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Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production
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Chapters
- 00:05Title
- 01:39Cytokines in γHV68-infected Mice: qPCR and ELISA
- 03:52MEF Cell Infection and Cytokine Quantification
- 05:17Lentivirus Production, Infection, and Generation of Stable Cell Lines
- 07:09Results: Antiviral Cytokine Production in vivo
- 08:07Conclusion
We describe a protocol to measure the antiviral cytokine production in mice infected with a model herpesvirus, murine gamma herpesvirus 68 (γHV68) that is closely-related to human Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV). Utilizing genetically modified mouse strains and mouse embryonic fibroblasts (MEFs), we assessed the antiviral cytokine production both in vivo and ex vivo. “Reconstituting” the expression of innate immune components in knockout embryonic fibroblasts by lentiviral transduction, we further pinpoint specific innate immune molecules and dissect the key signaling events that differentially regulate the antiviral cytokine production.
