Presentation by a major histocompatibility complex class I molecule of nucleoprotein peptide expressed in two different genes of an influenza virus transfectant.
Major histocompatibility (MHC) class I glycoproteins are specialized to present to CD8+ T cells, peptides that originate from proteins synthesized within the cytoplasm. Conventional killed vaccines are unable to get into the cell cytoplasm and therefore fail to expand the CD8+ T cell population. We have created a novel influenza transfectant virus, R10, which carries an immunogenic peptide from the nucleoprotein (NP) of PR8 influenza virus in its hemagglutinin (HA) and another similar peptide in its HK influenza virus NP. The two peptides are both presented by H-2Db and bind with approximately equal affinity. They can compete with one another for binding to H-2Db. Yet in cells infected with R10, both peptides are presented efficiently enough to expand the respective cytotoxic T lymphocyte (CTL) precursors in vivo and to serve as targets for CTL lysis in vitro. It has been proposed that proteins bearing signal sequences may be processed by a transporter-independent pathway. To investigate this, we infected the transporter-deficient cell line RMA-S with the R10 virus to see if the NP peptide expressed by the HA would be presented. The result shows that even the presence of a signal peptide in the HA does not overcome the lack of a transporter function, suggesting that the presentation of both peptides is dependent on functional transporter proteins. Our data also suggest the feasibility of creating by genetic engineering, recombinant vaccines expressing multiple epitopes that can effectively stimulate a cellular immune response.
Isobe
H
H
Department of Microbiology, Mount Sinai School of Medicine, New York 10029-6574.
Moran
T
T
Li
S
S
Young
A
A
Nathenson
S
S
Palese
P
P
Bona
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000956:Antigens, Viral / Q000737:chemistry*; D001667:Binding, Competitive; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D000939:Epitopes; D006183:H-2 Antigens / Q000276:immunology*; D006389:Hemagglutinins, Viral / Q000276:immunology; D007111:Immunity, Cellular; D009980:Influenza A virus / Q000276:immunology*; D007252:Influenza Vaccines / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014758:Viral Core Proteins / Q000276:immunology*
D000956:Antigens, Viral; D000939:Epitopes; D006183:H-2 Antigens; D006389:Hemagglutinins, Viral; D007252:Influenza Vaccines; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D010455:Peptides; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014758:Viral Core Proteins