Unknown,Transcriptomics,Genomics,Proteomics

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Expression analysis of human PBMCs stimulated with Borrelia, MDP or Pam3cys.

ABSTRACT: Analysis examining the effect of the Crohn's disease-associated ATG16L1 polymorphism on the responses to stimulation components derived from various bacteria classes. Expression analysis was performed on RNA extracted from peripheral blood mononuclear cells (PBMCs) from healthy genotyped volunteers were stimulated for 24h with RPMI alone, or RPMI containing heat-killed Borrelia burgdorferi , muramyl dipeptide (MDP) or Pam3Cys.

ORGANISM(S): Homo sapiens

SUBMITTER: Aylwin Ng

PROVIDER: E-GEOD-68765 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Rationale: Previous work demonstrated that pre-exposure to ozone primes innate immunity and increases TLR4-mediated response to subsequent stimulation with lipopolysaccharide (LPS). To further explore the pulmonary innate immune response to ozone exposure, we investigated the effect of ozone in combination with Pam3CYS, a synthetic TLR2/TLR1 agonist. Methods: Bronchoalveolar lavage (BAL) and lungs were harvested from C57Bl/6 mice after exposure to ozone or filtered air followed by saline or Pam3CYS 24 hours later. Cells and cytokines in the BAL, surface expression of TLRs on macrophages, and lung RNA genomic expression profiles were examined. Results: We demonstrate an increased BAL cell influx, increased IL-6 and KC, and decreased MIP-1α and TNF-α in response to Pam3CYS as a result of ozone pre-exposure. We also observed increased cell surface expression of TLR4, TLR2 and TLR1 on macrophages as a result of ozone alone or in combination with Pam3CYS. Gene expression analysis of lung tissue revealed a significant increase in expression of genes related to injury repair and cell cycle as a result of ozone exposure. When comparing Pam3CYS treated animals to saline treated animals with or without ozone, genes associated with inflammation were significantly increased. Potentially novel ozone exposure candidate genes (CCK, RELM-α and β) were identified. Conclusion: Our results extend previous findings with ozone/LPS to other TLRs PAMPs and suggest that ozone priming of innate immunity is a general mechanism. Gene expression profiling of lung tissue identified transcriptional networks and genes that contribute to the priming of innate immunity at the molecular level. Mice were exposed to either 2ppm ozone or filtered air (FA) for 3 hours. 24 hours following ozone exposure, mice from either the ozone or FA group were treated intratracheally with either 100ug of Pam3CYS in saline or saline alone. Animals were euthanized 4 or 24 hours post-Pam3CYS exposure. RNA from whole lung tissue from 4 animals per group was profiled.

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Expression analysis of human PBMCs stimulated with Borrelia, MDP or Pam3cys.

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