Fusion of Bacterial Flagellin to a Dendritic Cell-Targeting αCD40 Antibody Construct Coupled With Viral or Leukemia-Specific Antigens Enhances Dendritic Cell Maturation and Activates Peptide-Responsive T Cells
Copyright © 2020 Schmitt, Tahk, Lohner, Hänel, Maiser, Hauke, Patel, Rothe, Josenhans, Leonhardt, Griffioen, Deiser, Fenn, Hopfner and Subklewe..
Conventional dendritic cell (DC) vaccine strategies, in which DCs are loaded with antigens ex vivo, suffer biological issues such as impaired DC migration capacity and laborious GMP production procedures. In a promising alternative, antigens are targeted to DC-associated endocytic receptors in vivo with antibody-antigen conjugates co-administered with toll-like receptor (TLR) agonists as adjuvants. To combine the potential advantages of in vivo targeting of DCs with those of conjugated TLR agonists, we generated a multifunctional antibody construct integrating the DC-specific delivery of viral- or tumor-associated antigens and DC activation by TLR ligation in one molecule. We validated its functionality in vitro and determined if TLR ligation might improve the efficacy of such a molecule. In proof-of-principle studies, an αCD40 antibody containing a CMV pp65-derived peptide as an antigen domain (αCD40CMV) was genetically fused to the TLR5-binding D0/D1 domain of bacterial flagellin (αCD40.FlgCMV). The analysis of surface maturation markers on immature DCs revealed that fusion of flagellin to αCD40CMV highly increased DC maturation (3.4-fold elevation of CD80 expression compared to αCD40CMV alone) by specifically interacting with TLR5. Immature DCs loaded with αCD40.FlgCMV induced significantly higher CMVNLV-specific T cell activation and proliferation compared to αCD40CMV in co-culture experiments with allogeneic and autologous T cells (1.8-fold increase in % IFN-γ/TNF-α+ CD8+ T cells and 3.9-fold increase in % CMVNLV-specific dextramer+ CD8+ T cells). More importantly, we confirmed the beneficial effects of flagellin-dependent DC stimulation using a tumor-specific neoantigen as the antigen domain. Specifically, the acute myeloid leukemia (AML)-specific mutated NPM1 (mNPM1)-derived neoantigen CLAVEEVSL was delivered to DCs in the form of αCD40mNPM1 and αCD40.FlgmNPM1 antibody constructs, making this study the first to investigate mNPM1 in a DC vaccination context. Again, αCD40.FlgmNPM1-loaded DCs more potently activated allogeneic mNPM1CLA-specific T cells compared to αCD40mNPM1. These in vitro results confirmed the functionality of our multifunctional antibody construct and demonstrated that TLR5 ligation improved the efficacy of the molecule. Future mouse studies are required to examine the T cell-activating potential of αCD40.FlgmNPM1 after targeting of dendritic cells in vivo using AML xenograft models.
Medienart: |
E-Artikel |
---|
Erscheinungsjahr: |
2020 |
---|---|
Erschienen: |
2020 |
Enthalten in: |
Zur Gesamtaufnahme - volume:11 |
---|---|
Enthalten in: |
Frontiers in immunology - 11(2020) vom: 25., Seite 602802 |
Sprache: |
Englisch |
---|
Beteiligte Personen: |
Schmitt, Saskia [VerfasserIn] |
---|
Links: |
---|
Anmerkungen: |
Date Completed 21.06.2021 Date Revised 04.12.2021 published: Electronic-eCollection Citation Status MEDLINE |
---|
doi: |
10.3389/fimmu.2020.602802 |
---|
funding: |
|
---|---|
Förderinstitution / Projekttitel: |
|
PPN (Katalog-ID): |
NLM318477092 |
---|
LEADER | 01000caa a22002652 4500 | ||
---|---|---|---|
001 | NLM318477092 | ||
003 | DE-627 | ||
005 | 20231226202307.0 | ||
007 | cr uuu---uuuuu | ||
008 | 231225s2020 xx |||||o 00| ||eng c | ||
024 | 7 | |a 10.3389/fimmu.2020.602802 |2 doi | |
028 | 5 | 2 | |a pubmed24n1061.xml |
035 | |a (DE-627)NLM318477092 | ||
035 | |a (NLM)33281829 | ||
040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
041 | |a eng | ||
100 | 1 | |a Schmitt, Saskia |e verfasserin |4 aut | |
245 | 1 | 0 | |a Fusion of Bacterial Flagellin to a Dendritic Cell-Targeting αCD40 Antibody Construct Coupled With Viral or Leukemia-Specific Antigens Enhances Dendritic Cell Maturation and Activates Peptide-Responsive T Cells |
264 | 1 | |c 2020 | |
336 | |a Text |b txt |2 rdacontent | ||
337 | |a ƒaComputermedien |b c |2 rdamedia | ||
338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
500 | |a Date Completed 21.06.2021 | ||
500 | |a Date Revised 04.12.2021 | ||
500 | |a published: Electronic-eCollection | ||
500 | |a Citation Status MEDLINE | ||
520 | |a Copyright © 2020 Schmitt, Tahk, Lohner, Hänel, Maiser, Hauke, Patel, Rothe, Josenhans, Leonhardt, Griffioen, Deiser, Fenn, Hopfner and Subklewe. | ||
520 | |a Conventional dendritic cell (DC) vaccine strategies, in which DCs are loaded with antigens ex vivo, suffer biological issues such as impaired DC migration capacity and laborious GMP production procedures. In a promising alternative, antigens are targeted to DC-associated endocytic receptors in vivo with antibody-antigen conjugates co-administered with toll-like receptor (TLR) agonists as adjuvants. To combine the potential advantages of in vivo targeting of DCs with those of conjugated TLR agonists, we generated a multifunctional antibody construct integrating the DC-specific delivery of viral- or tumor-associated antigens and DC activation by TLR ligation in one molecule. We validated its functionality in vitro and determined if TLR ligation might improve the efficacy of such a molecule. In proof-of-principle studies, an αCD40 antibody containing a CMV pp65-derived peptide as an antigen domain (αCD40CMV) was genetically fused to the TLR5-binding D0/D1 domain of bacterial flagellin (αCD40.FlgCMV). The analysis of surface maturation markers on immature DCs revealed that fusion of flagellin to αCD40CMV highly increased DC maturation (3.4-fold elevation of CD80 expression compared to αCD40CMV alone) by specifically interacting with TLR5. Immature DCs loaded with αCD40.FlgCMV induced significantly higher CMVNLV-specific T cell activation and proliferation compared to αCD40CMV in co-culture experiments with allogeneic and autologous T cells (1.8-fold increase in % IFN-γ/TNF-α+ CD8+ T cells and 3.9-fold increase in % CMVNLV-specific dextramer+ CD8+ T cells). More importantly, we confirmed the beneficial effects of flagellin-dependent DC stimulation using a tumor-specific neoantigen as the antigen domain. Specifically, the acute myeloid leukemia (AML)-specific mutated NPM1 (mNPM1)-derived neoantigen CLAVEEVSL was delivered to DCs in the form of αCD40mNPM1 and αCD40.FlgmNPM1 antibody constructs, making this study the first to investigate mNPM1 in a DC vaccination context. Again, αCD40.FlgmNPM1-loaded DCs more potently activated allogeneic mNPM1CLA-specific T cells compared to αCD40mNPM1. These in vitro results confirmed the functionality of our multifunctional antibody construct and demonstrated that TLR5 ligation improved the efficacy of the molecule. Future mouse studies are required to examine the T cell-activating potential of αCD40.FlgmNPM1 after targeting of dendritic cells in vivo using AML xenograft models | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, Non-U.S. Gov't | |
650 | 4 | |a acute myeloid leukemia | |
650 | 4 | |a antibody | |
650 | 4 | |a dendritic cell | |
650 | 4 | |a flagellin | |
650 | 4 | |a neoantigen | |
650 | 4 | |a vaccine | |
650 | 7 | |a Antibodies |2 NLM | |
650 | 7 | |a CD40 Antigens |2 NLM | |
650 | 7 | |a Cancer Vaccines |2 NLM | |
650 | 7 | |a Epitopes |2 NLM | |
650 | 7 | |a FLG protein, human |2 NLM | |
650 | 7 | |a Filaggrin Proteins |2 NLM | |
650 | 7 | |a NPM1 protein, human |2 NLM | |
650 | 7 | |a Npm1 protein, mouse |2 NLM | |
650 | 7 | |a Nuclear Proteins |2 NLM | |
650 | 7 | |a Recombinant Fusion Proteins |2 NLM | |
650 | 7 | |a TLR5 protein, human |2 NLM | |
650 | 7 | |a Toll-Like Receptor 5 |2 NLM | |
650 | 7 | |a Viral Matrix Proteins |2 NLM | |
650 | 7 | |a cytomegalovirus matrix protein 65kDa |2 NLM | |
650 | 7 | |a Nucleophosmin |2 NLM | |
650 | 7 | |a 117896-08-9 |2 NLM | |
650 | 7 | |a Flagellin |2 NLM | |
650 | 7 | |a 12777-81-0 |2 NLM | |
700 | 1 | |a Tahk, Siret |e verfasserin |4 aut | |
700 | 1 | |a Lohner, Alina |e verfasserin |4 aut | |
700 | 1 | |a Hänel, Gerulf |e verfasserin |4 aut | |
700 | 1 | |a Maiser, Andreas |e verfasserin |4 aut | |
700 | 1 | |a Hauke, Martina |e verfasserin |4 aut | |
700 | 1 | |a Patel, Lubna |e verfasserin |4 aut | |
700 | 1 | |a Rothe, Maurine |e verfasserin |4 aut | |
700 | 1 | |a Josenhans, Christine |e verfasserin |4 aut | |
700 | 1 | |a Leonhardt, Heinrich |e verfasserin |4 aut | |
700 | 1 | |a Griffioen, Marieke |e verfasserin |4 aut | |
700 | 1 | |a Deiser, Katrin |e verfasserin |4 aut | |
700 | 1 | |a Fenn, Nadja C |e verfasserin |4 aut | |
700 | 1 | |a Hopfner, Karl-Peter |e verfasserin |4 aut | |
700 | 1 | |a Subklewe, Marion |e verfasserin |4 aut | |
773 | 0 | 8 | |i Enthalten in |t Frontiers in immunology |d 2010 |g 11(2020) vom: 25., Seite 602802 |w (DE-627)NLM215811453 |x 1664-3224 |7 nnns |
773 | 1 | 8 | |g volume:11 |g year:2020 |g day:25 |g pages:602802 |
856 | 4 | 0 | |u http://dx.doi.org/10.3389/fimmu.2020.602802 |3 Volltext |
912 | |a GBV_USEFLAG_A | ||
912 | |a GBV_NLM | ||
951 | |a AR | ||
952 | |d 11 |j 2020 |b 25 |h 602802 |