25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency
Abstract Background Ex vivo cultivation is a promising strategy for increasing the number of NK cells and enhancing their antitumor activity prior to clinical application. Recent studies show that stimulation with 25KDa branched polyethylenimine (25KbPEI) generates NK cells with enhanced antitumor a...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-05-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-023-01101-8 |
| _version_ | 1797827417202491392 |
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| author | Eun-Su Ko Seung Hee Choi Minwook Lee Kyung-Soon Park |
| author_facet | Eun-Su Ko Seung Hee Choi Minwook Lee Kyung-Soon Park |
| author_sort | Eun-Su Ko |
| collection | DOAJ |
| description | Abstract Background Ex vivo cultivation is a promising strategy for increasing the number of NK cells and enhancing their antitumor activity prior to clinical application. Recent studies show that stimulation with 25KDa branched polyethylenimine (25KbPEI) generates NK cells with enhanced antitumor activity. To better understand how 25KbPEI primes NK cells, we explored the mechanism underlying increase in production of IFN-γ. Methods Chemical priming was performed on NK-92MI cells by incubating them with 5 μg/ml of 25KbPEI. The production of IFN-γ was evaluated by RT-qPCR, ELISA, and Flow cytometry. By evaluating the effect of pharmacological inhibition of ERK/mTOR-eIF4E signaling pathways on IFN-γ translation, the function of these signaling pathways in IFN-γ translation was examined. To comprehend the level of 25KbPEI activity on immune-related components in NK cells, RNA sequencing and proteomics analyses were conducted. Results 25KbPEI enhances the production of IFN-γ by NK cells without transcriptional activation. Activation of ERK and mTOR signaling pathways was found to be associated with 25KbPEI-mediated calcium influx in NK cells. The activation of ERK/mTOR signaling was linked to the phosphorylation of 4E-BP1, which resulted in the activation of translation initiation complex and subsequent IFN-γ translation. Analysis of RNA sequencing and proteomics data revealed that the activity of 25KbPEI to improve translation efficiency in NK cells could be extended to additional immune-related molecules. Conclusions This study provides substantial insight into the process by which 25KbPEI primes NK cells. Our data demonstrated that the 25KbPEI mediated activation of ERK/mTOR signaling and subsequent stimulation of eIF4E is the primary mechanism by which the chemical stimulates translation of IFN-γ in NK cells. Video abstract |
| first_indexed | 2024-04-09T12:47:57Z |
| format | Article |
| id | doaj.art-0452aa39ef7d4e4d933c0eccf48ba0c1 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-04-09T12:47:57Z |
| publishDate | 2023-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-0452aa39ef7d4e4d933c0eccf48ba0c12023-05-14T11:22:50ZengBMCCell Communication and Signaling1478-811X2023-05-0121111310.1186/s12964-023-01101-825KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiencyEun-Su Ko0Seung Hee Choi1Minwook Lee2Kyung-Soon Park3Department of Biomedical Science, CHA UniversityDepartment of Biomedical Science, CHA UniversityDepartment of Biomedical Science, CHA UniversityDepartment of Biomedical Science, CHA UniversityAbstract Background Ex vivo cultivation is a promising strategy for increasing the number of NK cells and enhancing their antitumor activity prior to clinical application. Recent studies show that stimulation with 25KDa branched polyethylenimine (25KbPEI) generates NK cells with enhanced antitumor activity. To better understand how 25KbPEI primes NK cells, we explored the mechanism underlying increase in production of IFN-γ. Methods Chemical priming was performed on NK-92MI cells by incubating them with 5 μg/ml of 25KbPEI. The production of IFN-γ was evaluated by RT-qPCR, ELISA, and Flow cytometry. By evaluating the effect of pharmacological inhibition of ERK/mTOR-eIF4E signaling pathways on IFN-γ translation, the function of these signaling pathways in IFN-γ translation was examined. To comprehend the level of 25KbPEI activity on immune-related components in NK cells, RNA sequencing and proteomics analyses were conducted. Results 25KbPEI enhances the production of IFN-γ by NK cells without transcriptional activation. Activation of ERK and mTOR signaling pathways was found to be associated with 25KbPEI-mediated calcium influx in NK cells. The activation of ERK/mTOR signaling was linked to the phosphorylation of 4E-BP1, which resulted in the activation of translation initiation complex and subsequent IFN-γ translation. Analysis of RNA sequencing and proteomics data revealed that the activity of 25KbPEI to improve translation efficiency in NK cells could be extended to additional immune-related molecules. Conclusions This study provides substantial insight into the process by which 25KbPEI primes NK cells. Our data demonstrated that the 25KbPEI mediated activation of ERK/mTOR signaling and subsequent stimulation of eIF4E is the primary mechanism by which the chemical stimulates translation of IFN-γ in NK cells. Video abstracthttps://doi.org/10.1186/s12964-023-01101-825KDa branched polyethylenimineNatural killer cellCalciumInterferon-γTranslation efficiencyERK signaling |
| spellingShingle | Eun-Su Ko Seung Hee Choi Minwook Lee Kyung-Soon Park 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency Cell Communication and Signaling 25KDa branched polyethylenimine Natural killer cell Calcium Interferon-γ Translation efficiency ERK signaling |
| title | 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency |
| title_full | 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency |
| title_fullStr | 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency |
| title_full_unstemmed | 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency |
| title_short | 25KDa branched polyethylenimine increases interferon-γ production in natural killer cells via improving translation efficiency |
| title_sort | 25kda branched polyethylenimine increases interferon γ production in natural killer cells via improving translation efficiency |
| topic | 25KDa branched polyethylenimine Natural killer cell Calcium Interferon-γ Translation efficiency ERK signaling |
| url | https://doi.org/10.1186/s12964-023-01101-8 |
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