We have generated a recombinant influenza A virus with the HIV-1 p17Gag(rFlu-p17) gene inserted into the influenza virus neuraminidase (NA) gene. Expression of HIV-1 p17 protein was detected by conventional Western blot analysis and also by liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis of rFlu-p17 infected cells. The latter method does not depend on protein-specific antibody preparations and proved to be a powerful tool to detect proteins of interest. Next, antigen presentation of p17 expressed after infection of antigen-presenting cells was determined. Cloned p17-specific CD8+ T-cells were co-cultured with rFlu-p17 infected B-cells and produced IFN-γ upon stimulation. Furthermore, we showed that immunization with rFlu-p17 elicited a humoral immune response in mice. This study shows that replication-deficient rFlu-p17 is antigenic in vitro and immunogenic in vivo and warrants further development as a candidate vaccine vector.

HIV-1, Influenza, Viral vector
dx.doi.org/10.1016/j.vaccine.2009.07.032, hdl.handle.net/1765/24524
Erasmus MC: University Medical Center Rotterdam

de Goede, A.L, Boers, P.H.M, Dekker, L.J.M, Osterhaus, A.D.M.E, Gruters, R.A, & Rimmelzwaan, G.F. (2009). Characterization of recombinant influenza A virus as a vector for HIV-1 p17Gag. Vaccine, 27(42), 5735–5739. doi:10.1016/j.vaccine.2009.07.032