Involvement of an ATP-dependent peptide chaperone in cross-presentation after DNA immunization.
Immunization with plasmid DNA holds promise as a vaccination strategy perhaps useful in situations that currently lack vaccines, since the major means of immune induction may differ from more conventional approach. In the present study, we demonstrate that exposure of macrophages to plasmid DNA encoding viral proteins or OVA generates Ag-specific material that, when presented in vitro by dendritic cells to naive T cells, induces primary CTL response or elicits IL-2 production from an OVA peptide-specific T-T hybridoma. The immunogenic material released was proteinaceous in nature, free of apoptotic bodies, and had an apparent m.w. much larger than a 9-11-aa CTL-recognizable peptide. The macrophage-released factor(s) specifically required a hydrolyzable ATP substrate and was inhibited by procedures that removed or hydrolyzed ATP; in addition, anti-heat-shock protein 70 antiserum abrogated the activity to a large extent. These results indicate the possible involvement of a heat-shock protein 70-linked peptide chaperone in a cross-priming method of immune induction by DNA vaccination. Such a cross-priming process may represent a principal mechanism by which plasmid DNA delivered to cells such as myocytes effectively shuttle Ag to DC or other APC to achieve CTL induction in vivo.
Kumaraguru
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Department of Microbiology, University of Tennessee, Knoxville, TN 37996, USA.
Rouse
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RJ
Nair
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Bruce
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BD
Rouse
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D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D017951:Antigen Presentation / Q000235:genetics; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005260:Female; D008213:Lymphocyte Activation / Q000235:genetics; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018832:Molecular Chaperones / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D009842:Oligopeptides / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D010047:Ovalbumin / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments; D010957:Plasmids / Q000138:chemical synthesis / Q000276:immunology; D014162:Transfection / Q000276:immunology; D014407:Tumor Cells, Cultured; D019444:Vaccines, DNA / Q000138:chemical synthesis / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D004527:Egg Proteins; D018832:Molecular Chaperones; C110366:OVA-8; D009842:Oligopeptides; D010446:Peptide Fragments; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; D000255:Adenosine Triphosphate; D010047:Ovalbumin