|Application:||Gene expression microarray analysis|
|Number of samples:||9|
|Release date:||Jul 23 2018|
|Last update date:||Oct 22 2018|
|Diseases:||Breast Neoplasms, Neoplasms|
|Dataset link||The mRNA and lncRNA profiles of the T cells activated by breast tumor-specific DCs pulsed by MDA-MB-231 lysates at day 0, day 1 and day 6|
Mononuclear cells were obtained from peripheral blood of three HLA-A2+ healthy donors and cultured for 14 d in serum-free X-VIVO 20 medium (BioWhittaker, Walkersville, Maryland) with human GM-CSF (50 ng/ml; Peprotech) and IL-4 (20 ng/ml; Peprotech). Non-adherent DCs were enriched by depletion of contaminating T and B lymphocytes and pulsed for 20 h with lysates (200 μg protein/1 × 10^6 cells/ml) from MDA-MB-231 (HLA-A2+) that were lysed by 5 freeze/thaw cycles. T cells were incubated for 13 d in RPMI-1640 with 10% human AB serum (PromoCell) and IL-2 (25 U/ml; Peprotech) followed by incubation in the same medium with antigen-specific DCs (5:1) for 6 days. T cells were retrieved from the cocultures at day 0, day 1 and day6. Total RNA was extracted from T cells with indicated treatments (5 x 106 cells) using the TRIzol Reagent (Cat#A33251, Thermo Fisher). The quality of extracted RNA was assessed using the NanoDrop ND-2000 spectrophotometer.