A novel in vitro model for the study of human keratinocyte/leucocyte interactions under autologous conditions
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Published on: November 21, 2014
N Boyera1, D Cavey, F Delamadeleine
1CIRD GALDERMA, Valbonne, France.
Insights
Researchers developed a novel in vitro model for studying keratinocyte/leucocyte interactions. This system demonstrated that activated T lymphocytes induce intercellular adhesion molecule 1 (ICAM-1) and HLA-DR expression on keratinocytes.
Area of Science:
- Immunodermatology
- Cell Biology
- In vitro modeling
Background:
- Keratinocyte and leucocyte interactions are crucial in skin immunity.
- Existing in vitro models for studying these interactions are limited.
- A need exists for a convenient and reliable co-culture system.
Purpose of the Study:
- To develop a novel in vitro co-culture system for autologous human keratinocyte/leucocyte interactions.
- To investigate the effects of activated T lymphocytes on keratinocytes.
- To provide direct in vitro evidence of leucocyte-induced changes in keratinocytes.
Main Methods:
- Developed a co-culture system using outer root sheath cells for pure keratinocyte isolation.
- Employed a Transwell system for co-culturing keratinocytes and leucocytes.
- Utilized phytohaemagglutinin-activated T lymphocytes and monocytes for co-culture experiments.
Main Results:
- Activated T lymphocytes induced intercellular adhesion molecule 1 (ICAM-1) and HLA-DR expression on keratinocytes.
- Methyl-3H-thymidine incorporation in keratinocytes was inhibited by activated T lymphocytes.
- These effects were mediated by soluble factors and required lymphocyte activation.
Conclusions:
- The developed Transwell system is a novel and effective tool for studying keratinocyte/leucocyte interactions.
- This study provides the first direct in vitro evidence of leucocyte-induced ICAM-1 and HLA-DR expression on keratinocytes.
- The system facilitates research into immune responses in the skin.
Abstract:
Keratinocyte/leucocyte interactions have become an area of intense investigations in the last decade. However, few convenient in vitro models are available at present. We have therefore designed a novel in vitro system for autologous human keratinocyte/leucocyte co-culture. Non-invasive epidermal cell sampling was achieved by using outer root sheath cells from hair follicles. After one passage, pure keratinocyte cultures (no Langerhans cells or melanocytes) were obtained. Co-culture experiments were performed on a Transwell system: keratinocytes were grown on the porous cupula, and then laid on to wells containing leucocytes. Alternatively, leucocytes can be added to the cupula when contact interactions between the two cell types are to be investigated. Using this system, we demonstrated that Phaesolus vulgaris phytohaemagglutinin-activated T lymphocytes (with 10% monocytes) in the lower compartment induced intercellular adhesion molecule 1 (ICAM-1) and HLA-DR expression, and inhibited methyl-3H-thymidine incorporation in normal human autologous keratinocytes cultured on the cupula. These changes were mediated by soluble factors (no cell contacts between keratinocytes and leucocytes), and required lymphocyte activation. This is the first direct in vitro evidence for leucocyte-induced ICAM-1 and HLA-DR expression on keratinocytes. This system is a potential tool for the study of keratinocyte/leucocyte interactions.(ABSTRACT TRUNCATED AT 250 WORDS)
