移植肾间质中浸润细胞免疫标记蛋白的变化与急性细胞性排异的…

观察移植肾发生急性细胞性排异和无ＡＣＲ时，肾间质浸润细胞中一组标记蛋白的变化，及其与间质浸润ＣＤ４或ＣＤ８细胞的关系。方法 选择１７例肾移植患者，定期行移植肾活检，采用ＰＡＰ四层法对一组标记蛋白进行观察。结果 无ＡＣＲ的肾组织中，增殖细胞核抗原（ＰＣＮＡ）和ＤＲ蛋白仅轻度增加，原癌蛋白－２（Ｂｄ－２）、ＩＬ－２Ｒ和细胞间粘附分子－１（ＩＣＡＭ－１）的增加不明显。当移植肾出现ＡＣＲ时，上述标记蛋白均     

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。     

移植肾间质中浸润细胞免疫标记蛋白的变化与急性细胞性排异的关系

观察移植肾发生急性细胞性排异（ＡＣＲ）和无ＡＣＲ时，肾间质浸润细胞中一组标记蛋白的变化，及其与间质浸润ＣＤ４或ＣＤ８细胞的关系。方法选择１７例肾移植患者，定期行移植肾活检，采用ＰＡＰ四层法对一组标记蛋白进行观察。结果无ＡＣＲ的肾组织中，增殖细胞核抗原（ＰＣＮＡ）和ＤＲ蛋白仅轻度增加，原癌蛋白２（Ｂｃｌ２）、ＩＬ２Ｒ和细胞间粘附分子１（ＩＣＡＭ１）的增加不明显。当移植肾出现ＡＣＲ时，上述标记蛋白均明显增加，尤以ＰＣＮＡ和ＩＬ２Ｒ增加最为显著，并与间质浸润的ＣＤ８密切相关。结论移植肾的免疫组化检查对移植肾急性细胞性排异反应有一定的意义；ＩＬ２Ｒ对于ＡＣＲ的诊断及鉴别诊断具有一定的临床应用价值。     

同种异体移植肾组织中B7蛋白的表达及其意义

目的观察同种异体移植肾组织中Ｂ７蛋白表达的特点，以期阐明其在急性细胞性排异中可能的致病作用。方法用ＰＡＰ免疫组织化学方法对急性细胞性排异（ＡＣＲ），无急性细胞性排异（Ｎ－ＡＣＲ）和正常对照肾组织中Ｂ７蛋白的表达进行观察，并结合肾间质中浸润淋巴细胞数和肾小管上ＨＬＡ－ＤＲ抗原的表达进行分析。结果ＡＣＲ组肾间质ＣＤ４＋，ＣＤ８＋和Ｂ细胞数较Ｎ－ＡＣＲ组和正常对照明显增高，与此相一致的是肾间质表达Ｂ７－１和Ｂ７－２的细胞也较Ｎ－ＡＣＲ组增高，但Ｂ７－２增加更明显。ＡＣＲ组肾小管上皮细胞ＨＬＡ－ＤＲ及Ｂ７－１的表达较Ｎ－ＡＣＲ组明显增高。结论肾小管上皮细胞有可能通过ＨＬＡ－ＤＲ和Ｂ７－１表达的上调作为抗原提呈细胞主动参与上述免疫反应的发生。     

同种异体移植肾组织中PCNA蛋白检测诊断的意义

为了提高肾移植急性排斥反应的诊断和治疗效果，选择增殖细胞核抗原（ＰＣＮＡ）作为标记蛋白，观察１７例５９例次移植肾发生急性细胞性排斥（ＡＣＲ）和无ＡＣＲ时，其肾小管浸润细胞中该蛋白的变化，以及与间质浸润细胞的关系。结果表明，无ＡＣＲ的肾组织中，ＰＣＮＡ蛋白仅轻度增加；当移植肾出现ＡＣＲ时，ＰＣＮＡ的增加十分显著，并与间质浸润的ＣＤ８密切相关。结果认为：ＰＣＮＡ对于ＡＣＲ的诊断及鉴别诊断具有一定的临床应用价值。     

同种异体肾移植患者血清SIL－2R水平变化及其临床意义

作者采用双抗体夹心酶联免疫吸附法（ＥＬＩＳＡ）对２０例同种异体尸肾移植患者进行了８９例次可溶性白介素２受体（ＳＩＬ－２Ｒ）检查。结果表明：移植前明显高于正常对照组，Ｐ＜０．００１。移植后随着肾功能的恢复而接近正常，但仍轻度高于正常对照组，Ｐ＜０．０１。发生急性排斥反应时较稳定期明显升高，Ｐ＜０．００１，且其上升时间早于血肌酐上升２～７天。而发生环孢素Ａ肾中毒或急性肾小管坏死时，血清ＳＩＬ－２Ｒ水平则变化不明显，Ｐ＜０．０５。因此，ＳＩＬ－２Ｒ的测定可作为移植肾排斥反应诊断和鉴别诊断的重要非创伤性指标。     

移植肾白细胞介素2受体的表达及其临床意义

我们连续观察了７１例１７５次同种异体尸体肾移植患者移植肾白细胞介素２受体（ＩＬ－２Ｒ）表达的变化，探讨其在肾移植排斥反应诊断及鉴别诊断中的意义。一、临床资料１．病例选择：７１例异体肾移植患者，术后常规应用环孢素（ＣｓＡ）、泼尼松（Ｐｒｅｄ）和硫唑嘌呤...     

慢性排斥移植肾浸润细胞亚群及TNFα和IL-2R原位表达

目的探讨移植肾慢性排斥的细胞免疫学机制。方法对１８例肾移植术后慢性排斥病人移植肾进行活检,采用免疫组化技术检测移植肾内浸润炎性细胞亚群及ＴＮＦα和ＩＬ２Ｒ原位表达。结果慢性排斥移植肾间质中呈明显的单核细胞／巨噬细胞和淋巴细胞灶样浸润伴纤维化和小管萎缩,浸润细胞组成分别为ＣＤ＋３细胞（４６８±１９．０）％,ＣＤ＋４细胞（２４１±１７３）％,ＣＤ＋８细胞（２７３±６９）％,ＣＤ＋１４细胞（３３４±１９６）％,ＣＤ＋１９细胞仅（３２±１９）％,以Ｔ淋巴细胞和巨噬细胞为主。移植肾中ＴＮＦα和ＩＬ２Ｒ表达较正常肾脏明显增强,且主要分布于移植肾间质浸润炎性细胞。结论慢性排斥移植肾内浸润淋巴细胞和巨噬细胞处于活化状态,可能通过产生和释放各种细胞因子发挥其生物学作用,从而介导细胞免疫损伤,认为细胞免疫可能在慢性排斥发病中起作用。     

冷冻保存对带血管异体关节移植排斥反应的影响实验研究

目的 观察冷冻保存和环孢霉素Ａ（ＣｙＡ）对鼠带智力这异体关节移植斥反应的作用。方法 ４０只ＳＤ大鼠随机分成４组：１组、新鲜同系移植；２组、新鲜异体移植；３组，冷冻保存异体移植；４组、冷冻保存异体移植术后用ＣｙＡ。术后进行血管造影，测定ＩＬ０－２活性及Ｔ细胞亚群（ＣＤ４／ＣＤ８）的变化，按照Ｓａｋａｉ等评分标准进行组织这评分。结果 ２组通畅率为０，与科各组比较差异有显著性（Ｐ〈０．０１）。ＩＬ－２及     

动态监测血清、尿液和肾组织液中SIL－2R水平在肾移植早期的应用价值

为了了解可溶性白介素２受体（ＳＩＬ－２Ｒ）在肾移植早期的应用价值，动态监测４０例肾移植患者血清、尿液及肾组织液中可溶性白介素２受体水平变化，发现术前透析患者血清ＳＩＬ－２Ｒ水平较正常人高，术后迅速下降。急性排斥（ＡＲ）发生时，ＳＩＬ－２Ｒ水平均有不同程度的升高，以肾组织液中升高最明显，比临床症状和血清肌酐出现早１～５天。在急性ＣｓＡ中毒发生时血清和肾组织液中ＳＩＬ－２Ｒ水平明显升高，而急性感染发生时只在血清中明显升高。结果表明：动态监测血清、尿液ＳＩＬ－２Ｒ水平有助于预测和早期诊断ＡＲ，同时行肾组织液及血清、尿液ＳＩＬ－２Ｒ水平测定能较好地诊断与鉴别诊断ＡＲ、急性ＣｓＡ中毒和感染     

Frequency of T cell expressing Th1 and Th2 associated chemokine receptor in patients with renal allograft dysfunction

Acute rejection of human renal allografts is a frequent, serious posttransplantation complication, occurring in up to 50% of recipients. Leukocyte recruitment is a central feature of acute allograft rejection. Chemokine receptors are expressed on leukocytes in a cell type-specific manner. Recently CCR5+ and CXCR3+ cells have been observed in allograft biopsy specimens of patients undergoing acute cellular rejection (ACR). Herein we investigated the expression of Th1 (CCR5, CXCR3, and CCR2) and Th2 (CCR4, CCR3, and CCR8)-associated chemokine receptors on CD4 and CD8 T-cell populations. We sought to correlate chemokine receptor expression in peripheral blood T-cell subsets with the types of graft dysfunction (biopsy-proven rejections). In the peripheral blood CD4+ and CD8+ T-cell populations of patients with graft dysfunction, we observed a high frequency of Th1-associated chemokine receptors CCR5+ and CCR2+ but not CXCR3.     

CD40 expression on graft infiltrates and parenchymal CD154 (CD40L) induction in human chronic renal allograft rejection

BACKGROUND: CD40-CD154 (CD40L) costimulatory signaling plays a pivotal role in the effector mechanisms of transplant graft rejection. In animal models, CD40-CD154 blockade induces long-term graft acceptance concurrent with an absence of chronic rejection (CR) lesions. Given the critical importance of CD40-CD154 interactions in the development of chronic transplant allograft rejection, the relevance of in situ CD40 and CD154 expression was assessed in human chronic renal allograft rejection. METHODS: The expression of CD40, CD154, CD68, and T-cell receptor (TCR)alpha/beta was analyzed by immunohistochemistry. Serial cryostat sections of snap-frozen core renal allograft biopsies were obtained from 30 renal transplant patients. Biopsy specimens received diagnoses of CR (N = 23) according to the Banff classification and were compared with controls (N = 7) consisting of stable allografts and normal kidney tissue. RESULTS: Striking CD40 staining of graft cellular infiltrates (P = 0.016) was observed in renal allografts with CR compared with controls. The CD40+ cellular infiltrates in CR were predominantly TCR alpha/beta + T cells and some CD68+ macrophages. These findings were contrasted by the low-level CD40 expression detected in glomeruli and tubules of CR and controls. However, glomerular induction of CD154 was observed in CR allografts (P = 0.028) as compared with controls. CD154 immunoreactivity was demonstrated on glomerular endothelial, epithelial, and mesangial cells. Moderate CD154 expression was detected on tubular epithelial cells, and only weak CD154 immunoreactivity was observed on the infiltrates in isolated CR cases. CONCLUSION: In human chronic renal allograft rejection, CD40 is expressed on graft-infiltrating cells of the T cell and macrophage compartments. CD154 expression is induced on glomerular and tubular epithelial cells during CR, demonstrating another novel source of CD154 expression. The data substantiate the potential contributory role of an interaction between CD40+ graft-destructive effector T cells and macrophages with CD154+ renal allograft parenchymal cells in the development of chronic renal allograft rejection.     

Expression of vascular cell adhesion molecule-1 in human renal allografts.

The expression of vascular cell adhesion molecule-1 (VCAM-1) in 11 human renal allograft biopsies and 3 normal kidney specimens was investigated by immunocytochemistry. VCAM-1 expression was correlated with the degree of CD3+ T cell infiltration and the clinicopathologic diagnosis of acute rejection. CD3+ infiltrates were seen in all biopsies with rejection, but not in normal biopsies or one with acute tubular necrosis, and were accompanied by CD68+ monocyte/macrophage infiltrates. In normal biopsies, VCAM-1 was present on occasional tubules, where its expression was patchy and restricted to the basolateral surface of cells with slight cytoplasmic staining. The total number of tubules expressing VCAM-1 significantly increased in specimens infiltrated with CD3+ T cells. Moreover, in these infiltrated biopsy specimens, VCAM-1 was present throughout the cytoplasm of tubular cells concentrated on the basolateral surface. VCAM-1 was also observed on vascular endothelial cells where its expression correlated with the degree of CD3+ infiltrate. Mean scores (0 to 3+) for endothelial VCAM-1 expression increased from 0 (CD3+ score, 0) to a mean score of 2.25 in association with CD3+ T cell infiltrates (CD3+ score, 3). Endothelial VCAM-1 was predominantly on vessels in areas of infiltrate, including peritubular capillaries, venules, and arterioles, but was notably absent on glomerular endothelium. VCAM-1 also stained mesangial cells in an occasional CD3+ infiltrated specimen. It was concluded that the expression of VCAM-1 is increased on renal tubules and renovascular endothelium in rejecting renal allografts in association with CD3+ infiltrates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Function and surface phenotype of T lymphocytes infiltrating renal allografts in nonhuman primates treated with monoclonal antibodies

The phenotype and function of T lymphocyte cell lines established in vitro from kidney biopsies at the time of acute cellular rejection were studied using a nonhuman primate renal allograft model. Our objectives were to investigate the function and surface phenotype of cells that infiltrate renal allografts in animals that were untreated, that were given subtherapeutic cyclosporin, or that developed rejection after treatment with monoclonal antibodies to IL-2R B chain (CD25), immune cell adhesion molecule-1 (ICAM-1), or CD8. Lines from allograft biopsies and peripheral blood were expanded in vitro using solely human recombinant IL-2 and analyzed after 6-20 days in culture. We found that the large majority of cells cultured from cynomolgus allografts at the time of acute rejection or, when possible, assayed directly without culture, were CD3+4-8+ T lymphoblasts that possessed donor-specific cytolytic function and an NK-line, cytotoxic activity. In contrast, it was rarely possible to establish T cell lines exhibiting donor-specific cytotoxic activity from the blood except in the absence of immunosuppression or during CsA taper. A stable number of graft-derived CD4+8- cells was only observed in an unsuppressed animal 2 days after transplantation in the absence of manifest signs of rejection. Taken together, the above data indicate that similar T lymphocyte populations associated with allograft rejection are present in acutely rejecting allografts after the various types of immunosuppressive therapy. Since the infiltrating cells were similar to those obtained prior to therapy, recurrent rejection most likely represents cells that have escaped elimination. The T cells derived from monkey grafts differ from those from human renal allografts by the decreased frequency of CD4+ cells. Whether this difference is species-related or therapy-related is not known.     

MIC expression in renal and pancreatic allografts

BACKGROUND: MHC class I chain-related antigen A (MICA) and MHC class I chain-related antigen B (MICB) are HLA class I related products of polymorphic MHC genes. Constitutive expression in normal tissue is limited to gut epithelium but can be induced in other epithelial cells by stress. Specific antibodies against MICA have been reported in the serum of patients who had rejected kidney allografts, suggesting a potential role for these molecules in transplant immunopathology. However, expression of MICA and MICB in transplanted organs has not been demonstrated. In this study, we report the expression of MICA and MICB in renal and pancreatic allograft biopsies, which were obtained due to clinical signs of rejection. METHODS: A monoclonal antibody directed against MICA and MICB was used to perform indirect immunohistochemistry on formalin fixed, paraffin embedded needle biopsies of kidney and pancreas allografts. The results of staining were then compared to the standard light microscopic evaluation of the biopsies for rejection. RESULTS: A total of 53 individual renal transplant biopsies and 19 pancreas transplant biopsies were assayed for expression of MIC. Histologically, renal biopsies were diagnosed as no rejection, acute tubular necrosis (ATN), acute rejection (AR), chronic rejection (CR), and acute and chronic rejection (ACR). No staining was observed in 7 of 10 kidneys showing no rejection. All 11 of the kidney biopsies with AR were positive, as were the 11 ATN cases, 9 of the 11 kidney biopsies with CR, and 7 of the 10 with ACR. The acini of normal, nontransplanted, pancreas, control specimen were consistently negative; however, islets were positive in all specimens. The acini and islets of five histologically normal pancreas biopsies were positive, as were the four biopsies with AR, seven biopsies with CR, and two with ACR. CONCLUSIONS: MICA and MICB are expressed in epithelial cells in allografted kidney and pancreas that show histologic evidence of rejection and/or cellular injury. In addition to previous findings of alloantibodies against MICA, expression of these gene products may play a role in allograft rejection.     

Prognostic value of cytotoxic T-lymphocytes and CD40 in biopsies with early renal allograft rejection

After renal transplantation, different immunological and non-immunological factors lead to long-term allograft deterioration. Acute rejection episodes are one risk factor for chronic renal allograft dysfunction (CRAD). Following the current Banff classification the histological grade in acute rejection episodes is of limited prognostic value, therefore, additional morphological surrogate markers would be helpful. We investigated the biopsies of 91 patients with early acute rejection episodes for the immunohistochemical expression of key molecules (perforin, granzyme B, TIA-1, CD40) in the T cell-mediated rejection process. Staining results were correlated to long-term allograft outcome. Patients with greater than 2% of granzyme B or greater than 25% of CD40-positive cells in the interstitial infiltrate showed significantly shorter allograft survival. Patients with a CD40-positive vascular rejection or greater than 2% of granzyme B-positive cells in the interstitial infiltrate were significantly correlated with an earlier onset of CRAD. Our findings provide potential morphological surrogate markers in biopsies with early acute rejection episodes after renal transplantation. These could become part of combined clinical and histological algorithms, allowing patient-specific risk estimation and customized therapy options to be made.     

Expression of cyclooxygenase-1 and cyclooxygenase-2 in human renal allograft rejection – a prospective study

Cyclooxygenases (COX) are known to be involved in inflammatory kidney diseases. However, there are no data available about the expression of COX-1 and only preliminary reports about the expression of COX-2 in biopsies of patients undergoing acute renal allograft rejection. We conducted this prospective study to analyze the expression, distribution, and cellular localization of COX-1 and -2 and thus to elucidate the role of COX in human kidney transplantation. One hundred forty-four biopsies were included from patients without rejection and unaltered morphology (n = 60), with acute interstitial rejection (n = 7), with acute vascular rejection (n = 21), with chronic allograft nephropathy (n = 16), without rejection but with various other lesions (n = 40). COX-1 and -2 expression was localized in each biopsy by immunohistochemistry. We found a highly significant up-regulation of COX-1 in vessels and in infiltrating interstitial cells of patients with acute allograft rejection compared with biopsies with well-preserved tissue. Also, COX-2 expression was significantly elevated in infiltrating interstitial cells of biopsies with acute rejection. This is the first prospective study demonstrating a significant induction of both COX-1 and -2 in human allograft biopsies with acute rejection after renal transplantation.