Pre-transplant and post-transplant soluble CD30 for prediction and diagnosis of acute kidney allograft rejection

Background  Serum levels of soluble CD30 (sCD30) have been considered as a predictor of acute kidney allograft rejection. We have evaluated the pre-transplant and post-transplant levels of sCD30 with the aim of determining its value in predicting and diagnosing kidney rejection. Methods  We measured sCD30 serum levels before kidney transplantation, 5 days post-operatively, and at creatinine elevation episodes. The predictive value of sCD30 for diagnosing acute rejection (AR) within the first 6 post-operative months was assessed in 203 kidney recipients from living donors. Results  Pre-transplant and post-operative levels of serum sCD30 were 58.10 ± 52.55 and 51.55 ± 49.65 U/ml, respectively (P = 0.12). Twenty-three patients experienced biopsy-proven acute rejection, and 28 had acute allograft dysfunction due to non-immunologic diseases. The pre-transplant sCD30 level was not different between patients with and without AR. However, post-transplant sCD30 was higher in the AR group. The median serum level of post-transplant sCD30 was 52 U/ml in the AR group and 26.3 U/ml in a control group (P < 0.001). The relative changes of sCD30 on day 5 were higher in patients with AR (P = 0.003). Based on post-transplant sCD30 levels, we were able to differentiate between kidney recipients who experienced an AR within 6 months post-surgery and those without an AR (cutoff value 41 U/ml; sensitivity 70%; specificity 71.7%). The level of sCD30 during periods of elevated serum creatinine was not independently associated with the diagnosis of AR. Conclusion  Post-transplant sCD30 levels and their relative changes are higher in patients experiencing AR. We propose further studies on the post-transplant trend of this marker for the prediction of AR.     

Hepatocyte growth factor (HGF) as a rapid diagnostic marker and its potential in the prevention of acute renal rejection

Several investigators have reported that hepatocyte growth factor (HGF) may be related to the protection or reconstruction of the kidney during acute renal rejection. To address this question, we examined the relationship between HGF and acute rejection in the following two studies with rat renal transplantation models. In study 1, the relationship between serum HGF levels and acute renal rejection in iso-, allo- and allograft with cyclosporine (CsA)-treated models was examined. In study 2, the focus was whether or not the injection of recombinant HGF can prevent acute renal rejection. Our results demonstrated that HGF levels were rapidly increased during acute rejection and that recombinant HGF effectively protected the kidney from acute rejection. These results suggest that HGF may be induced as a counter-response to the renal injury and that it can be used as a reliable indicator for the diagnosis of acute rejection. We suggest that recombinant HGF suppresses the onset of the pathological changes of acute rejection.     

Pre- and post-transplant monitoring of soluble CD30 levels as predictor of acute renal allograft rejection

Identification of renal graft candidates at high risk of impending acute rejection (AR) and graft loss may be helpful for patient-tailored immunosuppressive regimens and renal graft survival. To investigate the feasibility with soluble CD30 (sCD30) as predictor of AR, sCD30 levels of 70 patients were detected on day 0 pre-transplant and day 1, 3, 5, 7, 10, 14, 21, and 30 post-transplant. AR episodes in 6 months were recorded and then patients were divided into Group AR (n=11) and Group UC (n=59). Results showed that the patients had higher pre-transplant sCD30 levels than healthy people. A significant decrease of sCD30 was observed on the first day post-transplant and continued until day 14 post-transplant. Soluble CD30 presented a stable level from day 14 to 30 post-transplant. Pre-transplant sCD30 levels of Group AR were much higher than those of Group UC (P<0.001). Patients of Group AR also had higher sCD30 levels than those of Group UC on day 1, 3, 5, 7, 10 and 14 (P<0.001). The sCD30 level presented a significantly delayed decrease in the patients of Group AR. Statistical results showed that the highest value of area under ROC curve (0.95) was obtained on day 5 post-transplant, suggesting that sCD30 levels on day 5 are of high predictive value. Therefore, sCD30 level may be a good marker of increased alloreactivity and of significant predictive value. It's necessary to monitor the variation of sCD30 in the early period post-transplant.     

肾移植患者急性排斥反应与sCD30的相关性

目的 研究检测肾移植患者手术前后血清溶解性CD30（sCD30）水平的临床意义。方法 采用酶联免疫吸附剂测定法（ELISA）检测69例肾移植患者术前及术后sCD30的水平，并分析sCD30与肾移植受者术后急性排斥发生的关系。结果 术前sCD30阳性患者11例，其中有6例发生急性排斥，sCD30阴性患者58例，发生急性排斥5例。两组相比排斥反应发生率差异有统计学意义（P〈0．01）。术后5dsCD30在发生排斥患者组中的水平与对照组间差异有统计学意义（P〈0．05），而术后1、3d水平两组间差异无统计学意义（P〉0．05）。结论 肾移植手术前后监测sCD30水平，特别是术前及术后第5天左右时的检测水平，对于评估和预测急性排斥反应发生的可能性，具有重要的参考价值。     

肾移植受者术前血清可溶性CD30水平对急性排斥的影响

目的 研究术前血清可溶性CD30(sCD30)水平对肾移植受者术后6个月内急性排斥及排斥类型的预测作用。方法 共纳入自1998年12月至2003年8月在本中心行同种异体肾移植手术且存有术前血标本的707例受者。 回顾性总结该组受者术后6个月内急性排斥的发生情况及其它临床资料,同时选取健康对照40例。用sCD30 ELISA 试剂盒复孔检测肾移植受者术前和健康对照血清sCD30水平。根据术前sCD30水平将肾移植受者分为低sCD30组、中间sCD30组和高sCD30组。结果 肾移植组术前sCD30水平明显高于健康对照。血管性、细胞性排斥及临界改变的发生率随着sCD30水平的升高而升高(P均 < 0.05),但低、中、高sCD30 3组急性排斥逆转率却分别为100%、90.6%和78.6%,低sCD30组、中间sCD30组与高sCD30组比较差异均有统计学意义(P均 < 0.05)。血管性排斥、细胞性排斥、临界改变和临床排斥的sCD30水平[(198.95±76.09)、(165.89±44.56)、(172.94±74.22)和(161.23±64.87) U/ml]和未排斥组[(133.76±61.95) U/ml]比较,差异均有统计学意义(P均 < 0.01)。多因素logistic回归分析显示,高sCD30、群体反应性抗体(PRA)阳性和巨细胞病毒(CMV)抗原阳性均为急性排斥的危险因素,优势比分别为2.683、2.384和2.065。结论 术前高sCD30水平预示术后急性排斥发生率的增高。     

尿液中性粒细胞明胶酶相关脂质运载蛋白和白细胞介素18水平检测在移植肾功能延迟恢复早期预测中的应用价值

目的 评价尿液中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和白细胞介素18(IL-18)水平检测在早期预测移植肾功能延迟恢复(DGF)中的价值.方法 采用ELISA法,连续监测86例肾移植患者术前和术后12、24 h尿液标本中NGAL,IL-18及视黄醇结合蛋白(RBP)水平,并分析其与DGF的关系. 结果 86例中发生DGF 15例.术后12 h DGF组尿NGAL(ng/mg)水平明显高于非DGF组(1712.7±474.6比863.1±199.8,P<0.001),尿IL-18(ng/mg)水平与非DGF组比较差异无统计学意义(29.2±4.1比28.7±4.2,P>0.05);术后24 h DGF组尿NGAL和IL-18水平均明显高于非DGF组(分别为2905.0±1108.1比911.8±221.0,211.3±34.0比86.9±22.8,均P<0.001);而尿RBP(μg/mmol)(92.7±17.1比92.8±13.8,P>0.05)和SCr值(μmol/L)(608.1±85.5比542.3±99.6,P>0.05)与非DGF组比较差异无统计学意义.ROC曲线下面积分析结果显示,术后12 h尿NGAL曲线下面积为0.90,cut-off值为996.5 ng/mg时,诊断敏感性90.2%,特异性82.6%;术后24 h IL-18曲线下面积为0.76,cut-off值为148.5 ng/mg时,诊断敏感性76.3%,特异性66.4%. 结论 尿NGAL及IL-18水平变化对早期预测DGF发生有重要的临床意义.     

人肝细胞生长因子修饰的BMSCs对大鼠同种异体肝移植免疫排斥的影响

目的观察肝细胞生长因子(hepatocyte growth factor,HGF)修饰的BMSCs对大鼠同种异体肝移植后免疫排斥反应的影响,探讨其诱导免疫耐受的机制。方法 3～4周龄清洁级雄性SD大鼠8只,体重75～85 g,用于分离培养BMSCs;成年清洁级雄性SD大鼠64只,体重200～250 g,作为供体;成年清洁级雄性Wistar大鼠64只,体重230～280 g,作为受体。建立稳定的同种异体肝移植动物模型并随机分为4组(n=16),A、B、C、D组分别立即经门静脉注入1 mL生理盐水、1 mL细胞密度为2×106个/mL的BMSCs、1 mL细胞密度为2×106个/mL的BMSCs/绿色荧光蛋白、1 mL细胞密度为2×106个/mL的BMSCs/hHGF。术后作实验动物生存曲线分析;术后7 d检测实验动物肝功能,并取肝脏组织行病理组织学观察,RT-PCR检测肝组织内hHGF mRNA表达,ELISA法检测血清中hHGF、IL-2、IL-4、IL-10、IFN-γ细胞因子的表达,TUNEL法检测肝组织内细胞凋亡情况,免疫组织化学方法检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在肝组织内的表达。结果 D组生存时间显著高于A、B、C组(P<0.01);B、C组生存时间明显高于A组(P<0.01),但B、C组间生存时间差异无统计学意义(P>0.05)。RT-PCR示检测D组肝组织内有hHGF mRNA表达,ELISA检测血清中hHGF水平达(6.2±1.0)ng/mL。相比B、C组,D组疗效更显著,肝功能明显改善,病理学无急性排斥反应或仅为轻度;IL-2、IFN-γ水平降低,IL-4、IL-10水平升高,细胞凋亡明显减少,PCNA表达显著升高,除IL-4水平比较差异无统计学意义(P>0.05)外,其余指标比较差异均有统计学意义(P<0.05)。结论通过门静脉输注BMSCs/hHGF至大鼠同种异体肝移植模型能成功诱导免疫耐受。与单纯应用BMSCs相比,BMSCs/hHGF在移植肝局部形成高浓度hHGF免疫抑制微环境,能更显著抑制急性排斥反应,延长移植受体存活时间。BMSCs/hHGF的免疫抑制作用与诱导Th1细胞向Th2细胞偏移、减少肝细胞凋亡、促进肝细胞增殖有关。     

The evidence and rationale for the perioperative use of loop diuretics during kidney transplantation: A comprehensive review

Purpose

Loop diuretics (LD) attenuate ischemic injury in nephrons. They are thought to decrease delayed graft function (DGF) during kidney transplantation (KT). This review aimed to summarize the current evidence for the perioperative use of LD during KT.

Enzymuria and low molecular weight protein excretion as the differentiating marker of complications in the early post kidney transplantation period

Renal function in the early post-transplantation period depends largely on factors affecting the kidney prior to implantation. Function of the graft may be also disturbed by the most common complications of the early post-operative period such as acute graft rejection (AGR), acute tubular necrosis (ATN) and may be modified by nephrotoxic action of cyclosporine A (CsA). Evaluation of excretion of enzymes and low molecular weight proteins (LMWP) may help in the differentiation of these complications. Aim Comparison of the urinary excretion of markers of tubular injury in patients with AGR, ATN, or patients with stable graft function (SGF) was made and differences between groups and correlations between markers and cold ischemia time (CIT), warm ischemia time (WIT) and blood trough level of cyclosporine A (CsA₀) were determined. Material and methods In 60 cadaveric renal allograft recipients in the early post-transplantation period urinary excretion of N-acetyl-β-d-glucosaminidase (NAG) and B isoenzyme (NAG-B), alanylaminopeptidase (AAP), γ-glutamyltransferase (GGT), α and π isoenzymes of glutathione S-transferase (α-GST, π-GST), retinol binding protein (RBP) and β2- microglobulin (β2M), were analyzed. Results NAG and NAB-B activities were higher in ATN (P<0.05, P<0.01) and in AGR (P<0.005, P<0.02) than in SGF. Excretion of π-GST was higher in AGR than in SGF (P<0.0002) or ATN (P<0.007). CIT and WIT in patients with ATN were higher (P<0.05) than in SGF group. In ATN patients, correlations of CIT with RBP (P<0.05) and π-GST (P<0.05), and WIT with RBP (P<0.05), and π-GST (P<0.001) were found. Conclusions High urinary NAG and NAG B excretion characterizes ATN and AGR patients. Evaluating urinary excretion of π-GST may be helpful in differentiating AGR from ATN. However, taking into account ischemia time is necessary in interpreting the π-GST value in early post transplant period.     

Soluble CD30 and ELISA‐detected human leukocyte antigen antibodies for the prediction of acute rejection in pediatric renal transplant recipients

Biomarker‐based post‐transplant immune monitoring for the prediction of impending graft rejection requires validation in specific patient populations. Serum of 28 pediatric renal transplant recipients within the framework of a well‐controlled prospective randomized trial was analyzed pre‐ and post‐transplant for soluble CD30 (sCD30), a biomarker reflecting mainly T‐cell reactivity, and anti‐human leukocyte antigen (anti‐HLA) antibody reactivity, a biomarker for B‐cell activation. A sCD30 concentration ≥40.3 U/ml on day 14 was able to discriminate between patients with or without biopsy‐proven acute rejection (BPAR) with a sensitivity of 100% and a specificity of 76%. Six of seven patients (86%) with BPAR showed a sCD30 above this cut‐off, whereas only 3/21 patients (14%) without BPAR had a sCD30 above this cut‐off (P = 0.004). For pre‐ and post‐transplant anti‐HLA class II reactivities by enzyme‐linked immunosorbent assay, a cut‐off value of 140 optical density was able to discriminate rejecters from nonrejecters with a sensitivity of 86% or 71% and a specificity of 81% or 90%, respectively. Withdrawal of steroids was associated with a approximately twofold higher serum sCD30 compared to controls, but did not affect anti‐HLA reactivities. An increased post‐transplant sCD30 serum concentration and positive pre‐ and post‐transplant anti‐HLA class II reactivities are informative biomarkers for impending BPAR in pediatric renal transplant recipients. (TWIST, Clinical Trial No: FG‐506‐02‐43)