Related Experiment Video
Updated: May 4, 2026
Isolation and Culture of Primary Mouse Keratinocytes from Neonatal and Adult Mouse Skin
Published on: July 14, 2017
Cathelicidin antimicrobial peptide LL-37 in cholesteatoma enables keratinocyte reactivity with cytosolic DNA
1Department of Otolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China.
Abstract:
The purpose of this study was to determine whether self-DNA can trigger the inflammatory response in cholesteatoma. Specimens were collected from nine patients with invasive cholesteatoma, nine patients with attic-type cholesteatoma (pars flaccida was perforated in five patients and intact in four) and four healthy skins. Expression and localization of LL-37 and interferon-alpha were detected by immunofluorescence and immunoblot analysis. Cultures of human cholesteatomatous keratinocytes were exposed to CpG DNA, LL-37 or CpG DNA complexed to LL-37 for 24 h. Expression of interferon-alpha was detected by RT-PCR. We detected abundant cytosolic DNA, increased LL-37 and interferon-alpha in keratinocytes in invasive cholesteatoma and attic-type cholesteatoma with pars flaccida perforation, but not in attic-type cholesteatoma with pars flaccida intact and normal skin. In cultured keratinocytes, LL-37-DNA complexes induced IFN-α expression. These data suggest that cytosolic DNA is an important disease-associated molecular pattern that triggers the inflammation response in cholesteatoma. Furthermore, LL-37 played an important role in DNA-triggered inflammation. Thus, we have identified a link between cytosolic DNA, LL-37 and autoinflammation in cholesteatoma, providing new potential targets for the treatment of this disease.
Insights
Self-DNA triggers inflammation in cholesteatoma by activating keratinocytes. The antimicrobial peptide LL-37 plays a key role in this DNA-triggered inflammatory response, suggesting new therapeutic targets.
Area of Science:
- Immunology
- Otolaryngology
- Dermatology
Background:
- Cholesteatoma is a destructive epithelial growth in the middle ear.
- The role of self-DNA in cholesteatoma-associated inflammation is not well understood.
Purpose of the Study:
- To investigate if self-DNA can initiate inflammatory responses in cholesteatoma.
- To explore the involvement of LL-37 and interferon-alpha in this process.
Main Methods:
- Immunofluorescence and immunoblot analysis of patient specimens.
- In vitro culture of cholesteatoma keratinocytes exposed to DNA and LL-37.
- RT-PCR to detect interferon-alpha expression.
Main Results:
- Abundant cytosolic DNA, increased LL-37, and interferon-alpha were found in invasive and perforated attic-type cholesteatoma keratinocytes.
- LL-37-DNA complexes induced interferon-alpha expression in cultured keratinocytes.
- These inflammatory markers were absent in intact attic-type cholesteatoma and healthy skin.
Conclusions:
- Cytosolic DNA acts as a disease-associated molecular pattern triggering inflammation in cholesteatoma.
- LL-37 is crucial in mediating DNA-induced inflammation.
- The interplay between cytosolic DNA, LL-37, and autoinflammation offers potential therapeutic targets for cholesteatoma.
