Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Bulk RNA-seq analysis of the macrophage response to Salmonella enterica subsp. Typhimurium (SL1344) exposure

ABSTRACT: A time course of the macrophage response to Salmonella exposure analyzing the effects of input cell number as a control for single cell studies Mouse macrophages were exposed to Salmonella enterica for different lengths of time. Libraries were constructed using either approximately 500,00 macrophages lysed directly on a tissue culture dish (bulk) or using only 150 cells isolated using FACS (sorted). All libraries were constructed in duplicate (bulk) or triplicate (sorted). All replicates are biological replicates

ORGANISM(S): Mus musculus

SUBMITTER: Deborah Hung

PROVIDER: E-GEOD-65530 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Pathogen Cell-to-Cell Variability Drives Heterogeneity in Host Immune Responses.

Avraham Roi R Haseley Nathan N Brown Douglas D Penaranda Cristina C Jijon Humberto B HB Trombetta John J JJ Satija Rahul R Shalek Alex K AK Xavier Ramnik J RJ Regev Aviv A Hung Deborah T DT

Cell 20150903 6

Encounters between immune cells and invading bacteria ultimately determine the course of infection. These interactions are usually measured in populations of cells, masking cell-to-cell variation that may be important for infection outcome. To characterize the gene expression variation that underlies distinct infection outcomes and monitor infection phenotypes, we developed an experimental system that combines single-cell RNA-seq with fluorescent markers. Probing the responses of individual macr ...[more]

PMID: 26343579

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Project description:The highly infectious bacterium Francisella tularensis is a facultative intracellular pathogen, whose virulence requires proliferation inside host cells, including macrophages. Although some Francisella determinants of intracellular growth have been identified, much remains to be understood about the pathogenesis of this organism. In particular, how Francisella responds to its intracellular environment could provide clues about its intracellular biology and reveal pathogenic determinants based on their intracellular expression profiles. Here we have performed a global transcriptional profiling of the highly virulent F. tularensis subsp. tularensis Schu S4 strain during its intracellular cycle within primary murine macrophages. Phagocytosed bacteria rapidly responded to their intracellular environment and progressively altered their transcriptional profile over time. Differential gene expression profiles were revealed that correlated with specific intracellular locations of the bacteria. Upregulation of general and oxidative stress response genes was a hallmark of the early phagosomal and late endosomal stages, while induction of a subset of transport and metabolic genes characterized the cytosolic replication stages. Expression of the Francisella Pathogenicity Island (FPI) genes, the functions of which are associated with intracellular proliferation, increased during the intracellular cycle. Similarly, 27 chromosomal loci putatively encoding hypothetical, secreted, outer membrane proteins or transcriptional regulators were identified as upregulated during the intracellular cycle. In-frame deletion of FTT0383, the Schu S4 ortholog of fevR, of FTT0369c or FTT1676 abolished the ability of Schu S4 to either survive or proliferate intracellularly, demonstrating that bacterial factors of intracellular pathogenesis can be identified based on their intracellular expression profile. In conclusion, establishing the intracellular transcriptome of Francisella has revealed important aspects of its intracellular biology and identified novel virulence determinants of this pathogen. Keywords: Time series Intracellular cycle within primary murine macrophages using the time series of zero, one, two, four, eight, twelve, sixteen and twenty-four hours

Dataset Information

Bulk RNA-seq analysis of the macrophage response to Salmonella enterica subsp. Typhimurium (SL1344) exposure

Publications

Pathogen Cell-to-Cell Variability Drives Heterogeneity in Host Immune Responses.

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