Project description:Rabies is a fatal zoonotic disease posing a threat to the public health globally. Rabies virus (RABV) is excreted in the saliva of infected animals, and is primarily transmitted through bite contact. Salivary glands play an important role for virus propagation. However, the significance of salivary glands is less studied in RABV pathogenic mechanisms. To identify functionally important genes in the salivary glands, we employed RNA sequencing (RNA-seq) to establish and analyze mRNA expression profiles in parotid tissue infected with two RABV strains, CVS-11 and PB4. We map the transcriptome changes in response to RABV infection in parotid tissue for the first time. This work provides new clues to the study of RABV-affected salivary gland function and RABV transmission mechanisms in parotid tissue. And the salivary gland-enriched transcripts could be potential targets of interest for rabies disease control.
Project description:Aedes aegypti mosquitoes were orally infected with DENV, ZIKV or CHIKV virus, and once virus infection reached salivary gland tissues, these glands were dissected out and i-TRAQ performed to identify differentially expressed proteins during virus infection.
Project description:In order to select genes that are differentially expressed in salivary glands during Ixodes ricinus infection by Bartonella henselae we compare the transcriptome of infected and non-infected salivary glands
Project description:Street-strain rabies virus primarily replicates in central nervous system without inducing significant immune response or structural damages on neurons, but the manifested symptoms of rabies indicate inherent neuronal dysfunctions in CNS. To understand the underlying state of rabies virus-infected neurons and find probable mechanisms for the neuronal dysfunction, we performed RNA-Seq at multiple time-points. This dataset provides RNA-Seq results of wild-type and mutant rabies virus-infected neuron transcriptome, with clear differential expressions between conditions. Through comparative analysis of different time-points, we have found that the matrix protein of rabies virus plays an important role in early suppression of host gene expression and maintaining control over immune response and other processes. The signaling pathways previously known to interact with rabies virus were confirmed to be modulated in this dataset, and contribute to neuronal function-associated processes. We have verified the regulation of gene expressions that could impact neuronal functions collectively, and demonstrated in calcium imaging that indeed the oscillation of calcium trace in neurons are influenced by rabies virus infection.
Project description:In order to select genes that are differentially expressed in salivary glands during Ixodes ricinus infection by Bartonella henselae we compare the transcriptome of infected and non-infected salivary glands I. ricinus from a pathogen free colony were infected -or not- with B. henselae at the larval and then the nymphal stage and salivary glands of the resulting females adult were dissected for RNA extraction. 454 sequencing was performed on pooled conditions in order to obtain a reference databank. Infected and non-infected samples were then sequenced separatly with illumina, blaston teh reference datbank and compared.
Project description:Cortical circuit tracing using modified rabies virus can identify input neurons making direct monosynaptic connections onto neurons of interest. However, challenges remain in our ability to establish the cell type identity of rabies-labeled input neurons. While transcriptomics may offer an avenue to characterize inputs, the extent of rabies-induced transcriptional changes in distinct neuronal cell types remains unclear and whether these changes preclude characterization of rabies-infected neurons according to established transcriptomic cell types is unknown. We used single-nucleus RNA sequencing to survey the gene expression profiles of rabies-infected neurons and assessed their correspondence with established transcriptomic cell types. We demonstrated that when using transcriptome-wide RNA profiles, rabies-infected cortical neurons can be transcriptomically characterized despite global and cell-type-specific rabies-induced transcriptional changes. Notably, we found differential modulation of neuronal marker gene expression, suggesting that caution should be taken when attempting to characterize rabies-infected cells with single genes or small gene sets.
Project description:Rabies virus (RABV) infection led to alteration of microRNA expression in NA and microglia. A sixteen-chip study was performed using total RNA isolated from NA and microglia infected with rabies virus CVS-11 (Challenge virus standard) or mock infected as a control. RNA was isolated from microglia at 12, 24, or 48 hpi and from NA at 24 hpi.
Project description:Acetylation is the PTM that strongly interlinked with glucose metabolism, so we performed Liquid chromatography–mass spectrometry (LC-MS) to detect acetylated proteins in RABV-infected mouse brains to further investigate the reason how Rabies virus (RABV) infection promotes glucose uptake. To reveal the changes in lysine acetylation due to RABV infection, the brains of mice infected with different RABV strains were harvested for a quantitative proteomic assay.
Project description:Recent work indicates that salivary glands are able to constitutively recruit CD8+ T cells and retain them as tissue resident memory T cells (TRM), independently of local infection, inflammation or antigen. To understand the mechanisms supporting T cell recruitment to the salivary gland, we compared T cell migration to the salivary gland in mice infected or not with murine cytomegalovirus (MCMV), a herpesvirus that infects the salivary gland and promotes the accumulation of salivary gland TRM. We found that acute MCMV infection increased rapid T cell recruitment to the salivary gland, but that equal numbers of activated CD8+ 44 T cells eventually accumulated in both infected and uninfected glands. T cell recruitment to uninfected salivary glands depended on chemokines and the integrin α4. Several chemokines were expressed in the salivary glands of both infected and uninfected mice and many of these could promote the migration of MCMV-specific T cells in vitro. MCMV infection increased expression of chemokines that interact with the receptors CXCR3 and CCR5, but neither receptor was needed for T cell recruitment to the salivary gland during MCMV infection. Unexpectedly however, the chemokine receptor CXCR3 was critical for T cell accumulation in uninfected salivary glands. Together, these data suggest that CXCR3 and the integrin α4 mediate T cell recruitment to uninfected salivary glands, but that redundant mechanisms mediate T cell recruitment after MCMV infection.
Project description:This experiment is part of the project that primarily aims to utilize 3D hydrogel-based hiPSC-derived neuronal model to study rabies virus infection in the central nervous system. Having established the optimal 3D neuronal model, we then investigated the growth kinetics of two strains of rabies virus (TH and CVS-11) and comparatively analyzed the 2D and 3D culture models. We performed a gene expression analysis using NanoString to determine whether changes in gene expression could explain the differences in virus growth kinetics of two strains of rabies virus observed between the 2D and 3D neuronal culture models. Gene expression analysis of the neuropathological pathway observed during rabies virus infection demonstrated a vast number of differentially expressed genes in the 3D model as compared to the 2D model.