OBJECTIVE To evaluate whether the IDO1 inhibitor 1-methyl-L-tryptophan(1-MT)combine calcium influx inhibitor carboxyamidotriazole(CAI)could further enhance the suppression of programmed death 1(PD-1)in CD8^+T cells an...OBJECTIVE To evaluate whether the IDO1 inhibitor 1-methyl-L-tryptophan(1-MT)combine calcium influx inhibitor carboxyamidotriazole(CAI)could further enhance the suppression of programmed death 1(PD-1)in CD8^+T cells and investigate the curative effect of the combined use.METHODS CD8^+T cells were isolated from normal mice spleen by negative selection using magnetic cell separation.The isolated CD8^+T cells were cultured in RPMI 1640 medium containing 10%FBS and 100 U·mL^(-1)IL-2 and activated by the addition of anti-CD3 and anti-CD28(1 g·L^(-1) each mabs).CD8^+T cells were pretreated for 48 h with drug and the fluo-3 as a marker of intracellular calcium concentration was detected by flow cytometry.The calcineurin(Ca N)levels were assayed with ELISA in CD8^+T cells after 48 h incubation with 10μm CAI.The nuclear translocations of NFAT and AHR were detected by immunofluorescent staining after 48 h of drug treatment.The expression of PD-1 in CD8^+T cells was analyzed by flow cytometry.RESULTS Intracellular fluorescent intensity was markedly debase due to CAI treatment(P<0.01).Meanwhile,the changes of CaN content had a resembled correlation(P<0.01).Immunofluorescence experiment showed that after combination therapy the transfer of NFAT and AHR in nuclear substantially reduced.Flow cytometry revealed that after the combination caused a significant decrease in PD-1 expression in CD8^+T cells.CONCLUSION CAI and 1-MT could inhibit markedly the expression of PD-1 in CD8^+T cells by inhibiting the nuclear translocation of NFAT and AHR,respectively and the combination of them has synergetic effect.展开更多
OBJECTIVE Programmed death ligand-1(PD-L1)and indoleamine 2,3-dioxygenase 1(IDO1)are immune checkpoints which can be induced by interferon-γ(IFN-γ)in the tumor microenvironment,leading to immune escape of tumors.Myr...OBJECTIVE Programmed death ligand-1(PD-L1)and indoleamine 2,3-dioxygenase 1(IDO1)are immune checkpoints which can be induced by interferon-γ(IFN-γ)in the tumor microenvironment,leading to immune escape of tumors.Myricetin(MY)is a flavonoid distributed in many edible and medicinal plants.The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells.METHODS Expressions of PD-L1 and major histocompatibility complex-I(MHC-I)were evaluated by flow cytometry and Western blotting,and the expression of IDO1 was measured by Western blotting.qRT-PCR was used to detect their mRNA levels.The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1.Molecular docking analysis,Western blotting and immunofluorescence were used for mechanism study.RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells,while didn't show obvious effect on the expression of MHC-I.In addition,MY restored the survival,proliferation,CD69 expression and interleukin-2(IL-2)secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system.Mechanistically,IFN-γup-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis,which was targeted and inhibited by MY.CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression,supporting the potential of MY in anti-tumor immunotherapy.展开更多
程序性细胞死亡受体1(programmed cell death 1,PD-1)单抗治疗肺腺癌可能引起罕见而严重的血液学不良反应,可出现重症贫血等表现。尽管糖皮质激素被推荐用于免疫相关不良事件的管理,但针对PD-1单抗诱发的重症贫血的治疗经验仍十分有限,...程序性细胞死亡受体1(programmed cell death 1,PD-1)单抗治疗肺腺癌可能引起罕见而严重的血液学不良反应,可出现重症贫血等表现。尽管糖皮质激素被推荐用于免疫相关不良事件的管理,但针对PD-1单抗诱发的重症贫血的治疗经验仍十分有限,其疗效和安全性尚未充分验证。本文报道了1例化疗联合PD-1单抗治疗后发生重症贫血的晚期肺腺癌患者,经过系列检查,考虑诊断炎症性贫血,经足量糖皮质激素治疗后,血红蛋白显著回升,以期为临床中这类罕见血液学毒性的识别和治疗提供新的见解。展开更多
目的:通过观察进展期食管胃交界部腺癌(AEGJ)患者拟行术前新辅助放化疗术后化疗联合腹腔热灌注化疗(HIPEC)后血免疫细胞及癌组织中PD-L1的表达,证实PD-L1对AEGJ患者预后的影响,探讨HIPEC治疗后免疫细胞水平与PD-L1表达的关系。方法:选取...目的:通过观察进展期食管胃交界部腺癌(AEGJ)患者拟行术前新辅助放化疗术后化疗联合腹腔热灌注化疗(HIPEC)后血免疫细胞及癌组织中PD-L1的表达,证实PD-L1对AEGJ患者预后的影响,探讨HIPEC治疗后免疫细胞水平与PD-L1表达的关系。方法:选取2018年1月至2019年6月河北北方学院附属第一医院收治的进展期AEGJ术前新辅助放化疗术后辅助化疗联合HIPEC患者46例。采用转录组测序技术检测AEGJ术后辅助HIPEC患者的血免疫细胞水平,免疫组化检测AEGJ组织中PD-L1表达,应用相应的统计学方法观察免疫细胞与PD-L1水平对AEGJ患者预后的预测情况,以及二者的相关性。结果:截至2022年6月30日,患者肿瘤特异生存率(CSS)为28.26%,无疾病复发率(RFS)为45.65%,中位CSS和RFS分别为31个月和24个月。PD-L1阳性率为52.17%,PD-L1阳性和PD-L1阴性患者的RFS率和CSS率分别为16.67%、77.27%和54.17%、90.91%,前者的中位RPS和CSS明显短于后者(12个月vs 37个月)和(24个月vs 37个月)。本研究共检测出22种免疫细胞,与CPS<1组相比,CPS≥1组中CD8^(+)T细胞、活化树突状细胞和静息NK细胞水平明显降低(均P<0.05),而M2型巨噬细胞在前者中显著低于后者(0.013±0.012 vs 0.033±0.003,P<0.001)。ROC曲线结果显示,CD8^(+)T细胞和M2型巨噬细胞均可预测AEGJ患者治疗的预后(AUC分别为0.690和0.698)。COX多因素回归分析提示CD8^(+)T细胞可能是AEGJ患者治疗后的保护因子[HR=0.590,95%CI(0.248~0.708)],而M2型巨噬细胞可能作为AEGJ患者治疗后的风险因素[HR=6.448,95%CI(1.440~9.165)]。PD-L1的表达与M2型巨噬细胞水平呈正相关(R=0.44,P<0.01),与CD8^(+)T细胞呈负相关(R=-0.47,P<0.05)。结论:HIPEC辅助治疗可能调控M2型巨噬细胞、CD8^(+)T细胞的功能,其中M2型巨噬细胞水平与PD-L1表达呈正相关,这种免疫细胞的增多可能是影响AEGJ术后辅助HIPEC预后的因素。展开更多
基金supported by National Natural Science Foundation of China(81402943)CAMS Major Collaborative Innovation Project(2016-I2M-1-011)PUMC Youth Fund(3332015168)
文摘OBJECTIVE To evaluate whether the IDO1 inhibitor 1-methyl-L-tryptophan(1-MT)combine calcium influx inhibitor carboxyamidotriazole(CAI)could further enhance the suppression of programmed death 1(PD-1)in CD8^+T cells and investigate the curative effect of the combined use.METHODS CD8^+T cells were isolated from normal mice spleen by negative selection using magnetic cell separation.The isolated CD8^+T cells were cultured in RPMI 1640 medium containing 10%FBS and 100 U·mL^(-1)IL-2 and activated by the addition of anti-CD3 and anti-CD28(1 g·L^(-1) each mabs).CD8^+T cells were pretreated for 48 h with drug and the fluo-3 as a marker of intracellular calcium concentration was detected by flow cytometry.The calcineurin(Ca N)levels were assayed with ELISA in CD8^+T cells after 48 h incubation with 10μm CAI.The nuclear translocations of NFAT and AHR were detected by immunofluorescent staining after 48 h of drug treatment.The expression of PD-1 in CD8^+T cells was analyzed by flow cytometry.RESULTS Intracellular fluorescent intensity was markedly debase due to CAI treatment(P<0.01).Meanwhile,the changes of CaN content had a resembled correlation(P<0.01).Immunofluorescence experiment showed that after combination therapy the transfer of NFAT and AHR in nuclear substantially reduced.Flow cytometry revealed that after the combination caused a significant decrease in PD-1 expression in CD8^+T cells.CONCLUSION CAI and 1-MT could inhibit markedly the expression of PD-1 in CD8^+T cells by inhibiting the nuclear translocation of NFAT and AHR,respectively and the combination of them has synergetic effect.
基金Science and Technology Development Fund,Macao SAR(0129/2019/A3176/2017/A3)and University of Macao(MYRG2018-00165-ICMS)。
文摘OBJECTIVE Programmed death ligand-1(PD-L1)and indoleamine 2,3-dioxygenase 1(IDO1)are immune checkpoints which can be induced by interferon-γ(IFN-γ)in the tumor microenvironment,leading to immune escape of tumors.Myricetin(MY)is a flavonoid distributed in many edible and medicinal plants.The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells.METHODS Expressions of PD-L1 and major histocompatibility complex-I(MHC-I)were evaluated by flow cytometry and Western blotting,and the expression of IDO1 was measured by Western blotting.qRT-PCR was used to detect their mRNA levels.The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1.Molecular docking analysis,Western blotting and immunofluorescence were used for mechanism study.RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells,while didn't show obvious effect on the expression of MHC-I.In addition,MY restored the survival,proliferation,CD69 expression and interleukin-2(IL-2)secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system.Mechanistically,IFN-γup-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis,which was targeted and inhibited by MY.CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression,supporting the potential of MY in anti-tumor immunotherapy.
文摘程序性细胞死亡受体1(programmed cell death 1,PD-1)单抗治疗肺腺癌可能引起罕见而严重的血液学不良反应,可出现重症贫血等表现。尽管糖皮质激素被推荐用于免疫相关不良事件的管理,但针对PD-1单抗诱发的重症贫血的治疗经验仍十分有限,其疗效和安全性尚未充分验证。本文报道了1例化疗联合PD-1单抗治疗后发生重症贫血的晚期肺腺癌患者,经过系列检查,考虑诊断炎症性贫血,经足量糖皮质激素治疗后,血红蛋白显著回升,以期为临床中这类罕见血液学毒性的识别和治疗提供新的见解。
文摘目的:通过观察进展期食管胃交界部腺癌(AEGJ)患者拟行术前新辅助放化疗术后化疗联合腹腔热灌注化疗(HIPEC)后血免疫细胞及癌组织中PD-L1的表达,证实PD-L1对AEGJ患者预后的影响,探讨HIPEC治疗后免疫细胞水平与PD-L1表达的关系。方法:选取2018年1月至2019年6月河北北方学院附属第一医院收治的进展期AEGJ术前新辅助放化疗术后辅助化疗联合HIPEC患者46例。采用转录组测序技术检测AEGJ术后辅助HIPEC患者的血免疫细胞水平,免疫组化检测AEGJ组织中PD-L1表达,应用相应的统计学方法观察免疫细胞与PD-L1水平对AEGJ患者预后的预测情况,以及二者的相关性。结果:截至2022年6月30日,患者肿瘤特异生存率(CSS)为28.26%,无疾病复发率(RFS)为45.65%,中位CSS和RFS分别为31个月和24个月。PD-L1阳性率为52.17%,PD-L1阳性和PD-L1阴性患者的RFS率和CSS率分别为16.67%、77.27%和54.17%、90.91%,前者的中位RPS和CSS明显短于后者(12个月vs 37个月)和(24个月vs 37个月)。本研究共检测出22种免疫细胞,与CPS<1组相比,CPS≥1组中CD8^(+)T细胞、活化树突状细胞和静息NK细胞水平明显降低(均P<0.05),而M2型巨噬细胞在前者中显著低于后者(0.013±0.012 vs 0.033±0.003,P<0.001)。ROC曲线结果显示,CD8^(+)T细胞和M2型巨噬细胞均可预测AEGJ患者治疗的预后(AUC分别为0.690和0.698)。COX多因素回归分析提示CD8^(+)T细胞可能是AEGJ患者治疗后的保护因子[HR=0.590,95%CI(0.248~0.708)],而M2型巨噬细胞可能作为AEGJ患者治疗后的风险因素[HR=6.448,95%CI(1.440~9.165)]。PD-L1的表达与M2型巨噬细胞水平呈正相关(R=0.44,P<0.01),与CD8^(+)T细胞呈负相关(R=-0.47,P<0.05)。结论:HIPEC辅助治疗可能调控M2型巨噬细胞、CD8^(+)T细胞的功能,其中M2型巨噬细胞水平与PD-L1表达呈正相关,这种免疫细胞的增多可能是影响AEGJ术后辅助HIPEC预后的因素。
