Invariant Natural Killer T cells (iNK-T cells) are a powerful regulatory immune cell that can recruit both innate and adaptive immune cells. Unlike conventional T cells (CD4+ and CD8α+), they recognize glycolipid antigens via the MHC-class-I like molecule, CD1d. A synthetically derived glycolipid from the marine sponge, Agelas mauritianus, alpha-Galactosylceramide (α-GalCer) potently activates iNK-T cells. Within a few hours after activation, iNK-T cells produce high quantities of TH1 and TH2 type cytokines, thus shape subsequent adaptive immunity towards inflammation (TH1) or immune-suppression (TH2). Structural modification of α-GalCer’s phytosphingosine chain, α-GalCer C20:2 has been shown to polarize iNK-T responses towards TH2 and help treat several autoimmune diseases. Recently, Liposomal formulation of α-GalCer (RGI-2001) has been shown to help ameliorate Graft-Versus-Host-disease (GVHD) via expansion of regulatory T cells. However, molecular and cellular mechanism by which how Liposomal α-GalCer modulate iNK-T responses is unknown. Our preliminary data suggests that both TH2 biasing α-GalCer C20:2 and Liposomal α-GalCer elicit TH2 polarized iNK-T responses. We hypothesize Liposomal α-GalCer will display unique antigen uptake, processing and presentation with CD1d similar to known TH2 biasing α-GalCer analogues, allowing for an immunosuppressive cytokine response, thus reducing pro-inflammatory disease complications. In this study, we investigated the cellular and molecular requirements of CD1d association with Liposomal α-GalCer through various in vitro assays to assess antigen uptake, processing, and presentation to iNK-T cells. We found that Liposomal α-GalCer share kinetics of antigen association with CD1d likely within non-lipid raft domain in the antigen presenting cells, similarly to α-GalCer C20:2, but require cellular internalization for CD1d association, similarly to TH1 biasing α-GalCer. Our findings suggest that Liposomal aGalCer requires early endosomal recycling for association with CD1d. In the future, we plan to investigate the visual and kinetic sites of α-GalCer:CD1d loading using confocal microscopy, and impact on immunologic synapse formation.
","abstract_html":"<p>Invariant Natural Killer T cells (iNK-T cells) are a powerful regulatory immune cell that can recruit both innate and adaptive immune cells. Unlike conventional T cells (CD4<sup>+ </sup>and CD8α<sup>+</sup>), they recognize glycolipid antigens via the MHC-class-I like molecule, CD1d. A synthetically derived glycolipid from the marine sponge, <em>Agelas mauritianus</em>, alpha-Galactosylceramide (α-GalCer) potently activates iNK-T cells. Within a few hours after activation, iNK-T cells produce high quantities of T<sub>H</sub>1 and T<sub>H</sub>2 type cytokines, thus shape subsequent adaptive immunity towards inflammation (T<sub>H</sub>1) or immune-suppression (T<sub>H</sub>2). Structural modification of α-GalCer’s phytosphingosine chain, α-GalCer C20:2 has been shown to polarize iNK-T responses towards T<sub>H</sub>2 and help treat several autoimmune diseases. Recently, Liposomal formulation of α-GalCer (RGI-2001) has been shown to help ameliorate Graft-Versus-Host-disease (GVHD) via expansion of regulatory T cells. However, molecular and cellular mechanism by which how Liposomal α-GalCer modulate iNK-T responses is unknown. Our preliminary data suggests that both T<sub>H</sub>2 biasing α-GalCer C20:2 and Liposomal α-GalCer elicit T<sub>H</sub>2 polarized iNK-T responses. We hypothesize Liposomal α-GalCer will display unique antigen uptake, processing and presentation with CD1d similar to known T<sub>H</sub>2 biasing α-GalCer analogues, allowing for an immunosuppressive cytokine response, thus reducing pro-inflammatory disease complications. In this study, we investigated the cellular and molecular requirements of CD1d association with Liposomal α-GalCer through various <em>in vitro</em> assays to assess antigen uptake, processing, and presentation to iNK-T cells. We found that Liposomal α-GalCer share kinetics of antigen association with CD1d likely within non-lipid raft domain in the antigen presenting cells, similarly to α-GalCer C20:2, but require cellular internalization for CD1d association, similarly to T<sub>H</sub>1 biasing α-GalCer. Our findings suggest that Liposomal aGalCer requires early endosomal recycling for association with CD1d. In the future, we plan to investigate the visual and kinetic sites of α-GalCer:CD1d loading using confocal microscopy, and impact on immunologic synapse formation.</p>","abstract_has_math":false,"creators":["Flegle, Alexandra S","http://www.orcid.org/0000-0002-4639-7889
ID: 0000-0002-4639-7889
"],"institution":null,"degree_name":"Masters of Science (MS)","degree_level":"Thesis (MS)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Jin Seon Im","Vahid Afshar-Kharghan","Greg Lizee"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2023,"date_issued":"2023-05-01T07:00:00Z","date_published":"2023-05-01T07:00:00Z","updated_at":"2026-07-24T05:49:16Z","subjects":["CD1d restricted-invariant Natural Killer T cells","α-Galactosylceramide","liposomes","synthetic glycolipids","Immune System Diseases","Medical Immunology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1262","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Jin Seon Im","Vahid Afshar-Kharghan","Greg Lizee"]},{"key":"dc:creator","label":"Author","values":["Flegle, Alexandra S","http://www.orcid.org/0000-0002-4639-7889
ID: 0000-0002-4639-7889
"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2023-04-25T07:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis (MS)"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Masters of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["CD1d restricted-invariant Natural Killer T cells","α-Galactosylceramide","liposomes","synthetic glycolipids","Immune System Diseases","Medical Immunology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1262"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Invariant Natural Killer T cells (iNK-T cells) are a powerful regulatory immune cell that can recruit both innate and adaptive immune cells. Unlike conventional T cells (CD4+ and CD8α+), they recognize glycolipid antigens via the MHC-class-I like molecule, CD1d. A synthetically derived glycolipid from the marine sponge, Agelas mauritianus, alpha-Galactosylceramide (α-GalCer) potently activates iNK-T cells. Within a few hours after activation, iNK-T cells produce high quantities of TH1 and TH2 type cytokines, thus shape subsequent adaptive immunity towards inflammation (TH1) or immune-suppression (TH2). Structural modification of α-GalCer’s phytosphingosine chain, α-GalCer C20:2 has been shown to polarize iNK-T responses towards TH2 and help treat several autoimmune diseases. Recently, Liposomal formulation of α-GalCer (RGI-2001) has been shown to help ameliorate Graft-Versus-Host-disease (GVHD) via expansion of regulatory T cells. However, molecular and cellular mechanism by which how Liposomal α-GalCer modulate iNK-T responses is unknown. Our preliminary data suggests that both TH2 biasing α-GalCer C20:2 and Liposomal α-GalCer elicit TH2 polarized iNK-T responses. We hypothesize Liposomal α-GalCer will display unique antigen uptake, processing and presentation with CD1d similar to known TH2 biasing α-GalCer analogues, allowing for an immunosuppressive cytokine response, thus reducing pro-inflammatory disease complications. In this study, we investigated the cellular and molecular requirements of CD1d association with Liposomal α-GalCer through various in vitro assays to assess antigen uptake, processing, and presentation to iNK-T cells. We found that Liposomal α-GalCer share kinetics of antigen association with CD1d likely within non-lipid raft domain in the antigen presenting cells, similarly to α-GalCer C20:2, but require cellular internalization for CD1d association, similarly to TH1 biasing α-GalCer. Our findings suggest that Liposomal aGalCer requires early endosomal recycling for association with CD1d. In the future, we plan to investigate the visual and kinetic sites of α-GalCer:CD1d loading using confocal microscopy, and impact on immunologic synapse formation.
"]},{"key":"dc:title","label":"Title","values":["Uncovering Novel Mechanism of Immune Modulation of Invariant Natural Killer T Cells By Liposomal Α-Galactosylceramide"]}]}],"canonical_facts":{"dc:contributor":["Jin Seon Im","Vahid Afshar-Kharghan","Greg Lizee"],"dc:creator":["Flegle, Alexandra S","http://www.orcid.org/0000-0002-4639-7889
ID: 0000-0002-4639-7889
"],"dc:date.available":["2023-04-25T07:00:00Z"],"dc:description.abstract":["Invariant Natural Killer T cells (iNK-T cells) are a powerful regulatory immune cell that can recruit both innate and adaptive immune cells. Unlike conventional T cells (CD4+ and CD8α+), they recognize glycolipid antigens via the MHC-class-I like molecule, CD1d. A synthetically derived glycolipid from the marine sponge, Agelas mauritianus, alpha-Galactosylceramide (α-GalCer) potently activates iNK-T cells. Within a few hours after activation, iNK-T cells produce high quantities of TH1 and TH2 type cytokines, thus shape subsequent adaptive immunity towards inflammation (TH1) or immune-suppression (TH2). Structural modification of α-GalCer’s phytosphingosine chain, α-GalCer C20:2 has been shown to polarize iNK-T responses towards TH2 and help treat several autoimmune diseases. Recently, Liposomal formulation of α-GalCer (RGI-2001) has been shown to help ameliorate Graft-Versus-Host-disease (GVHD) via expansion of regulatory T cells. However, molecular and cellular mechanism by which how Liposomal α-GalCer modulate iNK-T responses is unknown. Our preliminary data suggests that both TH2 biasing α-GalCer C20:2 and Liposomal α-GalCer elicit TH2 polarized iNK-T responses. We hypothesize Liposomal α-GalCer will display unique antigen uptake, processing and presentation with CD1d similar to known TH2 biasing α-GalCer analogues, allowing for an immunosuppressive cytokine response, thus reducing pro-inflammatory disease complications. In this study, we investigated the cellular and molecular requirements of CD1d association with Liposomal α-GalCer through various in vitro assays to assess antigen uptake, processing, and presentation to iNK-T cells. We found that Liposomal α-GalCer share kinetics of antigen association with CD1d likely within non-lipid raft domain in the antigen presenting cells, similarly to α-GalCer C20:2, but require cellular internalization for CD1d association, similarly to TH1 biasing α-GalCer. Our findings suggest that Liposomal aGalCer requires early endosomal recycling for association with CD1d. In the future, we plan to investigate the visual and kinetic sites of α-GalCer:CD1d loading using confocal microscopy, and impact on immunologic synapse formation.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1262"],"dc:subject":["CD1d restricted-invariant Natural Killer T cells","α-Galactosylceramide","liposomes","synthetic glycolipids","Immune System Diseases","Medical Immunology"],"dc:title":["Uncovering Novel Mechanism of Immune Modulation of Invariant Natural Killer T Cells By Liposomal Α-Galactosylceramide"],"thesis:degree_level":["Thesis (MS)"],"thesis:degree_name":["Masters of Science (MS)"]},"updated_at":"2026-07-24T05:49:16Z"}