|Application:||Gene expression microarray analysis|
|Number of samples:||6|
|Release date:||Feb 1 2008|
|Last update date:||Dec 6 2012|
|Diseases:||Bacterial Infections, Mycobacterium Infections, Nontuberculous, Tuberculosis, Eye Infections, Viral|
|Dataset link||Mycobacterium tuberculosis targets DC-SIGN for immunosuppression through the PD-1/PD-Ligand pathway|
Human PBMC-derived DCs were grown in 6-well plates and stimulated for 18 h with either mycobacterial ManLAM or recombinantly expressed ICAM-3. To ensure complete naïve status of DCs before stimulation, cells were left in wells from the day of PBMC isolation until end of stimulation. Change of medium was performed by carefully removing medium from wells without any manipulation of DCs. After stimulation, total RNA was isolated as described above and labeled with Fluorescent Direct Labeling Kit (Agilent Technologies) following manufacturer’s protocols. Microarray experiments were performed as two-color dye-reversal hybridizations. To compensate for dye-specific effects, a color swap was performed. Labeled RNA was hybridized on Whole Human Genome Oligo Microarray Kit 44k Format and scanned using an Agilent scanner according to manufacturer’s protocol (Agilent Technologies). Image analysis was performed using Feature Extractor software (Agilent Technologies). For data analysis Rosetta Resolver software was used (Rosetta Inpharmatics).