膀胱尿路上皮癌中PD-L1和PD-L2的表达及临床意义

目的探讨膀胱尿路上皮癌(urothelial bladder cancer, UBC)中程序性死亡配体-1(programmed death-ligand 1, PD-L1)以及程序性死亡配体-2(programmed death-ligand 2, PD-L2)的表达及临床意义。方法采用免疫组化法检测58例UBC组织中PD-L1、PD-L2的表达,分析两者表达与UBC临床病理特征的关系。结果肿瘤细胞PD-L1在高级别UBC、pT2-pT4 UBC中的表达高于低级别UBC、pTa-pT1 UBC(cut-off值为1%、10%、50%,P均<0.05)。cut-off值为5%时,肿瘤浸润性免疫细胞(tumor-infiltrating immune cell, TIIC)PD-L1在高级别UBC、pT2-pT4 UBC中的表达高于低级别UBC、pTa-pT1 UBC(P均<0.05);cut-off值为10%时,TIIC PD-L1在pT2-pT4 UBC中的表达高于pTa-pT1 UBC(P=0.014)。肿瘤细胞PD-L2表达与UBC组织学分级以及pT分期等临床病理参数均无关(P均>0.05)。cut-off值为50%时,TIIC PD-L2在pT2-pT4 UBC中的表达高于pTa-pT1 UBC(P=0.044)。肿瘤细胞PD-L1、PD-L2表达状态与UBC患者的总生存期(overall survival, OS)均无关(P均>0.05)。cut-off值为5%时,TIIC PD-L1高表达患者比低表达患者的OS缩短(P=0.011);TIIC PD-L2表达状态与患者的OS无相关性(P均>0.05)。结论 UBC肿瘤细胞及TIIC异常表达PD-L1及PD-L2,评估UBC中PD-L1、PD-L2的表达状态可为临床开展免疫治疗提供参考。     

肺癌中PD-L1表达和调节性T细胞浸润的关系及意义

目的：探讨非小细胞肺癌(non-small cell lung cancer, NSCLC)中程序性死亡受体配体1(programmed death receptor ligand 1, PD-L1)表达和调节性T细胞(regulatory T cell, Treg)浸润的关系及临床意义。方法采用免疫组化法检测78例NSCLC组织中PD-L1表达和Treg浸润情况,分析二者之间的相关性及与NSCLC临床病理特征的关系。结果肺癌组织中PD-L1的表达显著高于癌旁组织(52．5% vs 6．4%),肺癌组织中 Treg浸润亦明显高于癌旁组织[(18．63±16．67)个/HPF vs (2．96±2．97)个/HPF]。晚期肺癌组织中PD-L1的表达高于早期肺癌组织(70．0% vs 41．7%),晚期肺癌组织中Treg浸润亦明显高于早期肺癌组织(73．3% vs 35．4%),有淋巴结转移组中PD-L1表达和Treg浸润均明显高于无淋巴结转移组,PD-L1表达和Treg浸润程度与淋巴结转移及临床分期相关(P<0．05)。肺癌组织中PD-L1表达和Foxp3+Treg的浸润密度呈正相关(rs =0．611,F=78．82,P=0．023)。结论肺癌微环境中PD-L1表达和Treg浸润呈正相关,二者可能共同参与肺癌的进展和免疫逃逸。     

免疫检查点分子PD-L1和B7-H4在卵巢癌组织中的表达及意义北大核心

为探讨免疫检查点分子PD-L1和B7-H4在卵巢癌组织中的表达特点以及临床意义,收集46例手术切除的卵巢癌组织,通过免疫组化染色技术检测PD-L1和B7-H4分子的表达水平和定位,采用χ2检验分析其与样本临床参数之间的关系,Spearman法分析相关性。结果显示,PD-L1和B7-H4分子在卵巢癌组织内中/高表达所占百分比分别为67.4%(31/46)和82.6%(38/46),二者同时中/高表达率为58.7%(27/46)。膜PD-L1中/高表达与病理类型、分化程度有关(P<0.05),膜B7-H4中/高表达与病理类型有关(P<0.05);膜PD-L1与膜B7-H4的表达有相关性(r=0.553,P<0.05),浆PD-L1和浆B7-H4的表达有相关性(r=0.429,P<0.05)。该研究提示,卵巢癌组织中癌细胞膜表面PD-L1中/高表达提示肿瘤分化差,胞膜同时中/高表达PD-L1和B7-H4分子主要集中在卵巢透明细胞癌亚组,PD-L1与B7-H4在胞膜和胞浆的表达具有相关性,在免疫治疗卵巢癌时需考虑PD-L1和B7-H4抗体联合靶向干预。     

胃癌中CMTM6和PD-L1的表达及其临床意义

目的 探讨CMTM6和PD-L1在胃癌中的表达及其与临床病理特征、预后的关系。方法 收集103例胃癌患者的临床资料，采用免疫组化MaxVision两步法检测胃癌组织中CMTM6和PD-L1的表达，分析两者表达与临床病理特征的关系。结果 103例胃癌组织中CMTM6高表达70例(68.0%),低表达33例(32.0%);PD-L1高表达66例(64.1%),低表达37例(35.9%)。正常胃黏膜组织中CMTM6高表达39例(37.9%)、低表达64例(62.1%);PD-L1高表达35例(34.0%)、低表达68例(66.0%);两组相比差异有显著性(P<0.001)。CMTM6表达与肿瘤分化程度、浸润深度、脉管浸润、淋巴结转移、远处转移、TNM分期和PD-L1表达相关(P<0.05)。PD-L1表达与患者性别、脉管浸润、神经侵犯和TNM分期相关(P<0.05)。Kaplan-Meier生存分析表明，CMTM6高表达与胃癌患者不良预后相关(P=0.008)。Cox单因素生存分析显示，胃癌患者的预后与肿瘤最大径、浸润深度、脉管浸润、远处转移、淋巴结转移、TNM分期和CMT...     

PD-1及其配体在原发性干燥综合征患者唇腺中的表达及临床意义

目的:观察干燥综合征(Sjogren's syndrome,SS)患者唇腺活检组织中程序性死亡分子-1(programmed death-1,PD-1)、程序性死亡配体-1(programmed death ligand-1,PD-L1)、程序性死亡配体-2(programmed death ligand-2,PD-L2)的表达,以及其与临床病理特征的相关性.方法:对所有患者和正常对照组的唇腺活检标本进行HE染色观察淋巴细胞浸润程度,免疫组织化学观察PD-1,PD-L1和PD-L2的表达.结果:SS患者唇腺活检组织中PD-1,PD-L1和PD-L2的表达较正常对照组明显增强.SS患者唇腺活检标本淋巴细胞的浸润程度与PD-1和PD-L2的表达强度无明显相关性,与PD-L1的表达强度呈正相关.SS患者疾病活动度评分与PD-1,PD-L1及PD-L2的表达强度均无明显相关性.SS患者疾病损害评分与PD-1的表达强度无明显相关性,与PD-L1和PD-L2的表达强度呈负相关.结论:PD-1,PD-L1及PD-L2在SS患者唇腺活检组织中的表达明显增强,PD-1/PD-L信号通路可能在SS的免疫病理损害中起重要作用.     

弥漫大B细胞淋巴瘤中PD-1、PD-L1的表达

目的探讨弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中程序性死亡受体-1(programmed death-1,PD-1)及程序性死亡配体-1(programmed death-ligand 1,PD-L1)的表达及临床意义。方法采用免疫组化法检测184例DLBCL组织中PD-1、PD-L1的表达,分析两者表达与DLBCL临床病理特征的关系。结果 184例DLBCL中肿瘤细胞PD-1、PD-L1阳性率分别为1. 63%、43. 48%;微环境细胞PD-1、PD-L1阳性率分别为11. 41%、26. 09%。肿瘤细胞及微环境细胞PD-1的表达与患者性别、年龄、Hans分型、CD5、EBER、ALK以及CD30的表达差异均无显著性(P均 0. 05)。非生发中心B细胞样(nongerminal center B-cell-like,non-GCB)型DLBCL肿瘤细胞(47. 55%)及微环境细胞(31. 47%)中PD-L1的阳性率高于生发中心B细胞样(germinal center B-cell-like,GCB)型DLBCL肿瘤细胞(29. 27%)及微环境细胞(7. 32%)(P均0. 05); EBV+DLBCL肿瘤细胞(75. 00%)及微环境细胞(58. 33%)中PD-L1的阳性率高于EBV-DLBCL肿瘤细胞(40. 74%)及微环境细胞(24. 07%)(P均0. 05); CD30+DLBCL(66. 67%)微环境细胞中PD-L1的阳性率高于CD30-DLBCL微环境细胞(27. 00%)(P=0. 005)。结论 PD-L1在non-GCB型以及EBV+DLBCL肿瘤细胞及微环境细胞中有更高的阳性率;且PD-L1在CD30+DLBCL微环境细胞中有更高的阳性率。     

程序性死亡蛋白配体-1/程序性死亡蛋白-1在老年胃癌患者外周血CD8+T淋巴细胞中的表达及临床意义

目的 通过分析程序性死亡蛋白配体-1/程序性死亡蛋白-1(programmed death ligand-1/programmed death-1,PD-L1/PD-1)在老年胃癌患者外周血CD8+T淋巴细胞中表达情况,探讨其临床意义.方法 选择同一时期90例老年胃癌患者(具有不同临床分期)及90例老年健康体检者,分别采取新鲜外周血后经流式细胞仪检测血中CD8+T淋巴细胞表面PD-L1/PD-1表达情况,结合老年胃癌患者的临床分期,分析PD-L1/PD-1在不同胃癌分期中的表达意义.结果 PD-L1/PD-1在老年胃癌患者外周血CD8+T淋巴细胞呈高表达,相较于老年健康者差异具有统计学意义(t=7.043,P<0.05);外周血CD8+T淋巴细胞的PD-L1/PD-1阳性表达均随着临床分期的增加而增加,呈明显的正相关性(r=0.883,P<0.05);外周血CD8+T淋巴细胞的PD-1阳性表达随着临床分期的增加而增加,两者之间呈正相关性(r=0.811,P<0.05);临床分期较晚的老年胃癌患者外周血CD8+T淋巴细胞PD-L1/PD-1表达比其他相对较早的临床分期老年胃癌患者显著上升(t=4.377,P<0.05).结论 PD-L1/PD-1信号通路异常在老年胃癌患者病变的发生发展过程中发挥了重要作用,外周血CD8+T淋巴细胞PD-L1/PD-1的表达对评判患者的预后具有指导作用.     

弥漫性大B细胞淋巴瘤患者PD-L1的表达及其不良预后因素分析

目的 探究弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)患者程序性死亡配体-1(programmed death-ligand 1,PD-L1)的表达情况及其不良预后因素.方法 选择2013年6月至2017年6月于我院就诊的DLBCL患者60例作为研究对象,所有患者随访3年.检测患者肿瘤细胞、微环境细胞中PD-L1的表达情况,并检测患者ALK、CD5、CD30、C-Myc、EBER、BCL-2、BCL-6的表达情况.比较PD-L1表达阳性和阴性患者各项临床指标间的差异,并分析影响患者3年总生存率以及无进展生存率的不良预后因素.结果 弥漫性大B细胞淋巴瘤患者中,肿瘤细胞PD-L1表达阳性的患者中,non-GCB型、临床分期Ⅲ所占比例均显著大于阴性患者,3年总生存率以及无进展生存率均显著小于阴性组.肿瘤微环境细胞PD-L1阳性患者各项临床指标与阴性患者间差异均无统计学意义.年龄≥60岁、肿瘤细胞PD-L1阳性表达、BCL-2阳性表达、BCL-6阳性表达均是影响患者3年总生存率的不良影响因素;临床分期III型、BCL-2阳性表达均是影响患者3年无进展生存率的不良影响因素.结论 肿瘤细胞PD-L1阳性表达是影响DLBCL患者生存及不良预后的显著相关因素.     

肺腺癌中MSI、PD-L1的表达与临床病理特征及预后的相关性

目的探讨浸润性肺腺癌中MSI、PD-L1表达与临床病理特征及预后的相关性。方法采用免疫组化EnVision法检测292例浸润性肺腺癌中MSI及PD-L1表达,应用单因素、多因素进行PFS、OS生存分析。结果 MSI阳性常见于女性(P=0.011)、临床T分期早期(P=0.009)、PD-L1阴性(P=0.013)的肺腺癌。临床T分期早期(P=0.012)是肿瘤细胞MSI-H阳性的独立相关因素。PD-L1阳性多出现于临床T分期晚期(P=0.015)。MSI阳性组平均生存时间和中位生存时间均比阴性组长,MSI-H组平均生存时间和中位生存时间均比MSI-L+MSS组长。PD-L1表达与预后呈正相关(PFS:P=0.004;OS:P=0.005);PD-L1表达是肺腺癌患者的独立预后因子(PFS:P=0.028;OS:P=0.049)。结论肺腺癌中MSI及MSI-H表达均与临床T分期早期有相关性,提示MSI、MSI-H可作为肺腺癌发生过程中的早期分子事件,PD-L1阳性提示患者预后较好,MSI及MSI-H均与PD-L1阴性有相关性。     

PD-1/PD-L1信号通路与肿瘤免疫逃逸

PD-1及其配体PD-L1、PD-L2广泛表达于多种细胞表面,是一组与免疫负调控有关的共刺激分子。PD-1/PD-L1信号通路的激活有利于肿瘤逃脱机体免疫监视,但具体的肿瘤免疫逃逸机制尚未清楚。就抗PD-1和抗PD-L1抗体疗效而言,仍然需要对肿瘤免疫逃逸机制做更深入的研究。文章就现有研究成果作一综述如下。     

PD-L1分子与疾病免疫调节

PD-LI(曾命名为B7-H1)是迄今发现的B7家族中较新的共刺激分子,与T细胞上的配体(PD-1)结合后可调节T细胞的活化及分化.它通过激活初始T细胞、抑制活化的效应T细胞及调节细胞因子的分泌等而参与多种免疫过程.以PD-L1为靶点的免疫调节与动物器官移植排斥反应、自身免疫性疾病、慢性病毒感染、肿瘤免疫逃逸等疾病的发生、发展密切相关.本文就PD-LI在上述疾病中发生、发展及治疗中的作用做一综述.     

PD-L1 and PD-L2 modulate airway inflammation and iNKT-cell-dependent airway hyperreactivity in opposing directions

Interactions of the inhibitory receptor programmed death-1 (PD-1) with its ligands, programmed death ligand (PD-L)1 and PD-L2, regulate T-cell activation and tolerance. In this study, we investigated the role of PD-L1 and PD-L2 in regulating invariant natural killer T (iNKT)-cell-mediated airway hyperreactivity (AHR) in a murine model of asthma. We found that the severity of AHR and airway inflammation is significantly greater in PD-L2^−/− mice compared with wild-type mice after either ovalbumin (OVA) sensitization and challenge or administration of α-galactosylceramide (α-GalCer). iNKT cells from PD-L2^−/− mice produced significantly more interleukin (IL)-4 than iNKT cells from control mice. Moreover, blockade of PD-L2 interactions of wild-type iNKT cells in vitro with monoclonal antibodies (mAbs) resulted in significantly enhanced levels of IL-4 production. In contrast, PD-L1^−/− mice showed significantly reduced AHR and enhanced production of interferon-γ (IFN-γ) by iNKT cells. iNKT-deficient Jα18^−/− mice reconstituted with iNKT cells from PD-L2^−/− mice developed high levels of AHR, whereas mice reconstituted with iNKT cells from PD-L1^−/− mice developed lower levels of AHR compared with control. As PD-L2 is not expressed on iNKT cells but rather is expressed on lung dendritic cells (DCs), in which its expression is upregulated by allergen challenge or IL-4, these findings suggest an important role of PD-L2 on lung DCs in modulating asthma pathogenesis. These studies also indicate that PD-L1 and PD-L2 have important but opposing roles in the regulation of AHR and iNKT-cell-mediated activation.     

PD-L1和PD-L2在树突状细胞上的表达及其生物学意义

探讨小鼠髓系DC (CD8α )中PD L1和PD L2的表达及其在T淋巴细胞活化中的作用。采用mCD4 0L CHO和TNF α分别刺激凋亡肿瘤细胞负载DC 4 8h ;免疫荧光标记检测DC表型 ;RT PCR和realtime PCR检测PD L1和PD L2mRNA转录水平 ;ELISA测定IL 2的分泌水平 ;3 H TdR掺入试验和51Cr释放试验测定DC体外激发T细胞的增殖和细胞毒杀伤率。结果显示 :PD L1和PD L2随着DC的成熟呈上调表达 ,CD4 0配基化DC的PD L1和PD L2均为中度表达 ,TNF α激发的DC为高度表达 ,二者呈现差异性表达 (P <0 0 5 ) ;CD4 0配基化髓系DC分泌IL 2的量明显高于TNF α组 (P <0 0 5 ) ,体外刺激T增殖和激活CTL能力在CD4 0配基化DC组最高 (P <0 0 5 )。提示CD4 0配基化的小鼠髓系DC呈现PD L1和PD L2的中度表达 ,IL 2大量分泌 ,这些均有助于激发有效的特异性免疫应答     

人外周血淋巴细胞及单核细胞表达PD-L1和PD-L2的动力学研究

目的： 探讨PD-L1和PD-L2在正常人外周血静息和活化的B细胞、T细胞及单核细胞表面的表达规律。 方法： 利用荧光抗体染色和流式细胞术分析静息状态及经多克隆刺激剂脂多糖（LPS）和美洲商陆丝裂原（PWM）刺激6、24、48和72 h后表达PD-L1和PD-L2的B细胞和T细胞百分率，同时分析静息状态及经IFN-γ和LPS共同刺激24、48、72 和96 h后表达PD-L2的单核细胞百分率。 结果： 静息B细胞和T细胞表面不表达PD-L1，LPS和PWM刺激6 h后表达PD-L1的B细胞百分率均显著高于静息B细胞，24 h达到最高，分别为(46.26±10.71)%和(43.67±6.14)%，之后随时间延长而降低；表达PD-L1的T细胞百分率在LPS刺激下无显著变化，但PWM刺激6 h后百分率明显高于静息条件，24 h达到最高，为(25.42±9.23)%，之后下降。静息B细胞和T细胞不表达PD-L2，活化后PD-L2表达也无明显上调。另外，静息状态单核细胞表面不表达PD-L2，体外活化24 h后表达PD-L2的单核细胞百分率显著上调，48 h达到最高为(28.70±14.22)%，之后随时间而降低。 结论： 活化的淋巴细胞表达PD-L1，但不表达PD-L2，后者在活化的单核细胞表面表达，并且后者达到高峰的时间迟于前者。     

Endothelial expression of PD-L1 and PD-L2 down-regulates CD8+ T cell activation and cytolysis

Interactions between CD8+ T cells and endothelial cells are important in both protective and pathologic immune responses. Endothelial cells regulate the recruitment of CD8+ T cells into tissues, and the activation of CD8+ T cells by antigen presentation and costimulatory signals. PD-L1 and PD-L2 are recently described B7-family molecules which bind to PD-1 on activated lymphocytes and down-regulate T cell activation. We found that PD-L1 is expressed on interferon-gamma stimulated cultured human and mouse endothelial cells, while PD-L2 was found on stimulated human but not mouse endothelial cells. Expression was further up-regulated by TNF-alpha. Antibody blockade of endothelial cell PD-L1 and PD-L2 enhanced endothelial cell costimulation of PHA-activated human CD8+ T cells. Antibody blockade of mouse endothelial cell PD-L1 enhanced both IFN-gamma secretion and cytolytic activity of CD8+ T cells in response to endothelial cell antigen presentation. These results show that IFN-gamma activated endothelial cells can inhibit T cell activation via expression of the immunoinhibitory PD-L1 and PD-L2 molecules. Endothelial expression of PD-ligands would allow activation and extravasation of T cells without excessive vessel damage. Our findings highlight a potentially important pathway by which endothelial cells down-regulate CD8+ T cell-mediated immune responses.     

PD-L1 and PD-L2 have distinct roles in regulating host immunity to cutaneous leishmaniasis

To compare the roles of programmed death 1 ligand 1 (PD-L1) and PD-L2 in regulating immunity to infection, we investigated responses of mice lacking PD-L1 or PD-L2 to infection with Leishmania mexicana. PD-L1(-/-) and PD-L2(-/-) mice exhibited distinct disease outcomes following infection with L. mexicana. In comparison to susceptible WT mice, PD-L1(-/-) mice showed resistance to L. mexicana, as demonstrated by reduced growth of cutaneous lesions and parasite burden. In contrast, PD-L2(-/-) mice developed exacerbated disease with increased parasite burden. Host resistance to L. mexicana is partly associated with the development of a Th1 response and down-regulation of the Th2 response. Both PD-L1(-/-) and PD-L2(-/-) mice produced levels of IFN-gamma similar to WT mice. However, the development of IL-4-producing cells was reduced in PD-L1(-/-) mice, demonstrating a role for PD-L1 in regulating Th cell differentiation. This inadequate Th2 response may explain the increased resistance of PD-L1(-/-) mice. Although no alterations in Th1/Th2 skewing were observed in PD-L2(-/-) mice, PD-L2(-/-) mice exhibited a marked increase in L. mexicana-specific antibody production. Increased Leishmania-specific IgG production may suppress the healing response through FcgammaR ligation on macrophages. Taken together, our results demonstrate that PD-L1 and PD-L2 have distinct roles in regulating the immune response to L. mexicana.     

PD-1/PD-L pathway and autoimmunity

Programmed cell death 1 (PD-1) was isolated in 1992 by subtractive-hybridization technique, as a molecule whose expression is enhanced by apoptotic stimuli. Since then we have been analyzing the function of PD-1 in the regulation of immune responses. Generation of PD-1 deficient mice, pathophysiological analyses of autoimmune diseases in PD-1 deficient mice, identification of two ligands, and analyses of downstream events of PD-1 revealed that PD-1 prevents autoimmunity by inhibiting activation of self-reactive lymphocytes. These findings were further applied on human autoimmune diseases and single nucleotide polymorphisms (SNPs) on human PD-1 gene have been reported to link with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and type I diabetes.     

PD-1/PD-L pathway and autoimmunity

Programmed cell death 1 (PD-1) was isolated in 1992 by subtractive-hybridization technique, as a molecule whose expression is enhanced by apoptotic stimuli. Since then we have been analyzing the function of PD-1 in the regulation of immune responses. Generation of PD-1 deficient mice, pathophysiological analyses of autoimmune diseases in PD-1 deficient mice, identification of two ligands, and analyses of downstream events of PD-1 revealed that PD-1 prevents autoimmunity by inhibiting activation of self-reactive lymphocytes. These findings were further applied on human autoimmune diseases and single nucleotide polymorphisms (SNPs) on human PD-1 gene have been reported to link with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and type I diabetes.