Anti-respiratory syncytial virus activity evaluation via primary human airway epithelial cell culture
This study aims to establish the culture method of human primary airway epithelial cell(hAEC)and to investigate the anti-respiratory syncytial virus(RSV)activity and mechanism of 3-thioindole compound RSVA-4 and immunosuppressive metabolite 6-MM-PR using hAEC system,which intends to construct a cell model for RSV drug screening and efficacy evaluation.The respiratory tract epithelial cells from volunteers were collected and cultured,then the morphology,activity and purity were identified.The anti-RSV ac-tivity and cytotoxicity of RSVA-4 and 6-MMPR were further verified in hAEC system.The mechanism of RSVA-4 and 6-MMPR ac-counting for the suppression of RSV replication on hAEC was explored by using fluorescence real-time quantitative PCR(RT-qPCR)and time-of-addition assay.The survival rate of cultured hAEC was 93.51％as determined by trypan blue staining.The half maximal inhibitory concentrations(IC50)of RSV-A-4 and 6-MMPR were(207.30±4.77)µmol/L and(3 191.00±6.11)μmol/L,respective-ly.The half maximal cytotoxic concentration(CC50)of 6-MMPR was(95 526.00±10.97)µmol/L,while no toxicity of RSVA-4 was observed on hAEC.Mechanistically,RSV-A-4 and 6-MMPr inhibited RSV replication in the genome replication/transcription phase.The hAEC culture method was successfully established,which could be used to screen and evaluate the anti-RSV drugs in vitro.RS-VA-4 and 6-MMPR could effectively inhibit RSV replication at the cellular level.Altogether,the result could provide an experimental basis for the research and development of RSV drug and pathogenesis.
human airway epithelial cellhuman respiratory syncytial virusantiviral compoundantiviral activityantiviral mechanism
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维普
