title: Recombinant influenza virus expressing HIV-1 p24 capsid protein induces mucosal HIV-specific CD8 T-cell responses.
abstract: Influenza viruses are promising mucosal vaccine vectors for HIV but their use has been limited by difficulties in engineering the expression of large amounts of foreign protein. We developed recombinant influenza viruses incorporating the HIV-1 p24 gag capsid into the NS-segment of PR8 (H1N1) and X31 (H3N2) influenza viruses with the use of multiple 2A ribosomal skip sequences. Despite the insertion of a sizable HIV-1 gene into the influenza genome, recombinant viruses were readily rescued to high titers. Intracellular expression of p24 capsid was confirmed by in vitro infection assays. The recombinant influenza viruses were subsequently tested as mucosal vaccines in BALB/c mice. Recombinant viruses were attenuated and safe in immunized mice. Systemic and mucosal HIV-specific CD8 T-cell responses were elicited in mice that were immunized via intranasal route with a prime-boost regimen. Isolated HIV-specific CD8 T-cells displayed polyfunctional cytokine and degranulation profiles. Mice boosted via intravaginal route induced recall responses from the distal lung mucosa and developed heightened HIV-specific CD8 T-cell responses in the vaginal mucosa. These findings demonstrate the potential utility of recombinant influenza viruses as vaccines for mucosal immunity against HIV-1 infection.
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Hyon-Xhi
authors.initials: HX
authors.lastname: Tan
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Brad P
authors.initials: BP
authors.lastname: Gilbertson
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia; Laboratory of Infectious Diseases, NIAID, NIH, Bethesda, MD 20892, United States.
authors.forename: Sinthujan
authors.initials: S
authors.lastname: Jegaskanda
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Sheilajen
authors.initials: S
authors.lastname: Alcantara
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Thakshila
authors.initials: T
authors.lastname: Amarasena
authors.affiliation: School of Medicine, Deakin University, Geelong, Victoria, Australia; CSIRO Animal Health Laboratories, Geelong, Victoria, Australia.
authors.forename: John
authors.initials: J
authors.lastname: Stambas
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Julie L
authors.initials: JL
authors.lastname: McAuley
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia; Melbourne Sexual Health Centre, Alfred Hospital, Monash University Central Clinical School, Victoria, Australia; ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, The University of Melbourne, Victoria, Australia. Electronic address: skent@unimelb.edu.au.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Kent
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Victoria 3010, Australia.
authors.forename: Robert
authors.initials: R
authors.lastname: De Rose
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D005260:Female; D005818:Genetic Engineering; D016655:HIV Core Protein p24 / Q000276:immunology*; D015497:HIV-1; D006801:Humans; D018928:Immunity, Mucosal; D007117:Immunization, Secondary; D053118:Influenza A Virus, H1N1 Subtype; D053122:Influenza A Virus, H3N2 Subtype; D008168:Lung / Q000276:immunology; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000276:immunology; D014613:Vaccines, Attenuated / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014621:Vagina / Q000276:immunology
chemical_list: D016915:AIDS Vaccines; D016655:HIV Core Protein p24; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; C514799:p24 protein, Human Immunodeficiency Virus Type 1
keywords: CD8 T-cell; HIV; Influenza; Mucosal immunity; Prime-boost; Vaccine