血小板/内皮细胞粘附分子在急性排斥反应时表达的研究

应用免疫级化SP法和计算机图像分析系统观察分析18例急性排斥反应时移植肾活检标本中血小板／内皮细胞粘附分子（PECAM－1，CD31）的表达。结果显示急性排斥反应时肾小球毛细血管内皮细胞中PECAM－1阳性反应强度较正常明显减低或由阳性转为阴性，而在肾小管上皮中表达增强；PECAM－1与HLA－DR抗原在肾小球和肾小管中的表达呈平行关系。提示细胞粘附分子PECAM－1对移植肾急性排斥反应有一定诊断意义。     

直肠间质瘤PD-1/PD-L1表达与患者预后的相关性分析

分析直肠间质瘤PD-1/PD-L1表达与患者的预后转归的相关性。随访直肠间质瘤患者100例,分析PD-1/PD-L1在直肠间质肿瘤组织中的表达、直肠间质肿瘤组织中PD-1/PD-L1蛋白的表达与术后生存期间的关系,通过多因素分析直肠间质肿瘤组织中PD-1/PD-L1与临床病理特征的关系。结果显示,不同性别患者肿瘤切除部位和分化程度细胞PD-L1表达的差异有统计学意义(P<0.05),直肠间质肿瘤组织中PD-1/PD-L1蛋白的表达与术后生存期间的关系存在差异性,Kaplan-Meier生存曲线显示直肠间质肿瘤组织中PD-1表达与患者术后生存时间存在相关性。结果表明,直肠间质瘤PD-1/PD-L1表达与患者的预后转归之间存在紧密的关系。     

程序性死亡-1及其配体信号通路在移植免疫中的作用

程序性死亡-1(PD-1)为负性调控共刺激分子,系T淋巴细胞和B淋巴细胞表面受体,属于免疫球蛋白超家族Ⅰ型跨膜糖蛋白,在T、B淋巴细胞抗原受体激发后呈诱导性表达.PD-1与其2个配体PD-L1和PD-L2结合,在免疫应答中起着负性调控作用.目前,人们愈加关注PD-1/PD-L信号通路与临床疾病的关系,如移植后排斥反应、哮喘、1型糖尿病等~([1]).本文对PD-1/PD-L信号通路在移植免疫中的作用进行如下综述.     

慢性排斥移植肾中细胞间粘附分子-1与HLA-DR表达的关系

目的：探讨慢性排斥移植肾中细胞间粘附分子－1（ICAM－1）和HLA－DR表达与间质淋巴细胞浸润的关系。方法：对20例慢性排斥肾移植受者进行肾活检，采用免疫组化技术（ABC法）检测移植肾内ICAM－1和HLA－DR的表达。结果：ICAM－1在慢性排斥移植肾肾小管上皮细胞和间质微小动脉内皮细胞表达增强，而HLA－DR表达则普遍上调，尤其在远曲小管。此外，在ICAM－1和HLA－DR表达增强的局部血管周围和小管间质伴有大量淋巴细胞浸润。结论：慢性排斥移植肾中ICAM－1和HLA－DR表达增强可能在排斥反应中起诱导作用，尤其是间质炎细胞的浸润及抗原递呈，同时它们又可能使表达上调的细胞成为免疫反应效应支的靶细胞，从而参与慢性排斥的细胞免疫损伤及移植肾间质损害过程。     

白细胞介素6mRNA在移植肾中的表达及意义

目的 探讨肾移植急性排斥反应时白细胞介素6(IL-6)产生的主要来源,并为初步阐明肾移植急性排斥反应的分子学发病机制提供实验依据。方法 采用3＇IL-6寡核苷酸探针,运用原位杂交技术观察IL-6mRNA在移植肾中的表达。结果 (1)在急性排斥反应时,移植肾各部位表达IL-6mRNA明显增多,较环孢素A(CsA)中毒、稳定期移植肾及正常人均有显著升高。(2)在急性排斥反应时,肾小管上皮细胞表达IL-6mRNA强度较肾小球细胞、血管内皮细胞及间质细胞均有明显升高。(3)CsA中毒患者肾脏IL-6mRNA表达较稳定期移植肾及正常对照无明显增多。结论 肾移植急性排斥反应时,移植肾细胞可直接产生IL-6,移植肾中IL-6异常激活与表达同肾移植急性排斥反应的发生机制有密切的关系;肾小管表达IL-6的异常增多和活化揭示了肾小管上皮细胞在肾移植急性排斥反应的发病机理中具有重要意义。     

IgA肾病患者肾组织中增殖细胞核抗原表达

用免疫组化方法研究了２０例原发性ＩｇＡ肾病患者肾组织中增殖细胞核抗原（ＰＣＮＡ）表达。结果显示ＩｇＡ肾病患者肾组织中ＰＣＮＡ表达增加，肾小球内ＰＣＮＡ阳性细胞数、肾小管和间质中ＰＣＮＡ阳性细胞百分数均与肾组织学损害程度呈正相关；临床－病理研究显示肾小管中ＰＣＮＡ阳性细胞百分数分别与２４小时尿蛋白量、血清肌酐浓度（Ｓｃｒ）呈正相关；肾间质中ＰＣＮＡ阳性细胞百分数亦与２４小时尿蛋白量呈正相关。肾脏细胞增殖程度作为一项判断肾小球肾炎组织学损害程度和预后的指标值得进一步研究。     

PD-1和PD-L1表达与慢性HBV感染者肝脏病变程度的相关性

目的研究程序性死亡分子-1(programmed death-1,PD-1)及其主要配体(programmed death-1ligand,PD-L1)表达与肝脏病变程度的关系,并探讨其临床意义。方法不同临床类型慢性HBV感染者137例,含慢性乙型肝炎(CHB)患者102例,活动性肝硬化(LC)患者25例,慢加急性肝衰竭(ACLF)患者10例。其中,102例CHB患者中有22例行肝组织活检,炎症分级为G1～G4,纤维化程度分为S0～S4。健康对照10例。取新鲜血液分离外周血单个核细胞(PBMCs),流式细胞仪检测PBMCs表面PD-1和PD-L1阳性细胞比例以及人白细胞抗原(HLA)型别;对HLA-A2阳性者的PBMCs加入重组HBcAg体外培养7d,加入PE标记的HBV抗原表位肽-五聚体复合体(含HBV抗原表位肽为HB-cAg18-27),流式细胞仪计数CD8+T细胞数以及CD8和PD-1双阳性细胞。ELISA检测PBMCs培养上清液中IFN-γ。比较CHB、LC、ACLF三组间以及不同肝组织病变程度患者间PD-1、PD-L1、CD8+细胞数量和PD-1表达水平、IFN-γ水平的差异。结果在CHB、LC和ACLF组PD-1在PBMCs表面的表达阳性率分别为(55.4±16.5)%、(54.3±21.9)%、(45.5±26.5)%,与健康对照组比较,P值分别为0.000、0.001、0.071;三组PD-L1表达阳性率分别为(41.7±17.7)%、(33.4±17.9)%、(26.6±11.8)%,和对照组比较,P值分别为0.006、0.252、0.854。62例HLA-A2阳性患者的CD8+T细胞比例分别为(1.6±1.0)%、(3.1±1.6)%和(3.3±1.7)%,LC组和ACLF组明显高于CHB组(P0.05);PD-1阳性细胞占CD8+T细胞的比例各组间比较无统计学差异;三组PBMCs培养上清液中IFN-γ含量比较无统计学差异。不同肝组织炎症和纤维化程度的CHB患者间PD-1、PD-L1表达虽无明显统计学差异,但PD-L1在炎症G1和纤维化S0的患者中的表达较其他组患者高。结论慢性HBV感染者PD-1和PD-L1表达明显上调。肝脏病变程度与PD-1表达水平无关,但与HBV特异性CD8+T细胞数量及PD-L1表达相关。     

移植肾活组织检查191例的病理学研究

目的：研究供肾可能携带的病变及移植肾在术后出现合并症时间相应的病理组织学变化。方法对191例移植肾进行了活组织检查（以下简称“活检”,其中术中活检52例,术后活检139例,并进行病理学诊断与分类。结果（1）52例术中活检,40例（76．92％）正常,余12例（23．07％）供肾携带病变,其中细小动脉硬化3例（5．77％）、间质炎症4例（7．69％）、局部肾小管轻度萎缩3例（5．77％）、肾小管上皮细胞变性2例（3．84％）。（2）139例术后活检中明确诊断的134例（96．40％）,其中正常19例（13．66％）,超急性排斥反应（SAR）1例（0．71％）疑为急性排斥反应／临界性变化（S／B）15例（10．795）,急性排斥反应（AR）23例（16．54％、慢性移植肾肾病（CNA）37例（26．61％）、环孢素肾毒性损伤（CsA-NT)29例（20．80％）、急性肾小管坏死（ATN）8例（5．75％）、复发性肾炎（RN）2例（1．43％）、难以明确诊断的5例（3．59％）。（3）免疫细胞化学色显示AR时,侵入肾小管上皮内的Leu-7阳性细胞数增加,间质内CD68阳性细胞数明显增加。结论活检是诊断术后多种合并症的有效手段;CAN可在术后3个月发生;侵入肾小管上皮的Leu－7阳性细胞是协助诊断AR的有效指标。     

乙型肝炎病毒相关性肾炎患者肾组织Notch1受体表达与临床病理的相关性

目的 观察Notch1受体在乙型肝炎病毒相关性肾炎（HBV-GN）患者肾组织中的表达分布特点,探讨其与肾组织病理及临床表现的相关性。 方法 以2008年至2010年经肾活检确诊的HBV-GN患者48例为研究对象。用免疫组化方法观察HBV-GN肾组织中Notch1受体的分布特征;免疫荧光双标观察Notch1受体与HBsAg的分布关系;分析其与病理类型、肾小球病变、肾小管病变及临床指标的关系。 结果 Notch1受体主要分布在肾小管上皮细胞及间质区域,呈棕红色颗粒状,肾小球内也有少量表达。HBV-GN组患者肾组织Notch1受体阳性积分显著高于血清HBsAg阳性或阴性的原发性肾小球肾炎患者及正常肾组织对照。系膜增生性肾小球肾炎（MsPGN）组和膜增生性肾小球肾炎（MPGN）组Notch1受体阳性积分较高,但各病理类型组间Notch1积分差异无统计学意义。Notch1受体分布与HBsAg分布一致,其强度与肾间质纤维化（r = 0.473,P = 0.001）、小管萎缩（r = 0.690,P = 0.000）、炎细胞浸润（r = 0.616,P = 0.000）等肾小管间质病变呈正相关;患者肾功能与Notch1受体阳性积分呈负相关（r = -0.393,P = 0.006)。 结论 HBV-GN患者肾组织Notch1受体的表达增强,主要分布在肾小管上皮细胞及间质,与HBsAg分布一致;其表达水平与肾间质病变和肾功能变化相关。Notch1受体表达异常可能参与了HBV-GN的肾脏病理的进展。     

移植肾BCL-2及UCHL1的表达及临床意义

目的 探讨细胞凋亡与排斥反应,ＢＣＬ－２及ＵＣＨＬ１表达之间的关系,方法 收集１０例手术切除的无功能移植肾病理标本作为实验组,相应的移植术前活检组织作为对照组。采用免疫组织化学染色技术观测ＢＣＬ－２及ＵＣＨＬ１的移植肾的表达,并观察其组织切片的形态学变化。结果 对照组５０％以上的肾小管细胞ＢＣＬ－２呈阳性表达,实验组ＢＣＬ－２的表达明显减少（Ｐ〈０．０１）,尤其是当发生急性排斥反应时;ＵＣＨＬ１的     

纳米刀消融联合程序性死亡蛋白-1/程序性死亡蛋白配体-1治疗胰腺癌及其免疫学机制研究进展

胰腺癌特殊的肿瘤微环境不利于免疫治疗,而纳米刀消融可在一定程度上逆转免疫抑制,使其成为目前唯一适用于胰腺癌的消融治疗方法。动物研究结果证实,纳米刀消融联合程序性死亡蛋白-1（PD-1）/程序性死亡蛋白配体-1（PD-L1）治疗胰腺癌可延长患者生存时间。本文对纳米刀消融联合PD-1/PD-L1治疗胰腺癌及其免疫学机制研究进展进行综述。     

Research progress regarding programmed cell death 1/programmed cell death ligand 1 inhibitors combined with targeted therapy for treating hepatocellular carcinoma

In recent years, a number of targeted therapeutic agents have achieved success in phase III trials in patients with advanced hepatocellular carcinoma (HCC), including sorafenib, lenvatinib, and regorafenib. Immunotherapy is considered to be an effective treatment for advanced HCC. Immune checkpoint inhibitors targeting programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) are important antitumor immunotherapy agents that represent breakthroughs in the treatment of advanced HCC. However, treating advanced HCC is still a great challenge, and the need for new treatments remains urgent. This review briefly summarizes the research progress in the use of PD-1/PD-L1 inhibitors combined with targeted therapy for treating HCC.     

PD-1/PD-L1与肿瘤相关巨噬细胞在肿瘤发生中的相互作用研究进展

程序性死亡蛋白1（PD-1）/程序性死亡配体1（PD-L1）和肿瘤微环境中的肿瘤相关巨噬细胞（TAMs）与肿瘤的发生密切相关。TAMs又主要分为抗肿瘤作用的M1型和促肿瘤作用的M2型,而肿瘤组织中浸润的巨噬细胞主要以M2-TAMs为主。近年的研究进展主要为PD-1/PD-L1轴抑制TAMs的抗肿瘤作用,TAMs分泌的细胞因子、趋化因子、外泌体等参与调节PD-1/PD-L1的表达等方面。笔者就PD-1/PD-L1与TAMs在肿瘤发生中的作用进行综述,同时也对两者之间可能存在的相互作用进行了归纳,探讨靶向调控TAMs的功能对PD-1/PD-L1单抗治疗肿瘤效果的影响。     

Prognostic value of PD-1 and PD-L1 expression in patients with metastatic clear cell renal cell carcinoma

Objective

In renal cell carcinoma (RCC), several prognostic biomarkers have been identified and are under investigation. Several reports have shown that the expression of programmed death 1 (PD-1) and its ligand PD-L1 is associated with poor outcome for patients with RCC. The present study is aimed at evaluating the expression of PD-1 and PD-L1 and to investigate their clinical and prognostic significance in patients with clear cell RCC (CCRCC) having received molecular targeted therapies. In addition, we also evaluated the relationship between the expression of PD-1 and PD-L1 and intratumoral tumor infiltrating lymphocytes (TILs).

A protective role for programmed death 1 in progression of murine adriamycin nephropathy

Programmed death 1 (PD-1) is a novel member of the CD28/cytotoxic T-lymphocyte-associated protein-4 superfamily, which plays an important role in the regulation of activated T cells. However, it is not clear how PD-1 is expressed in normal and diseased kidney, nor if it has a role in progression of chronic renal disease. PD-1 expression and the effect of monoclonal anti-PD-1 antibody (Ab) were examined in murine adriamycin nephropathy (AN). BALB/c mice were divided into three groups: (a) normal mice, (b) adriamycin (ADR) with control immunoglobulin (Ig)G (ADR-IgG), and (c) ADR with anti-PD-1 Ab (ADR-Ab). AN was induced by a single intravenous injection of ADR. Anti-PD-1 Ab was given by intraperitoneal injection on alternate days from day 0 to day 10, or to day 18. Animals were killed at week 4. Renal function, histological change, and cytokine expression were examined. PD-1 mRNA was detected in kidney tissue of mice with AN in a dose- and time-dependent manner. PD-1 was mainly expressed on injured tubule cells and some interstitial cells, which co-stained with alpha-smooth muscle actin in AN, but not in normal kidney. Anti-PD-1 treatment up to day 18, but not to day 10, worsened glomerular and tubulointerstitial injury. The ratio of urinary protein/creatinine was significantly higher in ADR-Ab mice than ADR-IgG mice. The number of macrophages was significantly increased in ADR-Ab mice compared with ADR-IgG mice. Blockade of PD-1 worsened progressive renal histopathological and functional injury in murine AN. This suggests a possible protective role for PD-1 in chronic renal disease, and its potential as a treatment to slow disease progression.     

Gene transfer of programmed death ligand-1.Ig prolongs cardiac allograft survival

BACKGROUND: The CD28 homologue programmed death-1 (PD-1) and its ligands, PD-L1 and PD-L2 (which are homologous to B7), constitute an inhibitory pathway of T cell costimulation. The PD-1 pathway is of interest for immune-mediated diseases given that PD-1-deficient mice develop autoimmune diseases. We have evaluated the effect of local overexpression of a PD-L1.Ig fusion protein on cardiac allograft survival. METHODS: Adenovirus-mediated PD-L1.Ig gene transfer was performed in F344 rat donor hearts placed in the abdominal position in Lewis recipients. Inflammatory cell infiltrates in the grafts were assessed by immunohistochemistry. RESULTS: Allografts transduced with the PD-L1.Ig gene survived for longer periods of time compared with those receiving noncoding adenovirus or virus dilution buffer alone: median survival time (MST), 17 (range: 16-20) days vs. 11 (8-14) and 9 (8-13) days, respectively (P < 0.001). PD-L1.Ig gene transfer combined with a subtherapeutic regimen of cyclosporin A (CsA) was superior to CsA alone: MST, 25 (15-42) vs. 15 (13-19) days (P < 0.05). PD-L1.Ig gene transfer was associated with decreased numbers of CD4 cells and monocytes/macrophages infiltrating the graft (P < 0.05). CONCLUSIONS: Localized PD-L1.Ig expression in donor hearts attenuates acute allograft rejection in a rat model. The effect is additive to that of a subtherapeutic regimen of CsA. These results suggest that targeting of PD-1 by gene therapy may inhibit acute cardiac allograft rejection in vivo.     

Discordance of high PD-L1 expression in primary and metastatic urothelial carcinoma lesions

Immune checkpoint inhibitors (ICI) targeting PD-(L)1 are effective in select patients with advanced urothelial carcinoma (UC). High PD-L1 expression enriches for response to ICIs; however, the predictive value of PD-L1 expression is limited, which may be due in part to dynamic expression of PD-L1 in the tumor environment. We sought to characterize PD-L1 expression in primary UC and paired metastatic lesions to gain insight into the potential discordance of tumor PD-L1 expression during the metastatic process. Materials and methods: Immunohistochemical staining for PD-L1 using the SP-142 antibody was performed on primary tumors and matched metastatic specimens in 77 evaluable subjects with advanced UC. Immunohistochemical staining was scored for the percentage of cells positive (<5%, ≥5%) in tumor cell (TC) and immune cell (IC) compartments. Correlation of PD-L1 expression in TCs and ICs was estimated using Spearman's correlation coefficients (rho, ρ). Cohen's kappa statistics (κ) were utilized to assess the agreement in PD-L1 expression between groups. Results: High (≥5%) PD-L1 expression in primary and metastatic biopsies, respectively, was observed in 6.0% and 7.7% of TCs and in 14.5% and 11.5% of ICs. IC PD-L1 expression in primary tumors was not correlated with IC PD-L1 expression in paired metastatic lesions (ρ?=?0.05, P?=?0.67) and there was poor agreement in high expression rates between primary and metastatic lesions in the IC compartment (κ= 0.086). Conclusion: High PD-L1 IC expression is temporally and spatially discordant between primary and metastatic UC lesions. Future studies of PD-(L)1 targeted therapies in patients with metastatic UC may benefit from use of fresh biopsies of metastatic lesions to define PD-L1 expression when feasible.