Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells
Summary: Immune synapse (IS) formation determines T cell antitumor activity. Here, we present a protocol for characterizing the IS formation between chimeric antigen receptor (CAR) T cells and tumor cells by measuring the IS size and calcium flux by live-cell imaging. We describe steps for CAR T cel...
| المؤلفون الرئيسيون: | , , , , , , , , , , , , |
|---|---|
| التنسيق: | مقال |
| اللغة: | English |
| منشور في: |
Elsevier
2024-12-01
|
| سلاسل: | STAR Protocols |
| الموضوعات: | |
| الوصول للمادة أونلاين: | http://www.sciencedirect.com/science/article/pii/S2666166724005872 |
| _version_ | 1826968832792592384 |
|---|---|
| author | Jorge Ibanez Nikhil Hebbar Unmesha Thanekar Zhongzhen Yi Haley Houke Meghan Ward Chris Nevitt Liqing Tian Stephen C. Mack Heather Sheppard Jason Chiang M. Paulina Velasquez Giedre Krenciute |
| author_facet | Jorge Ibanez Nikhil Hebbar Unmesha Thanekar Zhongzhen Yi Haley Houke Meghan Ward Chris Nevitt Liqing Tian Stephen C. Mack Heather Sheppard Jason Chiang M. Paulina Velasquez Giedre Krenciute |
| author_sort | Jorge Ibanez |
| collection | DOAJ |
| description | Summary: Immune synapse (IS) formation determines T cell antitumor activity. Here, we present a protocol for characterizing the IS formation between chimeric antigen receptor (CAR) T cells and tumor cells by measuring the IS size and calcium flux by live-cell imaging. We describe steps for CAR T cell manufacturing, sample preparation, image acquisition, and data analysis.For complete details on the use and execution of this protocol, please refer to Chockley et al.,1 Ibanez et al.,2 and Zoine et al.3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2025-02-18T04:18:14Z |
| format | Article |
| id | doaj.art-8b928a25c09a49fd8380df49b4e3b51d |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2025-02-18T04:18:14Z |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8b928a25c09a49fd8380df49b4e3b51d2024-11-20T05:08:09ZengElsevierSTAR Protocols2666-16672024-12-0154103422Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cellsJorge Ibanez0Nikhil Hebbar1Unmesha Thanekar2Zhongzhen Yi3Haley Houke4Meghan Ward5Chris Nevitt6Liqing Tian7Stephen C. Mack8Heather Sheppard9Jason Chiang10M. Paulina Velasquez11Giedre Krenciute12Department of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Developmental Neurobiology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Pathology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Pathology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USADepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA; Corresponding authorDepartment of Bone Marrow Transplantation and Cellular Therapy, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA; Corresponding authorSummary: Immune synapse (IS) formation determines T cell antitumor activity. Here, we present a protocol for characterizing the IS formation between chimeric antigen receptor (CAR) T cells and tumor cells by measuring the IS size and calcium flux by live-cell imaging. We describe steps for CAR T cell manufacturing, sample preparation, image acquisition, and data analysis.For complete details on the use and execution of this protocol, please refer to Chockley et al.,1 Ibanez et al.,2 and Zoine et al.3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724005872cancerimmunologymicroscopy |
| spellingShingle | Jorge Ibanez Nikhil Hebbar Unmesha Thanekar Zhongzhen Yi Haley Houke Meghan Ward Chris Nevitt Liqing Tian Stephen C. Mack Heather Sheppard Jason Chiang M. Paulina Velasquez Giedre Krenciute Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells STAR Protocols cancer immunology microscopy |
| title | Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells |
| title_full | Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells |
| title_fullStr | Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells |
| title_full_unstemmed | Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells |
| title_short | Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells |
| title_sort | protocol for live cell imaging of immune synapse formation and activation of car t cells against cancer cells |
| topic | cancer immunology microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724005872 |
| work_keys_str_mv | AT jorgeibanez protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT nikhilhebbar protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT unmeshathanekar protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT zhongzhenyi protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT haleyhouke protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT meghanward protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT chrisnevitt protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT liqingtian protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT stephencmack protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT heathersheppard protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT jasonchiang protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT mpaulinavelasquez protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells AT giedrekrenciute protocolforlivecellimagingofimmunesynapseformationandactivationofcartcellsagainstcancercells |