title: Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef.
abstract: Transfection with synthetic mRNA is a safe and efficient method of delivering antigens to dendritic cells for immunotherapy. Targeting antigens to the lysosome can sometimes enhance the CD4+ T-cell response. We transfected antigen-presenting cells (APCs) with mRNA encoding Gag-p24 and cytoplasmic, lysosomal, and secreted forms of Nef. Antigen-specific cytotoxic T cells were able to lyse the majority of transfected targets, indicating that transfection was efficient. Transfection of APCs with a Nef construct bearing lysosomal targeting signals produced rapid and prolonged antigen presentation to CD4+ and CD8+ T cells. Polyclonal CD4+ and CD8+ T-cell lines recognizing multiple distinct epitopes were expanded by coculture of transfected dendritic cells with peripheral blood mononuclear cells from viremic and aviremic HIV-infected subjects. Importantly, lysosome-targeted antigen drove a significantly greater expansion of Nef-specific CD4+ T cells than cytoplasmic antigen. The frequency of recognition of CD8 but not CD4 epitopes by mRNA-expanded T cells was inversely proportional to sequence entropy and was similar to ex vivo responses from a large chronic cohort. Thus human dendritic cells transfected with mRNA encoding lysosome-targeted HIV antigen can expand a broad, polyclonal repertoire of antiviral T cells, offering a promising approach to HIV immunotherapy.
authors.affiliation: Partners AIDS Research Center, Massachusetts General Hospital and Harvard Medical School, 149 13 St, Charlestown, MA 02129, USA. dkavanagh@partners.org
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Kavanagh
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Kaufmann
authors.forename: Sherzana
authors.initials: S
authors.lastname: Sunderji
authors.forename: Nicole
authors.initials: N
authors.lastname: Frahm
authors.forename: Sylvie
authors.initials: S
authors.lastname: Le Gall
authors.forename: David
authors.initials: D
authors.lastname: Boczkowski
authors.forename: Eric S
authors.initials: ES
authors.lastname: Rosenberg
authors.forename: David R
authors.initials: DR
authors.lastname: Stone
authors.forename: Mary N
authors.initials: MN
authors.lastname: Johnston
authors.forename: Bradford S
authors.initials: BS
authors.lastname: Wagner
authors.forename: Mohammad T
authors.initials: MT
authors.lastname: Zaman
authors.forename: Christian
authors.initials: C
authors.lastname: Brander
authors.forename: Eli
authors.initials: E
authors.lastname: Gilboa
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Walker
authors.forename: Nina
authors.initials: N
authors.lastname: Bhardwaj
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003593:Cytoplasm / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D015693:Gene Products, nef / Q000235:genetics / Q000276:immunology*; D006678:HIV / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D008247:Lysosomes / Q000276:immunology; D021381:Protein Transport / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology; D014162:Transfection / Q000379:methods; D054311:nef Gene Products, Human Immunodeficiency Virus
chemical_list: D018984:Epitopes, T-Lymphocyte; D015693:Gene Products, nef; D012333:RNA, Messenger; D054311:nef Gene Products, Human Immunodeficiency Virus