Retinoic acid-inducible gene I (RIG-I) is a cytosolic viral RNA receptor. Upon viral infection, the protein recognizes and then recruits adapter protein mitochondrial antiviral signaling (MAVS) protein, initiating the production of interferons and proinflammatory cytokines to establish an antiviral state. In the present study, we identify zinc finger protein 205 (ZNF205) which associates with RIG-I and promotes the Sendai virus (SeV)-induced antiviral innate immune response. Overexpression of ZNF205 facilitates interferon-beta (IFN-β) introduction, whereas ZNF205 deficiency restricts its introduction. Mechanistically, the C-terminal zinc finger domain of ZNF205 interacts with the N-terminal tandem caspase recruitment domains (CARDs) of RIG-I; this interaction markedly promotes K63 ubiquitin-linked polyubiquitination of RIG-I, which is crucial for RIG-I activation. Thus, our results demonstrate that ZNF205 is a positive regulator of the RIG-I-mediated innate antiviral immune signaling pathway.
Immune checkpoint inhibitors（ICIs） have improved durable clinical responses and longer survival time in patients with cancer. However, biomarkers of long-term survival are poorly understood to predict patients who will benefit from ICIs treatment. Here, by analyzing bulk RNA-seq of two ICIs-treated discovery cohorts（160patients）, we found that CHIT1 is significantly higher expressed in patients who acquire long-term overall survival（OS）. Further, we demonstrate that higher CHIT1 markedly prolonged progression-free survival（PFS） and OS in the two ICIs-treated discovery and five validation cohorts（198 patients）. Moreover, baseline of chitotriosidase greater than15 ng/ml in serum of anti-PD-1 treated patients gained longer PFS（12.4 months vs 9.5 months, P=0.039, HR=0.2,95% CI=0.04 to 1.07）. In addition, gene set enrichment analysis（GSEA） indicates that CHIT1 is positively related to hot tumor immune microenvironment（TIME） in the above cohorts and TCGA data. Together, our data suggests that CHIT1 is a novel potential biomarker for predicting the long-term OS in ICIs-treated cohorts.
目的 研究mTORC1抑制剂通过肿瘤浸润T淋巴细胞生物学活性影响KRAS基因突变型结肠癌的进展.方法 建立KRAS突变型结肠癌小鼠模型,将小鼠随机分为4组:对照组,KRAS突变组,mTORC1抑制剂组和KRAS突变+mTORC1抑制剂组;流式细胞术检测肿瘤组织中肿瘤浸润淋巴T细胞的数量;ELISA法检测小鼠血清中IL-2、IL-10、IFN-γ、TGF-β的表达情况;另外构建KRAS突变型结肠癌细胞系HT29,将细胞分为4组:HT-29组,KRAS突变组,mTORC1抑制剂组和KRAS突变+mTORC1抑制剂组;CCK-8法检测结肠癌细胞的增殖情况;流式细胞术检测其凋亡情况;Western blot法检测p53、Bcl-2、Bax、cleaved caspase3蛋白的表达水平.结果 与KRAS突变组结肠癌小鼠相比较,KRAS突变+mTORC1抑制剂组小鼠肿瘤组织中CD4+T细胞、CD8+T细胞数量增加(P＜0.01,P＜0.001),Foxp3+Treg细胞的数量减少(P＜0.000 1),小鼠血清中IL-2、IFN-γ的水平升高(P＜0.000 1,P＜0.001),IL-10、TGF-β的水平降低(均P＜0.001);另外,与KRAS突变型HT29组细胞相比,KRAS突变+mTORC1抑制剂组细胞的增殖能力减弱(P＜0.05),并且凋亡情况显著提高(P＜0.000 1),并且Bcl-2蛋白水平降低(P＜0.001),Bax、cleavedcaspase3、p53蛋白水平显著升高(P＜0.01,P＜0.001,P＜0.001).结论 mTORC1抑制剂通过影响肿瘤浸润淋巴细胞的分布情况以及通过p53/caspase相关通路抑制KRAS基因突变型结肠癌的进展.
目的 研究培菲康通过下调Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路减轻哮喘小鼠气道炎症及气道重塑的机制.方法 以卵白蛋白和氢氧化铝混悬液致敏、激发建立急性哮喘小鼠模型,随机分为4组(每组12只):模型组、培菲康低剂量(0.1g/kg)组、培菲康高剂量(0.2g/kg)组、培菲康(0.2g/kg)+脂多糖(TLR4激活剂,0.4 mg/kg)组,另取12只小鼠以等剂量生理盐水处理作为对照组以培菲康、LPS对小鼠分组进行干预后,评测各组小鼠哮喘症状评分;以瑞氏染色分类计数各组小鼠支气管肺泡灌洗液(BALF)中炎症细胞;以HE染色、Masson染色检测各组小鼠肺组织病理形态和气道重塑,并评测小鼠气管壁厚度和气道平滑肌厚度;以酶标仪检测各组小鼠BALF中炎症相关因子白细胞介素(IL)-17、肿瘤坏死因子-α(TNF-α)、IL-4及IL-10水平;以蛋白免疫印迹检测各组小鼠肺组织TLR4/NF-κB信号通路相关蛋白表达.结果 与对照组相比,模型组小鼠肺组织出现严重病理损伤和气道重塑,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和I L-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平显著升高(P＜0.05),抗炎因子IL-10水平显著降低(P＜0.05);与模型组相比,培菲康低、高剂量组小鼠肺组织病理损伤和气道重塑均减轻,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κBp65/NF-κB p65及TLR4蛋白表达水平均降低(P＜0.05),抗炎因子IL-10水平均升高(P＜0.05);与培菲康低剂量组相比,培菲康高剂量组小鼠肺组织病理损伤和气道重塑减轻,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平降低(P＜0.05),抗炎因子IL-10水平升高(P＜0.05);与培菲康高剂量组相比,培菲康+LPS组小鼠肺组织病理损伤和气道重塑加重,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平升高(P＜0.05),抗炎因子IL-10水平降低(P＜0.05).结论 培菲康可通过抑制TLR4/NF-κB信号而减轻哮喘小鼠气道炎症,缓解其肺组织病理损伤和气道重塑.
目的 研究N-乙酰半胱氨酸酰胺(NACA)对脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症的保护作用及作用机制,为NACA治疗脓毒症提供理论依据.方法 CCK-8法检测NACA和N-乙酰半胱氨酸(NAC)对RAW264.7细胞活力的影响;采用ELISA检测NACA和NAC对LPS诱导的RAW264.7细胞上清中TNF-α和IL-6水平;qRT-PCR检测NACA对LPS诱导的RAW264.7 细胞中 TNF-α、IL-6、IL-1β、ATF4、CHOP 的 mRNA 表达的影响;Western blot 检测 NACA 对 LPS 诱导的 RAW264.7 细胞中TLR4/NF-κB及PERK/ATF4/CHOP信号通路中关键蛋白表达的影响.结果 0.1～5 mmol/L的NACA和NAC对RAW264.7细胞活力无明显的影响(P＞0.05).1 mmol/L和5 mmol/L的NACA和NAC可显著降低LPS诱导的RAW264.7细胞上清中TNF-α和IL-6的水平(P＜0.01),且NACA的降低效果优于统计量的NAC.1 mmol/L和5 mmol/L的NACA也可显著降低LPS诱导的 RAW264.7 细胞中 TNF-α、IL-6、IL-1β、ATF4、CHOP mRNA 的表达(P＜0.05、P＜0.01).Western blot结果表明,1 mmol/L和5 mmol/L 的 NACA 可显著降低 TLR4、p-IκB、p-65、p-PERK、p-eIF2α、ATF4、Bip 和 CHOP 蛋白的表达(P＜0.05、P＜0.01).结论 NACA可能通过TLR4/NF-κB及PERK/ATF4/CHOP信号通路抑制LPS诱导的RAW264.7巨噬细胞的炎症反应,为NACA治疗脓毒症的抗炎性药物筛选提供了实验依据.
