可溶性CD44及其配体在肾移植患者急性排斥反应外周血中的表达及其意义

目的 探讨血清可溶性CD44分子及其配体在肾移植急性排斥反应时的变化及其意义.方法 收集123例肾移植患者外周血,酶联免疫吸附试验(ELISA)检测血清中sCD44水平,检测外周血透明质酸的水平.结果 13例围手术期急性排斥反应患者血清sCD44升高达(902.45±69.60)μg/L,和稳定组(355.70±82.01)μg/L比较,差异有统计学意义(P<0.05);15例出院后急性排斥反应患者血清sCD44升高至(1012.31±13.62)μg/L,和稳定组(315.38±26.72)μg/L比较,差异有统计学意义(P<0.05).结论 CD44分子与其配体相互作用在移植肾急性排斥反应中可能起到重要作用,有可能成为较好的早期诊断急性排斥的预测因子.     

可溶性CD20在肾移植排斥反应中的表达及其意义

目的 观察血清中可溶性CD20分子(sCD20)在肾移植急性排斥反应时的变化,探讨血清sCD20水平变化与移植肾预后的关系.方法 收集50例肾移植患者外周血,酶联免疫吸附测定法检测血清中sCD20水平.结果 25例急性排斥反应患者术前血清sCD20为(1750.60±359.59) ng/L,与稳定组(936.96±181.71) ng/L比较,差异有统计学意义(P＜0.05);移植术后急性排斥反应组血清sCD20为(2516.20 ±511.33) ng/L,与稳定组(988.40 ±215.78)ng/L比较,差异有统计学意义(P＜0.05);而且sCD20值水平较高的患者移植肾存活时间明显少于sCD20值水平较低的患者.结论 CD20分子在移植肾急性排斥反应和预测肾移植预后中可能起重要作用.     

排斥反应时血清可溶性白细胞相关免疫球蛋白样受体1的水平及意义

目的 探讨可溶性白细胞相关免疫球蛋白样受体1(sLAIR-1)与移植物排斥反应的相关性.方法 采用双抗体夹心酶联免疫吸附试验测定23例肝移植、139例肾移植患者的血清sLAIR-1水平,并以健康志愿者为对照.结果 健康志愿者的血清sLAIR-1水平为(4.3±2.3)μg/L,移植器官功能正常者(肝移植11例,肾移植87例)的sLAIR-1为(6.3±3.7)μg/L,二者比较,差异无统计学意义(P>0.05).肝移植后发生急性排斥反应的6例患者,其血清sLAIR-1水平为(47.2±25.9)μg/L;肾移植后后发生急性排斥反应的20例患者,其血清sLAIR-1水平(36.3±14.7)μg/L;移植肾功能丧失的5例患者,其血清sLAIR-1水平为(28.8±9.4)μg/L,上述三者的血清sLAIR-1水平均明显高于移植物功能正常者和健康志愿者(P<0.01).移植肝重度排斥反应的1例,其血清sLAIR-1高达117.3μg/L,为正常人的27倍.移植肝慢性排斥反应者(5例)和移植肾慢性排斥反应者(27例)的血清sLAIR-1水平分别为(16.1±6.4)μg/L和(13.1±5.5)μg/L,也明显高于移植物功能正常者和健康志愿者(P<0.05).结论 发生排斥反应者的血清sLAIR-1水平升高,其水平的升高可能是发生排斥反应的重要风险因素.     

可溶型LAIR分子与肾移植术后巨细胞病毒肺炎的关系

目的 探讨可溶型LAIR分子(sCD305,CD306)与肾移植术后巨细胞病毒(CMV)肺炎的关系. 方法肾移植受者血清19份,分为肺炎组(10份)和对照组(9份);以夹心ELISA方法定量检测血清中sCD305和CD306的浓度;用CHISS软件进行:检验. 结果 2组sCD305浓度均不符合正态分布,肺炎组波动于0.000～3.039 μg/L,对照组波动于0.000～8.375μg/L;肺炎组CD306浓度不符合正态分布,波动于0.000～0.017 μg/L,对照组符合正态分布.浓度为(0.046±0.035)μg/L;CD306在CMV肺炎患者血清内的表达降低,与对照组相比差异有统计学意义(P=0.000),而sCD305在2组的表达差异无统计学意义(P=0.316). 结论 CD306在肾移植术后发生CMV肺炎的患者血清内表达明显降低,CMV-PP65抗原检测联合CD306检测,有助于临床准确早期诊断CMV肺炎.     

肾移植排斥反应时血浆骨桥蛋白水平的变化及意义

目的 探讨肾移植排斥反应时血浆骨桥蛋白( OPN)水平的变化及意义.方法 对46例肾移植受者的临床资料及生物样本进行回顾性分析.根据移植肾组织学检查结果,46例受者被分为3组:移植肾功能稳定,且移植肾组织学检查未显示有排斥反应证据者16例,为非排斥组;移植肾组织学检查证实有急性细胞性排斥反应者22例,为急排组;移植肾组织学检查证实为慢性移植肾肾病(CAN)者8例,为慢排组.另以6名亲属活体供者作为对照组.于移植肾组织样本采集前抽取外周血,用人OPN酶联免疫吸附试验检测试剂盒测定血浆OPN水平,参照Banff 03标准对排斥反应进行分级.结果 对照组血浆OPN水平为(12.23±5.95)μg/L,非排斥组稍高,为(19.38±8.23)μg/L,两组间的差异无统计学意义( P＞0.05);慢排组血浆OPN水平为(27.77±12.27)μg/L,与非排斥组比较,差异无统计学意义(P＞0.05);急排组血浆OPN水平为(41.84±18.51)μg/L,与非排斥组比较,差异有统计学意义(P＜0.05),与慢排组比较,差异也有统计学意义(P＜0.05).急排组血浆OPN水平与排斥反应的级别具有正相关性(r=0.87,P＜0.05),发生Ⅰa级排斥反应和Ⅱb级排斥反应者间血浆OPN水平的差异有统计学意义(P＜0.05).结论 血浆OPN水平变化与急性排斥反应关系密切,其水平高低与排斥反应的级别呈正相关,可以作为诊断移植肾急性细胞性排斥反应、评估其严重程度的一个辅助指标.     

核心蛋白聚糖在移植肝慢性排斥反应中的表达及意义

目的 探讨核心蛋白聚糖(DCN)在移植肝慢性排斥反应(CR)患者肝组织及血清中的表达及意义.方法 以免疫组织化学(SP法)检测正常人16例、肝硬化20例、移植肝无CR 46例及合并CR 8例患者肝组织中DCN的表达情况,以酶联免疫方法(Elisa法)测定各组血清中DCN的含量情况.结果 移植肝并CR与无CR组织及正常组织中DCN均呈阴性表达,肝硬化组织中呈阳性表达,移植肝并CR阳性表达率、平均吸光度与肝硬化组织相比,有显著性差异(25%vs 55%,0.1249±0.0039 vs 0.2357±0.0396,P<0.01),与正常肝及移植肝无CR组相比,差异无统计学意义;移植肝并CR患者血清中DCN含量明显升高,与正常人、肝硬化及移植肝无CR组相比,有显著性差异(54.0833±6.0325μg/L vs 1.0232±0.9105 μg/L,12.6202±1.5370μg/L,17.7102±2.3562 μg/L,P<0.01),DCN血清浓度与排斥反应程度呈正相关.结论 DCN在移植肝并CR患者肝组织中呈阴性表达,而在血清中呈明显升高,提示其可能参与CR的发生、发展,同时血清DCN测定有可能成为移植肝CR早期诊断、发展及预后的非创伤性重要指标.     

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。     

血清中可溶性CD44v6含量和组织中CD44v6蛋白表达研究

目的　研究胃癌患者外周血中sCD44v6含量及其组织中CD44v6蛋白表达 ,探讨sCD44v6含量 /CD44v6蛋白与临床病理参数之间的关系。方法　应用酶联免疫吸附试验 (ELISA)检测胃癌患者 (术前 )及其中部分术后及健康对照组血清中sCD44v6含量并以 (S P)免疫组织化学法测定相应组织中CD44v6蛋白的表达。结果　 70例胃癌患者血清中sCD44v6含量 (2 .15± 0 .78)μg/L明显高于 16例正常对照组 (1.18± 0 .43 ) μg/L、14例根治性手术后血清中sCD44v6含量(1.2 1± 0 .3 9) μg/L比术前 (2 .67± 0 .83 ) μg/L明显下降 (P <0 .0 1) ,而 6例非根治性手术后(3 .2 9± 0 .41) μg/L比术前 (3 .61± 0 .49) μg/L下降不明显 (P >0 .0 5 )。结论　胃癌患者外周血中sCD44v6含量及组织中CD44v6蛋白表达的变化与转移、临床分期、病理分期有关 ,sCD44v6含量升高可作为胃癌患者淋巴结转移 (尤其是早期转移 )的监测指标。     

静脉注射人免疫球蛋白治疗肾移植后急性体液性排斥反应

目的 探讨静脉注射人免疫球蛋白(WIG)治疗肾移植术后急性体液性排斥反应(AHR)的临床效果.方法行尸体肾移植者252例,其中16例(6.3%,16/252)发生AHR,其移植肾Banff分级分别为1级3例,Ⅱ级9例,Ⅲ级4例.患者均采用他克莫司、霉酚酸酯及糖皮质激素预防排斥反应.采用IVIG治疗以逆转AHR,用量为20 g/d,连用3 d.术后1个月内发生AHR的12例因移植肾功能恢复延迟,进行血液透析,其中1例加用血浆置换治疗.观察AHR的逆转情况及受者体液免疫和细胞免疫的变化.结果 13例的AHR获得逆转,3例的移植肾功能丧失,行移植肾切除.治疗后,患者的抗HLA-I类、Ⅱ类抗体水平下降不明显(P>0.05);IgG水平明显上升,由(7.3±1.5)g/L升至(18.3±3.6)g/L(P<0.01);补体C3和C4水平分别为0.3±0.2)g/L和(0.1±0.1)g/L,较治疗前显著降低(P<0.01).IVIG治疗过程中,2例患者出现胸闷及体温升高,对症治疗后好转,患者治疗前后的肝功能均无显著变化.结论 IVIG对肾移植术后AHR的早期治疗有一定的效果,其机制可能与抗体封闭及对患者的体液免疫和细胞免疫的调节有关.     

细胞免疫能量测定在肾移植中的应用

目的 探讨适用于评价肾移植受者免疫状态的新方法.防止移植后的排斥反应和感染,提高人/肾生存质量和存活率.方法 应用ImmuKnow~(TM)-Cylex检测技术测定肾移植受者细胞免疫能量(三磷酸腺苷,ATP).收集62例肾移植受者术前(术前组)、术后稳定期(术后稳定组)、发生感染(术后感染组)和排斥反应(排斥反应组)等不同时期的肝素钠抗凝样本共150份.通过测量CD4+T淋巴细胞受刺激后释放的ATP浓度来判定细胞免疫力.并用秩和检验和两两比较统计方法,对结果进行分析和比较.结果 各组肾移植受者CD4+T淋巴细胞ATP浓度分别为:术前组(281.33±146.46)/μg/L、术后稳定组(310.19±147.12)/μg/L、术后感染组(142.41±118.26)μg/L、排斥反应组(332.77±154.44)μg/L;术后感染组ATP浓度显著低于其他各组.差异均有统计学意义(P<0.05).结论 LmmuKnow~(TM)-Cylex细胞免疫能量测定具有灵敏度高、特异性强、简便易操作等优点,适合于肾移植受者免疫状态的临床监测,特别是对术后感染有很好的预警作用,对指导感染后免疫抑制剂的个体化用药具有一定的参考价值.     

肾移植稳定病人血清可溶性E-选择素与淋巴细胞L-选择素的关系

目的　探讨肾移植稳定病人血清sE-选择素（sCD62E）浓度与其淋巴细胞表面配体L-选择素（CD62L）表达的关系和意义。方法　利用单克隆抗体-流式细胞仪荧光免疫技术和双抗体夹心酶联免疫法,测定10例移植稳定病人术后不同时间CD62L表达及sCD62E浓度。结果　术后CD62L表达率由(31.5±14.5)%渐升至（53.8±15.7）%,差异有非常显著性(P＜0.01),15d时明显高于术前和正常对照组（P＜0.05）;术后sCD62E浓度则由(31.7±10.0)μg/L渐降至(11.9±2.53)μg/L(P＜0.01),9d时明显低于术前组（P＜0.05）。sCD62E和CD62L之间呈负相关（r=-0.8183,P＜0.05）。结论　血清sCD62E浓度降低与淋巴细胞CD62L表达增加相关。术后监测sE-选择素浓度和CD62L表达将有助于肾移植受者免疫状态的判断。     

肾移植受者应用他克莫司治疗窗浓度的探讨

目的　寻求适合国人肾移植受者他克莫司 (FK5 0 6 )理想治疗窗浓度范围。方法　应用微粒子酶免疫分析法测定 5 8例肾移植患者口服FK5 0 6后 12h的血药谷浓度 ,并观察排斥反应的发生及药物的肾毒性。结果　FK5 0 6的血药浓度 ,术后 1个月为 (13.0± 2 .1) μg/L ,2～ 3个月为 (9.4±1.6 ) μg/L ,3个月以后为 (6 .5± 1.3) μg/L ,比较各时期全血FK5 0 6谷浓度 ,差异均有极显著性 (P <0 .0 1) ;术后发生急性排斥反应 3例次 ,肾毒性 4例次。结论　FK5 0 6具有良好的免疫抑制效果 ,其治疗窗浓度范围 ,术后第 1个月为 11～ 15 μg/L ,第 2～ 3个月为 8～ 11μg/L ;第 3个月后为 5～ 8μg/L ,此浓度范围既能达到满意的免疫抑制效果 ,又能减少FK5 0 6的肾毒性     

Soluble forms of CD95 and CD95 ligand after living related liver transplantation

BACKGROUND: Soluble forms of CD95 and CD95 ligand (sCD95 and sCD95L, respectively) can increase in the serum of patients with some inflammatory disease. In this study, we investigated the serum levels of sCD95 and sCD95L in liver transplantation recipients. METHODS: Serum levels of sCD95 and sCD95L in living related liver transplant recipients were analyzed by ELISA and their relation to the clinical findings estimated. RESULTS: Serum samples from the recipients did not show detectable levels of sCD95L but showed significantly increased levels of sCD95. The increase of sCD95 was positively associated with that of total-bilirubin and incidence of rejection, infection, and graft ischemia. CONCLUSIONS: The present results indicate an existence of sCD95 in the recipients of living related liver transplants. The increased serum levels of sCD95 may modify the immunological situation of the recipients after transplantation or represent the ongoing graft damage.     

Soluble CD30 and Cd27 levels in patients undergoing HLA antibody-incompatible renal transplantation

HLA antibody-incompatible transplantation has a higher risk of rejection when compared to standard renal transplantation. Soluble CD30 (sCD30) has been shown in many, but not all, studies to be a biomarker for risk of rejection in standard renal transplant recipients. We sought to define the value of sCD30 and soluble CD27 (sCD27) in patients receiving HLA antibody-incompatible transplants. Serum taken at different time points from 32 HLA antibody-incompatible transplant recipients was retrospectively assessed for sCD30 and sCD27 levels by enzyme-linked immunosorbent assay (ELISA). This was compared to episodes of acute rejection, post-transplant donor-specific antibody (DSA) levels and 12 month serum creatinine levels. No association was found between sCD27 and sCD30 levels and risk of acute rejection or DSA levels. Higher sCD30 levels at 4–6 weeks post-transplantation were associated with a higher serum creatinine at 12 months.Conclusion patients undergoing HLA antibody-incompatible transplantation are at a high risk of rejection but neither sCD30 (unlike in standard transplantation) nor sCD27 was found to be a risk factor. High sCD30 levels measured at 4–6 weeks post-transplantation was associated with poorer graft function at one year.     

Immunological monitoring after organ transplantation: potential role of soluble CD30 blood level measurement

Analysing the relevance of soluble CD30 (sCD30) in the bloodstream before and after transplantation may be important for the monitoring of transplant recipients. In this study, 27 patients (15 pediatric liver and 12 adult kidney graft recipients) were investigated. In the liver graft group, the patients who developed acute rejection during the first month (n=9) had a slightly higher sCD30 value on pre-transplantation baseline (day 0) and post-transplantation day 7, when compared to patients with normal graft function (n=6) (day 0: 102(1.6) U/ml versus 118(1.5) U/ml, p=0.52) and (day 7: 69(1.5) U/ml versus 83(1.6) U/ml, p=0.47). Increased serum sCD30 was shown to correlate with increased interleukin-10 circulating levels between day 0 and day 7 (r=0.53; p=0.04), whereas, no correlation could be evidenced between interferon-gamma (IFN-gamma) and sCD30 (r=0.02; p=0.47). Similarly, in the kidney transplantation group, no significant difference was found in sCD30 levels at day 0 in both groups with graft rejection or normal graft function (n=6) (85(1.3) U/ml versus 77(1.6) U/ml, p=0.66), but sCD30 decreased significantly at day 7 post-transplantation from baseline value in the rejection group (n=6) (77(1.6) versus 35(1.4); p=0.02). We conclude that increased serum sCD30 was correlated with increased IL-10 (interleukin-10) circulating levels, but not with IFN-gamma levels in the post-transplantation period. Neither pre-transplantation sCD30 nor sCD30 at day 7 post-transplantation could be correlated with acute rejection in liver graft recipient. The monitoring of sCD30 might constitute a tool to assess the risk of acute rejection in renal transplant but did not appear as a valuable mean for early immunological monitoring in the small group of liver allograft recipients patients analysed in this study.     

Plasma levels of soluble CD30 in kidney graft recipients as predictors of acute allograft rejection

In renal transplant recipients elevated soluble serum CD30 levels are associated with increased rejection and graft loss. We sought to determine the sCD30 plasma levels before and after kidney transplantation and to assess whether sCD30 was a predictive factor of immunological risk. sCD30 plasma levels were determined by an enzyme-linked immunosorbent assay assay in 52 kidney graft recipients before as well as 7, 15, and 21 days after transplantation. Eighteen patients developed acute allograft rejection (group I) and 34 patients showed uneventful courses (group II). Before transplantation sCD30 plasma levels were elevated in both groups (mean: 162.6 +/- 89.5 U/mL). After transplantation, group I recipients with acute rejection showed higher relative levels of plasma sCD30 on days 7 and 15 (120.8 +/- 74.6 U/mL and 210.6 +/- 108.7 U/mL respectively) compared with group II patients without rejection (95 +/- 45 U/mL and 59.4 +/- 31.6 U/mL), a difference that was significant for group I (P = .0003) and not significant for group II (P = .09). On day 21, sCD30 decreased in the two groups but remained higher among group I patients (120.6 +/- 92.7 U/mL). HLA antibodies were positive in 18 patients (34.6%) with 9 (50%) experiencing at last one episode of acute rejection. Among 34 patients negative for anti-HLA antibodies, nine displayed acute rejection only (26.4%), a difference that was not significant (P > .05). If we consider 100 U/mL as the minimum predictive level for allograft rejection, our results suggested that levels of sCD30 should be taken into consideration with the presence of HLA-antibodies detectable before and after transplantation, especially in patients with more than three HLA mismatches [RR = 3.20 (0.94 < RR < 10.91)]. These data suggested that measurement of plasma sCD30 is a useful procedure for the recognition of rejection in its earliest stages.     

High Pre-Transplant Soluble CD30 Levels are Predictive of the Grade of Rejection

In renal transplantation, serum soluble CD30 (sCD30) levels in graft recipients are associated with increased rejection and graft loss. We investigated whether pre-transplant sCD30 concentrations are predictive of the grade of rejection. Pre-transplant sera of 51 patients with tubulointerstitial rejection (TIR), 16 patients with vascular rejection (VR) and an age-matched control group of 41 patients with no rejection (NR) were analyzed for sCD30. The transplant biopsies were immunostained for C4d. The median sCD30 level was significantly elevated in the group with VR (248 Units (U)/mL, range: 92-802) when compared with TIR (103 U/mL, range: 36-309, p<0.001) and NR (179 U/mL, range: 70-343, p<0.03). Moreover, patients with TIR had significantly lower sCD30 levels compared to NR. Based on C4d staining, a TH2 driven process, the median sCD30 levels were significantly raised in C4d+ patients compared with C4d- group (177 U/mL vs. 120 U/mL, p<0.05). sCD30 levels measured at time of transplantation correlate with the grade of rejection. High pre-transplant levels are associated with antibody-mediated rejection which carries a poorer prognosis. sCD30 could be another tool to assess immunological risk prior to transplantation and enable a patient centered approach to immunosuppression.     

肾移植患者围手术期Toll样受体4与CD80测定及其意义

目的 研究肾移植患者围手术期外周血单核细胞Toll样受体4(TLR4)及CD80的变化规律及其临床意义. 方法 初次肾移植受者32例,术前群体反应抗体检测均阴性.流式细胞术测定受者肾移植术前及术后第1、4、7、14、21、28、35天外周血单核细胞TLR4、CD80表达的百分率;根据肾移植术后2周内发生排斥与否将患者分为排斥组(7例)和无排斥组(25例).10例PRA阴性健康成年志愿者标本为正常对照组.根据临床表现、实验室检查、移植肾多普勒超声检查、必要时行移植肾穿刺活检等综合判断有无急性排斥反应.应用SPSS 13.0统计软件处理数据. 结果 排斥组肾移植术前外周血单核细胞TLR4、CD80表达值为(8.03±0.84)%、(0.85±0.31)%,无排斥组为(6.14±0.85)%、(0.84±0.39)%,正常对照组为(6.37±0.56)%、(0.85±0.35)%;排斥组TLR4表达显著高于无排斥组和正常对照组(P<0.01),无排斥组与正常对照组比较差异无统计学意义(P=0.448);3组间CD80表达差异无统计学意义(P=0.995).患者单核细胞TLR4、CD80表达在移植术后第4天开始出现升高趋势,排斥组为(16.50±1.02)%、(7.82±1.66)%,无排斥组为(11.60±0.98)%、(2.26±0.96)%,较术前均显著升高(P<0.01);第7天左右达高峰,排斥组为(36.40±4.86)%、(9.53±1.97)%,无排斥组为(22.70±3.45)%、(1.87±0.72)%;随肾功能恢复逐渐回落,排斥组于第35天降至(7.10±0.82)%、(0.87±0.57)%,无排斥组第21天降至(7.20±0.76)%、(0.81±0.37)%;与无排斥组比较,排斥组TLR4、CD80表达峰值较高(P<0.01),且持续时间较长.结论 TLR4高表达增加肾移植术后急性排斥反应发生的风险;TLR4、CD80在肾移植术后早期上调,参与急性排斥反应的发生.