title: Archaeosomes induce long-term CD8+ cytotoxic T cell response to entrapped soluble protein by the exogenous cytosolic pathway, in the absence of CD4+ T cell help.
abstract: The unique ether glycerolipids of Archaea can be formulated into vesicles (archaeosomes) with strong adjuvant activity for MHC class II presentation. Herein, we assess the ability of archaeosomes to facilitate MHC class I presentation of entrapped protein Ag. Immunization of mice with OVA entrapped in archaeosomes resulted in a potent Ag-specific CD8(+) T cell response, as measured by IFN-gamma production and cytolytic activity toward the immunodominant CTL epitope OVA(257-264). In contrast, administration of OVA with aluminum hydroxide or entrapped in conventional ester-phospholipid liposomes failed to evoke significant CTL response. The archaeosome-mediated CD8(+) T cell response was primarily perforin dependent because CTL activity was undetectable in perforin-deficient mice. Interestingly, a long-term CTL response was generated with a low Ag dose even in CD4(+) T cell deficient mice, indicating that the archaeosomes could mediate a potent T helper cell-independent CD8(+) T cell response. Macrophages incubated in vitro with OVA archaeosomes strongly stimulated cytokine production by OVA-specific CD8(+) T cells, indicating that archaeosomes efficiently delivered entrapped protein for MHC class I presentation. This processing of Ag was Brefeldin A sensitive, suggesting that the peptides were transported through the endoplasmic reticulum and presented by the cytosolic MHC class I pathway. Finally, archaeosomes induced a potent memory CTL response to OVA even 154 days after immunization. This correlated to strong Ag-specific up-regulation of CD44 on splenic CD8(+) T cells. Thus, delivery of proteins in self-adjuvanting archaeosomes represents a novel strategy for targeting exogenous Ags to the MHC class I pathway for induction of CTL response.
authors.affiliation: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada. lakshmi.krishnan@nrc.ca
authors.forename: L
authors.initials: L
authors.lastname: Krishnan
authors.forename: S
authors.initials: S
authors.lastname: Sad
authors.forename: G B
authors.initials: GB
authors.lastname: Patel
authors.forename: G D
authors.initials: GD
authors.lastname: Sprott
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage*; D000818:Animals; D017951:Antigen Presentation; D001105:Archaea / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D003600:Cytosol / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D004527:Egg Proteins / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005995:Glyceryl Ethers / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000096:biosynthesis; D007156:Immunologic Memory; D007274:Injections, Intraperitoneal; D008081:Liposomes / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D008562:Membrane Glycoproteins / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000276:immunology* / Q000378:metabolism; D010446:Peptide Fragments; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012995:Solubility; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D004527:Egg Proteins; D005995:Glyceryl Ethers; D015395:Histocompatibility Antigens Class I; D018960:Hyaluronan Receptors; D008081:Liposomes; D008562:Membrane Glycoproteins; C110366:OVA-8; D010446:Peptide Fragments; D052899:Pore Forming Cytotoxic Proteins; C094417:archaeol lipid; D054353:Perforin; D010047:Ovalbumin