{"id":{"repo_id":"wustl","oai_identifier":"oai:openscholarship.wustl.edu:etd-1644"},"canonical_url":"https://search.dev.ndltd.org/etd/wustl/oai:openscholarship.wustl.edu:etd-1644","repository":{"repo_id":"wustl","name":"Washington University in St. Louis","base_url":"https://openscholarship.wustl.edu/do/oai/"},"display":{"title":"Type B Presentation from HEL Protein in the Context of Inflammation","abstract":"Peptides from the model antigen hen egg white lysozyme: HEL) are capable of binding MHC II in multiple conformations. Resting antigen presenting cells: APC) load peptide in early endosomal compartments where multiple conformations of a given pMHC pair are allowed, generating both type A and B pMHC complexes. Protein traffics to the late endosomal compartment and is loaded onto nascent MHC II in the presence of the editor H2-DM where type A pMHC conformers are selectively formed, generating exclusively type A pMHC from protein. Previous work demonstrated that type B pMHC are also generated when HEL is administered in vivo along with certain inflammatory stimulants. Using an in vitro system I established, I found that TLR ligands and type I interferons: IFN) act directly on dendritic cells: DC), allowing generation of type B pMHC from HEL. Both CD8alpha+ and CD8alpha- DC present type B pMHC with TLR stimulation, but the relative effectiveness varied based on the ligand used. Using a type I IFN receptor: IFNAR1) blocking antibody and DC from mice deficient in the receptor, I found that TLR-induced type I IFN production amplifies but is not required for TLR-induced type B presentation from HEL. While I have not determined a sub-cellular mechanism for TLR-induced type B presentation, I have excluded mechanisms and made several observations. DC deficient in H2-DM are capable of TLR-induced type B presentation, indicating that regulation of H2-DM is not a critical mechanism. DC do not release meaningful levels of peptides allowing generation of type A or B pMHC. Additionally, generation of type B pMHC from HEL is a delayed event, requiring eighteen hours for appearance at the cell surface. While surface MHC II levels only modestly increase, there is a significant increase in total pMHC complexes after TLR or type I IFN stimulation. I employed two-photon microcopy to observe type A and B T cells in intact lymph nodes. While there were no remarkable differences in T cell behavior between the two groups, I determined that DC present type A or both type A and B but never exclusively type B pMHC. Using mice that express HEL as a membrane-bound protein on APC: mHEL), I found that TLR stimulation increases type B presentation from membrane-bound protein. I found that while immunization of mHEL mice with CpG increases presentation of type B pMHC from pseudo-self HEL and primed anti-HEL CD4 T cells, no observable disease was detected. These results identified the cells and signals required for inflammation-associated type B presentation from HEL and established tools for future investigation of the sub-cellular mechanism controlling these events.","abstract_html":"Peptides from the model antigen hen egg white lysozyme: HEL) are capable of binding MHC II in multiple conformations. Resting antigen presenting cells: APC) load peptide in early endosomal compartments where multiple conformations of a given pMHC pair are allowed, generating both type A and B pMHC complexes. Protein traffics to the late endosomal compartment and is loaded onto nascent MHC II in the presence of the editor H2-DM where type A pMHC conformers are selectively formed, generating exclusively type A pMHC from protein. Previous work demonstrated that type B pMHC are also generated when HEL is administered in vivo along with certain inflammatory stimulants. Using an in vitro system I established, I found that TLR ligands and type I interferons: IFN) act directly on dendritic cells: DC), allowing generation of type B pMHC from HEL. Both CD8alpha+ and CD8alpha- DC present type B pMHC with TLR stimulation, but the relative effectiveness varied based on the ligand used. Using a type I IFN receptor: IFNAR1) blocking antibody and DC from mice deficient in the receptor, I found that TLR-induced type I IFN production amplifies but is not required for TLR-induced type B presentation from HEL. While I have not determined a sub-cellular mechanism for TLR-induced type B presentation, I have excluded mechanisms and made several observations. DC deficient in H2-DM are capable of TLR-induced type B presentation, indicating that regulation of H2-DM is not a critical mechanism. DC do not release meaningful levels of peptides allowing generation of type A or B pMHC. Additionally, generation of type B pMHC from HEL is a delayed event, requiring eighteen hours for appearance at the cell surface. While surface MHC II levels only modestly increase, there is a significant increase in total pMHC complexes after TLR or type I IFN stimulation. I employed two-photon microcopy to observe type A and B T cells in intact lymph nodes. While there were no remarkable differences in T cell behavior between the two groups, I determined that DC present type A or both type A and B but never exclusively type B pMHC. Using mice that express HEL as a membrane-bound protein on APC: mHEL), I found that TLR stimulation increases type B presentation from membrane-bound protein. I found that while immunization of mHEL mice with CpG increases presentation of type B pMHC from pseudo-self HEL and primed anti-HEL CD4 T cells, no observable disease was detected. These results identified the cells and signals required for inflammation-associated type B presentation from HEL and established tools for future investigation of the sub-cellular mechanism controlling these events.","abstract_has_math":false,"creators":["Strong, Beverly"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation","degree_discipline":"Biology and Biomedical Sciences: Immunology","degree_department":null,"school":null,"contributors":["Emil Unanue"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2012,"date_issued":"2012-01-01T08:00:00Z","date_published":"2012-01-01T08:00:00Z","updated_at":"2026-07-24T06:12:25Z","subjects":["Immunology","antigen presentation","dendritic cells","MHC","toll-like receptor ligands"],"languages":["English (en)"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier.doi","label":"DOI","values":["https://doi.org/10.7936/K75Q4T3T"],"render_values":[{"text":"https://doi.org/10.7936/K75Q4T3T","href":"https://doi.org/10.7936/K75Q4T3T","code":true}]}]},"links":{"outbound_url":"https://openscholarship.wustl.edu/etd/645","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Emil Unanue"]},{"key":"dc:creator","label":"Author","values":["Strong, Beverly"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2012-05-17T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biology and Biomedical Sciences: Immunology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Immunology","antigen presentation","dendritic cells","MHC","toll-like receptor ligands"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["English (en)"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://openscholarship.wustl.edu/etd/645"]},{"key":"dc:identifier.doi","label":"DOI","values":["https://doi.org/10.7936/K75Q4T3T"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Peptides from the model antigen hen egg white lysozyme: HEL) are capable of binding MHC II in multiple conformations. Resting antigen presenting cells: APC) load peptide in early endosomal compartments where multiple conformations of a given pMHC pair are allowed, generating both type A and B pMHC complexes. Protein traffics to the late endosomal compartment and is loaded onto nascent MHC II in the presence of the editor H2-DM where type A pMHC conformers are selectively formed, generating exclusively type A pMHC from protein. Previous work demonstrated that type B pMHC are also generated when HEL is administered in vivo along with certain inflammatory stimulants. Using an in vitro system I established, I found that TLR ligands and type I interferons: IFN) act directly on dendritic cells: DC), allowing generation of type B pMHC from HEL. Both CD8alpha+ and CD8alpha- DC present type B pMHC with TLR stimulation, but the relative effectiveness varied based on the ligand used. Using a type I IFN receptor: IFNAR1) blocking antibody and DC from mice deficient in the receptor, I found that TLR-induced type I IFN production amplifies but is not required for TLR-induced type B presentation from HEL. While I have not determined a sub-cellular mechanism for TLR-induced type B presentation, I have excluded mechanisms and made several observations. DC deficient in H2-DM are capable of TLR-induced type B presentation, indicating that regulation of H2-DM is not a critical mechanism. DC do not release meaningful levels of peptides allowing generation of type A or B pMHC. Additionally, generation of type B pMHC from HEL is a delayed event, requiring eighteen hours for appearance at the cell surface. While surface MHC II levels only modestly increase, there is a significant increase in total pMHC complexes after TLR or type I IFN stimulation. I employed two-photon microcopy to observe type A and B T cells in intact lymph nodes. While there were no remarkable differences in T cell behavior between the two groups, I determined that DC present type A or both type A and B but never exclusively type B pMHC. Using mice that express HEL as a membrane-bound protein on APC: mHEL), I found that TLR stimulation increases type B presentation from membrane-bound protein. I found that while immunization of mHEL mice with CpG increases presentation of type B pMHC from pseudo-self HEL and primed anti-HEL CD4 T cells, no observable disease was detected. These results identified the cells and signals required for inflammation-associated type B presentation from HEL and established tools for future investigation of the sub-cellular mechanism controlling these events."]},{"key":"dc:title","label":"Title","values":["Type B Presentation from HEL Protein in the Context of Inflammation"]}]}],"canonical_facts":{"dc:contributor":["Emil Unanue"],"dc:creator":["Strong, Beverly"],"dc:date.available":["2012-05-17T07:00:00Z"],"dc:description.abstract":["Peptides from the model antigen hen egg white lysozyme: HEL) are capable of binding MHC II in multiple conformations. Resting antigen presenting cells: APC) load peptide in early endosomal compartments where multiple conformations of a given pMHC pair are allowed, generating both type A and B pMHC complexes. Protein traffics to the late endosomal compartment and is loaded onto nascent MHC II in the presence of the editor H2-DM where type A pMHC conformers are selectively formed, generating exclusively type A pMHC from protein. Previous work demonstrated that type B pMHC are also generated when HEL is administered in vivo along with certain inflammatory stimulants. Using an in vitro system I established, I found that TLR ligands and type I interferons: IFN) act directly on dendritic cells: DC), allowing generation of type B pMHC from HEL. Both CD8alpha+ and CD8alpha- DC present type B pMHC with TLR stimulation, but the relative effectiveness varied based on the ligand used. Using a type I IFN receptor: IFNAR1) blocking antibody and DC from mice deficient in the receptor, I found that TLR-induced type I IFN production amplifies but is not required for TLR-induced type B presentation from HEL. While I have not determined a sub-cellular mechanism for TLR-induced type B presentation, I have excluded mechanisms and made several observations. DC deficient in H2-DM are capable of TLR-induced type B presentation, indicating that regulation of H2-DM is not a critical mechanism. DC do not release meaningful levels of peptides allowing generation of type A or B pMHC. Additionally, generation of type B pMHC from HEL is a delayed event, requiring eighteen hours for appearance at the cell surface. While surface MHC II levels only modestly increase, there is a significant increase in total pMHC complexes after TLR or type I IFN stimulation. I employed two-photon microcopy to observe type A and B T cells in intact lymph nodes. While there were no remarkable differences in T cell behavior between the two groups, I determined that DC present type A or both type A and B but never exclusively type B pMHC. Using mice that express HEL as a membrane-bound protein on APC: mHEL), I found that TLR stimulation increases type B presentation from membrane-bound protein. I found that while immunization of mHEL mice with CpG increases presentation of type B pMHC from pseudo-self HEL and primed anti-HEL CD4 T cells, no observable disease was detected. These results identified the cells and signals required for inflammation-associated type B presentation from HEL and established tools for future investigation of the sub-cellular mechanism controlling these events."],"dc:identifier":["https://openscholarship.wustl.edu/etd/645"],"dc:identifier.doi":["https://doi.org/10.7936/K75Q4T3T"],"dc:language":["English (en)"],"dc:subject":["Immunology","antigen presentation","dendritic cells","MHC","toll-like receptor ligands"],"dc:title":["Type B Presentation from HEL Protein in the Context of Inflammation"],"thesis:degree_discipline":["Biology and Biomedical Sciences: Immunology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T06:12:25Z"}