There is a need for alternative or adjunct therapies for influenza, because resistance to currently used antiviral drugs can emerge rapidly. In collaboration with CEL-SCI Corporation, we tested a ligand epitope antigen presentation system (LEAPS) technology as a new immune-based treatment for influenza virus infection in a mouse model. Influenza-J-LEAPS peptides were synthesized by conjugating the binding ligand derived from the β2 -microglobulin chain of the human MHC class I molecule (J-LEAPS) with 15 to 30 amino acidlong peptides derived from influenza virus NP, M, or HA proteins. DCs were stimulated with influenza-J-LEAPS peptides (influenza-J-LEAPS) and injected intravenously into infected mice. Antigen-specific LEAPS stimulated DCs were effective in reducing influenza virus replication in the lungs and enhancing survival of infected animals. Additionally, they augmented influenza-specific T cell responses in the lungs and reduced the severity of disease by limiting excessive cytokine responses, which are known to contribute to morbidity and mortality following influenza virus infection. Our data demonstrated that influenza-J-LEAPS pulsed DCs reduce virus replication in the lungs, enhance survival, and modulate the protective immune responses that eliminate the virus while preventing excessive cytokines that could injure the host. This approach shows promise as an adjunct to antiviral treatment of influenza virus infections. Rapid antigenic variation of HA, the major virion surface protein of influenza A virus, remains the principal challenge to the development of broader and more effective vaccines. Some regions of HA, such as the stem region proximal to the viral membrane, are nevertheless highly conserved across strains and among most subtypes. A fundamental question in vaccine design is the extent to which HA stem regions on the surface of the virus are accessible to broadly neutralizing antibodies. In collaboration with Sriram Subramaniams lab from NCI, we reported 3D structures derived from cryoelectron tomography of HA on intact 2009 pandemic H1N1 virions in the presence and absence of the antibody C179, which neutralizes viruses expressing a broad range of HA subtypes, including H1, H2, H5, H6, and H9. By fitting previously derived crystallographic structures of trimeric HA into the density maps, we deduced the locations of the molecular surfaces of HA involved in interaction with C179. Using computational methods to distinguish individual unliganded HA trimers from those that have bound C179 antibody, we demonstrated that ∼75% of HA trimers on the surface of the virus have C179 bound to the stem domain. Thus, despite their close packing on the viral membrane, the majority of HA trimers on intact virions are available to bind anti-stem antibodies that target conserved HA epitopes, establishing the feasibility of universal influenza vaccines that elicit such antibodies.

Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2013
Total Cost
$1,521,186
Indirect Cost
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State
Country
Zip Code
Lakdawala, Seema S; Wu, Yicong; Wawrzusin, Peter et al. (2014) Influenza a virus assembly intermediates fuse in the cytoplasm. PLoS Pathog 10:e1003971
O'Donnell, Christopher D; Wright, Amber; Vogel, Leatrice et al. (2014) Humans and ferrets with prior H1N1 influenza virus infections do not exhibit evidence of original antigenic sin after infection or vaccination with the 2009 pandemic H1N1 influenza virus. Clin Vaccine Immunol 21:737-46
Boonnak, Kobporn; Vogel, Leatrice; Orandle, Marlene et al. (2013) Antigen-activated dendritic cells ameliorate influenza A infections. J Clin Invest 123:2850-61
Harris, Audray K; Meyerson, Joel R; Matsuoka, Yumiko et al. (2013) Structure and accessibility of HA trimers on intact 2009 H1N1 pandemic influenza virus to stem region-specific neutralizing antibodies. Proc Natl Acad Sci U S A 110:4592-7