【病毒外文文献】2012 P115 Pathogenic Influenza A viruses and SARS-Coronaviruses modulate global interferon stimulated gene induction thr

nations of vaccine adjuvants were administered intranasally in a maximum volume of 50 ml Tissue samples were harvested from euthanized mice and processed for analysis of viral titers quantitative RT PCR or ELISA In survival experiments mice were sacrificed if their weight dropped more than 25 of their initial body weight as per institutional guidelines The animal experiments were performed according to the guidelines of the German animal protection law All animal protocols were approved by the relevant German authorities Flow cytometry Lung samples were enzymatically digested with 2 mg ml of Col lagenase D Roche Mediastinal lymph nodes were disrupted mechanically Tissue samples were processed for flow cytometry and red cells were lysed using BD Red Blood Cell Lysing Buffer Samples were acquired on a FACS canto II instrument and analyzed with FlowJO software Treestar Results Engagement of TLR3 and type I IFN signaling by candidate adjuvants of cold adapted influenza vaccines in mice increased vaccine induced cytokine produc tion early after mucosal administration including enhanced production of MCP1 IP 10 and G CSF This increase in vaccine induced early inflammatory responses resulted in robust cytotoxic T lymphocyte CTL activity rapid viral clearance and enhanced survival to influenza challenge Conversely vaccine adjuvants did not influence anti body titers in vaccinated individuals suggesting that the early events that take place during the innate phase of the immune response play a chief role on the establish ment and modulation of adaptive immunity Conclusion Our results indicated that specific adjuvants with the ability to enhance TLR3 and type I IFN signaling enhance cytokine responses associated to vaccine administration which resulted in improved CD8 T cell dependent recall responses Of note adjuvant vaccine regimens reduced the vaccine dose necessary to provide protection to lethal challenge These findings indicate that modulation of innate immunity via TLR ligands and type I IFN agonists may serve as a therapeutic strategy to enhance T cell memory maintenance after influenza vaccination and may provide means to reduce the amount of vaccine stock necessary to protect individuals at risk in the face of an influenza pandemic Disclosure of interest None declared http dx doi org 10 1016 j cyto 2012 06 205 P115 Pathogenic Influenza A viruses and SARS Coronaviruses modulate global inter feron stimulated gene induction through diverse mechanisms V D Menachery 1 A C Sims 1 A J Eisfeld 2 L E Gralinski 1 J Chang 3 K Waters 4 T O Metz 4 Y Kawaoka 2 M G Katze 3 R S Baric 1 1 Epidemiology University of North Carolina Chapel Hill NC United States 2 Influenza Research Institute University of Wisconsin Madison WI United States 3 Microbiology in contrast SARS CoV induces increased abundance of several ISG proteins but not until after peak viral titers have been achieved Together the data illustrate that these two virulent respiratory viruses control the global ISG response Further examination reveals ISG manipulation is absent in a less virulent IAV strain H1N1 2009 infection results in robust ISG transcript expression with minimal down regulation Similarly proteomics analysis finds increased abundance of ISG pro teins absent in H5N1 infection Transcriptional factor analysis found no direct antag onism however truncation of the C terminal portion of NS1 ablated ISG down regulation in H5N1 infections Meta analysis finds a similar ablation is observed in A549 cells infected H1N1 1918 when NS1 is manipulated Together the data strongly implicate a role for NS1 in mediating control of the entire global ISG response follow ing virulent IAV infections For SARS CoV the delayed ISG induction is maintained in both mutant and pre cursor viruses DORF6 a SARS CoV mutant lacking a major IFN antagonist fails to induce ISG induction any earlier Similarly Bat SRBD a synthetic precursor virus maintains the ability control the global ISG response suggesting this trait is main tained in pre cursors of the epidemic SARS CoV Additional work examines the ability of the SARS CoV to camouflage its viral intermediates as well as replication within double membrane vesicles as mechanisms mediating delayed ISG induction Conclusion Together the work highlights the contrasting approaches used by SARS CoV and IAV strains to limit global ISG induction In addition the results imply an important role for controlling ISG response in the context of virulent respiratory virus infection Further examination of these results will likely produce novel approaches to treat important respiratory virus infection Disclosure of interest None declared http dx doi org 10 1016 j cyto 2012 06 206 556 Abstract Cytokine 59 2012 553 556