title: Antigen delivery with poly(propylacrylic acid) conjugation enhances MHC-1 presentation and T-cell activation.
abstract: While many infectious diseases are controlled by vaccine strategies, important limitations continue to motivate the development of better antigen delivery systems. This study focuses on the use of a pH-sensitive polymeric carrier based on poly(propylacrylic acid) (PPAA) to address the need for more potent CD8 cytotoxic T-cell (CTL) responses. An MHC-1/CD8 CTL cell model system with ovalbumin as the protein antigen was used to test whether PPAA could enhance the delivery of ovalbumin into the MHC-1 display pathway. Ovalbumin was conjugated to poly(propylacrylic acid-co-pyridyldisulfide acrylate) (PPAA-PDSA) by disulfide exchange to make reversible conjugates that could be reduced by the glutathione redox system in the cytosol of antigen presenting cells. The PPAA-PDSA ovalbumin conjugates displayed the pH-sensitive membrane disruptive properties of the parent polymer as determined by their hemolysis activities (sharply active at the endosomal pH values of 6-6.5). The polymer-ovalbumin conjugates exhibited strong 22-fold increases in the MHC-1 presentation and ovalbumin-specific CTL activation compared to free ovalbumin. No CTL activation was observed with control conjugates of ovalbumin and poly(methylacrylic acid) (PMAA) that do not display membrane disruptive activies, suggesting that it is the membrane destabilizing properties of the polymer that result in increased MHC-1 display and CTL activation. Further mechanistic studies quantitated the time course of stable intracellular localization of radiolabeled conjugates. 52% of initially internalized PPAA-conjugated ovalbumin remained in the cells after 4 h, compared to less than 10% of ovalbumin or PMAA-ovalbumin. These results showing enhanced cytosolic delivery and MHC-1 presentation for the PPAA-antigen conjugates suggest that they warrant future characterization as a CD8-enhancing vaccine delivery system.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Flanary
authors.forename: Allan S
authors.initials: AS
authors.lastname: Hoffman
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Stayton
mesh_terms: D000179:Acrylates / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D000818:Animals; D017951:Antigen Presentation / Q000187:drug effects*; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D002250:Carbon Radioisotopes / Q000737:chemistry; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity; D004705:Endocytosis; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology; D005089:Exocytosis; D005978:Glutathione / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D008213:Lymphocyte Activation / Q000187:drug effects*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D014612:Vaccines / Q000276:immunology / Q000378:metabolism
chemical_list: D000179:Acrylates; D002250:Carbon Radioisotopes; D004220:Disulfides; D004337:Drug Carriers; D015395:Histocompatibility Antigens Class I; D011108:Polymers; D011948:Receptors, Antigen, T-Cell; D014612:Vaccines; C501628:poly(propyl acrylic acid); D010047:Ovalbumin; D005978:Glutathione