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Flow cytometry analysis of adhesion molecules on human Langerhans cells
J Vedel1, P Vincendeau, J H Bézian
1Service de Dermatologie, Hôpital des Enfants, Bordeaux, France.
Insights
Langerhans cells (LCs) change cell surface molecules for migration. Fresh LCs express CD11c and CD18, similar to macrophages, while CD58 appears after culture, aiding T-cell interactions.
Area of Science:
- Immunology
- Cell Biology
Background:
- Langerhans cells (LCs) are critical for skin immunity, migrating between the epidermis and lymph nodes to present antigens.
- This migration necessitates dynamic changes in cell-surface molecule expression.
Purpose of the Study:
- To investigate the expression patterns of adhesion molecules CD11/CD18 and CD58 on human Langerhans cells.
- To understand how these molecules change upon culture and their implications for LC function.
Main Methods:
- Enrichment of human epidermal Langerhans cells (LCs) using Ficoll-Hypaque gradient and Fc receptor panning.
- Immunophenotypic analysis of LCs using specific antibodies against CD1a, HLA-DR, CD11 subunits (CD11a, CD11b, CD11c), CD18, and CD58.
- Assessment of molecule expression on fresh LCs and after 48 hours in culture.
Main Results:
- Freshly isolated LCs expressed CD11c (weaker than CD18) and CD18, resembling tissue macrophages.
- CD11c expression diminished during culture.
- CD58 was not detected in fresh LCs but emerged after 2 days of culture.
- The expression of CD58 post-culture aligns with LC interactions with CD2-bearing T-lymphocytes.
Conclusions:
- Langerhans cells display macrophage-like surface markers (CD18, CD11c) in their native epidermal environment.
- The induction of CD58 upon culture suggests its role in facilitating T-cell interactions essential for antigen presentation in lymph nodes.
Abstract:
The Langerhans cell (LC) migrates between the epidermis and the regional lymph nodes to present antigens. This migration pattern requires the expression of a changing repertoire of cell-surface molecules. In this work, we have investigated the expression of the adhesion molecules CD 11/CD 18 and CD 58 on LCs. Human epidermal cell suspensions were enriched in LCs (mean enrichment 75%) using a two-step technique including a Ficoll-Hypaque gradient followed by Fc receptor panning with IgG-coated sheep erythrocytes. The number of cells obtained per experiment was 750,000 (extremes 280,000-1,800,000), and the following antibodies were tested on fresh suspensions and/or after 48 hours in culture: BB3 (antithyroglobulin negative control IgG2a), OKT6 (anti CD1a, Ortho), anti HLA-DR (Becton-Dickinson), MHM 24 (anti CD 11a, leukocyte typing workshop n(0)3), MO1 and 44 (anti CD 11b, leukocyte typing workshop n(0)3), anti CD 11c (Immunotech), 60.3 and MHM 23 (anti CD 18, leukocyte typing workshop n(0)2), TS2/9.1.1 (anti CD 58, leukocyte typing workshop n(0)3). We found that amongst CD 11 subunits, only CD 11c was expressed in fresh suspensions, but was weaker than CD 18, and disappeared with culture. CD 58 was not detected in fresh suspensions but appeared after 2 days of culture, confirming earlier work. Thus the LC exhibits cell surface characteristics similar to tissue macrophages (CD 18 and CD 11c) prior to culture. The expression of CD 58 after culture is in accordance with the interaction of LC with CD2 bearing T-lymphocytes during antigen presentation in peripheral lymph-nodes.
