tetano
Editor, Senior Moderator
Viruses. 2019 Oct 25;11(11). pii: E986. doi: 10.3390/v11110986. [h=1]High-Throughput MicroRNA Profiles of Permissive Madin-Darby Canine Kidney Cell Line Infected with Influenza B Viruses.[/h]
Saengchoowong S[SUP]1,[/SUP][SUP]2[/SUP], Khongnomnan K[SUP]3[/SUP], Poomipak W[SUP]4[/SUP], Praianantathavorn K[SUP]3[/SUP], Poovorawan Y[SUP]5[/SUP], Zhang Q[SUP]2[/SUP], Payungporn S[SUP]3,[/SUP][SUP]4[/SUP].
[h=3]Author information[/h] 1 Graduate Division, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 2 Department of Clinical Infection, Microbiology and Immunology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7BE, UK. 3 Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 4 Chulalongkorn University Center of Excellence in Systems Biology, Research Affairs, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 5 Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
[h=3]Abstract[/h] Victoria and Yamagata lineages of influenza B viruses are globally circulating in seasonal epidemics. Madin-Darby canine kidney (MDCK) cells are permissive for viral isolation and vaccine manufacture. Nevertheless, the interplay between influenza B viruses and host microRNAs has not been investigated in this cell line. Therefore, the present study aims at high-throughput analysis of canine microRNA profile upon infection of influenza B viruses. Briefly, MDCK cells were infected with Victoria or Yamagata lineage at MOI of 0.01. After being harvested at 6, 12 and 24 h post infection, microRNAs were subjected to high-throughput sequencing based on MiSeq platform (Illumina). The results demonstrated that five microRNAs including cfa-miR-197, cfa-miR-215, cfa-miR361, cfa-miR-1841, and cfa-miR-1842 were overexpressed in both Victoria and Yamagata lineage infections. Interestingly, computational prediction showed that karyopherin alpha 6 (KPNA6) was targeted by cfa-miR-197 and cfa-miR-215. Moreover, the binding sites of both microRNAs were assessed by 3'-UTR reporter assay. The results showed that only cfa-miR-197 could bind to the target sites of KPNA6, leading to suppressing luciferase activity. Additionally, silencing of KPNA6 was confirmed by overexpression of cfa-miR-197. This study provides canine microRNA responses to seasonal influenza B viruses, suggesting that virus-mediated microRNAs might play crucial roles in host gene regulation.
[h=4]KEYWORDS:[/h] Canis lupus familiaris; MDCK; Victoria; Yamagata; influenza B viruses; microRNAs; next-generation sequencing
PMID: 31717720 DOI: 10.3390/v11110986
Saengchoowong S[SUP]1,[/SUP][SUP]2[/SUP], Khongnomnan K[SUP]3[/SUP], Poomipak W[SUP]4[/SUP], Praianantathavorn K[SUP]3[/SUP], Poovorawan Y[SUP]5[/SUP], Zhang Q[SUP]2[/SUP], Payungporn S[SUP]3,[/SUP][SUP]4[/SUP].
[h=3]Author information[/h] 1 Graduate Division, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 2 Department of Clinical Infection, Microbiology and Immunology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7BE, UK. 3 Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 4 Chulalongkorn University Center of Excellence in Systems Biology, Research Affairs, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. 5 Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
[h=3]Abstract[/h] Victoria and Yamagata lineages of influenza B viruses are globally circulating in seasonal epidemics. Madin-Darby canine kidney (MDCK) cells are permissive for viral isolation and vaccine manufacture. Nevertheless, the interplay between influenza B viruses and host microRNAs has not been investigated in this cell line. Therefore, the present study aims at high-throughput analysis of canine microRNA profile upon infection of influenza B viruses. Briefly, MDCK cells were infected with Victoria or Yamagata lineage at MOI of 0.01. After being harvested at 6, 12 and 24 h post infection, microRNAs were subjected to high-throughput sequencing based on MiSeq platform (Illumina). The results demonstrated that five microRNAs including cfa-miR-197, cfa-miR-215, cfa-miR361, cfa-miR-1841, and cfa-miR-1842 were overexpressed in both Victoria and Yamagata lineage infections. Interestingly, computational prediction showed that karyopherin alpha 6 (KPNA6) was targeted by cfa-miR-197 and cfa-miR-215. Moreover, the binding sites of both microRNAs were assessed by 3'-UTR reporter assay. The results showed that only cfa-miR-197 could bind to the target sites of KPNA6, leading to suppressing luciferase activity. Additionally, silencing of KPNA6 was confirmed by overexpression of cfa-miR-197. This study provides canine microRNA responses to seasonal influenza B viruses, suggesting that virus-mediated microRNAs might play crucial roles in host gene regulation.
[h=4]KEYWORDS:[/h] Canis lupus familiaris; MDCK; Victoria; Yamagata; influenza B viruses; microRNAs; next-generation sequencing
PMID: 31717720 DOI: 10.3390/v11110986