Dataset Information

Figure 7. Long-lasting antitumor T-cell memory induced by a single Lenti-HPV-07 vaccination

ABSTRACT: Lenti-HPV-treated and cured mice which have been re-challenged with TC-1 cells (n = 5/group), received at day 286 post tumor engraftment an i.v. injection of an adjuvant-free mixture of E7HPV16:ETTDPDRAHYNIVTF and E7HPV16:PDRAHYNIVTFCCKC peptides (25 µg of each/mouse). Naïve control mice received the same i.v. treatment. At 48h post injection, splenocytes were stimulated in vitro with homologous or negative control peptides during 6h prior to IFN-g ICS and surface labeling.

ORGANISM(S): Mus musculus (mouse)

SUBMITTER:

PROVIDER: S-BSST1108 | biostudies-other |

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:To compare human memory CD4+ T cell subsets in peripheral blood (PB) and bone marrow (BM) of healthy individuals at transcriptional level, we analyzed the global gene expression of ex vivo PB CD69- as well as BM CD69- and CD69+ memory CD4+ T cells from 4 paired PB and BM samples. The gene expression of these subsets was additionally compared to the transcriptional profile of 8 PB samples taken ex vivo or stimulated with phorbol myristate acetate (PMA) and Ionomycin for 3 hours. Three ex vivo memory CD4+ T cell subsets (CD4+CD45RO+CD69+ and CD4+CD45RO+CD69- cells from bone marrow; CD4+CD45RO+CD69- cells from peripheral blood) were isolated from 4 paired bone marrow and blood samples (two males and two females) or from a different cohort of 8 blood samples (3 males and five females). The purity of sorted cells was higher than 95% as assessed by FACS. Subsequently, a fraction of purified cells from the 8 blood samples were stimulated with phorbol-myristic-acid/ionomycin (PMA/iono) for 3 h and used as high controls. Total RNA of each cell subset was extracted using a NucleoSpin RNA XS Kit (Macherey-Nagel) or RNeasy Mini kit (Qiagen). The integrity and amount of isolated RNA was assessed for each sample using an Agilent 2100 Bioanalyzer (Agilent, Waldbronn, Germany) and a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE). Double-stranded complementary RNA was synthesized from 1 M-BM-5g total RNA using Message AmpII Biotin (Ambion, USA). Fifteen micrograms of fragmented cRNA of each sample were hybridized to 28 HG-U133A plus 2.0 GeneChips (Affymetrix). Hybridization was performed in a Hybridization Oven 640, and chips were washed and stained in the Fluidics Station 400 (both Affymetrix). Finally, the arrays were scanned with a GeneChip Scanner 3000 using the GCOS software, version 1.4, both Affymetrix. All relevant GCOS data of quality checked microarrays were analyzed with High Performance Chip Data Analysis (HPCDA, unpublished), using the BioRetis database (www.bioretis-analysis.de), as described and validated previously.

Dataset Information

Figure 7. Long-lasting antitumor T-cell memory induced by a single Lenti-HPV-07 vaccination

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets