Introduction of protein or DNA delivered via recombinant Salmonella typhimurium into the major histocompatibility complex class I presentation pathway of macrophages.
Recombinant (r) Salmonella typhimurium aroA strains which display the hen egg ovalbumin OVA(257-264) peptide SIINFEKL in secreted form were constructed. In addition, attenuated rS. typhimurium pcDNA-OVA constructs harbouring a eukaryotic expression plasmid encoding complete OVA were used to introduce the immunodominant OVA(257-264) epitope into the major histocompatibility complex (MHC) class I presentation pathway. Both modes of antigen delivery (DNA and protein) by Salmonella vaccine carriers stimulated OVA(257-264)-specific CD8 T-cell hybridomas. An in vitro infection system was established that allowed both rSalmonella carrier devices to facilitate MHC class I delivery of OVA(257-264) by coexpression of listeriolysin (Hly) or by coinfection with rS. typhimurium Hlys (Hess J., Gentschev I., Miko D., Welzel M., Ladel C., Goebel W., Kaufmann S.H.E., Proc. Natl. Acad. Sci. USA 93 (1996) 1458-1463). Coexpression of Hly and coinfection with rS. typhimurium Hlys slightly improved MHC class I processing of OVA. Our data provide further evidence for the feasibility of attenuated, Hly-expressing rS. typhimurium carriers secreting heterologous antigens or harbouring heterologous DNA as effective vaccines for stimulating CD8 T cells in addition to CD4 T cells.
Catic
A
A
Department of Immunology, University Clinics Ulm, D-89070 Ulm, Germany.
Dietrich
G
G
Gentschev
I
I
Goebel
W
W
Kaufmann
S H
SH
Hess
J
J
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D001427:Bacterial Toxins; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005805:Genes, MHC Class I / Q000276:immunology; D005822:Genetic Vectors; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006460:Hemolysin Proteins; D008264:Macrophages / Q000276:immunology* / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012486:Salmonella typhimurium / Q000235:genetics*; D014169:Transformation, Bacterial; D019444:Vaccines, DNA
D001427:Bacterial Toxins; D000939:Epitopes; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D010047:Ovalbumin; C038705:hlyA protein, Listeria monocytogenes