人巨细胞病毒感染对肾移植受者血清白细胞介素－6水平的影响

应用ＰＣＲ检测ＨＣＭＶ－ＤＮＡ，ＥＬＩＳＡ检测ＨＣＭＶ－ＩｇＭ、ＩｇＧ，诊断肾移植受者ＨＣＭＶ感染，６５例受者中ＨＣＭＶ感染者３９例，非感染者２６例。应用ＭＴＴ法检测受者血清ＩＬ－６生物活性，阐明了ＨＣＭＶ感染对肾移植受者血清ＩＬ－６水平的影响。结果表明：感染与非感染组间血清ＩＬ－６水平差异无显著性（Ｐ＞０．０５）；６例原发性感染者血清ＩＬ－６水平随感染时间延长呈增高及降低双相改变，表明慢性迁延性感染者血清ＩＬ－６水平降低。临床工作中监测ＨＣＭＶ感染的肾移植受者血清ＩＬ－６水平变化具有重要意义。     

人巨细胞病毒感染对肾移植受者血清白细胞介素—6水平的影响

应用ＰＣＲ检测ＨＣＭＶ－ＤＮＡ，ＥＬＩＳＡ检测ＨＣＭＶ－ＩｇＭ，ＩｇＧ，诊断肾移植受者ＨＣＭＶ感染，６５例受者中ＨＣＭＶ感染者３９例，非感染者２６例。应用ＭＴＴ法检测受者血清ＩＬ－６生物活性，阐明了ＨＣＭＶ感染对肾移植受者血清ＩＬ－６水平的影响。结果表明：感染与非感染组间血清ＩＬ－６水平差异无显著性；６例原发性感染者血清ＩＬ－６水平随感染时间延长呈增高及降低双相改变，表明慢性迁延性感染者血清ＩＬ－     

自然流产夫妇巨细胞病毒抗体测定的临床意义

目的：探讨自然流产病人ＨＣＭＶ抗体测定的临床意义。方法；用市售ＨＣＭＶ－Ｉｇ的间接ＥＬＩＳＡ试剂盒测定病人血清中的ＨＣＭＶ－Ｉｇ滴度。结果：４２例ＲＳＡ病人、３３例早孕对照组、３７例正常对照组ＨＣＭＶ－ＩｇＧ，ＨＣＭＶ－ＩｇＭ抗体测定结果为：１０倍稀释的血清ＨＣＭＶ－ＩｇＧ在３组标本中的检出率分别为８８．１％，８７．９％和８３．５％，血清ＨＣＭＶ－ＩｇＭ抗体检出率分别为８３．３％、６６．７％和６７     

肾移植术后的巨细胞病毒感染及其对急性排斥反应的影响

目的 探讨肾移植受者术后活动性巨细胞病毒（ＣＭＶ）感染的发生率、感染的原因以及ＣＭＶ感染对急性排斥反应的影响。方法 检测１８７例肾移植受者和供者术前血清抗－ＣＭＶ抗体;受者术后定期检测体内ＣＭＶ ＤＮＡ、对ＣＭＶ ＤＮＡ阳性的部分患者给予抗ＣＭＶ治疗,并比较各组排斥反应的发生率。结果 无论是供者还是受者,术前如血清抗－ＣＭＶ抗体阳性,受者术后发生活动性ＣＭＶ感染者明显增多,这些患者急性排斥反应的发     

自然流产夫妇巨细胞病毒抗体测定的临床意义

目的：探讨自然流产病人ＨＣＭＶ抗体测定的临床意义。方法：用市售ＨＣＭＶ－Ｉｇ的间接ＥＬＩＳＡ试剂盒测定病人血清中的ＨＣＭＶ－Ｉｇ滴度。结果：４２例ＲＳＡ病人、３３例早孕对照组、３７例正常对照组ＨＣＭＶ－ＩｇＧ、ＨＣＭＶ－ＩｇＭ抗体测定结果为：１０倍稀释的血清ＨＣＭＶ－ＩｇＧ在３组标本中的检出率分别为８８．１％、８７．９％和８３．５％;血清ＨＣＭＶ－ＩｇＭ抗体检出率分别为８３．３％、６６．７％和６７．７％;血清１００倍稀释时,ＲＳＡ组两种类别ＨＣＭＶ抗体的阳性检出率分别为２８．６％和３８．１％,对照组的ＨＣＭＶ－ＩｇＧ检出率分别为１５．２％和２．７％;ＨＣＭＶ－ＩｇＭ分别为６．１％和２．７％;血清１０００倍稀释,ＨＣＭＶ的两种类别抗体仅实验组有较低的检出率（分别为９．５％和７．１％）外,两个对照组均未检出阳性病例。结论：早孕妇女反复自然流产与ＨＣＭＶ感染有密切关系,对早孕妇女进行ＨＣＭＶ抗体调查时,血清１∶１０００稀释对结果的判定有重要价值。     

聚合酶链反应检测肾移植受者尿中人巨细胞病毒

聚合酶链反应检测肾移植受者尿中人巨细胞病毒廖利民，石炳毅，梁春泉，庄玉辉人巨细胞病毒（ＨＣＭＶ）在免疫低下患者（如器官移植受者）中可导致严重的疾病。以往诊断ＨＣＭＶ感染的方法大多存在耗时长、灵敏度低等缺点。聚合酶链反应（ＰＣＲ）技术检测ＨＣＭＶ具有灵...     

全自动快速微粒子酶免疫检测法在肝移植中的应用

目的 探讨全自动快速微粒子酶免疫检测法（ＭＥＩＡ）在肝移植中的应用。方法 采用ＭＥＩＡ对１３例肝移植患者术 后不同时期血清中巨细胞病毒（ＣＭＶ）ＩｇＧ、ＩｇＭ的动态进行观察。结果 ＣＭＶ感染率在肝移植患者中高达３０－６５％。结论 ＭＥＩＡ具有特异性强、灵敏度高、重复性好、快速简便等优点,为目前最新最理想的ＣＭＶＩｇＧ、ＩｇＭ检测方法。     

用多聚酶链反应检测病毒DNA诊断肾移植受者术后人巨细胞病毒活性感染

应用多聚酶链反应（ＰＣＲ）技术，直接通过肾移植受者外周血白细胞检测病毒ＤＮＡ，诊断人类巨细胞病毒（ＨＣＭＶ）活性感染。结果术前所有受者及其供者ＤＮＡ检测均为阴性，术后共检出４例阳性受者。另有３例患者亦出现类似病毒感染的症状，ＤＮＡ检测为阴性，后经检查确诊为结核和霉菌感染。２例术后１年多出现发热的受者经ＤＮＡ检测，排除ＣＭＶ感染，确诊为慢性排斥反应和霉菌感染。以上结果表明，ＰＣＲ作为移植受者术后ＨＣＭＶ活性感染的诊断手段，简单快速，准确性高，成本低，并可用于鉴别其它细菌（如霉菌）感染或慢性排斥反应。     

用PCR检测肾移植受者人巨细胞病毒感染

用自行设计的引物，建立了聚合酶链反应（ＰＣＲ）检人巨细胞病毒（ＨＣＭＶ）ＤＮＡ的方法，经实验证明有高度的特异性和敏感性，对其它疱疹病毒和正常人基因组ＤＮＡ无交叉反应，ＨＣＭＶ ＡＤ１６９株ＤＮＡ ＥｃｏＲＩ酶切Ｊ片段的最小检出量为０．１ｆｇ，相当于６个基因拷贝。对２２例肾移植受者的血和尿标本进行ＰＣＲ检测，ＨＣＭＶ ＤＮＡ的阳性率为５９．１％。实验结果表现ＰＣＲ是一种敏感特异、简便快速、能早期诊断     

人巨细胞病毒感染对肾移植受者外周血T细胞亚群的影响及…

应用ＰＣＲ技术及ＥＬＩＳＡ法诊断ＨＣＭＶ感染，６５例肾移植受者中感染率为６０％。应用ＡＰＡＡＰ法检测其外周血Ｔ细胞亚群，发现ＨＣＭＶ感染导致ＣＤ４／ＣＤ８值显著降低。ＨＣＭＶ产生的超级免疫抑制导致了严重的临床后果：ＨＣＭＶ感染组革兰细菌、结核杆菌、霉菌、ＨＳＶ及ＶＺＶ等所致的机会感经明显高于ＨＣＭＶ未感染组，ＡＡＧ发生率明显增高。     

IgG alloantibody responses to donor-specific blood transfusion in different rat strain combinations as a predictor of renal allograft survival.

Donor-specific blood transfusion prolongs the survival of fully allogeneic ACI (RT1a) renal allografts in PVG (RT1c) recipients from 7-10 days to greater than 100 days. We have observed significant differences in the alloantibody (Ab1) responses to ACI renal allografts in control and DSBT-treated PVG recipients: DSBT is associated with decreased IgG and IgM alloantibody circulating in serum, deposited in the allograft, and produced in culture by splenocytes. In the present studies the effects of DSBT on alloantibody production and renal allograft survival were extended to examine other recipient strains: F344 (RT1lv1), BN (RT1n), W/F (RT1u) and LEW (RT1l). Animals of each recipient strain were injected i.v. with 0.5 ml of ACI blood alone or followed by a renal allograft. Studies on the kinetics of IgM and IgG alloantibody responses were performed by flow cytometry on lymphocytes from donor ACI, PVG, and PVG.R1 (RT1.Aa class I MHC antigen on PVG background) rats. In F344 and PVG rats, DSBT from ACI rats elicited a transient IgM response that peaked at day 7 and was not followed by a switch to IgG. In control PBS transfused F344 recipients, an ACI renal allograft stimulated both IgM and IgG alloantibody production. DSBT pretreatment significantly decreased circulating IgG alloantibody following ACI renal transplantation and prolonged graft survival in F344 recipients. In DSBT-treated F344 recipients that rejected ACI renal allografts acutely, small amounts of IgG (5-12 mode channel shift) were detected in sera harvested 7 days after transplantation, whereas almost no IgG was detected in the sera from DSBT treated F344 rats that accepted their renal allografts indefinitely. In contrast, DSBT alone from ACI to BN, W/F, or LEW strains elicited a transient IgM response that peaked at day 7 and was followed by a strong IgG response that peaked on days 10-14 and remained high through day 21. DSBT failed to prolong ACI renal allograft survival in any of these strains (survival less than 11 days in control and DSBT rats). The alloantibody response to DSBT in all five recipient strains examined was directed primarily to RT1.Aa class I MHC antigens, as determined by binding studies on lymphocytes from ACI, PVG and PVG.R1 rats and alloantibody blocking studies using biotinylated rat monoclonal antibodies to distinct epitopes of the RT1.Aa antigen. The relative magnitude of blocking of R2/10P and R2/15S binding by sera from BN, W/F, and LEW rats was: control allograft recipients greater than DSBT pretreated allograft recipients greater than DSBT alone.(ABSTRACT TRUNCATED AT 400 WORDS)     

Kidney transplantation across positive B and T cell crossmatches.

Cadaveric renal transplants were performed despite a positive conventional crossmatch (usually intermediate positive) resulting from donor-specific B cell lymphocytotoxins (both IgG and IgM) or IgM cold-reactive T cell lymphocytotoxins. Graft survival at 2 months was 72% in the 14 patients with B cell-specific antibodies and 71% in the 7 recipients with T cell antibodies. No correlation was observed between graft rejection and warm (mainly IgG) or cold (IgM) B cell-specific antibodies. These results indicate that not all positive crossmatches are a contraindication to transplantation. Attempts should be made to study the nature of the lymphocytotoxins before withholding the allograft from the recipient.     

A retrospective flow cytometric crossmatch study in transplant recipients with autoreactive lymphocytotoxic antibodies

Abstract Our previous data shows renal transplant recipients with autoreactive lymphocytotoxic antibodies to have a reduced transplant survival when compared to patients without autoantibodies. This could have been due to the presence of weak IgG antibodies inhibited by the dithiothreitol used to remove IgM antibodies in the pretransplant cytotoxicity cross-match. That possibility was investigated in a retrospective study of 52 recipients of 57 renal transplants who were recrossmatched using a more sensitive flow cytometry crossmatch (FCXM) to detect recipient IgG antibodies to donor T and/or B cell splenic lymphocytes. Fourteen of the 57 (24%) transplants failed. Six losses were within the 1st month posttransplant and four of these were immunological failures. None of the transplant failures had a positive pretransplant FCXM. These results showed that the recipients with autoantibodies did not have pretransplant IgG anti-donor antibodies. The transplant failures did not, therefore, relate to the presence of antibodies undetected by the dithiothreitol-treated cytotoxicity crossmatch.     

Donor Monoclonal Gammopathy May Cause Lymphoproliferative Disorders in Solid Organ Transplant Recipients

Prior research on donor monoclonal gammopathy of undetermined significance (MGUS) has been inadequate regarding the risk for lymphoproliferative disease in solid organ transplantation recipients. Seven organ recipients from two different donors developed lymphoproliferative disease. The origin of the malignancy was determined by use of microsatellite analysis, and the plasma of the two donors was analyzed with the use of electrophoresis. The clinical courses of the seven recipients were followed for 36–60 months. One donor transmitted lymphoplasmacytic lymphoma to two kidney recipients and MGUS to a liver recipient, all IgMκ. A second donor caused IgGλ myeloma in two kidney and one liver recipient, and IgGλ gammopathy in a heart recipient. Transplant nephrectomy was performed in three kidney recipients and remission was achieved. The fourth kidney recipient has kept the graft and the disease has progressed. The liver recipient died from myeloma. There were no clinical signs of lymphoproliferative disease in the donors, but retrospective serum analyses showed M‐components, IgMκ (37 g/L) and IgGλ (8 g/L). Donors with MGUS may cause donor‐transmitted malignancies via passenger lymphocytes/plasma cells in solid organ recipients. The results call for a large register study of the incidence of donor MGUS and lymphoproliferative disease in their recipients.     

高敏肾移植受者的二硫基苏糖醇—群体反应性抗体测定及其意义

为了进一步解释群体反应性抗体(PRA)的免疫球蛋白性质及其对异基因肾移植的潜在威胁。方法 借助二硫基苏糖醇(DTT)具有断裂Ig的二硫键,解聚大分子量的IgM分子的原理,将DTT引进PRA实验中,建立DTT-PRA分析方法,以分辨高PRA患者血清中的Ig类型。结果 发现在标准PRA阳性的701例受者中DTT-PRA阳性率为33.1％,依靠患者血清对DTT敏感与否可将PRA阳性患者分为单纯IgM、单纯IgG和IgG、IgM混合类型3种。结论 单纯IgM抗体与移植后排斥反应无关,而单纯IgG和IgC、IgM混合类型的HLA抗体将介导急性甚至超急排斥反应的发生。     

大鼠肾移植后抗波形蛋白抗体升高与慢性移植肾肾病的相关性

目的 研究大鼠肾移植术后抗波形蛋白抗体的表达水平与慢性移植肾肾病(CAN)的相关性.以及不同免疫抑制剂对其的影响.方法 选取近交系F344大鼠作为同系肾移植的供、受者(共9对),选取F344和Lewis大鼠分别作为同种肾移植的供、受者(共27对).同系移植组受者术后不给予免疫抑制剂;同种移植组受者术后10 d内给予环孢素A(CsA),然后将同种移植组受者随机平均分为生理盐水(NS)组、CsA组和霉酚酸酯(MMF)组(每组9只),分别给予NS、CsA和MMF灌胃.术后第4、8和12周时分别处死每组受者3只,观察CAN的进展、波形蛋白及其基因的表达以及抗波形蛋白抗体的水平,取正常大鼠(包括F344和Lewis大鼠)作为对照.结果 观察期内同系移植组未发生CAN;而同种移植组发生了CAN,且不断加重,其中CsA组和NS组的CAN病理改变非常明显,而MMF组明显较轻.术前所有受者血清中均未检测到抗波形蛋白IgM和IgG抗体,术后也未检测到抗波形蛋白IgM抗体;术后同系移植组仅检测到微量的抗波形蛋白IgG抗体,同种移植组检测到大量的抗波形蛋白IgG抗体.随着CAN的进展,同种移植中,CsA组和NS组血清抗波形蛋白IgG抗体的水平逐渐增高,而MMF组抗体水平的增高显著低于NS组(P<0.05),但仍显著高于同系移植组(P<0.05).结论 大鼠同种肾移植术后,受者体内可产生抗波形蛋白IgG抗体,且产生的时间早于CAN,抗波形蛋白IgG抗体水平也会随着CAN的进展而增高.MMF可抑制抗波形蛋白IgG抗体的产生,CsA无此作用.     

Fever, human herpesvirus-6 (HHV-6) seroconversion, and acute rejection episodes as a function of the initial seroprevalence for HHV-6 in renal transplant recipients

Human herpesvirus-6 (HHV-6) is an opportunistic viral pathogen of emerging clinical significance in immunocompromised patients. We performed a seroepidemiological survey to test the relation between seroprevalence among donors and recipients for HHV-6 at three endpoints. Before transplantation sera obtained from cadaveric donors and from potential recipients were tested for IgG antibodies against HHV-6 using an enzyme-linked immunoassay. The group of recipient sera, including samples obtained before as well as 2, 4, 12, and 48 weeks after transplantation, were tested for anti-HHV-6 IgM antibodies using an indirect immunofluorescence assay. The statistical analysis was performed with the Cox proportional hazards models. The HHV-6 seronegative group (n = 11) compared with the HHV-6 seropositive group (n = 109) showed twice the risk of HHV-6 IgM seroconversion (RR = 2.24; P < .04), with a greater risk of fever, namely 3.8, which was on the verge of statistical significance. The opposite trend toward an association with acute rejection episodes was observed among HHV-6 seronegative patients (RR = 1.81). The presence of IgG antibody in the sera of donors to IgG seropositive recipients had no association with the occurrence of IgM seroconversion. In contrast, IgM antibodies to HHV-6 appeared in four of five seronegative patients who received allografts from IgG seropositive donors. These preliminary data suggest that the effects seem to be the consequence of HHV-6 transmission through a renal allograft.     

Immunoglobulin-G subclass antidonor reactivity in transplant recipients.

Outcomes may differ after kidney transplantation compared to combined liver-kidney transplantation. In animal models, distinct patterns of antidonor immunoglobulin (Ig) G subclasses are associated with either rejection or transplant tolerance. Flow cytometry has increased the sensitivity of antidonor immunoglobulin detection. We compared antidonor IgG subclass responses in kidney transplant recipients to those in recipients of liver or multiorgan grafts. In this study of 19 organ (kidney, liver, pancreas) transplantations, recipient serum incubated with donor splenocytes was tested by flow cytometry for the presence of IgM, IgG, or IgG subclass 1-4. Sera before transplantation and 10 days and 100 days after transplantation were used. No differences were seen in antidonor IgM, IgG, or IgG subclass antibodies among recipients of kidney transplants and liver grafts or combination grafts, either before or after transplantation. IgG4 gradually but significantly increased after transplantation in all groups. High levels of antidonor IgG3 either before transplantation or produced after it were found in 3 kidney recipients who experienced acute rejection. No other patients experienced rejection, and no other increase in IgG3 was seen. In conclusion, antidonor IgG subclass profiles may be useful to distinguish populations at risk of rejection but they do not differentiate the immunological response after kidney transplantation from that after liver or combined transplantation. A late rise in antidonor IgG4 is consistent with decreased antidonor reactivity thought to occur late after transplantation.     

IgM and IgG alloantibody production by splenocytes and deposition in rat renal allografts are decreased by donor-specific blood transfusion

Untreated PVG (RT1c) rats reject ACI (RT1a) renal grafts in 6-8 days. Autologous (PVG) blood transfusions (ABT) do not alter ACI allograft rejection, but donor-specific blood transfusions (DSBT) 7 or 11 days prior to transplantation usually results in indefinite graft survival. We have reported previously that DSBT is associated with the development of antiidiotypic antibody and reduced circulating cytotoxic alloantibodies in this model. To further define the effects of DSBT on the alloantibody responses to renal allografts, we examined PVG rats that received DSBT or ABT prior to ACI renal transplantation. Antibody production by cells in the spleen was investigated by tissue culture techniques; circulating antibody titers were measured by antibody binding to target ACI lymphoblasts with flow cytometry; and antibodies bound to the ACI allograft were recovered by hypertonic acid elution and quantitated by flow cytometry and ELISA. Seven days after DSBT alone, circulating IgM alloantibodies to ACI reached peak titers. After renal allografting, serum IgM alloantibody titers decreased in DSBT-pretreated rats and little IgG could be detected. In contrast, renal allografts in ABT-pretreated rats elicited high titers of IgM and moderate titers of IgG in the circulation by 5-7 days posttransplantation. Spleens harvested one week posttransplantation from ABT-pretreated rats produced high titers (16-32) of IgM and IgG antibodies to ACI antigens, but no such antibody production was detected in spleens cultured from DSBT-pretreated rats. In addition, 4-32-fold more IgM and IgG was eluted from kidneys removed 6-7 days after grafting to ABT-treated rats than from allografts in DSBT-treated rats. IgG2a was the predominant subclass of IgG that bound to target ACI cells. No IgA was detected in graft eluates from any rat. Polyacrylamide gel electrophoresis demonstrated that the eluates contained predominantly IgM and IgG without significant contamination by other serum proteins. These data suggest that DSBT decreases the levels of IgM and IgG normally produced in the spleen and deposited in the graft following renal transplantation. Because IgM fixes complement and IgG (especially IgG2a) triggers ADCC, the reduced deposition of IgM and IgG in the graft may be of particular importance in DSBT enhancement.     

A role for chronic parvovirus B19 infection in liver dysfunction in renal transplant recipients?

BACKGROUND: Clinically, liver dysfunction in renal transplant recipients is related to hepatitis B virus (HBV) or hepatitis C virus (HCV) infection. The contribution of parvovirus B19 (B19) to liver disease in renal transplant recipients has not been studied. Here we present the association of liver dysfunction with or without the coinfection of B19, HBV, and HCV after renal transplantation. METHODS: We used enzyme-linked immunosorbent assay to identify B19, HBV, and HCV infections in serum samples taken from 144 renal transplant recipients before transplantation and at 12 and 24 months after transplantation. After each patient had fasted for 12 hr, blood was taken for measurement of aspartate aminotransferase and alanine aminotransferase monthly for at least 6 months. RESULTS: Liver dysfunction developed at the significantly higher incidence of 47% in the anti-HCV(+) patients compared with 6% in the noninfected group (P<0.0001). HBV infection had no impact on the incidence of liver dysfunction in renal transplant recipients. A higher incidence of liver dysfunction was found in 42% of B19 IgG(+)IgM(-) group patients compared with 13% of the B19 IgG(+)IgM(+) group (P=0.0051) and 9.5% of the B19 IgG(-)IgM(-) group (P=0.0003). A B19 polymerase chain reaction (PCR) assay revealed significantly higher liver dysfunction in 29% of B19 PCR(+) group patients compared with 13.6% of B19 PCR(-) patients (P=0.0419). Patients who were anti-HCV(+) and B19 PCR(+) had a significantly higher incidence of liver dysfunction than B19 PCR(-) patients (P=0.002). CONCLUSIONS: Chronic B19 infection and HCV infection, both separately and in combination, increase the incidence of liver dysfunction in renal transplant recipients. HBV infection does not seem to be independently or synergistically associated with liver dysfunction.