Work

OMIP‐119 : A 36‐Color Full‐Spectrum Flow Cytometry Panel for Deep Immunophenotyping of Peripheral Blood and Ex Vivo Expanded Human T Cells

Year: 2025

Type: article

Abstract: ABSTRACT The increasing uptake of adoptive CAR‐T and TCR‐T cell therapies into clinical practice has intensified the need for robust and detailed immunophenotyping of ex vivo expanded T cells. We have developed and optimized an extended 36‐color full‐spectrum flow cytometry panel suitable for in‐depth immunophenotyping of both peripheral blood and ex vivo expanded human T cells. Our panel allows for analysis of CD4 + and CD8 + memory subpopulations (Tn/Tscm, Tcm, Tem, and Temra), Treg subsets (CD45RA + and CD39 + ), and helper T cell subsets (Tfh, Th1, Th2, and Th17). Intracellular staining facilitates the evaluation of functional markers granzyme B (cytotoxicity) and Ki‐67 (proliferation). Our panel allows detailed investigation of T cell activation using markers carrying a range of expression kinetics. Exhaustion and senescence features, known to correlate with resistance to immunotherapies, can also be thoroughly evaluated. TCR staining with pHLA‐dextramer or anti‐TCR antibody is also included to allow evaluation of TCR specificity and/or transduction efficiency. A PD‐1 receptor occupancy component enables quantification of T cells bound to antibodies from immune checkpoint inhibitor (ICI) therapies. Overall, we have generated a fully optimized and fit‐for‐purpose extended T cell profiling panel with particular relevance to the field of immunotherapies including ICIs and adoptive T cell therapies.

Source: Cytometry Part A

DOI: 10.1002/cytoa.70001

Authors: Robert J. Harris, Benjamin Arkle, Elizabeth Evans, Charles H. V. Smith, Rebecca Teevan, Natalie Bath

Institution: Adaptimmune (United Kingdom)

Language: English

Topic: T-cell and B-cell Immunology

Subfield: Immunology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,970