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Assessment of the cytolytic potential of a multivirus-targeted T cell therapy using a vital dye-based, flow cytometric assay.
Koukoulias, Kiriakos; Papayanni, Penelope G; Jones, Julia; Kuvalekar, Manik; Watanabe, Ayumi; Velazquez, Yovana; Gilmore, Sarah; Papadopoulou, Anastasia; Leen, Ann M; Vasileiou, Spyridoula.
Afiliação
  • Koukoulias K; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Papayanni PG; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Jones J; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Kuvalekar M; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Watanabe A; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Velazquez Y; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Gilmore S; AlloVir, Waltham, MA, United States.
  • Papadopoulou A; Hematology Department- Hematopoietic Cell Transplantation Unit, Gene and Cell Therapy Center, "George Papanikolaou" Hospital, Thessaloniki, Greece.
  • Leen AM; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
  • Vasileiou S; Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital and Houston Methodist Hospital, Houston, TX, United States.
Front Immunol ; 14: 1299512, 2023.
Article em En | MEDLINE | ID: mdl-38187380
ABSTRACT
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 effectortarget co-culture duration enabled the reliable detection of both CD4- and CD8-mediated in vitro cytolytic activity against viral targets without non-specific effects.
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Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Vírus BK / Infecções por Vírus Epstein-Barr Limite: Humans Idioma: En Revista: Front Immunol / Front. immunol / Frontiers in immunology Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Vírus BK / Infecções por Vírus Epstein-Barr Limite: Humans Idioma: En Revista: Front Immunol / Front. immunol / Frontiers in immunology Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos