title: Induction in vitro of primary cytotoxic T-lymphocyte responses with DNA encoding herpes simplex virus proteins.
abstract: Vaccines which successfully protect against virus infections usually need to induce a broadly reactive immune response which includes the induction of cytotoxic T lymphocytes (CTL). In this study, we have used a convenient in vitro approach to investigate if plasmid DNAs encoding proteins of herpes simplex virus (HSV) are capable of inducing primary CD8+ CTL. Dendritic cells or macrophages were transfected with either plasmid DNA encoding glycoprotein B or DNA encoding the immediate-early protein ICP27. These antigen-presenting cells (APC) were then used to stimulate enriched populations of naive T cells in microcultures for 5 days in vitro. Antigen-specific CD8+ CTL which reacted both with specific protein-expressing targets and with syngeneic targets infected with HSV could be demonstrated. Dendritic cells, as APC, generated the maximal responses, but such cells needed to be transfected with DNA in the presence of a cationic lipid. However, macrophages could act as APC when they were exposed to purified DNA. HSV-primed splenocytes were also shown to generate specific CTL responses when they were stimulated with purified DNA encoding ICP27. The novel approach described in this paper promises to be extremely useful, since defining immunogenicity profiles and identifying epitopes on viral proteins should be easier and more convenient when working with DNA and investigating variables in vitro. This is particularly the case with complex viruses such as HSV, most of whose encoded proteins have yet to be isolated in sufficient quantity or purity to perform in vivo immunological studies.
authors.affiliation: Department of Microbiology, College of Veterinary Medicine, University of Tennessee, Knoxville 37996-0845.
authors.forename: R J
authors.initials: RJ
authors.lastname: Rouse
authors.forename: S K
authors.initials: SK
authors.lastname: Nair
authors.forename: S L
authors.initials: SL
authors.lastname: Lydy
authors.forename: J C
authors.initials: JC
authors.lastname: Bowen
authors.forename: B T
authors.initials: BT
authors.lastname: Rouse
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D016827:CD8 Antigens / Q000032:analysis; D003602:Cytotoxicity, Immunologic; D004279:DNA, Viral / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology; D005260:Female; D018259:Herpesvirus 1, Human / Q000276:immunology*; D007111:Immunity, Cellular; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D014614:Vaccines, Synthetic; D014765:Viral Vaccines / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D016827:CD8 Antigens; D004279:DNA, Viral; D014614:Vaccines, Synthetic; D014765:Viral Vaccines