RNA-Seq of Plasmodium chabaudi from BALB/c and C57BL/6 mice
ABSTRACT: Recent advances in high throughput sequencing methodologies allow the opportunity to probe in depth the transcriptomes of organisms including important human pathogens. In this project, we are using Illumina sequencing technology to analyze the transcriptome (RNA-Seq) of experimentally accessible stages of the mouse malaria parasite, P. chabaudi AS. The aim is to analyse cir gene expression during Plasmodium chabaudi infection and determine whether host genetic background can influence cir expression. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/. Abstract: Transcriptome sequencing of blood stage P. chabaudi AS parasites grown under different host genetic backgrounds.
INSTRUMENT(S): Illumina Genome Analyzer II, Illumina HiSeq 2000
Project description:During malaria infection is observed a robust immune response culminating on release of inflammatory mediators. This exacerbated immune response is involved in malaria symptoms and mortality. There are evidences that this response is mediated by innate immunity where pattern recognition receptors have a key role. We used microarrays to elucidate some pro-inflammatory genes that are differential expressed during P. chabaudi infection, a malarial murine model Spleen from C57BL/6 or MyD88 knockout mice non-infected or after 6 days post infection with 105 P. chabaudi infected red blood cells were harvested for RNA extraction and hybridization on Affymetrix microarrays. This time point was chose to coincides with rupture of red blood cells \since this event of the parasite life cycle is related with malaria outcomes.
Project description:Microarray profiling of whole blood of immunodeficient mouse strains at 24h after infection with Plasmodium chabaudi. Multiple independent replicates collected for each knockout. RNA isolated and amplified. Hybridized as two-color experiment against a common pooled reference.
Project description:Single-cell RNA sequencing (scRNA-seq) was used to study the various transcriptional states of individual CD4+ T cells during blood-stage Plasmodium chabaudi infection in mice. This is an experimental model of malaria in which CD4+ T cells are essential for controlling parasite numbers, and which is characterized by concurrent development of Th1 and Tfh cells. We have used Plasmodium-specific TCR transgenic CD4+ T cells to minimise the effects of TCR diversity on Th fate decisions. Activated antigen-specific cells were studied at days 0, 2, 3, 4 and 7. In addition, dendritic cells and monocytes were studied at days 0 and 3. Cell lysis, RT and cDNA preamplification was performed using Fluidigm C1 system.
Project description:Microarray profiling of amplified total splenic RNA isolated from TLR9-, TLR7-, UNC93b1- or MYD88- deficient mice or their C57BL/6 controls 24h after mock infection or infection with Plasmodium chabaudi. 4 independent replicates collected from each genotype/infection group, RNA isolated and amplified from whole spleen. Hybridized as a one color experiment.
Project description:Cerebral malaria (CM) is one of the most severe complications of malaria infection. There is evidence that repeated parasite exposure promotes resistance against CM, as indicated by the low incidence of CM in adults in malaria-endemic regions. However, the immunological basis of this infection-induced resistance remains poorly understood. Here, a microarray study done utilising the tractable Plasmodium berghei ANKA model of experimental cerebral malaria (ECM), we show that three rounds of infection and drug-cure protects against the development of ECM during a subsequent fourth infection.
Project description:The aim of the present study was to analyze the effect of protective vaccination on the miRNome of the liver during the crisis phase in vaccination-induced self-healing infections of Plasmodium chabaudi in comparison to lethal infections in non-protected Balb/c mice using Agilent’s microRNA expression microarrays.
Project description:The aim of the present study was to analyze the effect of protective vaccination on the transcriptome of the liver towards the end of the crisis phase in vaccination-induced self-healing infections of Plasmodium chabaudi in comparison to lethal infections in non-protected Balb/c mice using Agilent’s gene expression microarrays.