Characterization of recombinant influenza A virus as a vector for HIV-1 p17(Gag)

Anna Goede; Patrick Boers; Lennard Dekker; Ab Osterhaus; Rob Gruters; Guus Rimmelzwaan

doi:10.1016/j.vaccine.2009.07.032

Characterization of recombinant influenza A virus as a vector for HIV-1 p17(Gag)

Anna Goede, Patrick Boers, Lennard Dekker, Ab Osterhaus, Rob Gruters, Guus Rimmelzwaan

Research output: Contribution to journal › Article › Academic › peer-review

14 Citations (Scopus)

Abstract

We have generated a recombinant influenza A virus with the HIV-1 p17(Gag) (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-gamma 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. (C) 2009 Elsevier Ltd. All rights reserved.

Original language	Undefined/Unknown
Pages (from-to)	5735-5739
Number of pages	5
Journal	Vaccine
Volume	27
Issue number	42
DOIs	https://doi.org/10.1016/j.vaccine.2009.07.032
Publication status	Published - 2009

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EMC MM-03-44-06
EMC MM-04-27-01
EMC OR-01-34-01

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Access to Document

10.1016/j.vaccine.2009.07.032

http://hdl.handle.net/1765/24524

Cite this

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title = "Characterization of recombinant influenza A virus as a vector for HIV-1 p17(Gag)",

abstract = "We have generated a recombinant influenza A virus with the HIV-1 p17(Gag) (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-gamma 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. (C) 2009 Elsevier Ltd. All rights reserved.",

author = "Anna Goede and Patrick Boers and Lennard Dekker and Ab Osterhaus and Rob Gruters and Guus Rimmelzwaan",

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T1 - Characterization of recombinant influenza A virus as a vector for HIV-1 p17(Gag)

AU - Goede, Anna

AU - Boers, Patrick

AU - Dekker, Lennard

AU - Osterhaus, Ab

AU - Gruters, Rob

AU - Rimmelzwaan, Guus

PY - 2009

Y1 - 2009

N2 - We have generated a recombinant influenza A virus with the HIV-1 p17(Gag) (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-gamma 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. (C) 2009 Elsevier Ltd. All rights reserved.

AB - We have generated a recombinant influenza A virus with the HIV-1 p17(Gag) (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-gamma 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. (C) 2009 Elsevier Ltd. All rights reserved.

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