The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells
Developing new adjuvants/vaccines and better understanding their mode-of-action is an important task. To specifically improve birch pollen allergy treatment, we designed a fusion protein consisting of major birch pollen allergen Betv1 conjugated to the TLR5-ligand flagellin (rFlaA:Betv1). This study...
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MDPI AG
2021-12-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/12/3415 |
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| author | Yen-Ju Lin Adam Flaczyk Sonja Wolfheimer Alexandra Goretzki Annette Jamin Andrea Wangorsch Stefan Vieths Stephan Scheurer Stefan Schülke |
| author_facet | Yen-Ju Lin Adam Flaczyk Sonja Wolfheimer Alexandra Goretzki Annette Jamin Andrea Wangorsch Stefan Vieths Stephan Scheurer Stefan Schülke |
| author_sort | Yen-Ju Lin |
| collection | DOAJ |
| description | Developing new adjuvants/vaccines and better understanding their mode-of-action is an important task. To specifically improve birch pollen allergy treatment, we designed a fusion protein consisting of major birch pollen allergen Betv1 conjugated to the TLR5-ligand flagellin (rFlaA:Betv1). This study investigates the immune-modulatory effects of rFlaA:Betv1 on airway epithelial cells. LA-4 mouse lung epithelial cells were stimulated with rFlaA:Betv1 in the presence/absence of various inhibitors with cytokine- and chemokine secretion quantified by ELISA and activation of intracellular signaling cascades demonstrated by Western blot (WB). Either LA-4 cells or LA-4-derived supernatants were co-cultured with BALB/c bone marrow-derived myeloid dendritic cells (mDCs). Compared to equimolar amounts of flagellin and Betv1 provided as a mixture, rFlaA:Betv1 induced higher secretion of IL-6 and the chemokines CCL2 and CCL20 from LA-4 cells and a pronounced MAPK- and NF<i>κ</i>B-activation. Mechanistically, rFlaA:Betv1 was taken up more strongly and the induced cytokine production was inhibited by NF<i>κ</i>B-inhibitors, while ERK- and p38-MAPK-inhibitors only suppressed IL-6 and CCL2 secretion. In co-cultures of LA-4 cells with mDCs, rFlaA:Betv1-stimulated LA-4 cells p38-MAPK- and COX2-dependently secreted PGE<sub>2</sub>, which modulated DC responses by suppressing pro-inflammatory IL-12 and TNF-α secretion. Taken together, these results contribute to our understanding of the mechanisms underlying the strong immune-modulatory effects of flagellin-containing fusion proteins. |
| first_indexed | 2024-03-10T04:26:02Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T04:26:02Z |
| publishDate | 2021-12-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-ada92b2bd4b242b78ca3ffc836ea0bb82023-11-23T07:37:28ZengMDPI AGCells2073-44092021-12-011012341510.3390/cells10123415The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial CellsYen-Ju Lin0Adam Flaczyk1Sonja Wolfheimer2Alexandra Goretzki3Annette Jamin4Andrea Wangorsch5Stefan Vieths6Stephan Scheurer7Stefan Schülke8Molecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyMolecular Allergology, Paul-Ehrlich-Institut, 63225 Langen, GermanyDeveloping new adjuvants/vaccines and better understanding their mode-of-action is an important task. To specifically improve birch pollen allergy treatment, we designed a fusion protein consisting of major birch pollen allergen Betv1 conjugated to the TLR5-ligand flagellin (rFlaA:Betv1). This study investigates the immune-modulatory effects of rFlaA:Betv1 on airway epithelial cells. LA-4 mouse lung epithelial cells were stimulated with rFlaA:Betv1 in the presence/absence of various inhibitors with cytokine- and chemokine secretion quantified by ELISA and activation of intracellular signaling cascades demonstrated by Western blot (WB). Either LA-4 cells or LA-4-derived supernatants were co-cultured with BALB/c bone marrow-derived myeloid dendritic cells (mDCs). Compared to equimolar amounts of flagellin and Betv1 provided as a mixture, rFlaA:Betv1 induced higher secretion of IL-6 and the chemokines CCL2 and CCL20 from LA-4 cells and a pronounced MAPK- and NF<i>κ</i>B-activation. Mechanistically, rFlaA:Betv1 was taken up more strongly and the induced cytokine production was inhibited by NF<i>κ</i>B-inhibitors, while ERK- and p38-MAPK-inhibitors only suppressed IL-6 and CCL2 secretion. In co-cultures of LA-4 cells with mDCs, rFlaA:Betv1-stimulated LA-4 cells p38-MAPK- and COX2-dependently secreted PGE<sub>2</sub>, which modulated DC responses by suppressing pro-inflammatory IL-12 and TNF-α secretion. Taken together, these results contribute to our understanding of the mechanisms underlying the strong immune-modulatory effects of flagellin-containing fusion proteins.https://www.mdpi.com/2073-4409/10/12/3415flagellinBetv1epithelial cellsvaccinefusion protein |
| spellingShingle | Yen-Ju Lin Adam Flaczyk Sonja Wolfheimer Alexandra Goretzki Annette Jamin Andrea Wangorsch Stefan Vieths Stephan Scheurer Stefan Schülke The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells Cells flagellin Betv1 epithelial cells vaccine fusion protein |
| title | The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells |
| title_full | The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells |
| title_fullStr | The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells |
| title_full_unstemmed | The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells |
| title_short | The Fusion Protein rFlaA:Betv1 Modulates DC Responses by a p38-MAPK and COX2-Dependent Secretion of PGE<sub>2</sub> from Epithelial Cells |
| title_sort | fusion protein rflaa betv1 modulates dc responses by a p38 mapk and cox2 dependent secretion of pge sub 2 sub from epithelial cells |
| topic | flagellin Betv1 epithelial cells vaccine fusion protein |
| url | https://www.mdpi.com/2073-4409/10/12/3415 |
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