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Updated: Jun 25, 2026
Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions
Published on: April 20, 2016
Murine epidermal Langerhans cells and langerin-expressing dermal dendritic cells are unrelated and exhibit distinct
Keisuke Nagao1, Florent Ginhoux, Wolfgang W Leitner
1Dermatology Branch, National Cancer Institute, Center for Cancer Research, National Institutes of Health, Bethesda, MD 20892, USA.
Insights
Researchers identified distinct skin dendritic cell (DC) subsets, Langerhans cells (LC) and langerin(+) dermal DCs (dDC). These cells have specialized functions, impacting immune responses and vaccine development.
Area of Science:
- Immunology
- Dermatology
- Cell Biology
Background:
- Langerin(+) dermal dendritic cells (dDC) are a newly identified subset in murine dermis.
- The relationship between langerin(+) dDC and epidermal Langerhans cells (LC) is not fully understood.
- Distinguishing between these subsets is crucial for understanding skin immunity.
Purpose of the Study:
- To characterize the lineage and functional differences between langerin(+) dDC and LC.
- To investigate the distinct roles of these subsets in immune responses.
- To explore the implications for epicutaneous vaccine strategies.
Main Methods:
- Selective expression of EpCAM to distinguish LC from langerin(+) dDC.
- In vitro migration assays.
- In vivo repopulation studies after conditional depletion.
- Gene gun immunization studies.
Main Results:
- LC and langerin(+) dDC exhibit different migratory capabilities and repopulation kinetics.
- Langerin(+) dDC development does not require TGFbeta1, unlike LC.
- Langerin(+) dDC are essential for acute IgG2a/c and IgG2b production.
- LC play a role in regulating IgG1 and IgG2a antibody production.
Conclusions:
- LC and langerin(+) dDC are distinct subsets with specialized functions.
- LC are important immunoregulatory cells.
- The presence of at least three distinct skin DC subsets is relevant for epicutaneous vaccine design.
Abstract:
A new langerin(+) DC subset has recently been identified in murine dermis (langerin(+) dDC), but the lineage and functional relationships between these cells and langerin(+) epidermal Langerhans cells (LC) are incompletely characterized. Selective expression of the cell adhesion molecule EpCAM by LC allowed viable LC to be easily distinguished from langerin(+) dDC in skin and lymphoid tissue and ex vivo as well. Differential expression of EpCAM and langerin revealed the presence of at least 3 distinct skin DC subsets. We determined that LC and langerin(+) dDC exhibit different migratory capabilities in vitro and repopulate distinct anatomic compartments in skin at different rates after conditional depletion in vivo. Langerin(+) dDC, in contrast to LC, did not require TGFbeta1 for development. Carefully timed gene gun immunization studies designed to take advantage of the distinct repopulation kinetics of langerin(+) dDC and LC revealed that langerin(+) dDC were required for optimal production of beta-galactosidase-specific IgG2a/c and IgG2b in the acute phase. In contrast, immunization via LC-deficient skin resulted in persistent and strikingly reduced IgG1 and enhanced IgG2a Ab production. Our data support the concepts that LC and langerin(+) dDC represent distinct DC subsets that have specialized functions and that LC are important immunoregulatory cells. The presence of at least 3 functionally distinct skin DC subsets may have particular relevance for vaccines that are administered epicutaneously.
