Project description:Ribosome profiling (Ribo-Seq) (maps positions of translating ribosomes on the transcriptome) and RNA-Seq (quantifies the transcriptome) analysis of Rattus norvegicus cells infected with Moloney Murine Leukemia Virus (Mo-MuLV).
Project description:Histone modifcations and CTCF binding at the c-myb locus were compared in cell lines with c-myb expressing, which are myeloblatic M1 cells and leukemia cells with virus integration, VS. M1 cells without c-myb expression induced by IL-6. Distribution of active histone marks at the c-myb gene and the upstream regions are associated with active c-myb transcription. The enrichment of all of these active histone marks decreased with differentiation-induced down-regulation of c-myb, but increased and spread in tumor cells. ChIP-on-chip from murine myeloid cell line M1 and virus-induced myeloid leukemia cell lines for H3K4me3, H3K9/14ac, H3K4me1, H3K27me3, H3K9me3 and CTCF
Project description:It has been shown that antibodies can enhance Ebola virus (EBOV) infection at sub-neutralizing concentrations. The effect of antibodies on the infection of murine cell line (RAW 264.7) with MLV-EBOV, a murine leukemia virus pseudotype expressing surface glycoprotein (GP) of EBOV and green fluorescent protein (GFP) has been studied. Moreover, regions of EBOV GP targeted by antibodies enhancing the infection have been investigated. Serum and monoclonal antibodies (mAbs) were obtained from mice inoculated with MLV-EBOV. Plasma from patients and humanized anti-EBOV GP mAbs were obtained. The proportion of GFP-positive cells was determined by immunocytofluorometry. Trypsin-digested EBOV GP was incubated in affinity columns prepared with mAbs. Eluted peptides were analyzed by high-resolution mass spectrometry. The infection of cells with MLV-EBOV was enhanced by immune mouse serum, human plasma and mAbs. The infection was the result of interactions between MLV-EBOV /antibody complexes and FcγRs. Regions of EBOV GP, located in GP1 core, furin cleavage site, N-terminal part of GP2, and HR1 and HR2 domains are target of antibodies able to enhance the infection. In conclusion, the RAW 264.7 cell line associated with MLV-EBOV can be used to study the enhancing activity of murine and human antibodies and may help to select antibodies or vaccine formulations that do not lead to enhancement of EBOV infection.
Project description:Single-cell RNA-seq analysis of cortex and hippocampus tissue from murine males and females after recovery from West Niles virus encephalitis
Project description:Despite the advanced understanding of disease mechanisms, the current therapeutic regimens fail to cure most patients with acute myeloid leukemia (AML). In the present study, we address the role of protein synthesis control in AML leukemia stem cell (LSC) function and leukemia propagation. We apply a murine model of mixed-lineage leukemia-rearranged AML to demonstrate that LSCs synthesize more proteins per hour compared with the bulk of leukemia. Using a genetic model that permits inducible and graded regulation of ribosomal subunit joining, we show that defective ribosome assembly leads to a significant survival advantage by selectively eradicating LSCs but not normal hematopoietic stem and progenitor cells. Finally, transcriptomic and proteomic analyses identify a rare subset of LSCs with immature stem cell signature and high ribosome content that underlies the resistance to defective ribosome assembly. Collectively, our study unveils a critical requirement of high protein synthesis rate for LSC function, highlighting ribosome assembly as a therapeutic target in AML.
Project description:Infection of cells with murine hepatitis virus strain A59 (MHV-A59) results in massive amounts of viral RNA within infected cells. When applying transcriptional profiling by means of microarray analysis, this could potentially cause problems since high amounts of viral RNA could in theory interfere with the analysis. Since coronavirus RNAs are polyadenylated, the viral RNAs are also amplified by oligo(dT) primers (a standard procedure when processing RNA samples for array hybridization). In this experiment we compared two different array platforms for the potential interference of viral RNA, using MHV-A59 infection of LR7 (murine fibroblasts) as a model.