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A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
Published on: November 28, 2014
Detection of perforin and granzyme B mRNA expressing cells in lichen sclerosus
Robert E Hunger1, Marcel Brönnimann, Andreas Kappeler
1Department of Dermatology, University of Bern, Inselspital, 3010 Bern, Switzerland. robert.hunger@insel.ch
Abstract:
Granzyme B and perforin messenger RNA (mRNA) expression has been shown to be a specific in vivo activation marker for cytotoxic cells. The aim of this study was to assess the contribution of cell-mediated cytotoxicity in the pathogenesis of lichen sclerosus. In situ hybridization and immunohistochemistry were performed on serial tissue sections of lesional skin biopsies and normal skin as control. Immunohistochemical staining showed that the cellular infiltrate of diseased skin consisted predominantly of T cells (CD3+) and some B cells (CD20+). Among T cells CD4+ and CD8+ cells were found in about equal numbers. In normal skin samples perforin and granzyme B mRNA expressing cells were only rarely found. In contrast, in biopsies from diseased skin a high percentage of infiltrating cells expressed mRNA for perforin and granzyme B. The perforin and granzyme B expressing cells were found in the dermal infiltrate and intraepidermally in close proximity to keratinocytes suggesting in situ activation of these cells. These findings provide evidence that cell-mediated cytotoxicity plays a significant role in tissue destruction in lichen sclerosus.
Insights
Cell-mediated cytotoxicity, marked by granzyme B and perforin mRNA, is a key factor in lichen sclerosus pathogenesis. This study found high expression of these cytotoxic markers in affected skin, indicating their role in tissue damage.
Area of Science:
- Immunology
- Dermatology
- Molecular Biology
Background:
- Granzyme B and perforin mRNA are specific in vivo activation markers for cytotoxic cells.
- Cell-mediated cytotoxicity is implicated in various inflammatory skin conditions.
Purpose of the Study:
- To evaluate the role of cell-mediated cytotoxicity in the pathogenesis of lichen sclerosus.
- To investigate the expression of granzyme B and perforin mRNA in lesional skin of lichen sclerosus patients.
Main Methods:
- In situ hybridization and immunohistochemistry were performed on lesional and normal skin biopsies.
- Cellular infiltrates were characterized using T cell (CD3+, CD4+, CD8+) and B cell (CD20+) markers.
Main Results:
- Lesional skin showed a predominant infiltrate of T cells (CD3+) and some B cells (CD20+), with roughly equal numbers of CD4+ and CD8+ T cells.
- A high percentage of infiltrating cells in lichen sclerosus biopsies expressed mRNA for perforin and granzyme B.
- These cytotoxic marker-expressing cells were located in the dermal infiltrate and intraepidermally, near keratinocytes, suggesting in situ activation.
Conclusions:
- Cell-mediated cytotoxicity, evidenced by high perforin and granzyme B mRNA expression, plays a significant role in the tissue destruction observed in lichen sclerosus.
- The findings suggest that cytotoxic T cells are activated in situ within the skin lesions of lichen sclerosus.