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The gene expression profile of dendritic cells can be used as a predictor of function.

ABSTRACT: The human lung cancer cell line A549 was cultured in media alone, with cyclophosphamide (CPM), oxaliplatin (OXP) or DMSO alone for 72 hours. The exhausted culture media were aspirated and stored at -20C. Peripheral blood mononuclear cells (PBMCs) were isolated from four pathologically healthy donor buffy coats using Histopaque-1077. Following removal of red blood cell and platelet contamination, monocytes were isolated using magnetic beads coated with anti-CD14. Cells were resuspended and cultured in a DC-maturing medium for 7 days. Non- and loosely-adherent cells (the DC fraction) were harvested and purity assessed by CD11c/HLA-DR/CD14 immuno-discrimination by flow cytometry. 1x10^5 unstimulated DCs were cultured with tumour derived supernatants or culture media alone for 24 hours. DCs were then harvested and RNA isolated by Trizol. Total RNA was labelled and hybridised to Illumina Human HT-12v3 arrays. A separate aliquot of DCs from each patient was exposed to supernatant derived from tumour treated with OXP; changes in CD80, CD83 and CD86 were measured and DC populations showing a greater than two-fold up-regulation of these markers were classified as "GOOD_DC".

ORGANISM(S): Homo sapiens

SUBMITTER: Wai Liu

PROVIDER: E-MTAB-389 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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