Project description:miRNA profiling of resting and activated T cells Two condition experiment, resting versus activated T cells, measured pooled samples from three independent stimulations
Project description:Hemophagocytes are activated macrophages seen morphologically to have engulfed other hematopoietic cells. Their function is unknown. Attempts to induce these cells in vitro or purify them ex vivo have been unsuccessful. We isolated HPCs and resting splenic macrophages by laser capture microdissection. RNA was isolated from captured cells and microarray performed to compare the activation state of HPCs versus resting splenic macrophages.
Project description:Hemophagocytes are activated macrophages seen morphologically to have engulfed other hematopoietic cells. Their function is unknown. Attempts to induce these cells in vitro or purify them ex vivo have been unsuccessful. We isolated HPCs and resting splenic macrophages by laser capture microdissection. RNA was isolated from captured cells and microarray performed to compare the activation state of HPCs versus resting splenic macrophages. We treated WT mice with PBS or CpG1826 and an IL-10receptor blocking antibody. Cells were captured from splenic touch preps on PEN-coated slides, RNA isolated by WT-Ovation One-Direct Amplification System and assayed on Affymetrix mouse gene ST 1.0 chips.
Project description:Resting NK cells are notoriously difficult to transduce, especially using the VSVG lentivirus pseudotype. When NK cells are expanded and activated using the NKAES system, there is a modest improvement of transduction rates compared to their resting NK counterparts. Still, the transduction efficiency remains poor for robust development of NK cell-based cancer immunotherapy. We envisaged to have a genome-wide expression profile of resting and activated NK (NKAES) cells in order to improve currrent transduction protocols .
Project description:To compare up-regulation of genes following CpG activation, we performed microarray analysis of activated macrophages from B6 and F1(B6xMOLF) mouse strains. Cells were activated for 0, 2 and 4 hrs with 200nM of type B CpG. Levels of mRNA for many genes differened dramatically between the strains Peritoneal macrophages were elicited from pertoneal cavity of mice 3 days after thioglycollate injection. The cells were plated overnight at a density of 5 million cells per 100 mm dish and activated the next day with 200 nM CpG. Total RNA was isolated with TRIZOL followed by reverse transcrition, fragmentation and labeling accroding the manufacture's (Affymetrix) recommendations
Project description:This experiment compared the expression of mRNAs and noncoding RNAs between resting and activated CD8+ effector T cells, with the primary aim of identifying noncoding RNAs that were dynamically regulated during T cell activation.
Project description:CD4+ T cells (T helper cells) are cytokine-producing adaptive immune cells that activate or regulate the responses of different immune cells. They are known to play crucial roles in antibody class switching in B cells, neutrophil recruitment and activation of macrophages and CD8+ cytotoxic T cells. The activation and functional status of CD4+ T cells is important for adequate responses to pathogen infections but has also been associated with auto-immune disorders and survival in several cancers. In the current study, we carried out proteomic profiling of resting and activated primary human CD4+ T cells from healthy donors. In addition to identifying known markers of CD4+ T cell activation, we also identified protein kinases, protein phosphatases, and cytokines to be differentially expressed.