Corynebacterium tuberculostearicum, a human skin colonizer, induces the canonical nuclear factor-κB inflammatory signaling pathway in human skin cells

Immun Inflamm Dis. 2020 Mar;8(1):62-79. doi: 10.1002/iid3.284. Epub 2020 Jan 7.

Abstract

Introduction: Corynebacterium tuberculostearicum (C. t.) is a ubiquitous bacterium that colonizes human skin. In contrast to other members of the genus Corynebacterium, such as toxigenic Corynebacterium diphtheriae or the opportunistic pathogen Corynebacterium jeikeium, several studies suggest that C. t. may play a role in skin health and disease. However, the mechanisms underlying these effects remain poorly understood.

Methods: To investigate whether C. t. induces inflammatory pathways in primary human epidermal keratinocytes (HEKs) and human cutaneous squamous carcinoma cells (SCCs), cell culture, reverse transcription-polymerase chain reaction (PCR), enzyme-linked immunosorbent assay, immunofluorescence microscopy, Western blot, chromatin immunoprecipitation-PCR, small interfering RNA knockdown and luciferase reporter expression system were used.

Results: Herein, we demonstrate that C. t. upregulates the messenger RNA (mRNA) and protein levels of inflammatory mediators in two human skin cell lines, HEKs and SCCs. We further show activation of the canonical nuclear factor-κB (NF-κB) pathway in response to C. t. infection, including phosphorylation of the inhibitor of κB (IκB), the nuclear translocation of NF-κB subunit (NF-κB-P65 ) and the recruitment of NF-κB-P65 and RNA polymerase to the NF-κB response elements at the promoter region of the inflammatory genes. Lastly, the data confirm that C. t.-induced tumor necrosis factor mRNA expression in HEKs is toll-like receptor 2 (TLR2 ) dependent.

Conclusion: Our results offer a mechanistic model for C. t.-induced inflammation in human keratinocytes via TLR2 and activation of IκB kinase and downstream signaling through the canonical NF-κB pathway. Relevance to chronic inflammatory diseases of the skin and cutaneous oncology is discussed.

Keywords: Corynebacterium tuberculostearicum; IKK; NF-κB; cutaneous squamous carcinoma cells; inflammation; keratinocytes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Squamous Cell / microbiology
  • Cell Line
  • Corynebacterium
  • Corynebacterium Infections / genetics
  • Corynebacterium Infections / microbiology*
  • Humans
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / metabolism
  • Inflammation / microbiology*
  • Inflammation / pathology
  • Keratinocytes / microbiology
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Phosphorylation
  • RNA, Messenger / genetics
  • RNA, Small Interfering
  • Signal Transduction*
  • Transfection
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • NF-kappa B
  • RNA, Messenger
  • RNA, Small Interfering
  • Tumor Necrosis Factor-alpha
  • I-kappa B Kinase

Supplementary concepts

  • Corynebacterium tuberculostearicum