Transcription profiling of mouse lung trafficking of macrophages

ABSTRACT: Trafficking of monocytes into lung tissue and their differentiation into lung resident macrophages and dendritic cells is supposed to be regulated by the expression of specific gene clusters, which promote cell-cell interaction, migration and matrix degradation and the acquisition of tissue specific cellular phenotypes. Traffic related gene clusters include chemokines, integrins, and tissue-degrading matrix metallopeptidases, for all of which members have been shown to be functionally important. A complete picture, however, of the gene clusters that are regulated during in vivo migration and differentiation of peripheral blood monocytes under non-inflammatory conditions has not been obtained yet. Currently, adaptive changes of cellular phenotypes cannot be directly assessed by cell fate mapping during the slow trafficking of mononuclear phagocytes to lung tissue under steady-state conditions. Therefore, as an alternative approach to gain a better insight into the genetic programs that drive the mononuclear phagocyte migration and differentiation processes, the transcriptomes of circulating monocytes were compared with their lung tissue mononuclear phagocyte progeny. Experiment Overall Design: A total of 32 animals were used in this study. From each Animal, Monocytes, Macrophages and dendritic cells were isolated. Cells were sorted from blood-free lungs and peripheral blood. In order to get enough RNA for a labeling reaction, RNA of each 4 of these animals was pooled (same amount of RNA per animal). The different pools were subjected to microarray analysis comparing Monocytes, Macrophages and Dendritic Cells in a ring design with 4 hybridizations per comparison. There were 8 different pools per cell type, 4 of which were labeled with Cy3 and the other 4 were labelled with Cy5 (balanced dye-swap). Each possible combination of cell types was compared with 4 hybridizations, resulting in a total number of 12 hybridizations. Each hybridization used RNA from different animals. In each of these comparisons, each cell type was labelled twice with Cy3 and twice with Cy5.

ORGANISM(S): Mus musculus

SUBMITTER: Jochen Wilhelm 

PROVIDER: E-GEOD-13558 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress