DukeSpace will be down for maintenance at 9:15 AM EDT on Tuesday, July 29. Expected downtime is 20 minutes or less.

Show simple item record

dc.contributor.advisor Pizzo, Salvatore Vincent
dc.contributor.advisor Cianciolo, George J.
dc.contributor.advisor Abraham, Soman, N.
dc.contributor.advisor Staats, Herman F.
dc.contributor.advisor Dawson, Jeffrey R.
dc.contributor.author Anderson, Ryan Berger
dc.date 2007
dc.date.accessioned 2007-05-02T14:53:43Z
dc.date.available 2007-05-02T14:53:43Z
dc.date.issued 2007-05-02T14:53:43Z
dc.identifier.uri http://hdl.handle.net/10161/172
dc.description Dissertation
dc.description.abstract Bacterial DNA is immunostimulatory, and the motifs responsible for this activity are unmethylated CpG dinucleotides. Following cellular uptake, CpG-containing oligodeoxynucleotides (CpG ODN) are trafficked to the endosome where they bind Toll-like receptor 9 (TLR9) to initiate a signaling cascade that culminates in the release of numerous pro-inflammatory cytokines. Because of their immunostimulatory properties, CpG ODN are being clinically evaluated as treatments and vaccine adjuvants for infectious diseases, cancer, and allergic disorders. α2-Macroglobulin (α2M) is a human plasma protein that binds and modulates the activity of a variety of cytokines, growth factors, enzymes, and antigens. Upon proteolytic activation, α2M is converted to its receptor recognized form, α2M*, and rapidly binds to and is internalized by immune competent cells expressing the α2M* endocytic receptor, LRP, and is then trafficked to the endosome. Based on these interactions, α2M seems to play an important role at sites of infection and inflammation by controlling the level of proteinase activity, modulating cytokine signals, and enhancing antigen processing for the adaptive immune response. Here, we report the first evidence that α2M* binds and forms stable complexes with nucleic acids. We have characterized the mechanisms and stoichiometry of this interaction, examined the pH and temperature stability of these complexes, and identified structural variables in the nucleic acids, namely length, base composition, and chemical modifications, that affect the nature of this interaction. We hypothesized that CpG ODN incorporation into α2M* may alter their immunostimulatory properties. Murine macrophages (MΦs) treated with α2M*-ODN complexes respond more rapidly and produce a greater cytokine response than those treated with free CpG ODN alone. Treating human PBMCs with α2M*-ODN complexes likewise demonstrated their enhanced ability to elicit immune responses. This was due to more rapid uptake and CpG ODN protection from degradation by extracellular nucleases. Co-incorporation of both protein ligands and CpG ODN into α2M* yields ternary complexes; these may permit the simultaneous delivery of both protein antigens and adjuvants to immune competent cells, potentially greatly enhancing the adaptive immune response and protective immunity. Based on the findings that incorporation into α2M* confers enhanced immunostimulatory activity of CpG ODN, this technology may be exploited to improve CpG ODN-based therapeutics by increasing efficacy, minimizing side effects, reducing dosing requirements, and reducing cost. en
dc.format.extent 1502502 bytes
dc.format.mimetype application/pdf
dc.language.iso en_US en
dc.subject CpG-containing oligodeoxynucleotides (CpG ODN) en
dc.subject TLR9 en
dc.subject LRP en
dc.subject dendritic cells en
dc.title Incorporation of CpG Oligodeoxynucleotides into α2-Macroglobulin: Development of a Novel Vaccine Adjuvant Delivery Mechanism en
dc.type Dissertation en
dc.department Pathology

Files in this item

This item appears in the following Collection(s)

Show simple item record