【目的】克隆白来航鸡干扰素基因刺激因子基因(stimulator of interferon genes,STING)CDS区序列并进行生物信息学和组织表达分析,为阐明STING基因在抗病毒免疫应答中的作用奠定基础。【方法】采用PCR扩增并克隆白来航鸡STING基因CDS区...【目的】克隆白来航鸡干扰素基因刺激因子基因(stimulator of interferon genes,STING)CDS区序列并进行生物信息学和组织表达分析,为阐明STING基因在抗病毒免疫应答中的作用奠定基础。【方法】采用PCR扩增并克隆白来航鸡STING基因CDS区,测序后对其编码氨基酸序列进行相似性比对及系统进化树构建,利用生物信息学预测STING蛋白的理化特性及结构功能,并利用实时荧光定量PCR技术检测STING基因在鸡心脏、肝脏等14个组织中的表达情况。【结果】白来航鸡STING基因CDS区序列全长1140 bp,编码379个氨基酸。相似性比对和系统进化树分析结果表明,白来航鸡STING基因与原鸡的相似性最高(99.7%),亲缘关系最近,与冠小嘴乌鸦亲缘关系最远。STING蛋白为酸性、亲水性蛋白,分子质量为42.625 ku,等电点(pI)为6.67,不稳定系数为69.26,脂肪系数为105.01。该蛋白大部分在线粒体和内质网上合成,含有跨膜结构,不含信号肽。STING蛋白二级结构包括α-螺旋(54.62%)、延伸链(10.29%)、β-转角(3.43%)及无规则卷曲(31.66%)。蛋白互作分析表明,白来航鸡STING蛋白与NFKB1、DDX41、cGAS、TBK1等蛋白存在相互作用。实时荧光定量PCR结果显示,STING基因在白来航鸡组织中广泛表达,其中在肺脏中表达量最高,且显著高于其他组织(P<0.05);在胸肌中表达量最低。【结论】本研究成功克隆了白来航鸡STING基因,其CDS区序列全长1140 bp,编码379个氨基酸。白来航鸡STING蛋白为酸性、亲水性蛋白,含有跨膜结构。STING基因在白来航鸡肺脏中表达量最高。研究结果为深入探究白来航鸡STING基因编码蛋白功能提供了材料。展开更多
Oncolytic viruses have emerged as new powerful therapeutic agents for cancer therapy by specifically lysing cancer cells while activating innate immune responses at the same time.However,due to the thorny issues of sa...Oncolytic viruses have emerged as new powerful therapeutic agents for cancer therapy by specifically lysing cancer cells while activating innate immune responses at the same time.However,due to the thorny issues of safety concerns and host immune reaction,the clinical application of oncolytic viruses is still limited.Herein,we report a rationally designed oncolytic virus-like nanoparticles(OV-NPs)composed of stimulator of interferon genes(STING)-stimulating polymer loaded with therapeutic genes for cancer immunotherapy.After injection into tumor,the OV-NPs carrying OX40L plasmid could reprogram tumor cells to express OX40L immune checkpoint molecules and activate the STING pathway for cooperatively enhancing antitumor immunity,with a tumor suppression rate of 92.3%in B16F10 tumor model and 78.7%in MC38 tumor model without causing any toxicity.The OV-NPs could be further applied in carrying other plasmids(IL-12)and utilization in gene combination therapy.This study should inspire designing synthetic OV-NPs as alternative strategies for extending oncolytic virus application in cancer immunotherapy.展开更多
基金supported by the National Natural Science Foundation of China (22222509,52025035,51973215,22375198,52103194)Bureau of International Cooperation Chinese Academy of Sciences (121522KYSB20200029)+5 种基金Jilin Province Science and Technology DevelopmentPlan (YDZJ202101ZYTS131,20220402037GH,20210508049RQ)Jilin Provincial International Cooperation Key Laboratory of Biomedical Polymers (20210504001GH)Changchun Science and Technology Development Plan (21ZY09,21ZGY30)the China Postdoctoral Science Foundation (E21S2101)the Youth Talents Promotion Project of Jilin Province (QT202103)the Youth Innovation Promotion Association of Chinese Academy of Sciences (2020232)。
文摘Oncolytic viruses have emerged as new powerful therapeutic agents for cancer therapy by specifically lysing cancer cells while activating innate immune responses at the same time.However,due to the thorny issues of safety concerns and host immune reaction,the clinical application of oncolytic viruses is still limited.Herein,we report a rationally designed oncolytic virus-like nanoparticles(OV-NPs)composed of stimulator of interferon genes(STING)-stimulating polymer loaded with therapeutic genes for cancer immunotherapy.After injection into tumor,the OV-NPs carrying OX40L plasmid could reprogram tumor cells to express OX40L immune checkpoint molecules and activate the STING pathway for cooperatively enhancing antitumor immunity,with a tumor suppression rate of 92.3%in B16F10 tumor model and 78.7%in MC38 tumor model without causing any toxicity.The OV-NPs could be further applied in carrying other plasmids(IL-12)and utilization in gene combination therapy.This study should inspire designing synthetic OV-NPs as alternative strategies for extending oncolytic virus application in cancer immunotherapy.
