Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells
Among the polypeptides that comprise the T cell receptor (TCR), only CD3ζ is found in Natural Killer (NK) cells, where it transmits signals from activating receptors such as CD16 and NKp46. NK cells are potent immune cells that recognize target cells through germline-encoded activating and inhibitor...
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MDPI AG
2022-08-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/44/9/265 |
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| author | Khder H. Rasul Alamdar Hussain Hazel Reilly Maria Karvouni Carin I. M. Dahlberg Mustafa S. Al-Attar Arnika K. Wagner Evren Alici Dara K. Mohammad |
| author_facet | Khder H. Rasul Alamdar Hussain Hazel Reilly Maria Karvouni Carin I. M. Dahlberg Mustafa S. Al-Attar Arnika K. Wagner Evren Alici Dara K. Mohammad |
| author_sort | Khder H. Rasul |
| collection | DOAJ |
| description | Among the polypeptides that comprise the T cell receptor (TCR), only CD3ζ is found in Natural Killer (NK) cells, where it transmits signals from activating receptors such as CD16 and NKp46. NK cells are potent immune cells that recognize target cells through germline-encoded activating and inhibitory receptors. Genetic engineering of NK cells enables tumor-specific antigen recognition and, thus, has a significant promise in adoptive cell therapy. Ectopic expression of engineered TCR components in T cells leads to mispairing with the endogenous components, making a knockout of the endogenous TCR necessary. To circumvent the mispairing of TCRs or the need for knockout technologies, TCR complex expression has been studied in NK cells. In the current study, we explored the cellular processing of the TCR complex in NK cells. We observed that in the absence of CD3 subunits, the TCR was not expressed on the surface of NK cells and vice versa. Moreover, a progressive increase in surface expression of TCR between day three and day seven was observed after transduction. Interestingly, the TCR complex expression in NK92 cells was enhanced with a proteasome inhibitor (bortezomib) but not a lysosomal inhibitor (chloroquine). Additionally, we observed that the TCR complex was functional in NK92 cells as measured by estimating CD107a as a degranulation marker, IFNγ cytokine production, and killing assays. NK92 cells strongly degranulated when CD3ε was engaged in the presence of TCR, but not when only CD3 was overexpressed. Therefore, our findings encourage further investigation to unravel the mechanisms that prevent the surface expression of the TCR complex. |
| first_indexed | 2024-03-10T00:22:51Z |
| format | Article |
| id | doaj.art-7bfca1222273450aa573a8f6db2a9a01 |
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| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-03-10T00:22:51Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-7bfca1222273450aa573a8f6db2a9a012023-11-23T15:38:57ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452022-08-014493859387110.3390/cimb44090265Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer CellsKhder H. Rasul0Alamdar Hussain1Hazel Reilly2Maria Karvouni3Carin I. M. Dahlberg4Mustafa S. Al-Attar5Arnika K. Wagner6Evren Alici7Dara K. Mohammad8Center for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenDepartment of Biology, College of Science, Salahaddin University-Erbil, Erbil 44002, Kurdistan Region, IraqCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenCenter for Hematology and Regenerative Medicine (HERM), Department of Medicine Huddinge, Karolinska Institutet, SE-141 83 Stockholm, SwedenAmong the polypeptides that comprise the T cell receptor (TCR), only CD3ζ is found in Natural Killer (NK) cells, where it transmits signals from activating receptors such as CD16 and NKp46. NK cells are potent immune cells that recognize target cells through germline-encoded activating and inhibitory receptors. Genetic engineering of NK cells enables tumor-specific antigen recognition and, thus, has a significant promise in adoptive cell therapy. Ectopic expression of engineered TCR components in T cells leads to mispairing with the endogenous components, making a knockout of the endogenous TCR necessary. To circumvent the mispairing of TCRs or the need for knockout technologies, TCR complex expression has been studied in NK cells. In the current study, we explored the cellular processing of the TCR complex in NK cells. We observed that in the absence of CD3 subunits, the TCR was not expressed on the surface of NK cells and vice versa. Moreover, a progressive increase in surface expression of TCR between day three and day seven was observed after transduction. Interestingly, the TCR complex expression in NK92 cells was enhanced with a proteasome inhibitor (bortezomib) but not a lysosomal inhibitor (chloroquine). Additionally, we observed that the TCR complex was functional in NK92 cells as measured by estimating CD107a as a degranulation marker, IFNγ cytokine production, and killing assays. NK92 cells strongly degranulated when CD3ε was engaged in the presence of TCR, but not when only CD3 was overexpressed. Therefore, our findings encourage further investigation to unravel the mechanisms that prevent the surface expression of the TCR complex.https://www.mdpi.com/1467-3045/44/9/265T cell receptornatural killer cellsTCR positive natural killer cellsantigencancer cells |
| spellingShingle | Khder H. Rasul Alamdar Hussain Hazel Reilly Maria Karvouni Carin I. M. Dahlberg Mustafa S. Al-Attar Arnika K. Wagner Evren Alici Dara K. Mohammad Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells Current Issues in Molecular Biology T cell receptor natural killer cells TCR positive natural killer cells antigen cancer cells |
| title | Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells |
| title_full | Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells |
| title_fullStr | Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells |
| title_full_unstemmed | Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells |
| title_short | Assessment of T Cell Receptor Complex Expression Kinetics in Natural Killer Cells |
| title_sort | assessment of t cell receptor complex expression kinetics in natural killer cells |
| topic | T cell receptor natural killer cells TCR positive natural killer cells antigen cancer cells |
| url | https://www.mdpi.com/1467-3045/44/9/265 |
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