In vivo induction of cytotoxic T lymphocytes specific for a single epitope introduced into an unrelated molecule.
Cytotoxic T lymphocytes (CTLs) recognise antigenic peptides in the context of major histocompatibility complex (MHC) class I molecules on virus-infected cells. The formation and transportation of antigenic peptides to class I MHC in the cells are multi-step reactions known as antigen processing. In order to design a good DNA vaccine, it is important to dissect the specificity of antigen processing. Here we describe the construction of an epitope-based plasmid vector as a device to investigate antigen processing in transfected cells. The epitope-based plasmid vector was constructed by insertion of an epitope-encoding minigene into the lacZ gene. We used a CTL epitope on influenza A virus nucleoprotein (NP366-374 epitope) as a model. Upon transfection, the epitope-based plasmid vector induced the expression of NP epitope antigenically as well as immunogenically. Immunization of mice with plasmid-transfected cells was able to induce NP epitope-specific CTLs in vivo. Moreover, the plasmid vector functioned as a gene vaccine; NP epitope-specific CTLs were primed in vivo upon transfection of the vector into dermis by electroporation. The results suggest that this epitope-based DNA delivery system may provide a new strategy for in vivo induction of epitope-specific CTLs to investigate antigen processing and presentation.
Nomura
M
M
Division of Clinical Research and Radiation Health, National Institute of Radiological Sciences, Chiba, Japan.
Nakata
Y
Y
Inoue
T
T
Uzawa
A
A
Itamura
S
S
Nerome
K
K
Akashi
M
M
Suzuki
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D018274:Electroporation; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors / Q000276:immunology*; D008213:Lymphocyte Activation / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection / Q000276:immunology; D014407:Tumor Cells, Cultured
D000939:Epitopes