Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiling of macrophages from conventional or antibiotic-treated mice

ABSTRACT: Antibiotic-treated (ABX) mice exhibit an impaired innate and adaptive antiviral immune response and substantially delayed viral clearance following exposure to systemic LCMV or mucosal influenza virus. Genome-wide transcriptional profiling of macrophages isolated from ABX mice revealed decreased expression of genes associated with antiviral immunity. Moreover, macrophages from ABX mice exhibited defective responses to type I and type II IFNs and impaired capacity to limit viral replication. Collectively, these data indicate that commensal-derived signals provide tonic immune stimulation that establishes the activation threshold of the innate immune system required for optimal antiviral immunity. In this study, we performed gene expression profiling to compare the transcriptional signatures of macrophages isolated from mice exposed to commensal bacteria-derived signals to macrophages isolated from mice depleted of commensal bacteria. Peritoneal macrophages (CD3Îµ-, CD5-, CD19-, CD11b+, F4/80+) from naÃ¯ve CNV or ABX mice were sorted directly into TRIzol (Invitrogen) on a BD Aria (Beckson Dickson). Test sorts were â?¥95% pure. For microarray analysis, RNA was extracted from 3 sorted biological replicates of peritoneal macrophages from naÃ¯ve CNV or ABX mice. cDNA was amplified using NuGen WT Ovation Pico kit and hybridized to Affymetrix GeneChip Mouse Gene 1.0 ST microarrays

ORGANISM(S): Mus musculus

SUBMITTER: Michael Abt

PROVIDER: E-GEOD-74712 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Commensal bacteria calibrate the activation threshold of innate antiviral immunity.

Abt Michael C MC Osborne Lisa C LC Monticelli Laurel A LA Doering Travis A TA Alenghat Theresa T Sonnenberg Gregory F GF Paley Michael A MA Antenus Marcelo M Williams Katie L KL Erikson Jan J Wherry E John EJ Artis David D

Immunity 20120614 1

Signals from commensal bacteria can influence immune cell development and susceptibility to infectious or inflammatory diseases. However, the mechanisms by which commensal bacteria regulate protective immunity after exposure to systemic pathogens remain poorly understood. Here, we demonstrate that antibiotic-treated (ABX) mice exhibit impaired innate and adaptive antiviral immune responses and substantially delayed viral clearance after exposure to systemic LCMV or mucosal influenza virus. Furth ...[more]

PMID: 22705104

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Project description:Innate memory phenotype (IMP) CD4+ T cells are non-conventional Î±Î² T cells exhibiting features of innate immune cells, characterized as CD44high and CD62Llow in periphery. It is recently reported by our group that bone marrow chimeric mice lacking thymic MHCI expression develop predominantly IMP CD8+ T cells, while those lacking hematopoietic MHCI develop predominantly naÃ¯ve CD8+ T cells. Here we perform hirarchical clustering analysis and found that CD4+ T cells share similar property: chimeras lacking thymic MHCII gave rise to predominantly CD4+ T cells that resemble IMP CD4+ T cells observed in WT mice, and vice versa, chimeras lacking hematopoietic MHCII had a majority of naÃ¯ve-like CD4+ T cells resembling naÃ¯veCD4+ T cells seen in WT mice. We used microarrays to compare the global programme of gene expression to determine whether the hematopoietic MHCII selected CD4+ T cells are IMP, and whether the thymic MHCII selected CD4+ T cells are naÃ¯ve CD4+ T cells as observed in WT mice. Through hierarchical clustering and analysis of global gene differential expression, we determined that hematopoietic MHCII dependent IMP CD4+ T cells generated from WT bone marrow transplanted into irradiated MHCII-/- recipients, resemble IMP CD4+ T cells in WT mice, while naÃ¯ve CD4+ T cells generated from MHCII-/- bone marrow transplanted into irradiated WT recipients, resemble naÃ¯ve CD4+ T cells in WT mice. Cell Sorting was performed using a Cytopeia Influx Cell Sorter. Chimeric IMP (CD45.1+TCRÎ²+CD4+CD44highCD62Llow) CD4+ T cells were sorted from splenocytes of CD45.1+WTâCD45.2+MHCII-/- chimeras (WM IMP CD4), and chimeric naÃ¯ve (CD45.2+TCRÎ²+CD4+CD44lowCD62Lhigh) CD4+ T cells were sorted from splenocytes of CD45.2+MHCII-/- â CD45.1+WT chimeras (MW naÃ¯ve CD4) respectively, 8 weeks post transplantation. WT IMP (TCRÎ²+CD4+CD44highCD62Llow) and naÃ¯ve (TCRÎ²+CD4+CD44lowCD62Lhigh) CD4+ T cells were sorted from splenocytes of 8-week old WT mice.

Dataset Information

Gene expression profiling of macrophages from conventional or antibiotic-treated mice

Publications

Commensal bacteria calibrate the activation threshold of innate antiviral immunity.

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