GM-CSF Increases Mucosal and Systemic Immunogenicity of an H1N1 Influenza DNA Vaccine Administered into the Epidermis of Non-Human Primates

Background: The recent H5N1 avian and H1N1 swine-origin influenza virus outbreaks reaffirm that the threat of a worldwide influenza pandemic is both real and ever-present. Vaccination is still considered the best strategy for protection against influenza virus infection but a significant challenge is to identify new vaccine approaches that offer accelerated production, broader protection against drifted and shifted strains, and the capacity to elicit anti-viral immune responses in the respiratory tract at the site of viral entry. As a safe alternative to live attenuated vaccines, the mucosal and systemic immunogenicity of an H1N1 influenza (A/New Caledonia/20/99) HA DNA vaccine administered by particle-mediated epidermal delivery (PMED or gene gun) was analyzed in rhesus macaques. Methodology/Principal Findings: Macaques were immunized at weeks 0, 8, and 16 using a disposable single-shot particlemediated delivery device designed for clinical use that delivers plasmid DNA directly into cells of the epidermis. Significant levels of hemagglutination inhibiting (HI) antibodies and cytokine-secreting HA-specific T cells were observed in the periphery of macaques following 1-3 doses of the PMED HA DNA vaccine. In addition, HA DNA vaccination induced detectable levels of HA-specific mucosal antibodies and T cells in the lung and gut-associated lymphoid tissues of vaccinated macaques. Importantly, co-delivery of a DNA encoding the rhesus macaque GM-CSF gene was found to significantly enhance both the systemic and mucosal immunogenicity of the HA DNA vaccine. Conclusions/Significance: These results provide strong support for the development of a particle-mediated epidermal DNA vaccine for protection against respiratory pathogens such as influenza and demonstrate, for the first time, the ability of skindelivered GM-CSF to serve as an effective mucosal adjuvant for vaccine induction of immune responses in the gut and respiratory tract. © 2010 Loudon et al

Evasion of influenza A viruses from innate and adaptive immune responses

The influenza A virus is one of the leading causes of respiratory tract infections in humans. Upon infection with an influenza A virus, both innate and adaptive immune responses are induced. Here we discuss various strategies used by influenza A viruses to evade innate immune responses and recognition by components of the humoral and cellular immune response, which consequently may result in reduced clearing of the virus and virus-infected cells. Finally, we discuss how the current knowledge about immune evasion can be used to improve influenza A vaccination strategies

Antigen processing and MHC I-restricted presentation of influenza PA polymerase.

This laboratory has developed and is currently studying the cold-attenuated (ca) live influenza vaccine. Little is known of the immunogenicity of PA; however, PA, and only this polymerase, is recognized by MHC I-restricted T cells from mice with the H-2\sp{\rm b} haplotype. Furthermore, given that the cytotoxic T lymphocyte (CTL) response to the viral internal proteins is more important than that to the antigenically variable glycoproteins, and is crossreactive among influenza virus subtypes, then CTL recognition of PA is also likely to be directed against a conserved antigenic site(s). Therefore, study of CTL recognition of the ca PA may help to determine the antigen processing required for this functionally important protein to become immunogenic. The intracellular trafficking of PA, and its interaction with MHC I, was examined in cells which express complete MHC I molecules, and was compared to the behavior in cell variants unable to assemble MHC I. Effects of inhibitors of MHC I-restricted antigenic processing were revealed by immunofluorescence and immunoprecipitation. Immunological localization within cells was determined with monospecific sera to PA, antibodies to free and/or assembled MHC I, and antibodies to endosome/lysosome, Golgi and endoplasmic reticulum markers, in combination with subcellular fractionation. Direct examination of the binding of cell-associated MHC I with antigenic peptides, and their transport to the cell surface, was investigated using synthetic peptides of PA which contain putative CTL epitopes and MHC I-binding motifs. The results indicate that ca PA is antigenically processed and presented by a MHC I-restricted mechanism.Ph.D.Epidemiologic ScienceUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/103612/1/9332094.pdfDescription of 9332094.pdf : Restricted to UM users only

Mikyoung Kim: Monograph / Mikyoung Kim and Raphael Justewicz

Poster for a lecture and booksigning by Mikyoung Kim in the Bayard Ewing Building. Quotes from John Cage and John Beardsley printed onto poster.https://digitalcommons.risd.edu/risdposters/1309/thumbnail.jp