Hsp70 and nf-kb mediated control of innate inflammatory responses in a canine macrophage cell line

dc.contributor.authorLyu, Qingkang
dc.contributor.authorWawrzyniuk, Magdalena
dc.contributor.authorRutten, Victor P.M.G.
dc.contributor.authorSijts, Alice J.A.M.
dc.contributor.authorBroere, Femke
dc.date.accessioned2021-05-14T13:33:59Z
dc.date.available2021-05-14T13:33:59Z
dc.date.issued2020-09
dc.description.abstractThe pathogenesis of many inflammatory diseases is associated with the uncontrolled activation of nuclear factor kappa B (NF-κB) in macrophages. Previous studies have shown that in various cell types, heat shock protein 70 (Hsp70) plays a crucial role in controlling NF-κB activity. So far, little is known about the role of Hsp70 in canine inflammatory processes. In this study we investigated the potential anti-inflammatory effects of Hsp70 in canine macrophages as well as the mechanisms underlying these effects. To this end, a canine macrophage cell line was stressed with arsenite, a chemical stressor, which upregulated Hsp70 expression as detected by flow cytometry and qPCR. A gene-edited version of this macrophage cell line lacking inducible Hsp70 was generated using CRISPR-Cas9 technology. To determine the effects of Hsp70 on macrophage inflammatory properties, arsenite-stressed wild-type and Hsp70 knockout macrophages were exposed to lipopolysaccharide (LPS), and the expression of the inflammatory cytokines IL-6, IL-1β and tumor necrosis factor-α (TNF-α) and levels of phosphorylated NF-κB were determined by qPCR and Western Blotting, respectively. Our results show that non-toxic concentrations of arsenite induced Hsp70 expression in canine macrophages; Hsp70 upregulation significantly inhibited the LPS-induced expression of the pro-inflammatory mediators TNF-α and IL-6, as well as NF-κB activation in canine macrophages. Furthermore, the gene editing of inducible Hsp70 by CRISPR-Cas9-mediated gene editing neutralized this inhibitory effect of cell stress on NF-κB activation and pro-inflammatory cytokine expression. Collectively, our study reveals that Hsp70 may regulate inflammatory responses through NF-κB activation and cytokine expression in canine macrophages.en_ZA
dc.description.departmentVeterinary Tropical Diseasesen_ZA
dc.description.librarianpm2021en_ZA
dc.description.urihttp://www.mdpi.com/journal/ijmsen_ZA
dc.identifier.citationLyu, Q.; Wawrzyniuk, M.; Rutten, V.P.M.G.; van Eden, W.; Sijts, A.J.A.M.; Broere, F. Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line. International Journal of Molecular Sciences 2020, 21, 6464. https://doi.org/10.3390/ijms21186464.en_ZA
dc.identifier.issn1661-6596 (print)
dc.identifier.issn1422-0067 (online)
dc.identifier.other10.3390/ijms21186464
dc.identifier.urihttp://hdl.handle.net/2263/79915
dc.language.isoenen_ZA
dc.publisherMDPIen_ZA
dc.rights© 2020 by the authors. Licensee: MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.en_ZA
dc.subjectMacrophagesen_ZA
dc.subjectNF-κBen_ZA
dc.subjectCytokinesen_ZA
dc.subjectHeat shock protein 70 (Hsp70)en_ZA
dc.subjectNuclear factor kappa B (NF-κB)en_ZA
dc.titleHsp70 and nf-kb mediated control of innate inflammatory responses in a canine macrophage cell lineen_ZA
dc.typeArticleen_ZA

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