Identification of T:B Cell Multimers After Bispecific T Cell Engagement, Using Both Conventional and Imaging Flow Cytometry

Davis, JE; Ludford-Menting, M; Peng, H; Johnson, DN; Ritchie, DS; Koldej, R

Author(s): Davis, JE; Ludford-Menting, M; Peng, H; Johnson, DN; Ritchie, DS; Koldej, R;
Details: Publication Year 2026-06,Volume 109,Issue #6,Page 425-435
Journal Title: Cytometry Part A
Publication Type: Research article
Abstract: Bispecific T cell engagers bring together T cells with malignant B cell targets to enable synapse formation and disease eradication of hematological malignancies. The proportion of T cells and their ability to bind to their targets varies between patients and may be a biomarker of response. In this study, we developed a method to detect T and B cell conjugates, as measured by conventional flow cytometry, and confirmed using imaging flow cytometry. After bispecific antibody engagement, multiple T cells were bound to each target cell in complexes comprising CD4+, CD8+, and CD4+ and CD8+ T cells. Imaging flow cytometry confirmed that the number and size of multimers increased significantly in the presence of bispecific antibodies. Conventional cytometry misidentified the memory phenotype of T cells bound to target cells due to augmented co-expression of CD45RA and CCR7 cell surface memory markers. Imaging flow cytometry verified conventional flow cytometry data showing increased T:B synapse and multimer formation after incubation with bispecific T cell engagers. Therefore, both flow cytometry platforms are suitable for identification of T cell: target cell conjugates.
Publisher: Wiley
Keywords: *Flow Cytometry/methods; Humans; *Antibodies, Bispecific/immunology; *B-Lymphocytes/immunology; *T-Lymphocytes/immunology; Leukocyte Common Antigens/immunology; CD4-Positive T-Lymphocytes/immunology; Receptors, CCR7/immunology; CD8-Positive T-Lymphocytes/immunology; bispecific T cell engager; blinatumomab; conventional flow cytometry; imaging flow cytometry; multimer identification
Department(s): Haematology
Publisher's Version: https://doi.org/10.1002/cyto.a.70039
Open Access at Publisher's Site: https://doi.org/10.1002/cyto.a.70039
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2026-06-02 05:11:24

Last Modified: 2026-08-04 12:07:23