The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages
TLR5 ligand flagellin-containing fusion proteins are potential vaccine candidates for many diseases. A recombinant fusion protein of flagellin A and the major birch pollen allergen Bet v 1 (rFlaA:Betv1) modulates immune responses in vitro and in vivo. We studied the effects of rFlaA:Betv1 on bone ma...
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MDPI AG
2021-10-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/10/2614 |
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| author | Yen-Ju Lin Garibald Papp Csaba Miskey Anna Fiedler Alexandra Goretzki Sonja Wolfheimer Jennifer Zimmermann Peter Crauwels Zoltán Ivics Ger van Zandbergen Stefan Vieths Stephan Scheurer Stefan Schülke |
| author_facet | Yen-Ju Lin Garibald Papp Csaba Miskey Anna Fiedler Alexandra Goretzki Sonja Wolfheimer Jennifer Zimmermann Peter Crauwels Zoltán Ivics Ger van Zandbergen Stefan Vieths Stephan Scheurer Stefan Schülke |
| author_sort | Yen-Ju Lin |
| collection | DOAJ |
| description | TLR5 ligand flagellin-containing fusion proteins are potential vaccine candidates for many diseases. A recombinant fusion protein of flagellin A and the major birch pollen allergen Bet v 1 (rFlaA:Betv1) modulates immune responses in vitro and in vivo. We studied the effects of rFlaA:Betv1 on bone marrow-derived macrophages (BMDMs). BMDMs differentiated from BALB/c, C57BL/6, TLR5<sup>−/−</sup>, or MyD88<sup>−/−</sup> mice were pre-treated with inhibitors, stimulated with rFlaA:Betv1 or respective controls, and analyzed for activation, cytokine secretion, metabolic state, RNA transcriptome, and modulation of allergen-specific Th2 responses. Stimulation of BMDMs with rFlaA:Betv1 resulted in MyD88-dependent production of IL-1β, IL-6, TNF-α, IL-10, CD69 upregulation, and a pronounced shift towards glycolysis paralleled by activation of MAPK, NF<i>κ</i>B, and mTOR signaling. Inhibition of either mTOR (rapamycin) or SAP/JNK-MAPK signaling (SP600125) resulted in dose-dependent metabolic suppression. In BMDM and T cell co-cultures, rFlaA:Betv1 stimulation suppressed rBet v 1-induced IL-5 and IL-13 secretion while inducing IFN-γ production. mRNA-Seq analyses showed HIF-1a, JAK, STAT, phagosome, NLR, NF<i>κ</i>B, TNF, TLR, and chemokine signaling to participate in the interplay of cell activation, glycolysis, and immune response. rFlaA:Betv1 strongly activated BMDMs, resulting in MyD88−, MAPK−, and mTOR-dependent enhancement of glucose metabolism. Our results suggest macrophages are important target cells to consider during restauration of allergen tolerance during AIT. |
| first_indexed | 2024-03-10T06:39:42Z |
| format | Article |
| id | doaj.art-9be45d63424c4476bdedf46219cd4895 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T06:39:42Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-9be45d63424c4476bdedf46219cd48952023-11-22T17:46:24ZengMDPI AGCells2073-44092021-10-011010261410.3390/cells10102614The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in MacrophagesYen-Ju Lin0Garibald Papp1Csaba Miskey2Anna Fiedler3Alexandra Goretzki4Sonja Wolfheimer5Jennifer Zimmermann6Peter Crauwels7Zoltán Ivics8Ger van Zandbergen9Stefan Vieths10Stephan Scheurer11Stefan Schülke12Vice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMedical Biotechnology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyImmunology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMedical Biotechnology, Paul-Ehrlich-Institut, 63225 Langen, GermanyImmunology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyVice Presidents Research Group 1: Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyTLR5 ligand flagellin-containing fusion proteins are potential vaccine candidates for many diseases. A recombinant fusion protein of flagellin A and the major birch pollen allergen Bet v 1 (rFlaA:Betv1) modulates immune responses in vitro and in vivo. We studied the effects of rFlaA:Betv1 on bone marrow-derived macrophages (BMDMs). BMDMs differentiated from BALB/c, C57BL/6, TLR5<sup>−/−</sup>, or MyD88<sup>−/−</sup> mice were pre-treated with inhibitors, stimulated with rFlaA:Betv1 or respective controls, and analyzed for activation, cytokine secretion, metabolic state, RNA transcriptome, and modulation of allergen-specific Th2 responses. Stimulation of BMDMs with rFlaA:Betv1 resulted in MyD88-dependent production of IL-1β, IL-6, TNF-α, IL-10, CD69 upregulation, and a pronounced shift towards glycolysis paralleled by activation of MAPK, NF<i>κ</i>B, and mTOR signaling. Inhibition of either mTOR (rapamycin) or SAP/JNK-MAPK signaling (SP600125) resulted in dose-dependent metabolic suppression. In BMDM and T cell co-cultures, rFlaA:Betv1 stimulation suppressed rBet v 1-induced IL-5 and IL-13 secretion while inducing IFN-γ production. mRNA-Seq analyses showed HIF-1a, JAK, STAT, phagosome, NLR, NF<i>κ</i>B, TNF, TLR, and chemokine signaling to participate in the interplay of cell activation, glycolysis, and immune response. rFlaA:Betv1 strongly activated BMDMs, resulting in MyD88−, MAPK−, and mTOR-dependent enhancement of glucose metabolism. Our results suggest macrophages are important target cells to consider during restauration of allergen tolerance during AIT.https://www.mdpi.com/2073-4409/10/10/2614flagellinTLR5allergenBet v 1fusion proteinmetabolism |
| spellingShingle | Yen-Ju Lin Garibald Papp Csaba Miskey Anna Fiedler Alexandra Goretzki Sonja Wolfheimer Jennifer Zimmermann Peter Crauwels Zoltán Ivics Ger van Zandbergen Stefan Vieths Stephan Scheurer Stefan Schülke The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages Cells flagellin TLR5 allergen Bet v 1 fusion protein metabolism |
| title | The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages |
| title_full | The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages |
| title_fullStr | The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages |
| title_full_unstemmed | The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages |
| title_short | The Flagellin:Allergen Fusion Protein rFlaA:Betv1 Induces a MyD88− and MAPK-Dependent Activation of Glucose Metabolism in Macrophages |
| title_sort | flagellin allergen fusion protein rflaa betv1 induces a myd88 and mapk dependent activation of glucose metabolism in macrophages |
| topic | flagellin TLR5 allergen Bet v 1 fusion protein metabolism |
| url | https://www.mdpi.com/2073-4409/10/10/2614 |
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