 d时达高峰(P<0.05),而CD8+T细胞7 d内无明显变化,异种移植组CD4+和CD8+T细胞移植后7 d内均明显升高,且于移植后3 d时达高峰(P<0.05)。③免疫球蛋白水平:同基因移植组血清免疫球蛋白IgM和IgG水平在移植后0.5、1和3 d时未见明显变化,同种异基因移植组和异种移植组的IgM在移植后1 d时达高峰(P<0.05),而IgG在移植后3 d时达高峰(P<0.05)。④血清细胞因子水平:移植后7 d,同种异基因移植组和异种移植组的IFN-γ、TNF-α及IL-2血清浓度明显高于同基因移植组(P<0.05);异种移植组的IL-6血清浓度明显高于同基因移植组和同种异基因移植组(P<0.05)。结论无论同种还是异种肝细胞移植,初期均发生了强烈的CD4+及CD8+T细胞参与的细胞免疫和较强体液免疫反应。     

Alemtuzumab in the Treatment of Refractory Acute Rejection and Bronchiolitis Obliterans Syndrome After Human Lung Transplantation

Despite substantial improvements in early survival after lung transplantation, refractory acute rejection (RAR) and bronchiolitis obliterans syndrome (BOS) remain major contributors to transplant-related morbidity and mortality. We have utilized alemtuzumab, a humanized anti-CD52 antibody which results in potent lymphocyte depletion, in consecutive patients with RAR (n = 12) or BOS (n = 10). All patients failed conventional treatment with methylprednisolone and antithymocyte globulin and received strict infection prophylaxis. Alemtuzumab significantly improved histological rejection scores in RAR. Total rejection grade/biopsy was 1.98 +/- 0.25 preceding alemtuzumab versus 0.33 +/- 0.14 posttreatment, p-value <0.0001 (with a similar number of biopsies/patient per respective time interval). Freedom from BOS was observed in 65% of RAR patients 2 years after alemtuzumab treatment. Although there was no statistically significant change in forced expiratory volume in 1 second (FEV1) before and after alemtuzumab treatment in patients with BOS, a stabilization or improvement in BOS grade occurred in 70% of patients. Patient survival 2 years after alemtuzumab for BOS was 69%. Despite a dramatic decline in CD4 counts in alemtuzumab-treated patients, only one patient developed a lethal infection. Thus, we provide the first evidence that alemtuzumab is a potentially useful therapy in lung transplant recipients with RAR or BOS.