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Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
10:31

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

Published on: April 8, 2016

CD69, an early activation antigen on lymphocytes, is constitutively expressed by human epidermal Langerhans cells

T Bieber1, A Rieger, G Stingl

  • 1Department of Dermatology, University of Munich, Medical School, Germany.

The Journal of Investigative Dermatology
|May 1, 1992
PubMed

Insights

Skin Langerhans cells (LC) express CD69, a cell surface antigen. This CD69 expression on epidermal dendritic cells is unique and may indicate specific functional properties.

Area of Science:

  • Immunology
  • Dermatology
  • Cell Biology

Background:

  • Langerhans cells (LC) are critical immune cells within the epidermis.
  • CD69 is a cell surface marker found on various immune cells, including activated T and NK cells.
  • The expression and function of CD69 on LC are not fully understood.

Purpose of the Study:

  • To investigate the expression of CD69 on epidermal Langerhans cells (LC).
  • To characterize the CD69 molecule expressed by LC.
  • To explore factors influencing CD69 expression on LC in vitro.

Main Methods:

  • Immunohistochemical staining of skin cryosections with anti-CD69 monoclonal antibody (MoAb) Leu-23.
  • Flow cytometric analysis of LC-enriched epidermal cells (EC).
  • Immunoprecipitation and Endoglycosidase-F treatment to analyze the CD69 protein structure.
  • In vitro culture of LC-enriched EC with and without IFN-gamma.

Main Results:

  • Anti-CD69 MoAb Leu-23 reacted with a subpopulation of epidermal dendritic cells, identified as LC.
  • LC-enriched EC showed nearly all CD1a-bearing LC expressing CD69.
  • Immunoprecipitation revealed LC express a disulphide-linked dimer (26/30kDa) of CD69, differing slightly from T/NK cells.
  • Endoglycosidase-F treatment indicated a different glycosylation pattern in LC-derived CD69.
  • CD69 expression on LC decreased in culture but was sustained by IFN-gamma.

Conclusions:

  • Resident epidermal Langerhans cells express CD69.
  • The CD69 molecule on LC exhibits distinct biochemical properties compared to that on T/NK cells, likely due to glycosylation differences.
  • IFN-gamma can maintain CD69 expression on LC in vitro.
  • Further research is needed to link CD69 expression to specific functional properties of intraepidermal LC.

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