尿趋化因子IP-10和Fractalkine在移植肾急性排斥反应早期诊断中的应用

背景：相关研究已证实,趋化因子IP-10、Fractalkine在器官移植后急性排斥反应过程中发挥着重要的作用。目的：检测尿液中IP-10和Fractalkine水平变化,并结合肾组织活检病理,探讨尿液中IP-10和Fractalkine在移植肾急性排斥反应早期诊断中的意义。方法：106例同种异体肾移植患者,根据移植后临床表现、实验室检查及肾穿刺组织病理学检查结果,分为急性排斥反应组（n=16）和非急性排斥反应组（n=90）;另选择健康志愿者作为正常对照组。用双抗体夹心酶联免疫吸附试验检测尿IP-10和Fractalkine浓度变化。结果与结论：急性和非急性排斥反应组患者在移植后的尿IP-10及Fractalkine的表达水平均较移植前明显升高,但非急性排斥反应组在移植后7d呈下降趋势,至第11天降至移植前水平,而急性排斥反应组则持续高表达,IP-10在移植后第1天和Fractalkine在移植后第3天即与急性排斥反应组比较,差异有显著性意义（P〈0.05）。提示,肾移植后尿液中IP-10和Fractalkine水平的检测对于急性排斥反应发生的早期诊断和早期治疗具有重要意义。     

本期专题:肾移植与急性排斥反应的发生

1尿趋化因子IP-10和Fractalkine在移植肾急性排斥反应早期诊断中的应用,见2011年31期5765页2尿单核细胞趋化蛋白1与肾移植后急性排斥反应:有相关性吗?见2010年5期789页3尿CD54+淋巴细胞在肾移植急性排斥反应中的变化,见2011年53期9905页     

尿CD54~+淋巴细胞在肾移植急性排斥反应中的变化

背景:正常肾脏、肾小管上皮和血管内皮细胞仅有少量CD54表达,当发生急性排斥反应时,肾小管上皮细胞和血管内皮细胞CD54表达明显增加,同时大量白细胞浸润;间质浸润细胞和肾小管上皮细胞CD54表达增加。目的:探讨流式细胞仪检测尿CD54+淋巴细胞对移植肾急性排斥反应的诊断价值。方法:来自解放军成都军区总医院的肾移植后恢复正常者(n=18)、出现急性排斥反应者(n=8)、移植肾功不全者(n=9)以及健康志愿者(n=10)。流式细胞仪比较各组移植前后尿液中CD54+淋巴细胞比率变化。结果与结论:尿CD54+淋巴细胞在肾移植患者出现排斥反应时明显增加(P<0.01),抗排斥治疗后逐渐下降。移植肾功能正常者和移植肾功不全者CD54轻度升高。提示尿液中CD54+淋巴细胞水平能准确反映肾移植物移植后患者的免疫状态,可作为肾移植后急性排斥反应的特异标志。     

早期移植肾功能异常预警:尿中性粒细胞明胶酶相关脂笼蛋白和白细胞介素18有价值吗?

背景:移植肾功能延迟恢复是肾移植后常见的并发症,其预警诊断和病因诊断对指导早期干预治疗有重要价值,中性粒细胞明胶酶相关脂笼蛋白、白细胞介素18已被证实是一种诊断急性肾小管损伤的标志物,但其在早期移植肾功能异常的预测及病因诊断的意义方面尚未见报道.目的:评价尿中性粒细胞明胶酶相关脂笼蛋白(neutrophil gelatinase-associated lipocalin,NGAL)和白细胞介素18水平在早期移植肾功能异常预警诊断和病因诊断的价值.方法:应用ELISA方法检测71例患者肾移植后的第1个24 h尿NGAL和白细胞介素18水平.根据移植后肾功能恢复情况分为肾功能迅速恢复组(n=41)和早期肾功能异常组(n=30),将早期肾功能异常组按病因分为急性肾小管坏死组(n=18)和急性排斥反应组(n=12),按肾功能恢复形式分为缓慢恢复组(n=16)和延迟恢复组(n=14),观察尿液NGAL和白细胞介素18水平与肾功能恢复的关系.结果与结论:所有患者移植后尿NGAL和白细胞介素8均明显升高,但早期肾功能异常组明显高于迅速恢复组.急性排斥反应导致的肾功能异常组自细胞介素18明显高于急性肾小管坏死导致的肾功能异常组(p<0.01),两组尿NGAL水平差异无显著性意义(p>0.05).缓慢恢复组尿NGAL水平较延迟恢复组高(p<0.01),两组尿白细胞介素18水平差异无显著性意义(p>0.05).提示尿NGAL和白细胞介素18可作为预测早期移植肾功能异常的标志物,白细胞介素18有助于急性排斥反应病因诊断,而NGAL的升高除了诊断移植肾功能异常外,还可能成为预测肾功能恢复能力的指标.     

尿单核细胞趋化蛋白1与肾移植后急性排斥反应:有相关性吗?

背景:同前移植肾急性排斥反应依然是导致慢性排斥反应和移植物功能损伤的危险因素,如何无创、快速、准确地进行诊断并及时治疗尤为重要.目的:通过检测尿液中单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)水平变化,并结合部分肾组织活检病例,探讨尿MCP-1在肾移植后急性排斥反应早期诊断及治疗后的表达.方法:选择2008 10/2009-02在郑州人民医院肾病移植科住院治疗的慢性肾哀竭患者62例,均接受同种异体肾移植,肾功能稳定组为肾移植后肾功能稳定的42例患者,急性排斥组为肾移植后发牛急性排斥反应的20例患者,以同期在郑州人民医院体检中心检查肾功能正常且自愿留取尿样的10例健康人作为对照组.所用肾移植患者均给予常规免疫抑制方案,另外急性排斥组给予抗淋巴细胞免疫球蛋白或甲基强的松龙强化冲击治疗.应用双抗体夹心酶联免疫吸附试验检测尿MCP-1质量浓度变化.结果与结论:与对照组比较,肾功能稳定组尿MCP-1质量浓度无明显变化(P>0.05),急性排斥组尿MCP-1质量浓度显著升高(P<0.01).与治疗前比较,急性排斥组20例患者经强化治疗后尿MCP-1质量浓度均显著降低(P<0.01),其中17例临床症状缓解、辅助检查恢复正常,尿MCP-1质量浓度接近对照组,3例无效,尿MCP-1质量浓度高于对照组.肾穿刺活检8例,肾脏病理提示均为移植肾急性排斥反应,与急性排斥组治疗前尿MCP-1质量浓度基本相似(P>0.05).提示尿液中MCP-1水平可早期无创性诊断肾移植后急性排斥反应,可无创性临测治疗效果,其与肾移植后急性排斥反应时肾脏病理损伤可能存在相关性.     

尿HLA-DR在肾移植急性排斥中的检测及其临床意义

目的探讨流式细胞术检测尿淋巴细胞人类白细胞抗原(HLA-DR)对移植肾急性排斥反应的诊断价值。方法 36例肾移植患者按照移植肾功能正常、术后急性排斥反应、其他原因引起移植肾功不全等标准分为A~C组,另选取10例健康志愿者作为对照(D组),采用流式细胞术检测尿中HLA-DR阳性细胞表达率。结果与A、C、D组相比,尿淋巴细胞HLA-DR在排斥反应时表达明显增加(P<0.01),抗排斥治疗后逐渐下降。与D组相比,A组和C组HLA-DR表达率轻度升高(P<0.05),但两组间差异无统计学意义(P>0.05)。结论流式细胞术动态检测尿液中HLA-DR阳性率,能准确反映肾移植术后患者的免疫状态,可作为急性排斥反应的特异标志。     

Luminex技术检测肾移植患者血清IL-17预测急性排斥反应的回顾性研究

目的应用Luminex液相芯片技术检测肾移植患者血清IL-17的表达水平,探讨其与移植肾早期急性排斥反应的关系。方法选取2009年1月~2011年10月期间38例肾移植患者和20例健康对照者;根据预后将肾移植患者分为急性排斥组(18例)和非排斥组(20例),分别检测两组肾移植患者及健康对照者血清IL-17的表达情况,分析血清IL-17与移植肾患者发生急性排斥反应的相关性。结果肾移植患者术前血清IL-17的平均水平为1.3±1.9 pg/ml,显著低于健康对照_组(6.9±8.5 pg/ml),差异有统计学意义(t=3.968,P0.001);肾移植患者急性排斥组血清在术后第7天IL-17水平为3.4±5.8 pg/ml,高于非排斥组(0.5±0.4 pg/ml),差异有统计学意义(t=2.242,P=0.031)。Kaplan-Meier法生存分析显示肾移植术后非排斥反应患者血清IL-17高值水平的生存率显著低于低值水平(P0.001)ROC曲线显示,将肾移植患者术后第4天与第7天血清IL-17水平联合分析,预测其急性排斥反应的灵敏度和特异度分别为66.67%和10.0%。结论肾移植患者术后4~7天血清IL-17水平的动态升高趋势可预测其早期急性排斥反应发生的可能性。     

肾移植术后尿液DYZ-1检测的临床意义

背景:对移植受者尿液成分进行定期监测可以作为判断移植肾功能、状态的一种方法.目的:分析肾移植受者尿液中供者细胞出现与急性排斥反应的关系及临床意义.方法:选取60例供者为男性、受者为女性的肾移植受者,分为移植后早期检测组40例、急性排斥组10例、移植后功能稳定组10例.定期提取尿液中细胞DNA,利用PCR检测Y染色体上特异的基因片段DYZ-1.结果与结论:在手术第1日受者的尿液中可检测到供者细胞DNA,随着时间的延长,尿液中供者细胞DNA量逐渐减少,至术后1个月,有8例受者的尿液中供者细胞DNA消失,其中1例发生急性排斥反应;另外32例受者的尿液中仍有供者细胞DNA的出现,其中7例发生了急性排斥反应;存活3个月以上发生急性排斥反应10例患者,7例尿液标本中能检测到供者细胞DNA,对急性排斥者进行治疗后1个月,71.4%转为阴性;而在稳定期的10例肾功能良好受者,仅1例受者尿液中DYZ-1基因阳性.提示对肾移植受者尿中供者细胞DNA的进行定期检测,可以作为监测急性排斥反应发生及预后的一种手段.     

PRA在肾移植中的应用

目的探讨肾移植患者血清中群体反应性抗体（PRA）水平与术后急性排斥反应发生率以及患者1、2年存活率之间的关系。方法采用ELISA方法检测62例肾移植患者血清PRA水平,根据PRA的检测结果将患者分为致敏组和非致敏组,观察两组患者移植手术后发生急性排斥反应的情况,并随访PRA对肾移植患者移植效果的影响。结果致敏组患者急性排斥反应发生率较非致敏组增高,差异有统计学意义;且PRA水平与急性排斥反应发生率成正相关;致敏组患者移植后1、2年移植肾存活率较非致敏组降低,差异有统计学意义（P〈0.05）。结论术前检测患者血清PRA,对预测移植风险、供体选择、避免急性排斥反应的发生以及提高移植肾长期存活都具有重要的意义。     

CRTER关注“肾移植排斥反应中特异性抗体及细胞因子的作用”内容（本刊学术部）

肾移植排斥反应中特异性抗体补体与移植排斥反应尿单核细胞趋化蛋白1与肾移植后急性排斥反应：有相关性吗？肾移植患者群体反应性抗体水平与移植肾急性排斥的关系 肾移植急性排斥反应时Bcl一2／Bax及Fas／FasL蛋白表达的变化     

丹参注射液在肾移植后急慢性排斥反应中的作用

背景:临床观察,肾移植后发生急、慢性排斥反应时单用甲基强的松龙针冲击,尿蛋白、血肌酐值改善不明显,若加用丹参注射液静脉滴注,可使移植肾功能恢复更迅速。目的:观察丹参注射液对肾移植后急慢性排斥反应肾功能恢复的影响。方法:肾移植后发生急性排斥反应患者180例,慢性排斥反应患者140例,分别随机分成两组,对照组单纯应用常规甲基强的松龙冲击治疗3d,治疗组在此基础上加用丹参注射液静脉滴注15d。结果与结论:与治疗前相比,急慢性排斥反应两组肾功能恢复指标均有明显改善,差异有显著性意义(P<0.05);治疗组比对照组肾功能指标改善更明显(P<0.05)。且治疗组延长了凝血酶原时间和活化部分凝血活酶时间(P<0.05)。结果证实了肾移植患者发生急慢性排斥反应后在常规处理基础上加用丹参注射液能明显改善移植肾功能。     

Plasma and urine fibronectin concentration in kidney-transplanted patients

Plasma and urine fibronectin concentration was determined by electroimmunoassay and ELISA-method in patients who received renal transplantation. The plasma fibronectin concentration decreased both after the transplantation, in relation to the acute rejection of the graft, and in the relation to immunosuppressive therapy. Urine fibronectin excretion increased in relation to the kidney transplantation and acute rejection of the graft. In association with improved kidney function, the urine fibronectin excretion decreased. It is suggested that it might be of clinical importance to determine the excretion of fibronectin into the urine in patients undergoing kidney transplantation.     

Presence of donor- and recipient-derived DNA in cell-free urine samples of renal transplantation recipients: urinary DNA chimerism.

BACKGROUND: Previous studies have indicated that microchimerism is present in body tissues, peripheral blood, and plasma of recipients after organ transplantation. We hypothesize that donor-derived DNA may also be present in cell-free urine of renal transplant recipients and that the concentrations of urine DNA may be correlated with graft rejection. METHODS: Thirty-one female patients who had renal transplantation were enrolled in the study. In women with male organ donors, the SRY gene on the Y chromosome was used as a marker for donor-derived DNA. Real-time quantitative PCR for the SRY and beta-globin genes was carried out on cell-free urinary DNA from these patients. Serial urine samples from a female renal transplant recipient undergoing an acute rejection episode were also collected and analyzed with the beta-globin quantitative PCR system. RESULTS: SRY sequences were detected in the urine of 14 of 17 female patients with male organ donors. None of the 14 patients with female organ donors had detectable SRY sequences in urinary DNA. The median fractional concentration of donor-derived DNA was 8.7% (interquartile range, 1.9-26.4%). During the acute rejection episode, urinary concentrations of the beta-globin gene were markedly increased, with the concentrations returning rapidly to normal following antirejection treatment. CONCLUSIONS: Our results demonstrate that urinary DNA chimerism is present following renal transplantation. The measurement of urinary DNA using quantitative PCR may be useful for the diagnosis and monitoring of graft rejection.     

Aminoaciduria as a marker of acute renal transplant rejection--a patient study

Over 12 months, urine samples were systematically collected from 40 children who underwent renal transplantation for the treatment of end-stage renal disease. Sequential determinations of the excretion of individual amino acids relative to that of creatinine were carried out on 15 subjects. Nine of these (including three who sustained episodes of acute rejection) retained a native kidney in-situ, while in six patients (including three who underwent an episode of acute rejection) both native kidneys had been removed. In both subgroups, the amino acid/creatinine ratios of early morning urine samples were higher shortly before clinical manifestations of acute rejection became evident than in patients who, following renal transplantation, had stable kidney function, chronic graft rejection, or acute tubular necrosis, with one exception: a patient with one native kidney in-situ in whom acute tubular necrosis developed immediately after transplantation. The amino acids showing the greatest increase included Thr, Ser, Gly, and Ala. These values fell dramatically immediately prior to the clinical episode of acute rejection, with Thr, Ala, and Phe showing the most consistent changes. These alterations in urinary amino acid excretion occurred several days before changes in urinary protein excretion or the serum concentrations of urea and creatinine, and may have a role to play in the monitoring of renal transplant recipients.     

尿液DNA的检测在肾移植术后急性排斥反应诊断中的应用

目的　探讨肾移植受者尿液中供者细胞DNA的检测在诊断急性排斥反应的应用及其价值。方法　以供者为男性、受者为女性的35例肾移植受者为研究对象,定期收集尿液标本,从中提取DNA ,利用聚合酶链反应检测Y染色体上特异的基因片段DYZ -1。结果　手术第1日受者的尿液中即有供者细胞出现,随着时间的推移,尿液中供者细胞DNA的基因表达强度逐渐减弱,直至术后1个月,只有5例(14 . 3% )受者的尿液中供者细胞DNA的基因表达消失,其中1例(2 0 % )发生急性排斥反应;另外30例(85 . 7% )受者的尿液中仍有供者细胞DNA的基因表达,其中7例(2 3 .3% )发生了急性排斥反应;存活3个月以上发生急性排斥反应9例患者,7例(77 .8% )的尿液标本中能检测到供者细胞DNA ,抗排斥治疗结束后1个月,71 4 %转为阴性;9例肾功能良好的稳定期受者,仅1例(11. 1% )的受者尿液中DYZ -1基因阳性。结论　长期存活的肾移植受者尿中供者细胞DNA的检测可以作为诊断急性排斥反应一种方法。     

肾小管泌氢功能检测在肾移植对抗排斥反应治疗效果中的应用

背景：移植后的急性排斥是肾移植术后的主要并发症，也是导致慢性排斥反应和移植物失功最重要的危险因素，因此，了解肾小管泌氢功能能否早期反映移植物的功能情况有重要意义。目的：观察肾移植患者术后肾小管泌氢功能，并进行监测，探讨其在移植物急慢性排斥中的作用。设计：病例-对照观察。单位：解放军济南军区总医院泌尿外科。对象：选择2000—05／2005—06解放军济南军区总医院泌尿外科连续实施肾脏移植26例患者，男16例，女10例；年龄21～58岁，平均35岁。原发病均为慢性肾小球肾炎，慢性肾功能衰竭，全部为尸肾移植。供受者均血型相同、淋巴细胞毒试验阴性。其中1例为第2次移植。所有患者对检测项目知情同意。方法：依据典型的临床表现，彩色多普勒超声及血流变化诊断患者排斥反应，16例患者未发生排斥反应为稳定组，10例发生排斥反应的患者为排斥组，排斥组根据排斥情况分为排斥前期、排斥期及恢复期。对所有患者术前及术后1周起每周1次，连续10周分别以化学清洁玻璃瓶留取晨起中段尿测定尿可滴定酸、NH4^＋和净酸水平评估肾小管泌氢功能。主要观察指标：两组患者尿可滴定酸、NH4^＋和净酸水平。结果：纳入患者26例均进入结果分析。排斥反应组排斥前期患者肾小管泌氢功能各检测值显示泌氢功能开始建立，并趋向正常，排斥期患者肾小管泌氢功能检测值均显示明显下降，与排斥前期及稳定组比较均有统计学显著性差异（P〈0．01）。恢复期患者排斥反应治疗后监测的结果显示泌氢功能恢复较快。肾小管泌氢功能总体连续观察结果显示总体恢复的不均衡性，大部分病例恢复时间从1～10周不等，平均恢复期限约6周，2例10周内未恢复，4例严重排斥者中3例治疗后泌氢功能恢复缓慢。结论：肾小管泌氢功能可弥补血清肌酐不能良好反映肾小管功能的不足，能在连续观察中对移植肾急性排斥反应的诊断，特别是对抗排斥治疗的效果判断和预后评估中作为有价值的指标。     

Proinflammatory functions of vascular endothelial growth factor in alloimmunity

Vascular endothelial growth factor (VEGF), an established angiogenesis factor, is expressed in allografts undergoing rejection, but its function in the rejection process has not been defined. Here, we initially determined that VEGF is functional in the trafficking of human T cells into skin allografts in vivo in the humanized SCID mouse. In vitro, we found that VEGF enhanced endothelial cell expression of the chemokines monocyte chemoattractant protein 1 and IL-8, and in combination with IFN-gamma synergistically induced endothelial cell production of the potent T cell chemoattractant IFN-inducible protein-10 (IP-10). Treatment of BALB/c (H-2d) recipients of fully MHC-mismatched C57BL/6 (H-2b) donor hearts with anti-VEGF markedly inhibited T cell infiltration of allografts and acute rejection. Anti-VEGF failed to inhibit T cell activation responses in vivo, but inhibited intragraft expression of several endothelial cell adhesion molecules and chemokines, including IP-10. In addition, whereas VEGF expression was increased, neovascularization was not associated with acute rejection, and treatment of allograft recipients with the angiogenesis inhibitor endostatin failed to inhibit leukocyte infiltration of the grafts. Thus, VEGF appears to be functional in acute allograft rejection via its effects on leukocyte trafficking. Together, these observations provide mechanistic insight into the proinflammatory function of VEGF in immunity.     

Steroid-resistant late acute rejection after a living donor liver transplantation: case report and review of the literature

The majority of acute cellular rejection occurs in the first few months after liver transplantation. It has been, however, reported that some recipients experience late acute rejection, which occurs more than 3 months after transplantation. We herein report a case of late acute rejection that occurred nearly 10 years after liver transplantation. The patient is a 27-year-old male who underwent a living donor liver transplantation when he was 17 years old. At 9 years 6 months after transplantation, the patient presented with the elevated serum levels of liver enzymes and total bilirubin. A liver biopsy showed acute cellular rejection. Steroid bolus therapy was not effective, but we successfully used deoxyspergualin as a rescue therapy. Late acute cellular rejection that occurs nearly 10 years after transplantation has so far been rarely reported. It is generally believed that late acute rejection may be more resistant to treatment and be associated with a higher rate of graft loss, as well being associated with the development of chronic ductopenic rejection. In this report, we have shown that deoxyspergualin is safe and effective for treatment of steroid-resistant late acute rejection, preventing from graft loss of chronic rejection.