摘要
目的分析柯萨奇病毒B组2型(coxsackievirus B2,CVB2)病毒蛋白转染上皮细胞后诱导的免疫应答信号分子特征。方法构建CVB2结构蛋白重组真核表达质粒转染16HBE细胞,收集转染后的16HBE细胞培养上清和裂解物。利用RT-Q-PCR技术检测转染后的16HBE细胞中与固有免疫应答相关的信号分子的mRNA表达水平。利用ELISPOT检测转染后的16HBE细胞上清和裂解物与T细胞共培养后该T细胞的增殖效应。结果CVB2结构蛋白VP1~VP4转染16HBE细胞后与固有免疫相关的信号分子如TAK、NIK、IKKα、IFN-β等基因表达量均发生了不同程度的上调;CVB2 VP1~VP4转染后的16HBE细胞上清和裂解物与T细胞共培养发现,均能刺激T细胞增殖。结论CVB2结构蛋白VP1~VP4可以不同程度地激活与固有免疫相关信号分子的表达,并且对适应性免疫的激活具有促进作用。
Objective To investigate the molecular characteristics of immune response signaling molecules induced by transfection of coxsackievirus B2(CVB2)structural proteins into epithelial cells.Methods Recombinant eukaryotic expression plasmids containing the coding regions of CVB2 structural proteins VP1-VP4 were constructed and then transfected into 16HBE cells.Culture supernatants and cell lysates of the transfected 16HBE cells were collected.Expression of signaling molecules involved in innate immune responses in transfected 16HBE cells at mRNA level was detected by RT-Q-PCR.The proliferation of T cells co-cultured with culture supernatants and cell lysates of the transfected 16HBE cells was analyzed by ELISPOT.Results Expression of innate immunity-related signaling molecules such as TGF-β-activated kinase(TAK),NF-κB-inducing kinase(NIK),IκB kinaseα(IKKα)and IFN-βat mRNA level was up-regulated in 16HBE cells transfected with CVB2 structural proteins VP1-VP4.Both culture supernatants and cell lysates of the transfected 16HBE cells enhanced the proliferation of T cells.Conclusions CVB2 structural proteins VP1-VP4 could enhance the expression of innate immunity-related signaling molecules to varying degrees and promote the activation of adaptive immunity.
作者
牟唐维
吴化叶
柳蕾
王建斌
张莹
李琦涵
Mou Tangwei;Wu Huaye;Liu Lei;Wang Jianbin;Zhang Ying;Li Qihan(Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Disease,Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College,Kunming 650118,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2019年第5期321-326,共6页
Chinese Journal of Microbiology and Immunology
基金
中国医学科学院医学与健康科技创新工程重大协同创新项目(2016-I2M-1-019)
云南省重大专项(2016ZF002)
北京协和医学院2018年度研究生创新基金(2018-1001-09).
