Project description:The whole proteome analysis of the Pseudomonas sp. FIP_A4 strain in presence and absence of fipronil was conducted to evaluate the differentially expressed enzymes that can play role in fipronil degradation.
Project description:We performed MIcroarray experiments on a total of 12 immune organs, including spleen (SP), bone marrow (BM), lymph node (LN) and Peripheral blood mononuclear cell (PBMC) and three tumor tissues including MC38, EL4 and LLC. 12 mice from four immune organs, including the spleen (SP), bone marrow (BM), lymph nodes (LN), and Peripheral blood mononuclear cell (PBMC), were isolated using routine experimental methods.9 mice were randomly divided into 3 groups and implanted subcutaneously(s.c.) with EL4 (2.0*10^5 /mice), MC-38 (1.0*10^6 /mice), and LLC-JSP-t2 (2.0*10^5 /mice) cells, respectively. All mice were sacrificed and tumor tissues were collected when the tumor volume reached 100 mm 3 . The tissues were ground up with liquid nitrogen and the lysates were stocked using Trizol for subsequent array analysis. RNA isolation and hybridization were performed by Capitalbio Technology Corporation according to the standard Affymetrix protocol .
Project description:Irradiated granulocyte macrophage-colony stimulating factor (GM-CSF)-transduced autologous tumor cells induce substantial antitumor immunity through the maturation and migration of dendritic cells (DCs). However, little is known about the key molecules involved in GM-CSF-sensitized DCs (GM-DCs) in tumor draining lymph nodes (TDLNs). We initially confirmed that mice subcutaneously injected with poorly immunogenic syngeneic Lewis lung carcinoma (LLC) cells transduced with Sendai virus encoding GM-CSF (LLC/SeV/GM) significantly rejected the tumor growth. Using microarray expression profiling, we obtained a large number of gene expression data files from GM-DCs and control DCs in TDLNs, and subjected them to network-based cluster analysis and unexpectedly unraveled the expression levels of type I IFNs-related genes specifically expressed in plasmacytoid DCs (pDC) were robustly up-regulated in GM-DCs. In vivo depletion assay showed that pDC-depleted mice treated with subcutaneous LLC/SeV/GM cells abrogated the antitumor effects observed in control mice. Moreover combination use of imiquimod for TLR7 triggering on pDC with irradiated LLC/SeV/GM cells induced a significant therapeutic antitumor effect with marked induction of CD9+ pDC with antitumor phenotype, whereas other control mice groups had only minimal to-modest antitumor responses, implicating that this combined vaccine strategy using imiquimod could be promising for improvement of GM-CSF-induced antitumor immunity. Mouse GM-CSF induced gene expression in mature dendritic cells in tumor draining lymph nodes from C57/BL6N female mouse was measured at 2 days after s.c. tumor challenge with GM-CSF gene-transduced LLC cells (LLC/SeV/GM) or control cells (LLC, LLC/SeV/GFP).
