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Extracellular vesicles derived from Malassezia furfur stimulate IL-6 production in keratinocytes as demonstrated in in vitro and in vivo models

Yu-Jing Zhang, Yang Han, Yu-Zhe Sun, Hang-Hang Jiang, Min Liu, Rui-Qun Qi, Xing-Hua Gao
Journal of Dermatological Science 2019 March 7

BACKGROUND: Malassezia is one of the commensal microorganisms colonized on human skin and has been shown to be related to several inflammatory cutaneous disorders. Previous studies indicated that Malassezia. sympodialis (M. sympodialis) can produce extracellular vesicles, however, the immunoregulatory function of Malassezia extracellular vesicles on keratinocytes has not been studied.

OBJECTIVE: To investigate the extracellular vesicular production capability of Malassezia. furfur (M. furfur) and examine their immunoregulatory effects both in vitro and in vivo.

METHODS: Extracellular vesicles derived from M. furfur were isolated by sequential ultracentrifugation procedure. Their structure and diameter were determined by negative stain TEM and NTA, respectively. Confocal microscopy was used to visualize the internalization of these nanoparticles into HaCaT cells and mice epidermal keratinocytes. The expressions of inflammatory cytokines were screened using PCR Array assay and validated in vitro by qPCR and ELISA assays. In vivo cytokine production was measured by the IHC method. The role of NF-κB in such process was evaluated in HaCaT cells by western blot assay.

RESULTS: Our results showed that M. furfur produced ovoid-shaped nanoparticles, which could be then internalized into HaCaT cells, as well as mice epidermal keratinocytes. IL-6 expression was significantly enhanced in response to extracellular vesicular stimulation both in vitro and in vivo, in which process the activation of NF-κB was involved.

CONCLUSION: M. furfur has the ability to release extracellular vesicles, which can be internalized into keratinocytes and promote the production of IL-6 with the involvement of NF-κB dependent pathway. Such findings reveal some important new insights into Malassezia pathogenesis and therapy.


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