Project description:Rodent malaria parasite RNA hybridized on Illumina Mouse WG-6 v2.0 Expression BeadChip To investigate whether parasite RNA interfere with mouse beadchip analaysis. Malaria parasite resides in red blood cell, therefore RNA isolated from whole infected blood contains host RNA as well as parasite RNA
Project description:The Zika outbreak, spread by the Aedes aegypti mosquito, highlights the need to create high-quality assemblies of large genomes in a rapid and cost-effective fashion. Here, we combine Hi-C data with existing draft assemblies to generate chromosome-length scaffolds. We validate this method by assembling a human genome, de novo, from short reads alone (67X coverage, Sample GSM1551550). We then combine our method with draft sequences to create genome assemblies of the mosquito disease vectors Aedes aegypti and Culex quinquefasciatus, each consisting of three scaffolds corresponding to the three chromosomes in each species. These assemblies indicate that virtually all genomic rearrangements among these species occur within, rather than between, chromosome arms. The genome assembly procedure we describe is fast, inexpensive, accurate, and can be applied to many species.
Project description:The purpose of this research is to identify and evaluate the global gene expression of the rodent malaria parasites Plasmodium yoelii, Plasmodium berghei and Plasmodium chabaudi blood-stage parasites and specifically compare the blood stage gene expression profiles of samples derived from previous studies on Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi
Project description:We use the Plasmodium berghei rodent model to characterize the proteome of the final phase of liver stage development, the merosomes, packets of hepatic merozoites that bud from the host hepatocyte to initiate the blood stage of malaria. Plasmodium berghei WT ANKA strain was used to infect HepG2 hepatoma cells. Samples were fractionated by strong cation exchange, and nano-LC Orbitrap mass spectrometry was used to perform untargeted proteomic profiling of 3 biological replicates. Data was processed using MaxQuant and LFQ. Additional searches were performed to identify peptides from cleaved acetylated PEXELs (protein export elements) to identify proteins putatively exported to the host hepatocyte during liver stage development.
Project description:Pulmonary manifestations are life-threatening complications of malaria. To elucidate the immune landscape underlying malaria-associated lung pathology, we performed comprehensive transcriptomic analyses using a rodent malaria model. We identified that IFN-γ signaling specifically in T cells is a critical regulator of lung pathology. Our data demonstrate that the disruption of IFN-γ signaling in T cells results in reduced pulmonary parasite load and attenuated lung injury by enhancing T cell-monocyte interactions. Notably, this enhanced interaction promotes the expansion of a specific proinflammatory monocyte subset characterized by CD8 and Ly6C expression. This CD8+ Ly6C+ monocyte population exhibits significantly higher phagocytic capacity compared to its CD8− counterpart. Our study highlights the complex immune network induced by Plasmodium infection and reveals the essential role of the T cell-IFN-γ signaling axis in modulating monocyte-mediated parasite clearance and lung pathology.
Project description:PBMCs were obtain from mild malaria and cerebral malaria patient at the onset of P. falciparum infection. Patient recruitment took place in two hospital centers in Senegal Total RNAs from PBMCs of mild and cerebral malaria patients were profiled after hybridization with Agilent SurePrint G3 Human GE 8x60K Microarray to identify blood biomarkers for cerebral malaria phenotype Plasmodium falciparum malaria remains a major health problem in Africa. The mechanisms of pathogenesis leading to the severe form of the disease are not fully understood. Blood transcriptional profiles were investigated in patient with cerebral malaria, noncerebral malaria , and mild malaria by using the microarray technology. We identified a set of 447 genes that was differentially expressed between the three patient groups after a false discovery rate of 10%. Since the cerebral patients displayed a particular transcriptional pattern, we focused our analysis on the differences between cerebral malaria patients and mild malaria patients. We further found 849 genes differentially expressed after a false discovery rate of 10%. We validated our results by using the qPCR method for 5 genes. The enrichment analysis of their functional annotation indicates that genes involved in A, B, C pathways play a role in the occurrence of cerebral malaria. These results provide new insight into the potential effect of the dysregulation of gene expression and specific pathways. Host genetic variation may partly explain such alteration of gene expression. Further studies are required to investigate this in African populations.
Project description:Single-cell RNA-sequencing is revolutionising our understanding of seemingly homogeneous cell populations but has not yet been widely applied to single-celled organisms. Transcriptional variation in unicellular malaria parasites from the Plasmodium genus is associated with critical phenotypes including red blood cell invasion and immune evasion, yet transcriptional variation at an individual parasite level has not been examined in depth. Here, we describe the adaptation of a single-cell RNA-sequencing (scRNA-seq) protocol to deconvolute transcriptional variation for more than 500 individual parasites of both rodent and human malaria comprising asexual and sexual life-cycle stages. We uncover previously hidden discrete transcriptional signatures during the pathogenic part of the life cycle, suggesting that expression over development is not as continuous as commonly thought. In transmission stages, we find novel, sex-specific roles for differential expression of contingency gene families that are usually associated with immune evasion and pathogenesis.
Project description:Malaria-associated lung pathology, a severe and life-threatening manifestation of Plasmodium infection, involves complex immune remodeling within the pulmonary microenvironment. To resolve the spatial architecture of this immune response, we performed spatial transcriptomic analysis on lung tissues from a Plasmodium berghei NK65-induced rodent model. Our study identifies IFN-γ signaling in T cells as a pivotal regulator of the lung's spatial immune landscape. By utilizing conditional knockout (cKO) mice lacking Ifngr1 specifically in T cells, we observed that the blockade of T cell-intrinsic IFN-γ signaling attenuates lung pathology. Spatially, this protection is characterized by an enriched colocalization and enhanced interaction between T cells and monocytes. This robust interaction drives the expansion of a unique proinflammatory monocyte subset defined by CD8 and Ly6C expression within specific pulmonary niches. These CD8+ Ly6C+ monocytes exhibit significantly enhanced phagocytic capacity and elevated MHCII expression in situ. Our spatially resolved findings illustrate that the T cell-IFN-γ signaling axis dictates the spatial organization of T cell-monocyte communication, highlighting this interaction as a potential therapeutic target for mitigating malaria-induced lung injury.
Project description:Insect hemocytes mediate important cellular immune responses including phagocytosis and encapsulation, and also secrete immune factors such as opsonins, melanization factors, and antimicrobial peptides. In Anopheles, they contribute to the defense against malaria parasite invasion during the early sporogonic cycle. We used microarrays to identify if and to what degree circulating hemocytes have altered global expression profiles after infection with the rodent malaria parasite, Plasmodium berghei