Project description:description Blastocystis sp. is a highly prevalent anaerobic eukaryotic parasite of humans and animals. The genome of several representatives has been sequenced revealing specific traits such as an intriguing 3’-end processing of primary transcripts. We have acquired a first high-throughput proteomics dataset on the difficult to cultivate ST4 isolate WR1 and detected 2,761 proteins. We evidenced for the first time by proteogenomics a functional termination codon derived from transcript polyadenylation for seven different key cellular components.
Project description:Human immunodeficiency virus (HIV) infection is a chronic condition, where viral DNA integrates into the genome. The fate of the provirus determines the infection course. Latently infected cells form a persistent, heterogeneous reservoir. The reservoir that reinstates an active infection comprises cells with intact provirus that can be reactivated. We confirmed that latent cells from patients exhibited active transcription throughout the provirus. To find transcriptional determinants, we characterized the establishment and maintenance of latency during proviral chromatin maturation in primary CD4+ T-cells for four months after HIV infection. As heterochromatin (marked with H3K9me3 or H3K27me3) gradually stabilized, the provirus became less accessible and lost activation potential. In a subset of infected cells, active marks (i.e., H3K27ac) remained detectable, even after prolonged proviral silencing. After T-cell activation, the proviral activation occurred uniquely in cells with H3K27ac-marked proviruses. Our observations suggested that, after transient proviral activation, cells were actively returned to latency.
Project description:Chikungunya virus (CHIKV) infection is characterized by alterations in gene expression profile on host cells that consequently lead to an immune response. Here, we used RNA sequencing to analyze the mRNA expression profile in human monocyte-derived macrophages (MDMs) infected with a Colombian clinical isolate of CHIKV at 6 and 24 hpi. analyze the mRNA expression profile in the human monocyte-derived macrophages infected at 6 and 24 hrs with a Colombian clinical isolate of Chikungunya virus.
Project description:In rainbow trout, type A spermatogonia can be split into SP cells and non-SP cells by the ability to exclude Hoechst 33342 dye (H33342). The H33342 fluorescence of SP cells are lower than that of non-SP cells, after H33342 staining. To investigate whether SP cells were transcriptomically distinct from non-SP cells, we compared the transcriptome of these cells. We used fluorescence-activated cell sorting (FACS) to isolate SP cells and non-SP cells from the type A spermatogonia in rainbow trout.
Project description:Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide each year. Among the PPV strains identified so far, only PPV-C and PPV-CR are able to infect cherries under natural conditions. Herein, we evaluated the impact of strains differing by the pathogenic potential on herbaceous host Nicotiana benthamiana. At first glance, a faster accumulation of PPV capsid protein was noticed by semi-quantitative DAS-ELISA on tobacco leaves infected by PPV-CR (the RU-30sc isolate) in contrast to PPV-C (BY-101 isolate). This result was correlated perfectly with observed phenotypic symptoms. To assess the host response to infection more deeply, a comprehensive proteomic profiling was performed using reverse phase UHPLC followed by label-free mass spectrometry quantification. Thirty-one unique plant proteins were identified as significantly altered due to the infection. Precise evaluation of particular amount shifts of identified proteins in relation to infection has revealed that the aggressive PPV-CR isolate has a stronger influence on the abundance of photosynthesis-related proteins, mainly from Calvin cycle, as the mild PPV-C. This observation was accompanied by a significant reduction of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of remaining identified proteins, indicates activation of repair mechanism, stimulation of photosynthetic capacity, and modifications in amino acid and carbohydrate metabolism to affect plant growth and initiate energy formation via gluconeogenesis. Furthermore, we speculate that accumulation of H2O2, in PPV-CR infected leaves may activate glutathione synthesis, which plays a crucial role in further plant defense and development.
Project description:Virus and host factors contribute to cell-to-cell variation in viral infection and determine the outcome of the overall infection. However, the extent of the variability at the single cell level and how it impacts virus-host interactions at a systems level are not well understood. To characterize the dynamics of viral transcription and host responses, we used single-cell RNA sequencing to quantify at multiple time points the host and viral transcriptomes of human A549 cells and primary bronchial epithelial cells infected with influenza A virus. We observed substantial variability of viral transcription between cells, including the accumulation of defective viral genomes (DVGs) that impact viral replication. We show a correlation between DVGs and viral-induced variation of the host transcriptional program and an association between differential induction of innate immune response genes and attenuated viral transcription in subpopulations of cells. These observations at the single cell level improve our understanding of the complex virus-host interplay during influenza infection.
Project description:The underlying molecular mechanisms of pathogenesis and outcome of disease to different pathotypes of H5N1 influenza infection in ducks remain unclear. For that, we studied genome wide host gene expression of lung tissues infected with A/duck/India/02CA10/2011(AD2011) H5N1 virus and A/duck/Tripura/103597/2008 (AD2008) H5N1 virus in ducks using custom designed microarray. AD2011 is highly pathogenic whereas AD2008 is low pathogenic to ducks. Comparative analysis of differentially expressed genes revealed that 688 genes were commonly expressed, 877 and 1556 genes are uniquely expressed to infection with AD2011 and AD2008 virus isolate, respectively. The up-regulation of cytokines genes OAS, IL1B, IL17, IFITM2, CCL4, CXCR4, STAT3, TGFB1 and TGFB2 in the lungs tissues may cause high mortality in ducks infected with AD2011 virus. The expression of important antiviral immune genes IFIT5, IFITM5, RSAD2, EIF2AK2 (PKR), Mx, β-defensins, TRIM23 and SLC16A3 to AD2008 infection, but not in AD2011 infection, cause the host may fine-tune their innate immune responses and prevent from cytokines storms and tissue damage. Several immune related Gene ontology (GO) terms and immune pathways activated were qualitatively similar but quantitatively different to both virus infections. Based on these findings, we conclude that subtle differences in host immune responses may determine the different outcome of H5N1 infection in ducks. Agilent Custom Duck Gene Expression 8X60k (AMADID: G4102A_059612) designed by Genotypic Technology Private Limited , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)