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Establishment of an Anti-acne Vulgaris Evaluation Method Based on TLR2 and TLR4-mediated Interleukin-8 Production
Pichit Suvanprakorn1, Titiporn Tongyen1, Ornjira Prakhongcheep2,3
1Pan Rajdhevee Group Public Co., Ltd., Pathumthani, Thailand.
Background/Aim:
To date, no cell-based assay that focuses on the prime cause of acne initiation through activation of toll-like receptor2 and 4 and interleukin-8 (IL-8) production exists. Herein, we present an assay that evaluates acne by determining TLR2 and 4 expression and activation.
Materials And Methods:
Viability of keratinocytes was determined by the MTT assay. IL-8 was evaluated by ELISA. Immunocytochemistry was performed for determining receptor expression.
Results:
Lipoteichoic acid (LTA), peptidoglycan (PGN) and lipopolysaccharide (LPS) induced IL-8 production. Pre-treatment of cells with TLR2 and TLR4 inhibitors, before stimulation, reduced IL-8 production. Zinc gluconate was used for verification. Zinc can significantly suppress IL-8 production in the system. Treatment of cells with LTA+PGN or LPS resulted in increased TLR2 and TLR4 expression on the cell surface. This effect was prevented by zinc treatment.
Conclusion:
The measurement of IL-8 and TLR2 and TLR4 levels can be used for the evaluation of anti-acne treatment.
Insights
A new cell-based assay measures toll-like receptor 2 and 4 (TLR2/4) activation and interleukin-8 (IL-8) production to evaluate acne treatments. Zinc gluconate effectively reduced IL-8 production, indicating its potential as an anti-acne agent.
Area of Science:
- Dermatology
- Immunology
- Cell Biology
Background:
- Acne initiation involves toll-like receptor 2 and 4 (TLR2/4) activation and subsequent interleukin-8 (IL-8) production.
- Existing cell-based assays do not comprehensively evaluate these key acne pathogenesis factors.
Purpose of the Study:
- To develop and validate a novel cell-based assay for evaluating acne initiation.
- The assay quantifies TLR2 and TLR4 expression and activation, alongside IL-8 production.
Main Methods:
- Keratinocyte viability assessed via MTT assay.
- Interleukin-8 (IL-8) levels measured using ELISA.
- Toll-like receptor (TLR) expression determined by immunocytochemistry.
Main Results:
- Lipoteichoic acid (LTA), peptidoglycan (PGN), and lipopolysaccharide (LPS) stimulated IL-8 production.
- TLR2 and TLR4 inhibitors reduced IL-8 production, confirming pathway involvement.
- Zinc gluconate significantly suppressed IL-8 production and prevented TLR2/4 upregulation induced by LTA+PGN or LPS.
Conclusions:
- The developed assay effectively measures key indicators of acne initiation.
- Quantifying IL-8, TLR2, and TLR4 levels provides a reliable method for evaluating anti-acne treatments.
