Association of PD-L1 and HIF-1α Coexpression with Poor Prognosis in Hepatocellular Carcinoma

OBJECTIVE: To investigate the correlation between the expression of PD-L1 and HIF-1α in hepatocellular carcinoma (HCC) tissue and further analyze the association with clinical parameters and the prognostic value of coexpression in HCC patients. METHODS: We assessed the expression of PD-L1 and HIF-1α by immunohistochemistry in tumor tissue from 90 HCC patients who underwent curative hepatectomy. The results were validated in an independent cohort of additional 90 HCC patients. RESULTS: PD-L1 and HIF-1α exhibited in tumor tissue high expression rates of 41.11% (37/90) and 43.33% (43/90), respectively, and their expressions were positively correlated (r = 0.563, P < .01). High expression of PD-L1 was significantly associated with low albumin levels (P < .05); high expression of HIF-1α was significantly correlated with high alpha-fetoprotein (AFP) levels and low albumin levels (P < .05); high expression of both PD-L1 and HIF-1α was also significantly associated with high AFP levels and low albumin levels (P < .05). High expression of PD-L1, HIF-1α, as well as both PD-L1 and HIF-1 α was respectively significantly associated with worse overall survival (OS) and disease-free survival (DFS) (P < .05). Patients with co-overexpression of PD-L1 and HIF-1α had the worst prognosis compared with other groups. Additionally, multivariate Cox regression models suggested that high expression of PD-L1, HIF-1α, as well as both PD-L1 and HIF-1α was an independent prognostic factor for OS and DFS (P < .05). Furthermore, the positive correlation and prognostic values of PD-L1 and HIF-1α were validated in an independent data set. CONCLUSION: We demonstrated that HCC patients with co-overexpression of PD-L1 and HIF-1α in tumor tissue had a significantly higher risk of recurrence or metastasis and death compared with others. Therefore, more frequent follow-up is needed for patients with co-overexpression of PD-L1 and HIF-1α. At the same time, a combinational therapy with HIF-1α inhibitors in conjunction with PD-L1 blockade may be beneficial for HCC patients with co-overexpression in the future.     

全球PD-1/PD-L1单克隆抗体市场竞争格局

目的:从产品开发角度分析PD-1/PD-L1单克隆抗体的发展现状和未来趋势。方法:检索科睿唯安(Clarivate Analytics)的Cortellis数据库的数据,利用定量分析法和对比分析法对检索结果进行分析。结果:目前已有5种PD-1/PD-L1单克隆抗体上市、4种PD-1/PD-L1单克隆抗体处于预注册及6种PD-1/PD-L1单克隆抗体处于临床Ⅲ期,未来市场上的PD-1/PD-L1单克隆抗体将呈现快速增长趋势。此外,PD-1/PD-L1单克隆抗体的商业交易也越来越多,目前共发生包括药物开发及商业化许可、专利资产出售及早期药物研发合作等10余起交易,其中药物开发及商业化许可是最主要的交易模式。结论:虽然PD-1/PD-L1单克隆抗体市场尚处于起步阶段,但随着未来技术的不断发展改进,相信未来有更多的PD-1/PD-L1单克隆抗体上市,为癌症及其他疾病的治疗提供新的契机。     

Phosphorylated RB Promotes Cancer Immunity by Inhibiting NF-κB Activation and PD-L1 Expression

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PD-L1降解途径与肿瘤免疫治疗

程序性死亡受体配体1 (programmed death ligand 1, PD-L1)是肿瘤免疫检查点阻断治疗中的重要靶点,其在多种细胞中均有表达。肿瘤细胞可通过高表达PD-L1来增强程序性死亡受体1 (programmed death 1, PD-1)抑制信号,从而促进肿瘤免疫逃逸。近年来,以抗PD-1/PD-L1抗体为代表的肿瘤免疫治疗给癌症治疗带来了革命性的变化。然而,肿瘤免疫治疗仅能对部分患者产生持久的疗效,多数患者对肿瘤免疫治疗的应答短暂或没有应答。研究发现, PD-L1的降解对肿瘤免疫治疗应答至关重要。本文综述了PD-L1的溶酶体降解途径、蛋白酶体降解途径及PD-L1降解与肿瘤免疫治疗的相互作用,旨在为进一步增强肿瘤免疫治疗的应答率和应答范围提供研究思路。     

PD-1/PD-L1免疫检查点抗体药物制剂稳定性开发

近年来抗体药物在生物医药领域发展迅速。随着抗体疗法种类的不断增加和PD-1/PD-L1靶点蛋白结构的确证,临床上,越来越多针对PD-1/PD-L1免疫检查点的单克隆抗体药物被不断开发并应用于治疗死亡率高、治愈率低的多种癌症中。但是抗体药物制剂开发水平还需进一步提高,一方面同一靶点的抗体产品同质化严重,另一方面抗体药物的理化性质比小分子药物复杂,因此需要针对不同单克隆抗体的药物特性,筛选出适应于临床应用的稳定蛋白制剂处方。概括了不同抗体药物制剂处方成分(缓冲液成分、药物辅料)的作用,结合PD-1/PD-L1靶点介绍了抗体药物制剂稳定性开发的一般策略以及CDE相关的审评要点。     

PD-1/PD-L1免疫检查点抗体药物制剂稳定性开发

近年来抗体药物在生物医药领域发展迅速。随着抗体疗法种类的不断增加和PD-1/PD-L1靶点蛋白结构的确证,临床上,越来越多针对PD-1/PD-L1免疫检查点的单克隆抗体药物被不断开发并应用于治疗死亡率高、治愈率低的多种癌症中。但是抗体药物制剂开发水平还需进一步提高,一方面同一靶点的抗体产品同质化严重,另一方面抗体药物的理化性质比小分子药物复杂,因此需要针对不同单克隆抗体的药物特性,筛选出适应于临床应用的稳定蛋白制剂处方。概括了不同抗体药物制剂处方成分(缓冲液成分、药物辅料)的作用,结合PD-1/PD-L1靶点介绍了抗体药物制剂稳定性开发的一般策略以及CDE相关的审评要点。     

PD-1/PD-L1抗体在肿瘤临床治疗中的应用

PD-1是主要表达在活化T细胞上的免疫抑制性受体,与表达在肿瘤细胞表面的配体PD-L1结合,通过降低T细胞的免疫应答,使肿瘤免疫逃逸。靶向PD-1/PD-L1这一信号通路的单克隆抗体类药物的研发是当前抗肿瘤治疗领域的研究热点。目前,已有多项临床研究提示,PD-1及PD-L1抗体药物治疗肿瘤的疗效显著且不良反应较小,有望改善部分晚期肿瘤患者的预后。     

PD-1/PD-L1与Treg细胞相关性的研究新进展

PD-1和PD-L属于B7家族的共刺激分子,介导免疫反应的负性调节信号。Treg细胞是一个具有免疫调节作用的T细胞亚群,在机体的免疫耐受和免疫稳定中具有重要作用。本文就PD-1/PD-L1与Treg细胞的免疫调节作用及相关性研究进展作简要综述。     

PD-L1对Jurkat细胞活性及凋亡的影响

目的:探讨PD-L1(Programed Death Ligand-1)表达的调控以及其对Jurkat细胞增殖分化及凋亡的影响.方法:通过IFN-γ在体外诱导Jurkat细胞使PD-L1表达上调,并设计针对PD-L1的siRNA对Jurkat细胞进行转染抑制PD-L1的表达,通过RT-PCR、PCR测定Jurkat细胞分泌IL-2的变化;流式细胞术测定Jurkat细胞的凋亡情况.结果:运用β-actin进行半定量后,根据相应的模板量进行PCR,在设计的3条针对PD-L1的siRNA(siRNA-A,siRNA-B,siRNA-C)中选出一条可以高效抑制PD-L1表达的即siRNA-A,siRNA-A终浓度80nM转染Jurkat细胞24小时后PD-L1的表达明显抑制.使用IFN-γ终浓度2000U/ml作用于Jurkat细胞24小时后PD-L1表达明显增高,通过RT-PCR、PCR观察到PD-L1表达高低与Jurkat细胞分泌IL-2水平呈负相关,流式细胞术显示PD-LI表达增强能降低Jurkat细胞凋亡.结论:体外合成的针对PD-L1基因特异性siRNA,转染Jurkat细胞后可特异性抑制PD-L1的表达.IFN-γ在体外可刺激Jurkat细胞PD-L1表达增高.PD-L1信号在体外对Jurkat细胞分泌IL-2呈负相关,并且PD-L1能抑制Jurkat细胞凋亡.     

PD-1及其配体PD-L在HIV感染中的作用

共刺激信号不仅可以协助激活T细胞,而且可以通过负性共刺激信号下调免疫反应,从而精确地调节应答的程度和持续时间。以PD-1/PD-L为代表的负性共刺激途径,在免疫学的试验研究和临床免疫治疗中均有重要意义,近年来该途径被认为与移植排斥反应、自身免疫性疾病、肿瘤免疫逃逸,尤其是慢性病毒感染如艾滋病等疾病密切相关。本文主要介绍PD-1及其配体PD-L的结构、表达特性及其在HIV感染中的作用,探讨其在艾滋病病情发展中的作用机制和治疗方面的应用前景。     

系统性红斑狼疮患者外周血B淋巴细胞PD-L1的表达

目的 探讨系统性红斑狼疮（systemic lupus erythematosus,SLE）患者外周血B淋巴细胞PD-L1的表达及其在SLE发病中的意义.方法 应用免疫荧光标记和流式细胞仪技术检测SLE患者外周血B淋巴细胞上PD-L1的表达水平.结果 活动期SLE患者外周血B淋巴细胞CD19＋、CD19＋/PD-L1＋的表达水平明显高于SLE非活动期患者（P＜0.05）和正常对照组（P＜0.05）.结论 活动期SLE患者B淋巴细胞的PD-L1表达异常增加,其可能在SLE发病机制中起重要作用.     

PD-L1 Expression and Survival among Patients with Advanced Non–Small Cell Lung Cancer Treated with Chemotherapy

BACKGROUND: Recent clinical trial results have suggested that programmed cell death ligand 1 (PD-L1) expression measured by immunohistochemistry may predict response to anti–programmed cell death 1 (PD-1) therapy. Results on the association between PD-L1 expression and survival among patients with advanced non–small cell lung cancer (NSCLC) treated with chemotherapy are inconsistent. MATERIAL AND METHODS: We evaluated the relationship between PD-L1 expression and overall survival (OS) among 204 patients with advanced NSCLC treated at Aarhus University Hospital, Aarhus, Denmark, from 2007 to 2012. PD-L1 expression was measured using a prototype immunohistochemistry assay with the anti–PD-L1 22C3 antibody (Merck). PD-L1 strong positivity and weak positivity were defined to be traceable to the clinical trial version of the assay. RESULTS: Twenty-five percent of patients had PD-L1 strong-positive tumors, and 50% had PD-L1 weak-positive tumors. No statistically significant association was found between PD-L1 expression and survival; adjusted hazard ratio of 1.34 (95% confidence interval, 0.88-2.03; median OS, 9.0 months) for the PD-L1 strong-positive group and 1.07 (0.74-1.55; median OS, 9.8 months) for the PD-L1 weak-positive group compared with the PD-L1–negative group (median OS, 7.5 months). No association was seen between PD-L1 expression and OS when PD-L1 expression levels were stratified by median or tertiles. CONCLUSIONS: In concordance with previous studies, we found PD-L1 measured by immunohistochemistry to be frequently expressed in patients with advanced NSCLC. However, PD-L1 expression is not a strong prognostic marker in patients with advanced NSCLC treated with chemotherapy.     

PD-L1通过STAT3抑制血管内皮细胞增殖迁移

[目的]探讨PD-L1通过STAT3抑制血管内皮细胞增殖迁移的影响。[方法]实验分组:A:pcDNA3.1+siRNA-Control(NC组);B:pcDNA3.1+siRNA-PD-L1(si-PD-L1组);C:STAT3+siRNA-Control(STAT3组);D:STAT3+siRNA-PD-L(STAT3+si-PD-L1组),按照分组做细胞转染。通过MTT、Cell-LightTMEd U细胞增殖检测转录因子STAT3、siRNA-PD-L1对血管内皮细胞增殖率的影响;通过细胞划痕实验检测0、24、36 h融合率、Western Blot检测转录因子STAT3、siRNA-PD-L1对血管内皮细胞增殖标志物PCNA表达水平的影响。[结果]C组HUVEC细胞的增殖率比A组显著增高60%,共转pcDNA3.1-STAT3+siRNA-PD-L1后D组细胞增殖率比C组显著下降52%。划痕实验划痕融合率C组比A组显著增高(p0.05),D组与C组相比降低(p0.01)。PCNA表达C组显著高于其他组(p0.05),D组与C组相比显著降低40%(p0.01)。[结论]STAT3显著促进HUVECs增殖迁移能力,而干扰PD-L1以后,可下调细胞增殖迁移。     

PD-1/PD-L1在脓毒症发生发展过程中的作用研究

脓毒症是机体应对感染而发生的一种全身性炎性反应,且发病机制复杂,免疫调节紊乱是最显著的特点之一。程序性死亡分子-1(programmed death-1,PD-1)及其配体(programmed death ligand-1,PD-L1)是介导机体免疫反应的负性调节信号,在脓毒症的发生发展过程中扮演着重要角色。因此,对PD-1/PD-L1在脓毒症的发生发展及治疗中所发挥的作用进行简短归纳。     

PD-L1对细菌脓毒症免疫抑制的影响及其机制*

目的:探讨在脂多糖(LPS)所致细菌脓毒症免疫抑制中树突状细胞(DCs)程序性细胞死亡配体-1(PD-L1)的表达情况及其相关信号通路。方法:细菌脂多糖刺激骨髓来源树突状细胞诱导淋巴细胞免疫抑制模型,实验分为5组:对照组(Con)、脂多糖组(LPS)、2-(4-吗啉基)-8-苯基-4H-1-苯并吡喃-4-酮+脂多糖组(LY294002+LPS)、吡咯烷二硫代甲酸铵盐+脂多糖组(PDTC+LPS)和脂多糖+封闭PD-L1组(LPS+αPD-L1)。小鼠骨髓来源单核细胞用含粒细胞巨噬细胞集落刺激因子(rmGM-CSF 10 ng/ml)和白介素4(rmIL-4 1 ng/ml)的10%胎牛血清1640培养基于CO₂培养箱37℃静置培养4 d后,LPS(10 ng/ml)处理DCs静置12 h获得PD-L1高表达的DCs作为免疫抑制刺激细胞。通路抑制剂LY294002(10 μmol/L)、PDTC (20 μmol/L)作用1 h阻断PI3K和NF-κB信号。采用流式细胞分析、激光共聚焦显微成像检测LPS诱导树突状细胞PD-L1表达及磷脂酰肌醇3激酶/丝氨酸苏氨酸激酶B (PI3K/AKT) 信号通路活化情况;BrdU细胞增殖实验和γ-干扰素酶联免疫斑点实验检测LPS诱导树突状细胞PD-L1表达上调对抗原特异性T细胞增殖反应及细胞毒性T细胞杀伤作用的影响。结果:与对照组比较,LPS组DCs表面PD-L1阳性细胞百分比升高(P＜0.01),PD-L1荧光信号强度增强,且主要分布于细胞表面和细胞质,DCs介导的T细胞增殖水平降低(P＜0.01),γ-干扰素斑点形成细胞数下降(P＜0.01)。与LPS组比较,LY294002+LPS组、PDTC+LPS组和LPS+αPD-L1组PD-L1荧光信号强度降低,T细胞增殖水平升高(P＜0.01),γ-干扰素斑点形成细胞数上升(P＜0.01),改善树突状细胞介导的T细胞免疫抑制现象。 结论:PD-L1是介导脂多糖所致细菌脓毒症免疫抑制的关键分子,PI3K信号、NF-κB信号也参与此免疫抑制过程。