Macrophage-glia interactions regulate immune-damage to enteric neurons during West Nile virus infection [Dataset]
Description
In this study, we defined the neuron-glia-macrophage interactions after West Nile virus (WNV) infection; this model neurotropic virus causes GI tract dysmotility in mice via injury of enteric neurons through a T cell-mediated cytolytic mechanism. Our experiments demonstrate that resident macrophages play a critical role in regulating the glial immune response to viral infection in the intestinal muscularis, and that their depletion markedly exacerbates neuronal injury. Data were generated using flow cytometry, fluorescent activated cell sorting, qRT-PCR, immunofluorescent confocal microscopy, RNA sequencing, fluorescent in situ hybridization. There are 4 main Figures and 4 Supplementary Figures. Data were obtained using C57BL6/J mice and Ai3 (YFP) (B6.Cg-Gt(ROSA)26Sortm3(CAG-EYFP)Hze/J, Cx3cr1CreERT2 (B6.129P2(C)-Cx3cr1tm2.1(cre/ERT)Jung/J), Plp1CreERT2 B6.Cg-Tg(Plp1-cre/ERT)3Pop/J); NuTRAP (B6;129S6-Gt(ROSA)26Sortm2(CAG-NuTRAP)Evdr/J) mice; DTA (B6.129P2-Gt(ROSA)26Sortm1(DTA)Lky/J mice.
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We dissected muscularis externa of small intestine from mice infected subcutaneously with West Nile virus. We analyzed the virus titers by qRT-PCR, neuronal damage by immunofluorescent staining for neuronal markers followed by confocal imaging. The cell infiltrates after infection were assessed by flow cytometry. Sorted resident macrophages were analyzed by RNA sequencing. Response of enteric glia was assessed by RNA sequencing after enrichment with immunoprecipitation. Macrophages, T cells and neutrophils were depleted by injections of monoclonal antibodies. Enteric glia were genetically ablated using tamoxifen inducible Cre mouse line.
Institutions
- Washington University in St. LouisMO, Saint Louis
- Cleveland ClinicOH, Cleveland
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Funders
- National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthUnited StatesGrant ID: R01DK122790
- National Institute of Arthritis and Musculoskeletal and Skin DiseasesNational Institutes of HealthUnited StatesGrant ID: K08AR084597
- National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthUnited StatesGrant ID: T32DK077653
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| Keywords | West Nile virus, GI tract, inflammation, neurons, glia, macrophages, neutrophil, T cell, immune response |