Hsp70 and nf-kb mediated control of innate inflammatory responses in a canine macrophage cell line
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Date
Authors
Lyu, Qingkang
Wawrzyniuk, Magdalena
Rutten, Victor P.M.G.
Sijts, Alice J.A.M.
Broere, Femke
Journal Title
Journal ISSN
Volume Title
Publisher
MDPI
Abstract
The 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.
Description
Keywords
Macrophages, NF-κB, Cytokines, Heat shock protein 70 (Hsp70), Nuclear factor kappa B (NF-κB)
Sustainable Development Goals
Citation
Lyu, 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.