Delivery of a viral antigen to the class I processing and presentation pathway by Listeria monocytogenes.
Listeria monocytogenes is a facultative intracellular pathogen that grows in the cytoplasm of infected host cells. We examined the capacity of L. monocytogenes to introduce influenza nucleoprotein (NP) into the class I pathway of antigen presentation both in vitro and in vivo. Recombinant L. monocytogenes secreting a fusion of listeriolysin O and NP (LLO-NP) targeted infected cells for lysis by NP-specific class I-restricted cytotoxic T cells. Antigen presentation occurred in the context of three different class I haplotypes in vitro. A hemolysin-negative L. monocytogenes strain expressing LLO-NP was able to present in a class II-restricted manner. However, it failed to target infected cells for lysis by CD8+ T cells, indicating that hemolysin-dependent bacterial escape from the vacuole is necessary for class I presentation in vitro. Immunization of mice with a recombinant L. monocytogenes strain that stably expressed and secreted LLO-NP induced NP-specific CD8+ cytotoxic T lymphocytes. These studies have implications for the use of L. monocytogenes to deliver potentially any antigen to the class I pathway in vivo.
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia 19104-6076.
G
G
Ikonomidis
Y
Y
Paterson
F J
FJ
Kos
D A
DA
Portnoy
D000818:Animals; D000956:Antigens, Viral / Q000008:administration & dosage* / Q000276:immunology; D001427:Bacterial Toxins; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D017931:DNA Primers; D005822:Genetic Vectors; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000276:immunology; D006460:Hemolysin Proteins; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D008089:Listeria monocytogenes / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000096:biosynthesis / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D011995:Recombination, Genetic; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000956:Antigens, Viral; D001427:Bacterial Toxins; D017931:DNA Primers; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D009698:Nucleoproteins; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D014765:Viral Vaccines; C038705:hlyA protein, Listeria monocytogenes