肾移植急性排斥反应时B7-2/CD28信号通路的表达*☆

背景：以往动物研究表明,在器官移植急性排斥反应时共刺激分子的表达与急性排斥反应密切相关。 目的：观察急性排斥反应时患者移植肾脏组织和外周血中B7-2/CD28信号通路的表达。 方法：对53例同种异体肾移植患者于移植前1 d、移植后1,3,7,14,21,28 d分别取外周血以及在临床诊断急性排斥反应当天和抗排斥治疗1周后额外采血,用流式细胞仪检测共刺激分子B7-2/CD28在外周血淋巴细胞中的表达;同时,行经皮肾穿刺活检供肾修整结束时、移植后7 d、1个月、6个月、1年或以上获取活检肾脏组织,用免疫组织化学方法检测活检组织中B7-2/CD28的表达情况。 结果与结论：移植后1,3 d内所有患者外周血中CD28+,CD4+/CD28+,CD8+/CD28+细胞比率均有显著下降(P < 0.05),一二周后恢复到术前水平;移植后7 d未发生急性排斥反应的患者肾脏组织B7-2阳性表达率显著上升(P < 0.05),1个月后下降至移植前水平(P > 0.05)。移植后发生急性排斥反应的患者外周血CD28+,CD4+/CD28+,CD8+/CD28+细胞比率及肾脏组织B7-2阳性表达率明显上升(P < 0.05),经抗排斥治疗1周后均好转。结果证实,在肾移植后出现急性排斥反应时,肾脏组织以及外周血中共刺激分子B7-2/CD28的表达上调与急性排斥反应的发生密切相关。       

Peritubular capillary C4d deposition in chronic allograft dysfunction

Peritubular capillary (PTC) C4d staining represents a marker for acute humoral rejection, however, the impact of positive staining on chronic allograft dysfunction has received little attention. Ninety-three renal allograft biopsies from 93 patients were selected from a total of 174 renal allograft biopsies, which were obtained 6 months or more after transplantation (median: 89 months). Fresh frozen renal tissue was stained with monoclonal antibody against C4d. Sixteen of 93 biopsies showed C4d staining in PTC. C4d staining was positive in 40% of acute rejection cases (n=15) and 21% of chronic rejection cases (n=24). When the samples were divided according to C4d positivity, the C4d (+) group had a higher proportion of acute rejection than the C4d (-) group. However, no significant difference was observed between the two groups in terms of the prevalence of chronic rejection. Degrees of histological injury including tubulitis, interstitial inflammation and interstitial fibrosis were not significantly different between C4d (+) and C4d (-) groups. However, the 2-year graft survival rate after biopsy was lower in the C4d (+) group than in the C4d (-) group (24.8% versus 59.0%, p=0.1255). C4d staining in PTC is associated with late acute rejection, but not with chronic rejection based on conventional morphologic criteria in patients with chronic allograft dysfunction.     

HLA PROFILE OF ETHNIC PAKISTANI POPULATION GROUPS

Acute rejection is a major determinant of chronic allograft dysfunction and graft survival. This study evaluated the effect of basiliximab (Simulect^®), a 156-kDa chimeric monoclonal antibody (human and murine) directed against the alpha chain of the interleukin (IL)-2 receptor of human lymphocytes, on acute rejection in pediatric renal transplantation. Data were collected from two pediatric renal transplantation centers. Forty transplantations (22 males and 18 females; mean age 14.8±3.6 years) were performed between 1996 and 2001. Twelve of the grafts came from cadaveric donors and 28 from living-related donors. Twenty-four of the patients were on hemodialysis, 15 were on peritoneal dialysis, and one case was a pre-emptive transplantation. All patients were placed on triple-drug immunosuppression [prednisolone + (azathioprine or mycophenolate mofetil) +(cyclosporine or tacrolimus)]. Basiliximab was also administered in 17 cases. The respective rates of biopsy-proven acute rejection in the basiliximab group and the standard-regimen group were 0% vs. 17.4% ( P >0.05) at 1 month post-transplantation; 0% vs. 26.1% ( P <0.05) at 3 months; and 0% vs. 26.1% ( P <0.05) at 6 months. Thirty and 16 patients had completed 1- and 3-year follow ups, respectively, at the time of writing; the 1- and 3-year graft survival rates were 96% (29/30) and 81% (13/16), respectively.
Basiliximab significantly reduced the rates of acute rejection at 3- and 6 months post-pediatric renal transplantation. It was well tolerated by all patients, and caused no significant adverse effects. The effect of basiliximab on long-term graft survival and chronic allograft dysfunction deserves further investigation.     

Reduced CD40L expression on ex vivo activated CD4+ T-lymphocytes from patients with excellent renal allograft function measured with a rapid whole blood flow cytometry procedure

BACKGROUND: The CD40-CD40L (CD154) costimulatory pathway plays a critical role in the pathogenesis of kidney allograft rejection. In renal transplant biopsies, CD4+CD40L+ graft-infiltrating cells were detected during chronic rejection in contrast to acute rejection episodes. Using a rapid noninvasive FACS procedure, we were able to demonstrate CD40L upregulation in peripheral blood of patients with chronic renal allograft dysfunction. MATERIALS AND METHODS: Whole blood from recipients of renal allografts was stimulated with PMA and ionomycin and measured by flow cytometry. Patients were assigned to three groups based on transplant function. Group 1: 26 patients with excellent renal transplant function; group 2: 28 patients with impaired transplant function; group 3: 14 patients with chronic allograft dysfunction and group 4: 8 healthy controls. RESULTS: The median percentage +/- SEM of CD4+/CD40L+ cells stimulated ex vivo at 10 ng/ml PMA was as follows: group 1: 28.3 +/- 4.1%; group 2: 18.4 +/- 2.4%; group 3: 50.1 +/- 5.0% and group 4: 40.4 +/- 3.4%. Subdivisions of groups 2 and 3 resulted in different CD40L expression patterns. Patients with increased serum creatinine since the initial phase after transplantation (groups 2a and 3a) revealed a higher percentage of CD4+CD40L+ cells than patients showing a gradual increase over time (groups 2b and 3b). Consequently, patients of group 3a exhibited a significantly reduced transplant function compared with those of group 3b. CONCLUSION: After PMA + ionomycin stimulation, patients with excellent kidney graft function displayed significantly reduced expression of CD40L surface molecules on CD4+ cells early after transplantation. Those with a chronic dysfunction of the renal graft showed significantly more CD4+ cells expressing CD40L compared to the other transplanted groups. These results demonstrate that the percentage of CD4+CD40L+ cells stimulated ex vivo in peripheral blood may be a valuable marker for chronic allograft nephropathy.     

Interleukin 2 receptor expression in renal biopsies and the diagnosis of acute allograft rejection

The aim of the study is to test the diagnostic usefulness of assessing interleukin 2 receptor (IL2R) expression in infiltrating lymphocytes in renal biopsies from patients with suspected acute renal allograft rejection and to compare the NIH-CCTT and the Banff 97 systems of classifying the histopathologic changes in acute renal allograft rejection. The expression of interleukin 2 (IL2) and IL2R, as shown immuno-histochemically, is the final step in T cell mediated acute renal allograft rejection. Renal biopsies obtained from 40 patients clinically suspected to have early acute allograft rejection were examined histologically to diagnose acute allograft rejection and classified by the two systems. Frozen sections of the biopsies were stained with specific antibody for the presence of IL2R. 31 of the 40 patients were histologically and clinically confirmed to have acute allograft rejection. There was significant correlation with this diagnosis and the demonstration of IL2R on infiltrating lymphocytes. The CCTT system of grading correlated better with the presence of IL2R and the confirmed diagnosis of acute allograft rejection. The immunohistochemical demonstration of IL2R is a useful adjunct in the evaluation of biopsies suspected to show changes of acute cellular rejection. Since IL2 expression reflects the relative proportion of activated lymphocytes in the cellular infiltrate, it is proposed that the degree of IL2 expression may reflect the response of the use of monoclonal antibodies (Humanised/Chimaerised) as anti rejection therapy.     

NEW ADVANCES IN THE MOLECULAR BIOLOGY OF ISCHEMIA/REPERFUSION: MAPK PATHWAYS

Elevated levels of intact parathyroid hormone (iPTH) are not uncommon after renal transplantation, and this disturbance may affect renal allograft function. This study investigated whether iPTH levels were related to histopathological findings of acute rejection in kidney graft biopsies. Thirty-eight renal transplant recipients (29 males, nine females; mean age 29.5±10.3years) were studied. Renal allograft biopsy was performed in each case to investigate increased creatinine levels. Lymphocyte and macrophage infiltration of the interstitium was evaluated immunohistochemically with monoclonal antibodies to CD3 and CD68. Expression of HLA-DR and fibronectin were also studied. The proportion of interstitial cell staining was graded semiquantitatively. Serum calcium, phosphorus and iPTH levels before and after the renal transplantation were compared. Patients were grouped according to their stabilized iPTH levels at 3 months post-transplantation. Group I ( n =13) exhibited persistently elevated iPTH levels (levels elevated since surgery), and Group II ( n =25) showed persistently low-normal iPTH levels. The mean age of the Group I patients was significantly lower, and their serum calcium levels post-transplantation were significantly higher than those in Group II ( P =0.02 and P =0.03, respectively). Persistent elevation of iPTH was strongly associated with interstitial T-cell density, macrophage density, and glomerular fibronectin expression in the renal allograft biopsies ( P <0.05). Also, the time to fibrosis was significantly shorter in Group I ( P =0.02). The study revealed that PTH has a potent immunomodulatory effect, and that persistent hyperparathyroidism has a significant impact on renal allograft outcome. The data indicate that the serum iPTH level is a valuable indicator of immune response, and that regular monitoring of serum iPTH levels is essential after renal transplantation.     

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。     

Gammadelta T cells and interleukin-6 levels could provide information regarding the progression of human renal allograft

We have determined the percentage of alphabeta and gammadelta T cells by flow cytometry as well as serum interleukin-6 (IL-6) and soluble interleukin-6 receptor (sIL-6R) levels by enzyme-linked immunosorbent assay in kidney allograft recipients with acute, chronic or stable graft evolution. The percentage of CD4 and CD8 T cells in transplanted patients was lower than in the control group (P < 0.001) with the exception of CD8 gammadelta T cells from patients with stable evolution (P > 0.05). The serum levels of IL-6 and sIL-6R in acute and chronic rejection were higher than in the controls (P < 0.05). No differences in IL-6 levels were observed between the stable evolution and the control groups (P > 0.05). The levels of sIL-6R were higher in stable evolution patients than in the controls (P < 0.05) and no differences were observed between the chronic and stable evolution patients (P > 0.05). IL-6 decreased in patients with a favourable evolution, increased in those with an increased renal dysfunction and was maintained when the renal dysfunction was not modified. These results suggest that gammadelta T cells could participate in renal allograft maintenance and that IL-6 but not sIL-6R serum levels may provide a prognostic marker for measuring the evolution of kidney allograft.     

The value of electron microscopy in the diagnosis of chronic renal allograft rejection.

The main causes of the late dysfunction of renal allografts are chronic rejection and chronic transplant nephropathy. Both are clinicopathologic entities, with a similar clinical presentation, but different histologic appearances. Chronic rejection is characterized by the presence of alloantigen-induced lesions (transplant arteriopathy and transplant glomerulopathy), and chronic transplant nephropathy by nonspecific sclerosing changes. The incidence of transplant arteriopathy and transplant glomerulopathy is relatively low. Electron microscopy (EM) may overcome the limitations in the histologic diagnosis of chronic rejection, because it verifies alloantigen-induced chronic microvasculopathy in the peritubular capillaries (transplant capillaropathy), and identifies transplant glomerulopathy more precisely than does light microscopy. To assess the value of EM in chronic rejection diagnosis, a retrospective search for transplant capillaropathy and transplant glomerulopathy was performed in a consecutive series of 91 biopsies performed > or = 6 months after implantation (median: 26 months, range 6-186) and the diagnoses were reclassified on the basis of the ultrastructural findings. The definitions used were: transplant capillaropathy: a peritubular capillary profile with seven or more circumferential basement membrane layers, or at least three profiles with five or six circumferential layers; ultrastructurally verified transplant glomerulopathy: thickening of the capillary wall in at least three loops in consequence of the widening of the subendothelial space by abnormal basement membrane material, and the formation of a new layer(s) of basal lamina; and chronic rejection: the presence of transplant capillaropathy and/or transplant glomerulopathy and/or transplant arteriopathy. Histologically, chronic transplant nephropathy, chronic rejection, chronic cyclosporine nephrotoxicity, glomerulonephritis, acute rejection, "suspicious" for acute rejection, and "others" were diagnosed in 37%, 34%, 21%, 19%, 57%, 30%, and 5% of the specimens, respectively. The results of EM increased the diagnosis of chronic rejection to 69% of the cases, and decreased chronic transplant nephropathy to 15%. The individual incidence of transplant capillaropathy and transplant glomerulopathy was 79% and 57%, respectively, and their cumulative incidence was 92%. Five biopsies exhibited merely transplant arteriopathy. A late dysfunction typically had more than one cause; the most frequent combination was chronic rejection and acute rejection. In conclusion, the EM search for transplant capillaropathy and transplant glomerulopathy doubled the frequency of the diagnosis of chronic rejection. Currently, the evaluation of renal allograft biopsies from recipients with a late dysfunction relies on standard light microscopy. Because light microscopy per se proved to be insensitive in the diagnosis of chronic rejection, incorporation of EM into the evaluation of late dysfunction biopsies is strongly recommended.     

移植肾穿刺20例活组织病理检查的临床价值

背景：根据临床表现、无创性辅助检查及临床经验来判断肾移植后移植肾功能异常的原因常常会陷入困境,经皮肾穿刺活组织检查则可提供重要的循证学依据。 目的：探讨移植肾穿刺活组织检查的临床意义。 方法：回顾性分析20例移植肾穿刺活检病理资料及相应调整治疗后的临床结果资料。 结果与结论：20例患者病理诊断为急性排斥反应7例(35%),可疑性急性排斥反应2例(10%),可疑性急性排斥反应加免疫抑制剂中毒1例(5%),免疫抑制剂中毒3例(15%),慢性移植肾肾病3例(15%),未见明确异常3例(15%),肾小管坏死1例(5%)。相应调整治疗方案后,移植肾功能恢复正常9例(45%),移植肾功能好转7例(35%),肾功能无明显变化3例(15%),肾功能轻度恶化1例(5%)。结果可见肾穿刺活组织检查对肾移植后肾功能异常的病因诊断有重要的价值,有利于指导治疗方案的调整。     

494例次移植肝穿刺活检病理组织学分析

目的 通过对354例(494例次)移植肝穿刺活检组织进行病理分析，观察移植肝的组织学变化，探讨其出现肝功能不全的原因。方法 移植肝穿刺活检组织经10％中性福尔马林固定，快速石蜡连续切片，常规HE染色。部分病例做VG、Masson、PAS、网状纤维组织化学和免疫组织化学染色，抗体为HBsAg、HBcAg、HcVAg、CMV、CD8、CD4、CK19。对排斥反应病例，依照国际统一的BANFF标准进行急性排斥反应分级，应用排斥活动指数(RAI)进行排斥反应程度评分。结果 急性细胞性排斥反应最常见，180例(50．85％)，慢性排斥反应11例(3．11％)，再灌注缺血损伤20例(5．65％)，胆汁淤滞及急慢性小胆管炎64例(18．08％)，药物性肝损害18例(5．08％)，移植肝无功1例(0．28％)，CMV感染24例(6．78％)，乙肝病毒再感染及乙肝复发27例(7．63％)，丙型肝炎复发2例(0．56％)，原发性硬化性胆管炎复发1例(0．28％)，难以诊断6例(1．69％)。结论 移植肝穿刺活检对移植术后并发症的诊断及选择治疗方案具有重要价值。     

肾移植及移植后的血管并发症

背景：尽管肾移植手术技术已相当成熟,但各移植中心难免会出现术中及术后的血管并发症。 目的：分析肾移植过程中及移植后血管并发症的诊断与处理。 方法：回顾性分析 11 例肾移植过程中、移植后血管并发症资料。移植过程中并发症：动脉吻合口狭窄2例,肾静脉横断1例,髂外动脉硬化斑块剥脱堵塞肾动脉3例,移植肾静脉扭曲1例;移植后并发症：肾外型假性动脉瘤2例,吻合口狭窄1例,移植肾动脉压迫梗阻1例。 结果与结论：肾移植中吻合口狭窄2例移植后恢复良好,分别于移植后6年及11年移植肾失功;肾静脉横断1例随访12年移植肾失功;髂外动脉硬化斑块剥脱堵塞肾动脉3例,1例移植肾当即失功,另2例分别随访6年及2年,现移植肾功能良好;移植肾静脉扭曲1例移植后出现移植肾功能延迟恢复,1个月后因重症肺部感染死亡。移植后发生的血管并发症中肾外型假性动脉瘤2例移植肾均失功;吻合口狭窄1例经置入支架后肾功能恢复正常,已随访18个月;移植肾动脉压迫梗阻1例移植后出现移植肾功能延迟恢复,3周后因重症肺部感染死亡。结果说明肾移植过程中及移植后发生血管并发症,只要诊断准确、处理及时,可取得比较满意的治疗效果。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

同种异体尸体移植肾组织中Bcl—2蛋白的检测及意义

选择Ｂｃｌ－２蛋白质作为标记蛋白，着重观察移植肾组织结构和肾功能正常时，以及发生急性排异时其肾组织浸润细胞中该蛋白阳性细胞的变化，结果表明，未发生ＡＣＲ的肾组织结构正常的标本中，Ｂｃｌ－２阳性细胞数仅在术后３个月时高于术前水平；移植肾发生ＡＣＲ时，Ｂｃｌ－２阳性细胞增加明显，是未发生ＡＣＲ肾组织的７．６９倍。对移植肾间质Ｂｃｌ－２阳性细胞数与其浸润的ＣＤ４或ＣＤ８细胞数回归相关性分析发现，移植肾总     

尸肾移植急性排斥反应与患者淋巴细胞CD62L、CD11a、CD4、CD8表达的关系

为探讨同种异体尸肾移植排斥反应病人淋巴细胞CD62L、CD11a表达与T细胞亚群及CD4/CD8的关系和意义。利用单克隆抗体 流式细胞仪荧光免疫技术 ,测定 10例肾移植排斥反应病人术后不同时间外周血淋巴细胞CD62L、CD11a、CD4、CD8表达并计算CD4/CD8。结果 ,肾移植病人排斥反应时其CD62L (4 6 1± 18 7vs 31 3± 10 5 ,P <0 0 1)、CD11a (4 9 5±2 0 2vs 31 9± 12 4,P <0 0 1)、CD4(2 4 4± 7 7vs 17 9± 7 4,P <0 0 1)、CD8(14 7± 2 9vs 10 4± 3 2 ,P <0 0 5 )表达均较排斥前明显增加 ,抗排斥治疗后CD11a (14 8± 6 2vs 49 5± 2 0 2 ,P <0 0 1)、CD4(15 8± 6 4vs 2 4 4± 7 7,P <0 0 5 )和CD4/CD8(1 2 8± 0 6vs 1 73± 0 79,P <0 0 5 )均明显下降。CD62L变化和CD8呈明显正相关 (r=0 9779,P <0 0 5 )。认为淋巴细胞CD62L、CD11a、CD4、CD8表达及CD4/CD8与肾移植排斥反应密切相关。免疫抑制剂 ,尤其甲基强的松尤能明显抑制淋巴细胞CD11a、CD4表达和CD4/CD8比值可能是其发挥抗排斥作用的重要机制。     

Proteomic profiling of renal allograft rejection in serum using magnetic bead–based sample fractionation and MALDI-TOF MS

Proteomics is one of the emerging techniques for biomarker discovery. Biomarkers can be used for early noninvasive diagnosis and prognosis of diseases and treatment efficacy evaluation. In the present study, the well-established research systems of ClinProt Micro solution incorporated unique magnetic bead sample preparation technology, which, based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), have become very successful in bioinformatics due to its outstanding performance and reproducibility for discovery disease-related biomarker. We collected fasting blood samples from patients with biopsy-confirmed acute renal allograft rejection (n = 12), chronic rejection (n = 12), stable graft function (n = 12) and also from healthy volunteers (n = 13) to study serum peptidome patterns. Specimens were purified with magnetic bead–based weak cation exchange chromatography and analyzed with a MALDI-TOF mass spectrometer. The results indicated that 18 differential peptide peaks were selected as potential biomarkers of acute renal allograft rejection, and 6 differential peptide peaks were selected as potential biomarkers of chronic rejection. A Quick Classifier Algorithm was used to set up the classification models for acute and chronic renal allograft rejection. The algorithm models recognize 82.64% of acute rejection and 98.96% of chronic rejection episodes, respectively. We were able to identify serum protein fingerprints in small sample sizes of recipients with renal allograft rejection and establish the models for diagnosis of renal allograft rejection. This preliminary study demonstrated that proteomics is an emerging tool for early diagnosis of renal allograft rejection and helps us to better understand the pathogenesis of disease process.     

肾移植后吗替麦考酚酯联合低剂量他克莫司加皮质激素的应用

背景：足量吗替麦考酚酯联合低剂量他克莫司和皮质激素可能是目前针对肾移植受者的理想治疗方案,该方案因其具有低肾毒性以及较少的不良反应和较强的免疫抑制作用已在临床上开始逐渐普及。 目的：以吗替麦考酚酯联合标准剂量他克莫司加皮质激素为对照,评估吗替麦考酚酯联合低剂量他克莫司加皮质激素在肾移植患者中的疗效和安全性。 方法：210例首次接受单一器官同种异体移植的肾移植成人受者被随机分配到他克莫司标准剂量组(n=104)和他克莫司低剂量组(n=106),并接受12个月的治疗。主要疗效指标包括肾移植后第12个月慢性移植物损伤指数(CADI)以及肾小球滤过率;次要疗效指标主要包括急性排斥反应发生率、治疗失败率以及患者和移植肾的存活率等;同时对新发移植后糖尿病,新发高血压,新发高血脂等安全性指标进行评价。 结果与结论：两组绝大多数患者使用了足量的吗替麦考酚酯(1.5 g/d及以上)。在他克莫司剂量方面,他克莫司标准剂量组大多数受试者的实际血药浓度水平偏低,与低剂量组的实际血药浓度水平类似,由此反映了吗替麦考酚酯联合低剂量他克莫司和皮质激素方案已广泛为目前临床医师接受和使用。因此,两组也表现出类似的疗效和安全性：他克莫司标准剂量组和低剂量组肾移植后12个月肾脏病理改变的平均CADI评分分别为1.82分和2.13分(P=0.081 3),平均肾小球滤过率分别为77.08 mL/min和       80.12 mL/min(P=0.794 9),急性排斥反应发生率分别为2.6%和5.2%(P=0.681 2),患者和移植肾存活率分别高达100%和99.1%(P=1.000 0)。在安全性方面,他克莫司标准剂量组和低剂量组新发移植后糖尿病的比例分别为2.9%和1.9%,新发高血脂的比例分别为2.9%和3.8%。结果显示在吗替麦考酚酯联合他克莫司和皮质激素的肾移植免疫抑制治疗方案中,足量吗替麦考酚酯的使用,可以减少他克莫司的剂量,在保持较强的免疫抑制作用即成功地降低急性排斥反应发生率的同时,显著减少他克莫司所致的肾毒性、高血脂和新发糖尿病等不良反应,较好地达到了疗效和毒性间的平衡。     

Transcriptional Profiles in Urine During Acute Rejection, Bacteriuria, CMV Infection and Stable Graft Function After Renal Transplantation

Acute rejection remains an important cause of renal allograft dysfunction and the need for accurate diagnosis is essential to treat transplant recipients successfully. Molecular markers in urine may serve as a diagnostic tool in acute rejection, but controversy still exists regarding the uniqueness of these biomarkers. We measured mRNA of perforin (PRF), granzyme B (GZMB) and granulysin (GNLY) normalized to cyclophilin B in urine specimens from 24 renal allograft recipients with acute rejection, 12 with bacteriuria, 11 with cytomegalovirus (CMV) infections and 17 controls with stable graft function. Measurements were performed using a real-time polymerase chain reaction assay. mRNA levels (means [95% CI]) for all three markers were significantly higher in recipients with acute rejection compared with controls: PRF (0.23 [0.12–0.42] versus 0.04 [0.02–0.07] P  < 0.001), GZMB (0.14 [0.09–0.23] versus 0.05 [0.03–0.08] P  = 0.003), GNLY (0.24 [0.14–0.41] versus 0.06 [0.03–0.11] P  = 0.001). GZMB and GNLY levels during acute rejection were significantly higher when compared with bacteriuria ( P  = 0.011 and P  = 0.005 respectively), and PRF level during acute rejection was significantly elevated compared with CMV infection ( P  = 0.015). No significant difference was found when comparing marker levels during bacteriuria and CMV infection to controls. Urinary mRNA levels of PRF, GZMB and GNLY are significantly elevated during acute rejection but not during bacteriuria or CMV infections when compared with recipients with stable graft function. The ability to differentiate acute rejection from bacteriuria and CMV infections was only present for some of the markers, that is why careful consideration should be given before applying this technique to clinical practice.     

Characterization of ectopic lymphoid structures in different types of acute renal allograft rejection

We hypothesize that T cells such as interleukin (IL)‐21⁺B cell lymphoma 6 (BCL6)⁺ T follicular helper cells can regulate B cell‐mediated immunity within the allograft during acute T cell‐mediated rejection; this process may feed chronic allograft rejection in the long term. To investigate this mechanism, we determined the presence and activation status of organized T and B cells in so‐called ectopic lymphoid structures (ELSs) in different types of acute renal allograft rejection. Biopsies showing the following primary diagnosis were included: acute/active antibody‐mediated rejection, C4d⁺ (a/aABMR), acute T cell‐mediated rejection grade I (aTCMRI) and acute T cell‐mediated rejection grade II (aTCMRII). Paraffin sections were stained for T cells (CD3 and CD4), B cells (CD20), follicular dendritic cells (FDCs, CD23), activated B cells (CD79A), immunoglobulin (Ig)D, cell proliferation (Ki67) and double immunofluorescent stainings for IL‐21 and BCL6 were performed. Infiltrates of T cells were detected in all biopsies. In aTCMRI, B cells formed aggregates surrounded by T cells. In these aggregates, FDCs, IgD and Ki67 were detected, suggesting the presence of ELSs. In contrast, a/aABMR and aTCMRII showed diffuse infiltrates of T and B cells but no FDCs and IgD. IL‐21 was present in all biopsies. However, co‐localization with BCL6 was observed mainly in aTCMRI biopsies. In conclusion, ELSs with an activated phenotype are found predominantly in aTCMRI where T cells co‐localize with B cells. These findings suggest a direct pathway of B cell alloactivation at the graft site during T cell mediated rejection.     

肾移植受者血清sFas和sFasL变化及在早期急性排异反应预测中的应用

背景：细胞凋亡在移植免疫和移植物功能丧失发生过程中起十分重要的作用,其中Fas/FasL系统被认为是细胞凋亡参与肾移植的急性排异反应过程的主要途径之一。 目的：分析肾移植受者术后血清sFas和sFasL水平变化及其在预测早期急性排异反应中的应用价值。 方法：肾移植受者80例分为肾功能稳定组(49例)、急性排斥反应组(23例)和环孢素A中毒组(8例)。另选择性别、年龄与肾移植受者相匹配的健康体检者50例为对照组。肾移植受者术后均常规使用环孢素A+硫唑嘌呤+泼尼松三联免疫抑制治疗。发生急性排斥反应时给予每日甲基强的松龙6~8 mg/kg冲击治疗,3 d为1个疗程。采用ELISA法检测患者手术前后的血清sFas和sFasL水平。 结果与结论：肾移植组患者手术前的血清sFas和sFasL水平均明显高于对照组(P  < 0.05)。急性排斥反应组血清sFas、sFasL水平高于相同时间段肾功能稳定组(P < 0.05)。环孢素A中毒的肾移植患者术后各时间点血清sFas、sFasL水平变化与肾功能稳定组基本相同,差异无显著性意义。提示动态监测血清sFas、sFasL水平可能对早期诊断及鉴别诊断肾移植急性排斥反应具有重要参考价值。     

Participation of RANTES and T-cell apoptosis in human renal allograft

The aim of this study was to evaluate the serum RANTES (Regulated upon Activation of Normal T cell Expressed and Secreted) levels and the expression of CCR5, as well as the percentage of apoptotic cells, in peripheral T lymphocytes from renal transplanted patients with acute rejection (AR), chronic rejection (CR) or stable evolution (SE). RANTES serum levels were determined by enzyme-linked immunoadsorbent assay and CCR5 expression, as well as the percentage of apoptotic lymphocytes, on a FACScan flow cytometer. After staining with different antibodies, the cells were subjected to three-colour flow cytometric analysis. Data analysis was performed using winmdi 2.5 software. The serum RANTES level and percentages of CCR5/CD4 and CCR5/CD8 T lymphocytes in CR, AR and SE were lower than that in the control group (P <0.05). The level of CD4 and CD8 T lymphocytes in early apoptosis was higher in AR patients than in CR, SE or C groups (P <0.05). In the case of late apoptosis, the percentage of apoptotic/necrotic cells was higher in the CR than AR, SE or C groups (P <0.05). The RANTES serum levels and the percentage of peripheral CCR5 T lymphocytes would not indicate the renal allograft state. The increase of early apoptotic T lymphocytes could be a marker of AR process and could also indicate the initial step in reducing the cytotoxic T lymphocytes, thus favouring the graft evolution.