title: Clustered epitopes within the Gag-Pol fusion protein DNA vaccine enhance immune responses and protection against challenge with recombinant vaccinia viruses expressing HIV-1 Gag and Pol antigens.
abstract: We have generated a codon-optimized hGagp17p24-Polp51 plasmid DNA expressing the human immunodeficiency virus type 1 (HIV-1) Gag-Pol fusion protein that consists of clusters of highly conserved cytotoxic T lymphocyte (CTL) epitopes presented by multiple MHC class I alleles. In the hGagp17p24-Polp51 construct, the ribosomal frameshift site had been deleted together with the potentially immunosuppressive Gag nucleocapsid (p15) as well as Pol protease (p10) and integrase (p31). Analyses of the magnitude and breadth of cellular responses demonstrated that immunization of HLA-A2/K(b) transgenic mice with the hGagp17p24-Polp51 construct induced 2- to 5-fold higher CD8+ T-cell responses to Gag p17-, p24-, and Pol reverse transcriptase (RT)-specific CTL epitopes than the full-length hGag-PolDeltaFsDeltaPr counterpart. The increases were correlated with higher protection against challenge with recombinant vaccinia viruses (rVVs) expressing gag and pol gene products. Consistent with the profile of Gag- and Pol-specific CD8+ T cell responses, an elevated level of type 1 cytokine production was noted in p24- and RT-stimulated splenocyte cultures established from hGagp17p24-Polp51-immunized mice compared to responses induced with the hGag-PolDeltaFsDeltaPr vaccine. Sera of mice immunized with the hGagp17p24-Polp51 vaccine also exhibited an increased titer of p24- and RT-specific IgG2 antibody responses. The results from our studies provide insights into approaches for boosting the breadth of Gag- and Pol-specific immune responses.
authors.affiliation: Department of Immunology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Bolesta
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Gzyl
authors.forename: Andrzej
authors.initials: A
authors.lastname: Wierzbicki
authors.forename: Dariusz
authors.initials: D
authors.lastname: Kmieciak
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Kowalczyk
authors.forename: Yutaro
authors.initials: Y
authors.lastname: Kaneko
authors.forename: Alagarsamy
authors.initials: A
authors.lastname: Srinivasan
authors.forename: Danuta
authors.initials: D
authors.lastname: Kozbor
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D017124:Conserved Sequence; D016207:Cytokines / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D017931:DNA Primers; D000939:Epitopes / Q000032:analysis*; D015750:Genes, gag; D015751:Genes, pol; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D017232:Oncogene Proteins v-mos / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D019444:Vaccines, DNA; D014616:Vaccinia virus / Q000276:immunology
chemical_list: D016207:Cytokines; D017931:DNA Primers; D000939:Epitopes; D015789:HLA-A2 Antigen; D007074:Immunoglobulin G; D017232:Oncogene Proteins v-mos; D019444:Vaccines, DNA