免疫检查点及PD-1/PD-L1抑制药在肿瘤免疫诊断治疗中的应用

程序性死亡受体-1(PD-1)/程序性死亡配体-1(PD-L1)单克隆抗体为代表的免疫检查点抑制药在肿瘤治疗领域取得了巨大的成功。故对免疫检查点抑制药在肿瘤免疫中的作用进行概述,并分析了免疫检查点抑制药的研究热点PD-1/PD-L1通路在抗肿瘤免疫的治疗意义,再对目前PD-1/PD-L1抑制药在诊断、治疗和预后中的应用调查,在此基础上分析PD-1/PD-L1抑制药进一步应用需要研究的问题,以期为PD-1/PD-L1抑制药进一步应用提供研究参考。     

靶向USP2下调PD-L1表达的丹酚酸B抗肿瘤免疫作用研究

以程序性死亡配体1/程序性死亡受体1 (programmed cell death ligand 1/programmed cell death protein 1,PD-L1/PD-1)免疫检查点阻断治疗为代表的肿瘤免疫疗法在临床上取得了令人鼓舞的治疗效果。小分子肿瘤免疫治疗药物与免疫检查点抗体药物的联用,为肿瘤治疗提供了新策略。因此,研发能阻断PD-L1/PD-1免疫检查点相互作用的小分子药物是下一代肿瘤免疫疗法的新方向。本研究对丹酚酸B (salvianolic acid B, SAB)下调肿瘤细胞中PD-L1表达发挥抗肿瘤作用及机制进行了研究。利用Western blot、流式细胞术、PD-1/PD-L1相互作用分析SAB对结肠癌细胞RKO和前列腺癌细胞PC3细胞内和膜表面PD-L1蛋白表达水平的影响;荧光定量PCR检测SAB对PD-L1 mRNA的影响;细胞阻抗法和结晶紫法检测SAB对人类外周血单核细胞(peripheral blood mononuclear cell,PBMC)杀伤肿瘤细胞的效果;表面等离子共振技术分析SAB与泛素羧基末端水解酶2 (ubiquitin c...     

肿瘤免疫检查点PD-1/PD-L1抑制剂的中药活性成分研究进展

肿瘤免疫治疗已成为新型的癌症治疗手段,有望彻底消除肿瘤。免疫检查点抑制剂,特别是程序性死亡受体-1（PD-1）和程序性死亡受体-配体1（PD-L1）抗体在多种实体瘤的治疗中取得很好的临床疗效,但是生物制剂存在免疫原性强、价格昂贵等缺点,因此,寻找免疫检查点小分子抑制剂成为未来肿瘤免疫疗法的新挑战。本文将综述近年发现的抑制PD-1/PD-L1表达的中药活性小分子及其对肿瘤免疫微环境的调控作用。     

运用抗PD-L1抗体治疗晚期癌症患者的安全性与疗效评价

靶向单克隆抗体在肿瘤的诊断与治疗领域不断获得进展,并显示出较好的前景。通过抗原-抗体反应激活机体的免疫系统杀灭肿瘤细胞是众多学者共同关注的方向。T细胞应答是控制肿瘤的发生与发展的最重要免疫应答,因此怎样激活T细胞发挥杀灭肿瘤细胞作用具有极其重要的研究价值。现有的研究发现,肿瘤细胞能够在自身表面表达一种称为程序性死亡配体-1(PD-L1)的蛋白,这种蛋白能够与机体T细胞上PD-1蛋白质结合,这种结合能够抑制T细胞发现肿瘤细胞、     

程序性死亡受体1的检测方法及其在乳腺癌中表达的研究进展

近年来,肿瘤的免疫治疗已成为继传统手术、放疗、化疗、内分泌治疗和靶向药物治疗手段之后新的治疗方法,尤其以程序性死亡受体1(PD-1)/程序性死亡配体1(PD-L1)位靶点的免疫检查点抑制治疗使得非小细胞肺癌、黑色素瘤等恶性实体肿瘤患者获得了有效持久的临床获益。PD-L1的检测是免疫检查点抑制剂治疗的关键环节,但在乳腺癌中的其检测方法和判定标准尚未统一。就PD-L1的检测方法以及其在乳腺癌各分子分型中的表达进行综述。     

程序性死亡分子1及其配体与自身免疫病的研究进展

程序性死亡分子1 (programmed death-1,PD-1)及其配体(programmed death-1 ligand,PD-L)是一对新近研究较多的负性共刺激分子.PD-1与其配体结合后,传导的信号能抑制T淋巴细胞增殖,在调节T细胞活化和免疫耐受过程中发挥关键作用,从而对防止自身免疫病的发生、发展具有重要作用.PD-1/PD-L在免疫调节中的作用使其有望成为治疗免疫性疾病新的靶向分子.     

抗程序性死亡蛋白1及其配体抗体治疗经典霍奇金淋巴瘤进展

肿瘤细胞可利用抑制性免疫检查点蛋白程序性死亡蛋白1（programmed death-1,PD-1）及程序性死亡蛋白配体1（programmed death ligand-1,PD-L1）逃脱T细胞介导的细胞毒作用,阻断PD-1/PD-L1通路的治疗性抗体对越来越多的实体瘤产生持续的临床效应。此文综述PD-1/PD-L1信号通路阻断剂在经典霍奇金淋巴瘤免疫治疗中的作用,并总结使用该治疗手段的部分临床研究。     

来那度胺抑制PD1/PD-L1信号调节淋巴细胞对HepG2的免疫杀伤作用

目的 探究来那度胺抑制PD1/PD-L1信号增强淋巴细胞杀伤HepG2的能力。方法 提取人淋巴细胞与HepG2共培养,流式细胞术检测HepG2的凋亡,CCK-8检测HepG2的活力,Western blot检测PD-L1/PD-1表达,Elisa法检测IL-2、IL-12、TNF-α的表达。构建HepG2的荷瘤小鼠模型,来那度胺干预后,计算抑瘤率,Western blot检测小鼠肿瘤组织中PD-1和PD-L1的表达,qPCR检测肿瘤组织中IL-2、IL-12和TNF-α的mRNA表达。结果 来那度胺可以增加淋巴细胞对于HepG2的杀伤能力,HepG2凋亡率显著增高,PD-1和PD-L1表达下降。IL-2、IL-12和TNF-α表达增高。来那度胺对于荷瘤小鼠有明显的肿瘤抑制作用,可以降低肿瘤组织中PD-1和PD-L1表达,IL-2、IL-12和TNF-α的mRNA表达增高。结论 来那度胺抗肿瘤作用与其免疫调节有关,来那度胺可以通过抑制PD-1/PD-L1信号增强淋巴细胞对HepG2的杀伤作用。     

2型糖尿病患者单核细胞表面PD-L1的表达及其生物学意义

目的 探讨2型糖尿病(T2DM)合并血管病变患者外周单核细胞表面负性协同刺激分子程序性死亡配体-1(PD-L1)的表达变化及其生物学作用.方法 应用流式细胞术分析单纯糖尿病组(42例)、糖尿病合并大血管病变组(48例)、糖尿病合并急性冠脉综合症组(35例)和健康对照组(48例)外周血中单核细胞表面PD-L1分子的表达水平,同时检测各T淋巴细胞亚群表面程序性死亡受体1(PD-1)的表达;采用ELISA法检测各组血清中肿瘤坏死因子α(TNF-a)、γ-干扰素(IFN-γ)等细胞因子浓度.结果 与健康对照组相比,糖尿病三个组患者外周单核细胞表面PD-L1表达均显著增加(P<0.05),并与血管病变的严重程度呈正相关,而CD4+和CD8+T淋巴细胞表面PD-1受体的表达亦呈上调性表达(P<0.05).随着血管病变的进展,糖尿病患者外周1NF-α和IFN-γ水平呈增加的趋势(P<0.05).结论 T2DM患者外周单核细胞表面PD-L1表达上调在DM及合并大血管病变、急性冠脉综合症的发生和发展中发挥着一定的作用.     

程序性死亡因子-1（PD-1）抑制药新药Nivolumab的临床研究进展

摘 要 目的： 介绍程序性死亡因子-1(PD-1)抑制药抗癌新药Nivolumab。方法: 根据文献,对Nivolumab的作用机制以及目前获批或进入III期的几个主要适应证的临床研究结果进行综述与评价。结果: Nivolumab可通过与PD-1的结合,阻断其与关键配体PD-L1和PD-L2的相互作用,恢复T细胞的抗肿瘤活性;已经完成或正在进行中的多项临床研究结果显示Nivolumab单用或者与化疗药物或CTLA-4抑制药Ipilimumab合用,对于黑色素瘤、非小细胞肺癌以及肾细胞癌的效果优于目前临床应用的化疗药物,Nivolumab总体耐受性好,3～4级的不良事件主要为免疫介导肺炎,肝功能紊乱,疲劳等。结论:Nivolumab通过肿瘤免疫效应可改善对黑色素瘤、非小细胞肺癌以及肾细胞癌等多种肿瘤的治疗效果。     

PD-1/PD-L1信号通路在黑色素瘤中的研究进展

细胞程序性死亡因子1(programmed death-1,PD-1)及其配体1(programmed death-ligand 1,PD-L1)是一对共刺激分子,激活细胞程序性死亡因子1及其配体1信号通路可抑制肿瘤特异性T细胞活性,有助于肿瘤细胞逃避免疫监视。而阻断该通路可以激活机体抗肿瘤免疫应答,抑制肿瘤细胞的生长。近年来,针对细胞程序性死亡因子1及其配体1通路的免疫疗法在黑色素瘤的治疗中已取得显著效果。本文对细胞程序性死亡因子1及其配体1通路在黑色素瘤中的表达调控、细胞程序性死亡因子1及其配体1抑制剂在黑色素瘤治疗中的临床研究进行综述,讨论细胞程序性死亡因子1作为一种新的免疫疗法在临床的应用前景和不良反应。     

Gemcitabine and checkpoint blockade exhibit synergistic anti-tumor effects in a model of murine lung carcinoma

Lung cancer is leading cause of cancer death in the world. Chemotherapy is currently one of the standard treatments for lung cancer. Gemcitabine is a pyrimidine nucleoside drug which has been approved by FDA to treat lung cancer. However, acquired resistance inevitable develops after Gemcitabine treatment, limiting clinical efficacy. Lewis lung carcinoma (LLC) cells were treated with Gemcitabine and cell apoptosis and programmed cell death-ligand 1 (PD-L1) expression were analyzed by flow cytometry. LLC mouse model was established to analysis the proportion and programmed cell death-1 (PD-1) expression of CD8 + T cells. Anti-tumor effect by treating with Gemcitabine and anti-PD-1 antibody was measured through in vivo LLC mouse model. Gemcitabine treatment induces tumor cell apoptosis and PD-L1 expression. Further study showed that Gemcitabine treatment also increases CD8⁺ and CD4⁺ T cells proportion, PD-1 and PD-L1 expression in LLC mouse model. Combination therapy with Gemcitabine and αPD-1 not only has strong anti-tumor effect, but also could inhibit postsurgical recurrence of LLC. Our findings demonstrated that the combination therapy of Gemcitabine and αPD-1 is an effective therapeutic strategy for lung cancer.     

Mitomycin C synergizes with PD-L1 blockade in treatment of non-small cell lung cancer

OBJECTIVE Programmed death 1 ligand(PD-L1) checkpoint inhibitor was a promising therapy but the response rate was only about 20%. Chemotherapy was reported to final y kil tumor cel s by triggering immune response. To improve the response of PD-L1 blockade,we tried to find chemotherapeuticaldrugs to combine with PD-L1 antibody(Ab). METHODS Non-small cell lung cancer(NSCLC) cells were pre-treated with mitomycin C(MMC) and then co-cultured with PBMCs to explore the effect of the combination of MMC with PD-L1 Ab. Lewis lung cancer(LLC) cells were used to establish xenograft model in mice and subjected to MMC or PD-L1 Ab treatment alone or combo. RESULTS MMC increased the expressions of PD-L1 and MHC-Ⅰ in NSCLC cells in vitro and in vivo and enhanced the activity of lymphocytes against NSCLC in vitro.The mice treated with combo of PD-L1 Ab with MMC showed an improved overall survival and inhibition of the tumor growth compared to monotherapy,which was linked to the increases of lymphocyte infiltration and granzyme B release in LLC cell xenograftmodels in mice. Mechanically, MMC activated the ERK pathwayand subsequently activated c-Jun that bound with PD-L1 promoter and recruited its co-factor STAT3 to enhance PD-L1 expression. Meanwhile, the ERK pathway activated P65 to promote the MHC-I expression. CONCLUSION A combination of MMC with PD-L1 blockade may improve the antitumor response and offer a promising new treatment modality against NSCLC and encourage to further study to confirm in clinic.     

PD-1/PD-L1与CTLA-4联合治疗实体瘤患者有效性和安全性的Meta分析

目的: 系统评价PD-1/PD-L1与CTLA-4联合治疗对比PD-1/PD-L1单药治疗实体瘤患者的有效性和安全性,旨在为临床用药提供循证参考。方法: 计算机检索PubMed、Embase、Cochrane Library、中国知网、CBM、维普和万方数据,检索时限均为各数据库建库至2021年11月。根据纳入与排除标准筛选文献后,采用Cochrane 系统评价员手册 5.1.0 推荐的偏倚风险评估工具对纳入文献质量进行评价;采用Rev Man 5.4软件进行Meta分析;采用倒漏斗图进行发表偏倚分析。结果: 共纳入22项研究,共计8 535例患者。Meta分析结果显示,从生存指标方面分析,PD-1/PD-L1与CTLA-4联合治疗显著改善实体瘤患者的客观反应率(ORR)、疾病控制率(DCR)和无进展生存期(PFS),但总生存期(OS)改善无统计学差异。亚组分析结果显示,PD-1/PD-L1与CTLA-4联合治疗能显著改善黑色素瘤患者的ORR和PFS,而OS改善无统计学差异;能显著改善肺癌患者的PFS,而ORR和OS改善无统计学差异;显著改善其他实体瘤患者的ORR和DCR,而PFS和OS改善无统计学差异。从安全指标方面分析,PD-1/PD-L1与CTLA-4联合治疗显著增加实体瘤患者的所有级别和3~4级不良反应发生率、死亡率以及中止治疗率。亚组分析结果显示,PD-1/PD-L1与CTLA-4联合治疗能显著增加黑色素瘤和其他实体瘤患者的3~4级不良反应发生率和中止治疗率,显著增加肺癌患者所有级别和3~4级不良反应发生率、死亡率以及中止治疗率。结论: PD-1/PD-L1和CTLA-4联合治疗与PD-1/PD-L1单药治疗相比,能显著改善实体瘤患者的PFS、DCR和ORR,但是也显著增加实体瘤患者治疗相关不良反应发生率。     

PD0325901, an ERK inhibitor,enhances the efficacy of PD-1 inhibitor in non-small cell lung carcinoma

ERK pathway regulated the programmed death ligand-1 (PD-L1) expression which was linked to the response of programmed death-1 (PD-1)/PD-L1 blockade therapy. So it is deducible that ERK inhibitor could enhance the efficacy of PD-1 inhibitor in cancer immunotherapy. In this study, PD0325901, an oral potent ERK inhibitor, strongly enhanced the efficacy of PD-1 antibody in vitro and in vivo models in non-small cell lung carcinoma (NSCLC) cells. Mechanistically, PD0325901 or shRNA-ERK1/2 significantly downregulated the PD-L1 expression in NSCLC cells and increased the CD3⁺ T cells infiltration and functions in tumor tissue. There was a positive correlation between the p-ERK1/2 expression and PD-L1 expression in patients with NSCLC. And the patients with low p-ERK1/2 expression were observed a high response rate of PD-1/PD-L1 blockage therapy. Our results demonstrate that PD0325901, an ERK inhibitor, can enhance the efficacy of PD-1 blockage against NSCLC in vitro and in vivo models. And the combination of ERK inhibitor such as PD0325901 and PD-1/PD-L1 blockage is a promising regimen and encouraged to be further confirmed in the treatment of patients with NSCLC.     

胃癌组织中程序性死亡受体-1和程序性死亡-配体1表达与临床病理特征及预后的相关性

目的:探讨胃癌组织中程序性死亡受体-1(PD-1)和程序性死亡-配体1(PD-L1)表达情况和临床病理特征、预后相关性。方法:选取2016年2月~2018年2月某院诊治实施手术治疗胃癌患者103例临床资料行回顾性分析,分析癌组织和癌旁组织中PD-1、PD-L1表达阳性率、死亡和存活率,患者癌组织中PD-1、PD-L1表达阳性率,及不同病理分期中PD-1、PD-L1表达阳性率差异。结果:癌组织PD-1和PD-L1表达阳性率显著高于癌旁组织[40.78%vs.0.00%,63.11%vs.0.97%,P<0.05]。死亡患者癌组织中PD-1、PD-L1表达阳性率显著高于存活患者[62.50%vs.21.82%,87.50%vs.43.64%,P<0.05]。不同分化程度、不同TNM分期PD-1表达阳性率无统计学意义(P>0.05)。不同分化程度PD-L1表达阳性率无统计学意义(P>0.05);TNM分期中III^IV期患者PD-L1表达阳性率显著高于I^II期患者[78.85%vs.29.41%,P<0.05]。结论:胃癌组织中PD-1和PD-L1表达水平较高,PD-L1表达水平和病理特征有一定相关性,PD-1和PD-L1与患者预后有一定相关性。     

MicroRNA-497-5p down-regulation increases PD-L1 expression in clear cell renal cell carcinoma

Recent advances in immunotherapy are raising hope to treat clear cell renal cell carcinoma (ccRCC) with PD-L1 inhibitors, but only a small portion of patients are PD-L1 positive. The heterogeneous expression pattern of PD-L1 in patient population suggests that PD-L1 expression is under the control of diverse regulatory mechanisms. Although recent studies have identified numerous novel PD-L1 regulators, reports on microRNAs which modulate PD-L1 expression are much scarce. In this study, we confirmed that PD-L1 expression was up-regulated in ccRCC compared to paired normal tissues. Using miRDB and miRTarBase, 11 microRNAs were predicted to target PD-L1. After measuring the microRNA panel with TaqMan assays, we found that microRNA-497-5p down-regulation was associated with PD-L1 up-regulation. In TCGA-KIRC dataset, microRNA-497-5p down-regulation was also associated with PD-L1 up-regulation as well as shorter survival. We further validated that PD-L1 was a direct target of microRNA-497-5p in two RCC cell lines. In addition, microRNA-497-5p inhibited cell proliferation, clone formation and migration, while promoted apoptosis in in-vitro assays. Our study reveals a novel regulatory mechanism of PD-L1 expression and the potential of miR-497-5p as therapeutic target and biomarker deserves further investigation.     

Functional tumor specific CD8 + T cells in spleen express a high level of PD-1

The inhibitory effects of programmed cell death 1 (PD-1) receptor on tumor specific T cells were mainly investigated at tumor site. While PD-1 expression can be rapidly unregulated upon T cell activation at lymphoid tissues, little is known about where PD-1 signal exerts its inhibitory function in tumor-bearing host. To address this issue, we assessed the effects of PD-1 on vaccine induced activation of splenic CD8 + T cells in mice. The vaccine consisted of mice CD8 + T cell epitope peptide and poly IC. After vaccination, spleen or tumor tissues were dissociated, IFN-γ synthesis and PD-1 expression by CD8 + T cells were detected by flow cytometry. We found that CD8 + T cells could be successfully activated in spleen after immunization, characterized by the capability of producing IFN-γ when encountering relevant peptide. These activated splenic CD8 + T cells also expressed a high level of PD-1. Although PD-L1 expression in spleen parenchyma was also increased after vaccination, PD-1 blockade did not affect the activation of splenic CD8 + T cells, but enhanced the anti-tumor effects of peptide vaccine. This synergetic effect of peptide vaccine plus PD-1 blockade was coupled with increased aggregation of IFN-γ + CD8 + tumor infiltrated lymphocytes (TILs), rather than CD4 + TILs. The results indicated that for tumor-bearing host, PD-1 pathway exerted its inhibitory function at tumor site and PD-1 expression on the splenic CD8 + T cells correlated positively with IFN-γ synthesis.