Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration
Natural killer (NK) cell responses depend on the balance of signals from inhibitory and activating receptors. However, how the integration of antagonistic signals occurs upon NK cell–target cell interaction is not fully understood. Here we provide evidence that NK cell inhibition via the inhibitory...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.868496/full |
| _version_ | 1818240010772545536 |
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| author | David Tomaz David Tomaz Pedro Matos Pereira Pedro Matos Pereira Nadia Guerra Julian Dyson Keith Gould Ricardo Henriques Ricardo Henriques Ricardo Henriques |
| author_facet | David Tomaz David Tomaz Pedro Matos Pereira Pedro Matos Pereira Nadia Guerra Julian Dyson Keith Gould Ricardo Henriques Ricardo Henriques Ricardo Henriques |
| author_sort | David Tomaz |
| collection | DOAJ |
| description | Natural killer (NK) cell responses depend on the balance of signals from inhibitory and activating receptors. However, how the integration of antagonistic signals occurs upon NK cell–target cell interaction is not fully understood. Here we provide evidence that NK cell inhibition via the inhibitory receptor Ly49A is dependent on its relative colocalization at the nanometer scale with the activating receptor NKG2D upon immune synapse (IS) formation. NKG2D and Ly49A signal integration and colocalization were studied using NKG2D-GFP and Ly49A-RFP-expressing primary NK cells, forming ISs with NIH3T3 target cells, with or without the expression of single-chain trimer (SCT) H2-Dd and an extended form of SCT H2-Dd-CD4 MHC-I molecules. Nanoscale colocalization was assessed by Förster resonance energy transfer between NKG2D-GFP and Ly49A-RFP and measured for each synapse. In the presence of their respective cognate ligands, NKG2D and Ly49A colocalize at the nanometer scale, leading to NK cell inhibition. However, increasing the size of the Ly49A ligand reduced the nanoscale colocalization with NKG2D, consequently impairing Ly49A-mediated inhibition. Thus, our data shows that NK cell signal integration is critically dependent on the dimensions of NK cell ligand–receptor pairs by affecting their relative nanometer-scale colocalization at the IS. Our results together suggest that the balance of NK cell signals and NK cell responses is determined by the relative nanoscale colocalization of activating and inhibitory receptors in the immune synapse. |
| first_indexed | 2024-12-12T13:06:39Z |
| format | Article |
| id | doaj.art-94db0b8324664817a1f5cf288ba0a58a |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-12T13:06:39Z |
| publishDate | 2022-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-94db0b8324664817a1f5cf288ba0a58a2022-12-22T00:23:38ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-06-011310.3389/fimmu.2022.868496868496Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal IntegrationDavid Tomaz0David Tomaz1Pedro Matos Pereira2Pedro Matos Pereira3Nadia Guerra4Julian Dyson5Keith Gould6Ricardo Henriques7Ricardo Henriques8Ricardo Henriques9Department of Immunology, Wright-Fleming Institute, Imperial College London, London, United KingdomDepartment of Immunology, Hammersmith Hospital, Imperial College London, London, United KingdomMedical Research Council Laboratory for Molecular Cell Biology, University College London, London, United KingdomBacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, PortugalDivision of Cell and Molecular Biology, Faculty of Natural Sciences, Imperial College London, London, United KingdomDepartment of Immunology, Hammersmith Hospital, Imperial College London, London, United KingdomDepartment of Immunology, Wright-Fleming Institute, Imperial College London, London, United KingdomMedical Research Council Laboratory for Molecular Cell Biology, University College London, London, United KingdomDivision of Infection and Immunity, University College London, London, United KingdomOptical Cell Biology Lab, Instituto Gulbenkian de Ciência, Oeiras, PortugalNatural killer (NK) cell responses depend on the balance of signals from inhibitory and activating receptors. However, how the integration of antagonistic signals occurs upon NK cell–target cell interaction is not fully understood. Here we provide evidence that NK cell inhibition via the inhibitory receptor Ly49A is dependent on its relative colocalization at the nanometer scale with the activating receptor NKG2D upon immune synapse (IS) formation. NKG2D and Ly49A signal integration and colocalization were studied using NKG2D-GFP and Ly49A-RFP-expressing primary NK cells, forming ISs with NIH3T3 target cells, with or without the expression of single-chain trimer (SCT) H2-Dd and an extended form of SCT H2-Dd-CD4 MHC-I molecules. Nanoscale colocalization was assessed by Förster resonance energy transfer between NKG2D-GFP and Ly49A-RFP and measured for each synapse. In the presence of their respective cognate ligands, NKG2D and Ly49A colocalize at the nanometer scale, leading to NK cell inhibition. However, increasing the size of the Ly49A ligand reduced the nanoscale colocalization with NKG2D, consequently impairing Ly49A-mediated inhibition. Thus, our data shows that NK cell signal integration is critically dependent on the dimensions of NK cell ligand–receptor pairs by affecting their relative nanometer-scale colocalization at the IS. Our results together suggest that the balance of NK cell signals and NK cell responses is determined by the relative nanoscale colocalization of activating and inhibitory receptors in the immune synapse.https://www.frontiersin.org/articles/10.3389/fimmu.2022.868496/fullNK cell signalingnanoscale colocalizationNKG2DLy49ANK cell receptors and ligandsFRET |
| spellingShingle | David Tomaz David Tomaz Pedro Matos Pereira Pedro Matos Pereira Nadia Guerra Julian Dyson Keith Gould Ricardo Henriques Ricardo Henriques Ricardo Henriques Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration Frontiers in Immunology NK cell signaling nanoscale colocalization NKG2D Ly49A NK cell receptors and ligands FRET |
| title | Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration |
| title_full | Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration |
| title_fullStr | Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration |
| title_full_unstemmed | Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration |
| title_short | Nanoscale Colocalization of NK Cell Activating and Inhibitory Receptors Controls Signal Integration |
| title_sort | nanoscale colocalization of nk cell activating and inhibitory receptors controls signal integration |
| topic | NK cell signaling nanoscale colocalization NKG2D Ly49A NK cell receptors and ligands FRET |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.868496/full |
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