Delayed acute rejection episodes in cyclosporine-treated renal transplant recipients.

Of 109 cyclosporine-treated cadaveric renal allograft recipients, 45 were free of acute rejection in the first 4 weeks after transplantation. Eleven of 45 (24%) subsequently had delayed, biopsy-proven first rejection episodes 34-61 days after grafting, often after discharge from the hospital. Delayed rejectors had significantly higher plasma creatinine levels at all times during the first posttransplant month than 34 nonrejectors. Trough serum cyclosporine levels were similar in the two groups, although by the 4th week oral cyclosporine dose was significantly lower in the delayed rejection group. Two-thirds of those patients who had serum creatinine levels greater than or equal to 260 mumol/l at 2 weeks and greater than or equal to 225 mumol/l at 3 weeks had a delayed acute rejection episode. Renal transplant recipients treated with cyclosporine who have serum creatinine levels at or above these levels should be aggressively worked up and closely followed for the development of delayed acute rejection.     

ICAM-1和VCAM-1测定在肾移植急性排斥反应中的临床意义

目的探讨血清可溶性细胞间粘附分子１（ｓＩＣＡＭ１）和可溶性血管细胞粘附分子１（ｓＶＣＡＭ１）在肾移植急性排斥反应中的临床意义。方法采用酶联免疫吸附法（ＥＬＩＳＡ）动态监测５６例肾移植患者术后ｓＩＣＡＭ１和ｓＶＣＡＭ１水平的变化。结果肾移植患者术后ｓＩＣＡＭ１和ｓＶＣＡＭ１水平呈规律性变化,急性排斥反应组ｓＩＣＡＭ１为（３９０．６±９１．０）ｎｇ／ｍｌ,ｓＶＣＡＭ１为（１９５７．１±４０３．１）ｎｇ／ｍｌ,明显高于移植肾功能稳定组的（１３７．３±１６．８）ｎｇ／ｍｌ、（１１１８．４±２１０．４）ｎｇ／ｍｌ和ＣｓＡ肾中毒组的（１３２．７±２４．８）ｎｇ／ｍｌ、（１２８５．８±２７０．５）ｎｇ／ｍｌ,差异有非常显著性（Ｐ＜０．０１）。结论ｓＩＣＡＭ１和ｓＶＣＡＭ１可以作为肾移植术后急性排斥反应的免疫学监测指标。     

Comparative evaluation of 'TaqMan' RT-PCR and RT-PCR ELISA for immunological monitoring of renal transplant recipients

By sequentially monitoring cytokine gene expression (using RT-PCR ELISA technology) in peripheral blood cells of renal transplant recipients in the early post-operatively period we have shown that expression patterns correlate with clinical events, namely acute allograft rejection. This strategy may have the potential of predicting acute rejection prior to clinical detection. Unfortunately, the technique used was time consuming and only semi-quantitative and, therefore, not suitable for clinical application. In this study, we have sought to confirm the results of the early work using a real time quantitative RT-PCR technique ('TaqMan'), which may be applicable in the clinical laboratory. 'TaqMan' primers and probes were designed for Interleukin (IL)-4 and IL-10 using Primer Express software. Cytokine gene expression for both cytokines was re-measured in stored cDNA samples from 27 non-rejectors and 14 patients experiencing an episode of biopsy proven acute rejection. Compared to pre-transplant levels, IL-4 gene expression fell significantly on post-operative days 2 and 7 before returning to baseline values by day 14 in the non-rejectors. In the rejectors, the initial significant fall was again seen, but with an earlier return to pre-transplant levels at the time of rejection diagnosis. This was followed by a further significant fall in levels 48 h after the initiation of anti-rejection therapy. These different patterns for rejectors and non-rejectors were seen using both techniques. For IL-10, gene expression increased significantly following transplantation throughout the study period when compared to baseline values. This pattern was seen using both techniques. In the rejectors, there were different patterns seen depending on the technique used. When using RT-PCR ELISA, the initial rise was again seen followed by a return to baseline values at the time of rejection diagnosis followed by a further significant rise in gene expression after the start of anti-rejection treatment. The pattern resembled those of the non-rejectors when expression was measured using 'TaqMan'. This study has confirmed that sequential monitoring of cytokine gene expression, measured in peripheral blood mononuclear cells, detects significant changes that correlate with clinical events in renal transplant recipients, including acute rejection, although not all changes detected with RT-PCR ELISA were confirmed. Therefore, real time quantitative RT-PCR technology may be useful in monitoring the immunological status of these patients in the early post-operative period.     

肾移植后测定血清可溶性细胞间粘附分子-1的临床意义

目的　探讨血清可溶性细胞间粘附分子 1(sICAM 1)在肾移植术后免疫学监测中的价值。方法　采用酶联免疫吸附法 (ELISA)动态监测 5 6例肾移植患者术后血清sICAM 1的变化。结果　肾移植患者术后发生排斥反应时 ,其sICAM 1水平 (390 .6 μg/L±91.0 μg/L)明显高于移植肾功能稳定者 (137.3μg/L±16 .8μg/L)和环孢素A(CsA)肾中毒者 (132 .7μg/L±2 4.8μg/L) ,差异有极显著性 (P <0 .0 1) ;抗排斥反应治疗有效后 ,sICAM 1逐渐降至正常水平 ;CsA肾中毒者的sICAM 1水平无明显变化。结论　肾移植术后动态监测患者血清sICAM 1水平的变化 ,有助于急性排斥反应的诊断、鉴别诊断及抗排斥治疗的疗效评价 ,可以作为肾移植术后急性排斥反应的免疫学监测指标。     

Immunomonitoring of renal transplant recipients in the early posttransplant period by sequential analysis of chemokine and chemokine receptor gene expression in peripheral blood mononuclear cells

INTRODUCTION: We sought to determine whether sequential changes in chemokine ligand/receptor gene expression in the early posttransplant period of human renal allografts can be detected in peripheral blood mononuclear cells (PBMCs) and whether any such changes are predictive of clinical events. METHODS: Blood samples from 106 renal transplant recipients and 29 donor nephrectomy patients were taken preoperatively and daily for 14 days. Within the study period 22 patients had biopsy-proven acute rejection. From each blood sample PBMCs were separated and gene expression levels for chemokines CCL3, CCL4, CCL5, CXCL10, and their receptors CCR1, CCR5, and CXCR3, were determined using real-time quantitative PCR. RESULTS: Different gene expression patterns were seen between the rejector and nonrejector groups with decreases in CCL4 and CCR5 expression on days 6 to 8 and increases in CCR1 expression on days 9 and 10 posttransplant. With CXCL10, decreases in expression were seen in the nonrejector group but increases were seen in the rejector group posttransplant. With data aligned to time of rejection diagnosis, statistically significant increases, that preceded the clinical detection of acute rejection were seen in CCR1 and CXCL10 expression. Both their expression levels returned to pretransplant baseline values after successful antirejection therapy. CONCLUSION: We have demonstrated that changes in chemokine receptor/ligand gene expression by sequential monitoring in PBMCs can be detected in the early posttransplant period. In particular, CCR1 and CXCL10, which showed increased expression prior to rejection and returned to baseline levels with antirejection therapy, may have potential use in immunomonitoring and as predictive factors of rejection prior to its clinical manifestation.     

肾移植后血清白细胞介素2、可溶性白细胞介素2受体和白细胞介素6的监测

动态监测６０例肾移植患者术后２个月内血清白细胞介素２（ＩＬ－２）、可溶性ＩＬ－２受体（ｓＩＬ－２Ｒ）和白细胞介素６（ＩＬ－６）的变化。结果发现发生急性排斥反应时，上述细胞因子的升高较临床诊断提早数天，并且显著高于环孢素Ａ肾中毒组；对甲泼尼龙敏感的排斥反应，抗排斥治疗数天后上述因子下降到排斥前水平。提示肾移植术后动态监测患者血清ＩＬ－２、ｓＩＬ－２Ｒ和ＩＬ－６有助于急性排斥反应的早期诊断、鉴别诊断、及时治疗和甲泼尼龙抗排斥的疗效评价。     

Intragraft FOXP3 mRNA expression reflects antidonor immune reactivity in cardiac allograft patients

BACKGROUND: Regulatory FOXP3+ T cells control immune responses of effector T cells. However, whether these cells regulate antidonor responses in the graft of cardiac allograft patients is unknown. Therefore, we analyzed the gene expression profiles of regulatory and effector T-cell markers during immunological quiescence and acute rejection. METHODS: Quantitative real-time polymerase chain reaction was used to analyze mRNA expression levels in time-zero specimens (n=24) and endomyocardial biopsies (EMB; n=72) of cardiac allograft patients who remained free from rejection (nonrejectors; n=12) and patients with at least one histologically proven acute rejection episode (rejectors; International Society for Heart and Lung Transplantation [ISHLT] rejection grade>2; n=12). RESULTS: For all analyzed regulatory and effector T-cell markers, mRNA expression levels were increased in biopsies taken after heart transplantation compared with those in time-zero specimens. Posttransplantation, the FOXP3 mRNA levels were higher in EMB assigned to a higher ISHLT rejection grade than the biopsies with grade 0: the highest mRNA levels were detected in the rejection biopsies (rejection grade>2; P=0.003). In addition, the mRNA levels of CD25, glucocorticoid-induced TNF receptor family-related gene, cytotoxic T lymphocyte-associated antigen 4, interleukin-2, and granzyme B were also significantly higher in rejecting EMB than in nonrejecting EMB (rejection grade相似文献   

肾移植后尿中可溶性细胞间粘附分子-1浓度监测的临床意义

目的：评价肾移植后患者尿中可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）浓度变化的临床意义。方法：动态检测２０例肾移植患者术后２个月内尿中ｓＩＣＡＭ－１浓度的变化。结果：急性排斥反应（ＡＲ）时,ｓＩＣＡＭ－１的升高较临床诊断提早数天,并且显著高于环孢素Ａ肾中毒组;对甲基氢化泼尼松敏感的排斥反应,抗排斥治疗数天后ｓＩＣＡＭ－１下降到排斥前水平。结论：肾移植术后动态监测受者尿中ｓＩＣＡＭ－１浓度有助于ＡＲ     

Serological responses to cytomegalovirus during renal transplant rejection.

We studied the role played by CMV in kidney transplant rejection by recording serological responses to CMV replication in cadaver graft recipients and recording clinical graft rejection by monitoring acute changes in renal function and the appearance of antidonor lymphocyte antibody (anti-Dab). If CMV plays a significant role in rejection, clinical rejection should correlate with CMV activity; if CMV does not play a role, clinical rejection would be likely to correlate with anti-Dab but not necessarily with CMV activity. We selected retrospectively 18 rejectors and 18 nonrejectors by clinical criteria and assayed for anti-Dab (by fluorescence-activated flow cytometry) and CMV antibody (by complement fixation and Western blot for IgG and IgM) over a 3-6-month period after transplantation. Primary CMV infection occurred in 8 of 12 (67%) CMV seronegative graft recipients and reactivation or reinfection occurred in 16 of 24 (67%); 12 of 14 (86%) rejectors developed anti-Dab compared with 2 of 18 (11%) nonrejectors (P less than 0.00001). Active CMV infection occurred in 11 of 18 (61.1%) rejectors and 13 of 18 (72.2%) nonrejectors (P = 0.36), and in 8 of 15 (53.3%) of those who developed anti-Dab and 12 of 17 (70.6%) of those who did not (P = 0.26). The results show no evidence to link CMV activity with kidney graft rejection.     

Low circulating regulatory T-cell levels after acute rejection in liver transplantation.

Immune regulatory CD4+CD25+ T cells play a crucial role in inducing and maintaining allograft tolerance in experimental models of transplantation (Tx). In humans, the effect of Tx and immunosuppression on the function and homeostasis of CD4+CD25+ regulatory T cells (Tregs) is not well characterized. In this study, the frequency of Tregs in liver transplant recipients was determined based on flow cytometric analysis of CD4, CD25, CD45RO, and cytotoxic T lymphocyte antigen (CTLA)-4 markers, and the suppressor activity of Tregs was assessed in a mixed-leukocyte reaction. A link between Tregs, acute rejection, and immune-suppressive treatment was investigated. Liver transplant recipients had significantly higher Treg levels in peripheral blood pre-Tx than healthy controls. After Tx, a significant drop in the Treg fraction was observed. This reduction of circulating Tregs was transient and was associated with immunosuppression. In recipients who did not develop rejection, a relative recovery of Treg levels was seen within the first year after Tx. Recipients who experienced an episode of steroid-treated acute rejection, however, had sustained low Treg levels. The suppressive activities of CD4+CD25+ Tregs from rejectors, nonrejectors, and healthy controls on proliferation and interferon (IFN)-gamma production were indistinguishable. In conclusion, the percentage of CD4+CD25+CD45RO+CTLA-4+ quadruple-positive Tregs in peripheral blood decreases significantly after liver Tx. Treatment with methylprednisolone during Tx and for acute rejection is associated with low circulating Tregs. Despite these quantitative differences between rejectors and nonrejectors, the suppressive quality of CD4+CD25+ Tregs is identical in both groups.     

Quantitative detection of changes in cytokine gene expression in peripheral blood mononuclear cells correlates with and precedes acute rejection in renal transplant recipients

Immunological monitoring of transplant recipients is an attractive concept. Cytokines provide an obvious focus for research, as they are central to the human immune response. This study aimed to identify cytokines whose sequential gene expression differentiated rejectors from non-rejectors immediately following renal transplantation. Forty-five renal transplant recipients (15 rejectors) and 13 living donors were recruited. Total RNA was extracted from the peripheral blood mononuclear cells and reverse transcribed. Cytokine gene expression levels of IL-4, IL-10, TNF-alpha and TGF-beta1 were measured using TaqMan. IL-10 expression increased significantly following donor surgery. IL-4 and TNF-alpha patterns clearly differentiated between rejectors and non-rejectors. In the rejectors significant increases occurred more than 48 h before clinical graft dysfunction. Negative predictive values were 76% and 80% for IL-4 and TNF-alpha, respectively. This study has identified two cytokines (IL-4 and TNF-alpha) whose gene expression patterns differentiate between rejecting and non-rejecting renal transplant recipients making immunological monitoring possible.     

IL—2R在急性细胞性排异反应的同种异体尸体移植肾组织中的检测…

为探讨白但纱－２受体在同种异体肾移植急性细胞性排异临床诊断的作用，着重观察移植肾发生和无ＡＣＲ时，其 间质浸润细胞中ＩＬ－２Ｒ阳性细胞的变化，及其与间质浸润的淋巴细胞的关系。     

The role of interleukin-10 in lung transplantation

Interleukin-10 (IL-10) is a pleiotropic cytokine and its main function is to limit and terminate inflammatory responses. Lung transplantation is a relatively young clinical field compared to the transplantation of other solid organs and long-term survival is still limited. Complications after lung transplantation include ischemia-reperfusion injury immediately after transplantation, acute rejection and infection within the first year after transplantation and chronic allograft dysfunction in form of bronchiolitis obliterans thereafter. In the setting of lung transplantation two key functions of IL-10 might be of interest: (1) the inhibition of inflammatory immune responses; and (2) the inhibition of T-cell mediated immune responses. In animal models, it has been shown that exogenous IL-10 is able to prevent posttransplant ischemia-reperfusion injury as well as to decrease acute rejection. It was also effective in preventing airway obliteration in an animal model of posttransplant bronchiolitis obliterans. Beneficial effects of IL-10 may be found early and late after lung transplantation. Location of IL-10 expression as well as the timing of administration seems to be important for the desired effects.     

Monitoring anti-HLA Class I IgG antibodies in renal transplant recipients

Anti-HLA class I IgG antibodies play an important role in hyperacute rejection but the significance of its de novo appearance or increase in levels during the posttransplant period remains controversial. The purpose of this study was to determine the correlation between the anti-HLA class I IgG antibodies and posttransplant events during the first 4 months after renal transplantation. From 200 renal allograft recipients, 549 serum samples were retrospectively evaluated. Patients who experienced graft dysfunction confirmed by biopsy had three serum samples tested: before, during (within 24 hours), and after the event. The presence of anti-HLA antibodies was observed in recipients with chronic allograft nephropathy (60%); acute rejection (clinical criteria without biopsy 57.1%); rejection types IIA (7.1%), IIB (40%), and III (50%); borderline changes (42.8%); acute tubular necrosis (34.4%); infarction (25%); and no rejection (12.5%). We observed a high incidence of anti-HLA class I IgG antibodies during acute tubular necrosis, borderline changes, acute rejection types IIB and III, and chronic allograft nephropathy.     

Donor cytokine genotype influences the development of acute rejection after renal transplantation

BACKGROUND: Acute allograft rejection remains an important cause of morbidity after kidney transplantation, and has been shown to be a crucial determinant of long-term graft function. Although rejection is mediated by recipient lymphocytes, both donor and recipient factors contribute to the local environment that influences the nature, severity, and duration of the rejection response. Cytokines are a major determinant of this milieu, and this study sought to explore the impact of donor cytokine and cytokine receptor gene polymorphisms on acute rejection after renal transplantation. METHODS: A total of 145 cadaveric renal allograft donors were selected for analysis according to the presence or absence of graft rejection in the first 30 days after transplantation. DNA was genotyped for 20 polymorphisms in 11 cytokine and cytokine receptor genes using the polymerase chain reaction with sequence specific primers. Associations were assessed using contingency table analysis and the chi2 test, using a two-set design. RESULTS: A polymorphism at position -174 of the donor IL-6 gene was associated with the incidence (P=0.0002) and severity (P=0.000007) of recipient acute rejection. This finding was independent of HLA-DR matching. Acute rejection was not influenced by recipient IL-6 genotype, or by donor-recipient matching of IL-6 genotype. CONCLUSION: This study identifies donor IL-6 genotype as a major genetic risk factor for the development of acute rejection after renal transplantation. This provides evidence that donor-derived cytokines play a major role in determining outcome after transplantation, and will contribute to the development of therapeutic algorithms to predict individuals at particularly high risk of acute rejection.     

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。     

The effects of lipid-lowering agents on acute renal allograft rejection

BACKGROUND: Preliminary results from clinical trials suggest that 3-hydroxy-3-methylglutaryl co-enzyme A reductase inhibitors may help prevent acute renal allograft rejection. However, the mechanism for this putative effect of 3-hydroxy-3-methylglutaryl co-enzyme A reductase inhibitors, and whether it is independent of lipid-lowering per SE are unknown. METHODS: Immediately after renal transplantation we randomly allocated (proportioned 2:1:2) patients to: 1) simvastatin (10 mg/day, n=53), 2) simvastatin placebo plus gemfibrozil (dose adjusted for renal function, n=36), and 3) simvastatin placebo (n=52). RESULTS: Simvastatin, but not gemfibrozil, reduced total and low density lipoprotein cholesterol during the first 90 days posttransplant. There were no major adverse effects of therapy. However, there were no effects of treatment on acute rejection. Indeed, survival free of acute rejection at 90 days was 72% in the simvastatin group, 72% in the gemfibrozil group, and 77% in the placebo control group (P=0.771). A post hoc power analysis suggested that there was only a 7.5% chance that a true effect of simvastatin on acute rejection (versus placebo) was not detected, and a 2.5% chance that an effect of gemfibrozil on acute rejection (versus placebo) was not detected in this study. CONCLUSION: Lipid-lowering agents may not reduce the incidence of acute renal allograft rejection. However, additional studies are needed to confirm this observation. In the mean time, many if not most renal transplant recipients should be treated with HMG-CoA reductase inhibitors starting early posttransplant to prevent cardiovascular disease complications. The results of this study suggest that starting lipid-lowering therapy immediately after renal transplantation is both safe and effective in lowering total and low density lipoprotein cholesterol.