Interleukin 2 receptor expression on peripheral blood lymphocytes in association with renal allograft rejection

Periodic assay of IL-2 receptor expression on the surfaces of peripheral blood lymphocytes might provide information predictive of in vivo immunologic events. This study compares two methods of determining IL-2 receptor expression after renal transplantation in cynomolgus monkeys. The first utilized single color staining of peripheral blood mononuclear cells with mouse anti-human IL-2 receptor monoclonal antibody followed by a fluorescein-labeled goat anti-mouse IgG antibody. Epics C cell sorter windows were set to count cells of the size and granularity of normal lymphocytes. The second utilized two-color staining with fluorescein-labeled anti-IL-2 receptor antibody, combined with phycoerythrin-labeled anti-CD4 antibody or with phycoerythrin-labeled anti-CD8 antibody. Two-color staining allowed the sorter windows to be enlarged to count all mononuclear cells, regardless of size or granularity, without introducing the contaminating effects of monocytes. Data obtained from single-color staining showed no consistent or significant expression of the IL-2 receptor on peripheral lymphocytes in association with the rejection process. Data obtained from two-color staining revealed an increase of IL-2 receptor expression on peripheral T cells of at least 10% from the postoperative baseline, which preceded the creatinine rise from allograft rejection in 13 of 13 animals. Increases in IL-2 receptor expression on T cells were not specific to rejection, however. Some animals in which treatment produced a delay of rejection showed a transient rise in IL-2 receptor expression around post-transplant day 5, which was not followed by a rise in creatinine. The two-color staining technique described provides a sensitive means of detecting IL-2 receptor expression in vivo and documents the association of increases in IL-2 receptor expression on T cells with rejection.     

The association of HLA-DR antigens with acute steroid-resistant rejection and poor kidney graft survival

The association of an early steroid-resistant rejection (SRR) with HLA-DR antigens was analyzed in 410 kidney transplantations. A severe SRR leading to a poor (45%) 1-year graft survival (GS) occurred in 22 transplantations (5%). An acute reversible rejection (ARR) with a GS of 94% was found in 80 transplantations (20%). For the 308 (75%) transplantations with no early rejection episodes the GS was 91%. HLA-DR5 and -DR8 present in the donor as incompatible antigens were strongly associated with SRR. Further, a mismatched DR1 from the kidney donor predicted a rejection, either reversible or irreversible. These findings may have practical implications for an early diagnosis of SRR and for considering of rescue therapy whenever transplantation with disparities in these loci has been performed.     

Flow cytometric evaluation of cytotoxic peripheral blood lymphocytes in acute renal graft rejection.

The presence of the S6F1+ epitope on the surface of CD8+ lymphocytes is believed to be uniquely representative of cytotoxic subpopulations. A preliminary study was conducted to evaluate the CD8+ S6F1+ peripheral lymphocytes by flow cytometry in patients undergoing renal allograft biopsy for allograft dysfunction. Lymphocytes, obtained at the time of biopsy, were analyzed by flow cytometry with CD8-FITC/S6F1-RD1 as the test monoclonal antibody and MsIgG-RD1/MsIgG-FITC as internal control. A 100% increase in S6F1+ cells over internal control was considered to be positive result. The results were correlated with the histopathologic findings in 14 instances of allograft dysfunction occurring 26.5 +/- 11.6 days posttransplantation. The histopathologic diagnosis was acute cellular rejection in eight cases, acute tubular necrosis in four, and cyclosporine nephrotoxicity in two. Flow cytometric detection of an increase in S6F1+ cells yielded a sensitivity of 87.5% and a specificity of 83.3% for the diagnosis of acute rejection. It would appear that the use of a monoclonal antibody to detect increases in the number of CD8+ S6F1+ peripheral lymphocytes is a valuable test for the detection of acute allograft rejection in the initial period after transplantation.     

Renal tubular epithelial cells as immunoregulatory cells in renal allograft rejection

Kidney transplantation is the best therapeutic option for patients with end-stage kidney disease. However, the long-term survival of kidney transplants still remains relatively poor even under potent immunosuppression. Thus, it is necessary to further review the pathogenesis of renal allograft failure. Here, we discuss the potential impact of activated resident tubular epithelial cells (TECs) on infiltrating leukocyte responses on renal allograft failure. Immunohistochemical staining or in situ hybridization of renal allograft biopsies shows that activated TECs produce inflammatory cytokines and may act as nonprofessional antigen-presenting cells in the response to stimulation from leukocyte infiltration. Further experimental studies confirm that by a feedback loop, activated TECs positively or negatively regulate the destructive activity of infiltrating leukocyte through (1) alteration of leukocyte activation, proliferation, differentiation, and migration to the graft through secretion of cytokines and chemokines; and (2) direct regulation of infiltrating T-cell function through cell-cell contact. Specifically targeting kidney factors has significant impact on renal graft damage or kidney disease. This review suggests that graft TECs can regulate intragraft immune responses, and modulation of specific graft TEC responses as a therapeutic strategy may benefit long-term survival of kidney transplants.     

Mononuclear cells in renal allografts. Correlation with peripheral blood T lymphocyte subpopulations and graft prognosis

Cellular infiltrates in 20 renal allograft biopsies taken at the time of acute rejection episodes were analyzed with antibodies reactive with monocytes (BRL antihuman monocyte monoclonal antibody), T lymphocyte subpopulations (OKT- and Leu-series of monoclonal antibodies), and B lymphocytes (heterologous antihuman IgM antibodies). For demonstration of the various mononuclear cells, an indirect immunoperoxidase technique was used. The number of monocytes in the infiltrates varied from 10-20%; the number of B lymphocytes was always below 10%. The T lymphocytes accounted for 50-90% of the total number of mononuclear cells. The OKT4/OKT8 ratios for the T lymphocytes in the graft infiltrates were correlated with the peripheral blood OKT4/OKT8 ratios measured by indirect fluorescence and flow cytometry. The OKT4/OKT8 ratios for perivascular infiltrates correlated far better with peripheral blood OKT4/OKT8 ratios (r = 0.72) than did the OKT4/OKT8 ratios for interstitial infiltrates (r = 0.58). Low or inverted OKT4/OKT8 ratios and low or inverted Leu 3a/Leu 2a ratios were associated with a high risk of irreversible graft rejection (P less than 0.001 for perivascular infiltrates).     

移植肾排异反应中肾小管上皮细胞的表型转化

目的 观察人移植肾肾穿刺标本不同排异反应病变中肾小管上皮细胞表型转化状态,探讨排异反应与肾小管上皮细胞表型转化的相关性。 方法 免疫组织化学SP法检测55例移植肾穿刺不同病变组中肾小管上皮细胞α平滑肌肌动蛋白（α-SMA）的表达。 结果 各组萎缩病变的肾小管上皮细胞均有α-SMA阳性表达,表现为近基底膜处胞质阳性染色,提示出现了表型转化。在无萎缩病变的肾小管中,仍有部分肾小管细胞呈α-SMA阳性染色。7例基本正常病例组均无肾小管上皮细胞的表型转化。28例急性T细胞介导排异 IA级病例组中,1例肾小管上皮细胞α-SMA阳性表达率为25%~50%,3例为10%~25%。14例排异反应IB 级中,1例α-SMA阳性表达率达50%以上,2例25%~50%,2例10%~25%。 结论 随着急性T细胞介导性排异反应加重,肾小管上皮细胞发生表型转化的现象明显增强。     

肾移植急性排斥反应中Fas-L表达与肾小管上皮细胞凋亡的研究

目的 探讨异基大鼠肾移植急性排斥期肾小管上皮细胞凋亡与Ｆａｓ－Ｌ表达的变化。方法 选用近交系ＤＡ（ＲＴ１＾ａ）、ＬＥＷ（ＲＴ１＾ｌ）大鼠进行原位左肾移植。实验组：异基因移植组（ＤＡ→ＬＥＷ）;对照组：同基因移植组（ＬＥＷ→ＬＥＷ）。于手术后第２、４、６天分别取移植肾进行病理学检查及电镜扫描,采用末端脱氧核苷酸转移酶（ＴｄＴ）介导的脱氧核苷酸原位末端标记法（ＴＵＮＥＬ）检测移植肾脏中的凋亡细胞,逆转     

微小RNA-29b在肾小管上皮细胞的表达及调节上皮间质转分化的作用机制

目的研究血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)对大鼠肾小管上皮细胞(NRK-52E)中微小核糖核酸(miRNA)-29b(miR-29b)表达的影响,以及miR-29b在细胞间质转分化中的作用,为研究肾间质纤维化机制提供新思路。方法体外培养NRK-52E,将不加入AngⅡ的细胞作为对照组,加入AngⅡ的细胞为实验组,按加入不同浓度的AngⅡ(10~(-9)mol/L、10~(-7)mol/L)及AngⅡ刺激时间(12 h、24 h、48 h)的长短又分为实验亚组,同时设立miR-29b mimic转染组及阴性对照组。显微镜下观察对照组与实验组细胞的形态学改变,运用实时荧光定量聚合酶链反应检测rniR-29b表达,蛋白质免疫印迹试验检测Snail、E钙黏附素(E-cadherin)、α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)蛋白表达。两两比较用t检验。结果实验组中AngⅡ刺激12 h后,细胞形态呈梭形改变;随着AngⅡ刺激浓度的增加,α-SMA、Snail蛋白表达显著升高,E-cadherin表达量显著下降,与对照组相比,差异均有统计学意义(P0.05)。实验组中随着AngⅡ刺激时间的延长,细胞α-SMA、Snail和E-cadherin的表达呈时间依赖性,α-SMA、Snail的表达显著增加,E-cadherin表达量显著降低,与对照组相比,P0.05。同时AngⅡ呈浓度和时间依赖性下调NRK-52E细胞中miR-29b表达,与对照组相比,P0.05。此外,miR-29b mimic转染组和阴性对照组相比,α-SMA、Snail蛋白表达显著降低(P0.05),E-cadherin表达量显著升高(P0.05)。结论 AngⅡ可诱导NRK-52E细胞发生间质转分化,且miR-29b可抑制AngⅡ诱导的NRK-52E细胞发生间质转分化。     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4+ [(20.74±9.02)%] and CD8+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P < 0. 01 ). Fas-positive CD8+ ceils were reduced in invasive TCC as compared with superficial TCC [ ( 5. 83±3.95 ) % vs ( 5. 83±3.95 ) %, P < 0.05 ], but there was no difference between primary and recurrent cases ( P > 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC.     

膀胱尿路上皮癌患者外周血T淋巴细胞凋亡蛋白Fas的表达

目的 探讨膀胱尿路上皮癌与Fas介导的血T淋巴细胞凋亡的关系.方法 应用流式细胞仪检测52例膀胱癌患者和37例健康对照者外周血CD4Fas、CD8Fas阳性T细胞.结果 膀胱癌患者外周血CIM、CD8Fas阳性T细胞分别为(20.74±9.02)%、(7.51±5.93)%,对照组分别为(15.68±5.58)%、(4.27±3.08)%,肿瘤组较对照组明显增加(P<0.01).浸润组CD8Fas阳性T细胞数为(5.83±3.95)%,表浅组为(5.83±3.95)%,浸润组较表浅组明显降低(P<0.05).复发组CD4、CD8Fas阳性T细胞与初发组比较筹异无统计学意义(P>0.05).结论 血T淋巴细胞Fas表达的异常可能在膀胱癌的发生发展中起重要的作用.