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Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation
Published on: January 26, 2016
Murine epidermal Langerhans cells express significant amounts of class I major histocompatibility complex antigens
A Lenz1, C Heufler, H G Rammensee
1Department of Dermatology, University of Innsbruck, Austria.
Insights
Epidermal Langerhans cells (LC) express major histocompatibility complex (MHC) class I molecules on their surface. This finding challenges previous research suggesting LC are deficient in MHC class I expression, indicating a broader role in immune responses.
Area of Science:
- Immunology
- Cell Biology
- Dermatology
Background:
- Epidermal Langerhans cells (LC) are key antigen-presenting cells in the skin.
- Previous studies suggested LC lack major histocompatibility complex (MHC) class I antigen expression.
- This implied a specialization towards MHC class II-restricted antigen presentation.
Purpose of the Study:
- To re-evaluate the expression of MHC class I antigens on murine epidermal Langerhans cells.
- To distinguish MHC class I expression on LC from surrounding keratinocytes.
Main Methods:
- Utilized bone marrow chimeras to differentiate donor and recipient cells.
- Employed immunofluorescence microscopy with specific monoclonal antibodies.
- Analyzed epidermal cell suspensions using flow cytometry.
- Tested antibody-mediated depletion of LC.
Main Results:
- Donor-derived LC in chimeric mice stained positive for MHC class I antigens.
- LC from normal mice showed surface expression of MHC class I antigens.
- LC were significantly depleted from cell suspensions using anti-MHC class I antibodies and complement.
Conclusions:
- Murine epidermal Langerhans cells express MHC class I molecules on their surface.
- This contradicts prior findings of MHC class I deficiency on LC.
- LC possess a more comprehensive antigen-presenting capability than previously understood.
Abstract:
Epidermal Langerhans cells (LC) are leukocytes that express major histocompatibility complex (MHC) class II antigens and function as antigen-presenting and accessory cells. Caughman et al. [Caughman, S. W., Sharrow, S. O., Shimada, S., Stephany, D., Mizuochi, T., Rosenberg, A. S., Katz, S. I. & Singer, A. (1986) Proc. Natl. Acad. Sci. USA 83, 7438-7442] reported that LC are deficient in surface expression of MHC class I antigens, implying a specialization of these cells to class II-restricted antigen presentation. To readdress this obviously important issue, we have studied murine epidermal sheets prepared from B6 X BALB/c----B6 bone marrow chimeras 5 months after irradiation and bone marrow reconstitution. This enabled us to distinguish class I of LC from that of surrounding keratinocytes. When sheets were analyzed by immunofluorescence microscopy with monoclonal antibodies specific for donor class I antigens, donor-derived LC but not LC of recipient origin were stained. Appropriate controls for antibody isotype and MHC haplotype were negative. LC in epidermal cell suspensions, prepared from normal BALB/c and BALB/cBy mice (MHC haplotype d), were analyzed by flow cytometry as well as immunofluorescence microscopy. LC were stained by monoclonal antibodies to class I antigens of haplotype d, but not by isotype-matched control antibodies to class I antigens of haplotype k. We also found that LC were virtually depleted from epidermal cell suspensions by treatment with monoclonal antibodies to class I antigens of haplotype d and complement but not by treatment with control monoclonal antibodies and complement. Our data, therefore, show that LC express MHC class I molecules on their surface.
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