Do dermal dendritic cells phagocytose apoptotic keratinocytes in lupus erythematosus?
Thursday 16 April 2009
Project
Lupus erythematosus (LE) is an autoimmune disease with a broad clinical spectrum and immunological abnormalities. LE develops primarily in young women of child bearing age. The cause of LE is unknown, but research into the pathogenesis has deepened the insight. Especially studies on apoptosis and the clearance of apoptotic cells by phagocytosis in lupus have provided hypotheses on the development and course of the disease.The current hypothesis in the pathogenesis of LE is that macrophages are impaired in their phagocytic capabilities and the immune stimulatory dendritic cells take up apoptotic cells and induce inflammation. Whether the main dendritic cell in the skin, the Langerhans cell, also plays a role in cutaneous LE (within UV induced lesions) is sofar unclear.
In this project the UV light-induced apoptosis of keratinocytes and their subsequent phagocytosis by monocytes, macrophages, dendritic cells and neutrophils is examined. The efficiency of phagocytosis of apoptotic cells or fluorescent beads by different types of dendritic cells will be studied. This will be performed in the presence of serum of LE patients and controls. Different dendritic cell subtypes will be cultured from healthy and LE donors and tested for their phagocytic capacity.
Techniques
Cell isolation (monocytes)
Cell culture (HaCaT cellline, blood derived monocytes, dendritic cell differentiation)
Apoptosis assays (flow cytometry and immunohistochemistry)
Fluorescent labelling of cells
Phagocytosis assay (microscopic analysis and flow cytometry
Duration
Due to the duration of the in vitro maturation system a 9 months period is preferred.
Contact
Dr. Edward Knol, tel 088 75 562 67, e.f.knol@umcutrecht.nl
More info
Website Dermatology Utrecht
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