Project description:Local heart irradiation of 2, 8 or 16 Gy to ApoE-/- mice and follow up tp 20 and 40 weeks. Th goal of this study is to understand microvascular alterations of radiation-induced cardiactoxicity.
Project description:WT or CD27-/- OT-I T cells are stimulated in vitro with artficial antigen presenting cell (aAPC) lines expressing CD70 and/or CD80 for 2, 4, 8 or 14 hours.
Project description:Mesenteric, brachial and axilary lymph nodes from 4 C57Bl/6 females or 5 CD27KO females, both groups were 6 weeks old, were extracted. The different lymph nodes were pooled and a single cell suspension was created. The cell suspension from WT or CD27KO mice was stained with anti-TCRd-PE (GL3) and anti-CD3e-PE Cy7 (2C-11), and FACSorted for double-positive cells on a BD Aria, obtaining around 1 million cells of each sample. These cells were spun down and the pellet was frozen in liquid nitrogen and kept at -80 ºC before RNA extraction.
Project description:WT and CD137L-/- mice of 8-12 weeks old were immunized intra-peritoneally with 50 µg chicken gamma- globulin conjugated to 4-hydroxy-3-nitrophenylacetyl (NP-CG) in alum or infected intranasally with 25 hemagglutinin units of influenza virus strain A/NT/60/68 in 50 µl HBSS. Nine days after immunization or infection, B cells were enriched from pooled spleens and lymph nodes of 4 mice per test group by means of magnetic labelled bead cell separation (MACS) using anti-mouse BD ImagTM CD45R/B220-particles DM. The B220+ MACS-sorted populations were stained with anti-GL7-FITC and anti-CD19-PE to sort CD19+GL7+ GC and CD19+GL7- non-GC B lymphocytes by flow cytometry on a FACSAria (BD). Around 1 million cells of each sample were obtained. These cells were spun down and the pellet was frozen in liquid nitrogen and kept at -80 ºC before RNA extraction.
Project description:Wild type or Cd27-/- mice of 8-12 weeks old were immunized intranasally with 500 ug ovalbumin (OVA) protein and 1 ug cholera toxin in 50 ul HBSS. Spleen, draining lymph node (DLN) or lung were isolated at day 3, day 4 or day 8 after immunization. Single cell suspensions of indicated organs were prepared and OT-I T cells were isolated from this using flow cytometric cell sorting.
Project description:Gene expression array analysis on a series of ten different histological special types of invasive breast carcinomas (tubular, micropapillary, mucinous A, mucinous B, endocrine, apocrine, metaplastic, medullary, adenoid cystic, invasive ductal carcinoma with osteoclastic giant cells) and invasive lobular carcinoma.<br><br>Note: this experiment was reloaded into ArrayExpress in August 2010 to include mappings between raw and processed data files. It now includes additional dye-swap combined normalized data files.
Project description:Gene expression profiling to predict outcome after chemoradiation in head and neck cancer Purpose. The goal of the present study was to improve prediction of outcome after chemoradiation in advanced head and neck cancer using gene expression analysis. Materials and Methods. We collected 92 biopsies from untreated head and neck cancer patients subsequently given cisplatin-based chemoradiation (RADPLAT) for advanced squamous cell carcinomas (HNSCC). After RNA extraction and labeling we performed dye swap experiments using 35k oligo-microarrays. Supervised analyses were performed to create classifiers to predict local control, locoregional control and disease recurrence. Published gene sets with prognostic value in other studies were also tested. Results. Using supervised classification on the whole series, gene sets separating good and poor outcome could be found for all end-points. However, when splitting tumors into training and validation groups, no robust classifiers could be found. Also previously published signatures with prognostic value have been tested. Conclusion. Gene sets can be found with predictive potential for locoregional control after combined radiation and chemotherapy in HNSCC. How treatment-specific these gene sets are needs further study.