Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Analysis of bacterial and macrophage transcripts in ISRE-positive and negative infected macrophages

ABSTRACT: To identify bacterial transcripts that may be associated with type I IFN production in Salmonella enterica subsp typhimurium (SL1344) infected macrophages we transformed macrophages with an ISRE-GFP reporter construct and sorted separate populations of GFP positive and GFP negative infected macrophages. We then did whole transcriptome profiling, collecting both host and bacterial transcripts, for differential expression analysis Analysis of ISRE positive, negative, and mixed populations at two time points (unexposed and 24hours) in duplicate (biological replicates). A sample consisting of Salmonella prior to infection was also included

ORGANISM(S): Mus musculus

SUBMITTER: Deborah Hung

PROVIDER: E-GEOD-65531 | 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:Analysis of human primary macrophages after live Lactobacillus rhamnosus GG (LGG) or LC705 stimulation for 6h and 24h. The results reveal novel mechanisms for probiotics-induced activation of the healthy human innate immune system. Macrophages are phagocytic cells of the innate immune system that perform sentinel functions to initiate appropriate responses to surrounding stimuli. Macrophages that reside at mucosa encounter ingested bacteria. Our understanding of the responses elicited by nonpathogenic bacteria in human innate immune system is limited. Lactobacillus are nonpathogenic bacteria commonly used in food and as supplements with health-promoting probiotic potential. In this study, we have utilized global gene expression profiling to compare the responses of human primary macrophages to two closely related, well-characterized L. rhamnosus strains LGG and LC705. Our results demonstrate that live LGG and LC705 induced quantitatively different gene expression in macrophages. A gene ontology analysis revealed functional similarities and differences in responses to LGG and LC705. Both LGG and LC705 induced IL-1b production in macrophages that required caspase-1 activity. LC705 but not LGG induced a strong type I IFN-dependent gene activation that correlated with the ability of LC705 to prevent influenza A virus replication and production of viral proteins in macrophages. Differentiated 7d human primary macrophages from 18 healthy individuals were stimulated with either live L. rhamnosus GG (LGG) or LC705 for 6 h and 24 h in RPMI medium containing penicillin and streptomycin. As a control, macrophages were stimulated with the medium only. The experiment was performed three times, each time with cells from six different individuals (samples 1-3). For each experiment, macrophages were stimulated separately as described, and macrophages from different donors were pooled after each stimulation experiment. Extracted RNA was hybridized to AffymetrixM-BM-. U133-plus2.0 GeneChipM-BM-. arrays.

Dataset Information

Analysis of bacterial and macrophage transcripts in ISRE-positive and negative infected macrophages

Publications

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

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