title: Identification and visualization of CD8+ T cell mediated IFN-γ signaling in target cells during an antiviral immune response in the brain.
abstract: CD8(+) T cells infiltrate the brain during an anti-viral immune response. Within the brain CD8(+) T cells recognize cells expressing target antigens, become activated, and secrete IFNγ. However, there are no methods to recognize individual cells that respond to IFNγ. Using a model that studies the effects of the systemic anti-adenoviral immune response upon brain cells infected with an adenoviral vector in mice, we describe a method that identifies individual cells that respond to IFNγ. To identify individual mouse brain cells that respond to IFNγ we constructed a series of adenoviral vectors that contain a transcriptional response element that is selectively activated by IFNγ signaling, the gamma-activated site (GAS) promoter element; the GAS element drives expression of a transgene, Cre recombinase (Ad-GAS-Cre). Upon binding of IFNγ to its receptor, the intracellular signaling cascade activates the GAS promoter, which drives expression of the transgene Cre recombinase. We demonstrate that upon activation of a systemic immune response against adenovirus, CD8(+) T cells infiltrate the brain, interact with target cells, and cause an increase in the number of cells expressing Cre recombinase. This method can be used to identify, study, and eventually determine the long term fate of infected brain cells that are specifically targeted by IFNγ. The significance of this method is that it will allow to characterize the networks in the brain that respond to the specific secretion of IFNγ by anti-viral CD8(+) T cells that infiltrate the brain. This will allow novel insights into the cellular and molecular responses underlying brain immune responses.
authors: Board of Governors' Gene Therapeutics Research Institute, Cedars-Sinai Medical Center, Department of Medicine, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Mariana
authors: M
authors: Puntel
authors: Robert
authors: R
authors: Barrett
authors: Nicholas S R
authors: NS
authors: Sanderson
authors: Kurt M
authors: KM
authors: Kroeger
authors: Niyati
authors: N
authors: Bondale
authors: Mia
authors: M
authors: Wibowo
authors: Sean
authors: S
authors: Kennedy
authors: Chunyan
authors: C
authors: Liu
authors: Maria G
authors: MG
authors: Castro
authors: Pedro R
authors: PR
authors: Lowenstein
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D001921:Brain / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D005260:Female; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics; D019426:Integrases / Q000378:metabolism; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015398:Signal Transduction / Q000276:immunology*; D013045:Species Specificity; D013937:Thymidine Kinase / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D007371:Interferon-gamma; D013937:Thymidine Kinase; C045073:Cre recombinase; D019426:Integrases