title: Identification of luteolin as enterovirus 71 and coxsackievirus A16 inhibitors through reporter viruses and cell viability-based screening.
abstract: Hand, foot and mouth disease (HFMD) is a common pediatric illness mainly caused by infection with enterovirus 71 (EV71) and coxsackievirus A16 (CA16). The frequent HFMD outbreaks have become a serious public health problem. Currently, no vaccine or antiviral drug for EV71/CA16 infections has been approved. In this study, a two-step screening platform consisting of reporter virus-based assays and cell viability‑based assays was developed to identify potential inhibitors of EV71/CA16 infection. Two types of reporter viruses, a pseudovirus containing luciferase-encoding RNA replicons encapsidated by viral capsid proteins and a full-length reporter virus containing enhanced green fluorescent protein, were used for primary screening of 400 highly purified natural compounds. Thereafter, a cell viability-based secondary screen was performed for the identified hits to confirm their antiviral activities. Three compounds (luteolin, galangin, and quercetin) were identified, among which luteolin exhibited the most potent inhibition of viral infection. In the cell viability assay and plaque reduction assay, luteolin showed similar 50% effective concentration (EC50) values of about 10 μM. Luteolin targeted the post-attachment stage of EV71 and CA16 infection by inhibiting viral RNA replication. This study suggests that luteolin may serve as a lead compound to develop potent anti-EV71 and CA16 drugs.
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. xulinjlu@hotmail.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. suweiheng@163.com.
authors.forename: Weiheng
authors.initials: W
authors.lastname: Su
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. jun.jin@pasteur.fr.
authors.forename: Jun
authors.initials: J
authors.lastname: Jin
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. JiawenChen229@gmail.com.
authors.forename: Jiawen
authors.initials: J
authors.lastname: Chen
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. lixiaojun3345@163.com.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Li
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. zxy_better@163.com.
authors.forename: Xuyuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. sunmy990@163.com.
authors.forename: Meiyan
authors.initials: M
authors.lastname: Sun
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. sunbaoshiyang@163.com.
authors.forename: Shiyang
authors.initials: S
authors.lastname: Sun
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. fph121@163.com.
authors.forename: Peihu
authors.initials: P
authors.lastname: Fan
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. andonginlab@sina.cn.
authors.forename: Dong
authors.initials: D
authors.lastname: An
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. bszhfing@163.com.
authors.forename: Huafei
authors.initials: H
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. zxy_better@163.com.
authors.forename: Xiguang
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. weikong@jlu.edu.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.affiliation: College of Basic Medical Sciences, Dalian Medical University, Dalian 116000, China. tonghuima@dlmedu.edu.cn.
authors.forename: Tonghui
authors.initials: T
authors.lastname: Ma
authors.affiliation: School of Life Sciences, Jilin University, Changchun 130012, China. jiangcl@jlu.edu.cn.
authors.forename: Chunlai
authors.initials: C
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001688:Biological Products / Q000494:pharmacology; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D002522:Chlorocebus aethiops; D004770:Enterovirus / Q000187:drug effects* / Q000502:physiology; D029821:Enterovirus A, Human / Q000187:drug effects* / Q000502:physiology; D005419:Flavonoids / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D020128:Inhibitory Concentration 50; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D047311:Luteolin / Q000494:pharmacology*; D011794:Quercetin / Q000494:pharmacology; D012093:Replicon; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D001688:Biological Products; D036022:Capsid Proteins; D005419:Flavonoids; C485184:enhanced green fluorescent protein; C037032:galangin; D049452:Green Fluorescent Proteins; D011794:Quercetin; D008156:Luciferases; D047311:Luteolin