Publication Date
1-1-2023
Journal
Frontiers in Immunology
DOI
10.3389/fimmu.2023.1299512
PMID
38187380
PMCID
PMC10766817
PubMedCentral® Posted Date
12-22-2023
PubMedCentral® Full Text Version
Post-print
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 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.
Keywords
Humans, Epstein-Barr Virus Infections, Herpesvirus 4, Human, Adenoviridae, BK Virus, Cell- and Tissue-Based Therapy, viral infection, adoptive T cell immunotherapy, virus specific T cells (VSTs), potency assays, T cell cytotoxicity, chromium release assay, flow cytometric analysis, vital dye staining
Published Open-Access
yes
Recommended Citation
Koukoulias, Kiriakos; Papayanni, Penelope G; Jones, Julia; et al., "Assessment of the Cytolytic Potential of a Multivirus-Targeted T Cell Therapy Using a Vital Dye-Based, Flow Cytometric Assay" (2023). Faculty and Staff Publications. 4984.
https://digitalcommons.library.tmc.edu/baylor_docs/4984
Included in
Health Services Research Commons, Medical Cell Biology Commons, Medical Molecular Biology Commons, Medical Specialties Commons, Microbiology Commons