Differential Gene Expression in Cultured Human Langerhans Cells in Response to Phagocytic Stimulation
In order to identify genes whose differential expression in Langerhans cells (LC) is increased by a potent, specific stimulus that is associated with the antigen-processing and -presenting activity of these cells, we have begun the analysis of phagocytic stimulation of freshly isolated human LC in culture (cLC) using zymosan particles. Previously it has been shown that cLC actively take up zymosan, and that phagocytosis of these particles is mediated by the mannose/ß-glucan receptor(s)1. Genes that are preferentially expressed in cLC following phagocytic stimulation were identified by mRNA Representational Difference Analysis (RDA)2,3 and DNA sequence analysis of cloned RDA difference products (DP). In order to address the question of cell type-specific molecular activation patterns, a similar set of experiments with monocytes cultivated with GM-CSF and IL-4 (MoDC) was included in this analysis. In total, more than 150 recombinant plasmid clones containing DP were isolated, sequenced and compared with primate sequence databases and EST databases.
KeywordsSouthern Blot Analysis Differential Gene Expression Zymosan Particle Dendritic Cell Culture Fresh Surgical Specimen
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