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AD skin\u2010derived strains of <jats:italic>S.\u00a0aureus<\/jats:italic> (AD strain) are selectively internalized into keratinocytes (HaCaT cells) compared to standard strains. However, the mechanism of AD strain internalization by keratinocytes and effect of the skin environment on internalization remain unclear. HaCaT cells were exposed to heat\u2010killed AD or standard strains of fluorescently labeled <jats:italic>S.\u00a0aureus<\/jats:italic>, with or without interferon (IFN)\u2010\u03b3, interleukin (IL)\u20104, and IL\u201013 cytokines, for 24\u2009h. Filaggrin and fibronectin expression in HaCaT cells was knocked down using small interfering RNA. The amount of internalized <jats:italic>S.\u00a0aureus<\/jats:italic> was evaluated using a cell imaging system. The effects of INF\u2010\u03b3, IL\u20104, and <jats:italic>S.\u00a0aureus<\/jats:italic> exposure on mRNA expression in HaCaT cells were analyzed using single\u2010cell RNA sequencing. AD strains adhered to HaCaT cells in approximately 15\u2009min and were increasingly internalized for up to 3\u00a0h (2361\u2009\u00b1\u2009467 spots\/100 cells, mean\u2009\u00b1\u2009SD), whereas the standard strain was not (991\u2009\u00b1\u200971 spots\/100 cells). In the presence of IFN\u2010\u03b3, both the number of internalized strains and fibronectin expression significantly decreased compared to in the control, whereas Th2 cytokines had no significant effects. The number of internalized AD strains was significantly higher in filaggrin knockdown and lower in fibronectin knockdown HaCaT cells compared to in the control. RNA sequencing revealed that IFN\u2010\u03b3 decreased both fibronectin and filaggrin expression. Keratinocyte internalization of the AD strain may be predominantly mediated by the INF\u2010\u03b3\u2010fibronectin pathway and partially regulated by filaggrin expression.<\/jats:p>","DOI":"10.1111\/1346-8138.16546","type":"journal-article","created":{"date-parts":[[2022,8,19]],"date-time":"2022-08-19T09:24:15Z","timestamp":1660901055000},"page":"1148-1157","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Uptake of <i>Staphylococcus aureus<\/i> by keratinocytes is reduced by interferon\u2013fibronectin pathway and filaggrin expression"],"prefix":"10.1111","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9144-7358","authenticated-orcid":false,"given":"Ryu","family":"Miyake","sequence":"first","affiliation":[{"name":"Department of Dermatology Graduate School of Biomedical and Health Sciences Hiroshima University  Hiroshima 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