Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay
Reliable and sensitive characterization assays are important determinants of the successful clinical translation of immunotherapies. For the assessment of cytolytic potential, the chromium 51 (51Cr) release assay has long been considered the gold standard for testing effector cells. However, attaini...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-12-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1299512/full |
| _version_ | 1797382965105262592 |
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| author | Kiriakos Koukoulias Penelope G. Papayanni Julia Jones Manik Kuvalekar Ayumi Watanabe Yovana Velazquez Sarah Gilmore Anastasia Papadopoulou Ann M. Leen Spyridoula Vasileiou |
| author_facet | Kiriakos Koukoulias Penelope G. Papayanni Julia Jones Manik Kuvalekar Ayumi Watanabe Yovana Velazquez Sarah Gilmore Anastasia Papadopoulou Ann M. Leen Spyridoula Vasileiou |
| author_sort | Kiriakos Koukoulias |
| collection | DOAJ |
| description | Reliable and sensitive characterization assays are important determinants of the successful clinical translation of immunotherapies. For the assessment of cytolytic potential, the chromium 51 (51Cr) release assay has long been considered the gold standard for testing effector cells. However, attaining the approvals to access and use radioactive isotopes is becoming increasingly complex, while technical aspects [i.e. sensitivity, short (4-6 hours) assay duration] may lead to suboptimal performance. This has been the case with our ex vivo expanded, polyclonal (CD4+ and CD8+) multivirus-specific T cell (multiVST) lines, which recognize 5 difficult-to-treat viruses [Adenovirus (AdV), BK virus (BKV), cytomegalovirus (CMV), Epstein Barr virus (EBV), and human herpes virus 6 (HHV6)] and when administered to allogeneic hematopoietic stem cell (HCT) or solid organ transplant (SOT) recipients have been associated with clinical benefit. However, despite mediating potent antiviral effects in vivo, capturing in vitro cytotoxic potential has proven difficult in a traditional 51Cr release assay. Now, in addition to cytotoxicity surrogates, including CD107a and Granzyme B, we report on an alternative, vital dye -based, flow cytometric platform in which superior sensitivity and prolonged effector:target co-culture duration enabled the reliable detection of both CD4- and CD8-mediated in vitro cytolytic activity against viral targets without non-specific effects. |
| first_indexed | 2024-03-08T21:12:58Z |
| format | Article |
| id | doaj.art-05d14051cbc94363a2c29ed3b40a0427 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-08T21:12:58Z |
| publishDate | 2023-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-05d14051cbc94363a2c29ed3b40a04272023-12-22T04:26:24ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-12-011410.3389/fimmu.2023.12995121299512Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assayKiriakos Koukoulias0Penelope G. Papayanni1Julia Jones2Manik Kuvalekar3Ayumi Watanabe4Yovana Velazquez5Sarah Gilmore6Anastasia Papadopoulou7Ann M. Leen8Spyridoula Vasileiou9Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesAlloVir, Waltham, MA, United StatesHematology Department- Hematopoietic Cell Transplantation Unit, Gene and Cell Therapy Center, “George Papanikolaou” Hospital, Thessaloniki, GreeceCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesCenter for Cell and Gene Therapy, Baylor College of Medicine, Texas Children’s Hospital and Houston Methodist Hospital, Houston, TX, United StatesReliable and sensitive characterization assays are important determinants of the successful clinical translation of immunotherapies. For the assessment of cytolytic potential, the chromium 51 (51Cr) release assay has long been considered the gold standard for testing effector cells. However, attaining the approvals to access and use radioactive isotopes is becoming increasingly complex, while technical aspects [i.e. sensitivity, short (4-6 hours) assay duration] may lead to suboptimal performance. This has been the case with our ex vivo expanded, polyclonal (CD4+ and CD8+) multivirus-specific T cell (multiVST) lines, which recognize 5 difficult-to-treat viruses [Adenovirus (AdV), BK virus (BKV), cytomegalovirus (CMV), Epstein Barr virus (EBV), and human herpes virus 6 (HHV6)] and when administered to allogeneic hematopoietic stem cell (HCT) or solid organ transplant (SOT) recipients have been associated with clinical benefit. However, despite mediating potent antiviral effects in vivo, capturing in vitro cytotoxic potential has proven difficult in a traditional 51Cr release assay. Now, in addition to cytotoxicity surrogates, including CD107a and Granzyme B, we report on an alternative, vital dye -based, flow cytometric platform in which superior sensitivity and prolonged effector:target co-culture duration enabled the reliable detection of both CD4- and CD8-mediated in vitro cytolytic activity against viral targets without non-specific effects.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1299512/fullviral infectionadoptive T cell immunotherapyvirus specific T cells (VSTs)potency assaysT cell cytotoxicitychromium release assay |
| spellingShingle | Kiriakos Koukoulias Penelope G. Papayanni Julia Jones Manik Kuvalekar Ayumi Watanabe Yovana Velazquez Sarah Gilmore Anastasia Papadopoulou Ann M. Leen Spyridoula Vasileiou Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay Frontiers in Immunology viral infection adoptive T cell immunotherapy virus specific T cells (VSTs) potency assays T cell cytotoxicity chromium release assay |
| title | Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay |
| title_full | Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay |
| title_fullStr | Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay |
| title_full_unstemmed | Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay |
| title_short | Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay |
| title_sort | assessment of the cytolytic potential of a multivirus targeted t cell therapy using a vital dye based flow cytometric assay |
| topic | viral infection adoptive T cell immunotherapy virus specific T cells (VSTs) potency assays T cell cytotoxicity chromium release assay |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1299512/full |
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