As engineered immune cell therapies advance, researchers need tools capable of resolving complex T-cell behavior at the single-cell level. The Beacon® Discovery Optofluidic platform enables real-time functional profiling of individual immune cells through integrated measurements of cytotoxicity, cytokine secretion, activation state, and cell-cell interactions, helping provide deeper insight into immune response dynamics.

Allogeneic CAR T-cell therapy holds curative potential for patients with advanced B-cell malignancies following allogeneic hematopoietic stem cell transplantation (allo-HSCT), though broader clinical application remains constrained by the risk of graft-versus-host disease (GvHD). Dr. Inchingolo will present new findings investigating the mechanistic basis underlying the low incidence of acute GvHD previously observed in donor-derived anti-CD19 CAR T-cell clinical studies, including in patients receiving cells from HLA-mismatched unrelated donors.

Using the Beacon Discovery platform, donor-derived alloreactive CAR T cells were profiled at single-cell resolution through real-time measurements of cytotoxicity, cytokine secretion, and activation/exhaustion marker expression under CAR- and TCR-mediated stimulation conditions. The study demonstrated that while CAR-mediated cytotoxicity and killing kinetics remained preserved under dual-antigen stimulation, TCR-driven responses were markedly attenuated, with reduced cytotoxicity and prolonged time-to-kill compared with single TCR stimulation conditions.

By integrating single-cell transcriptomics with TCR sequencing of functionally defined exported alloreactive CAR T cells, this work aims to further characterize the transcriptional states and clonotypic dynamics associated with simultaneous CAR and TCR stimulation, helping guide the development of safer and more effective next-generation allogeneic CAR T-cell therapies.


KEY TAKEAWAYS

  • Learn how donor-derived allogeneic CAR T cells maintain potent tumor killing while suppressing harmful alloreactive responses linked to GvHD.
  • Discover how the Beacon Discovery Optofluidic Platform enables real-time single-cell analysis of cytotoxicity, cytokine secretion, activation state, and immune cell interactions.
  • See how integrated functional profiling and TCR sequencing reveal the mechanisms driving safer and more effective next-generation CAR T-cell therapies.
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Gabriele Inchingolo, Ph.D.
Ph.D. Candidate
University Hospital Regensburg
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About Speaker:
Gabriele Inchingolo, PhD, specializes in engineered T cell therapy for cancer, with a focus on the mechanisms governing T cell differentiation, exhaustion, and memory formation. Gabriele has presented his work at numerous international conferences and has been the recipient of multiple prestigious awards. He began his research career at the San Raffaele Scientific Institute (Milan, Italy) in Prof. Chiara Bonini’s laboratory, where he focused on engineering TCR-redirected memory stem T cells (TSCM) to resist tumor immunosuppressive mechanisms. He later transitioned to industry, joining EsoBiotec (Belgium, AstraZeneca) where he contributed to the development of in vivo CAR T cell therapies.

Gabriele obtained his PhD at the Leibniz Institute for Immunotherapy (Germany) under the supervision of Prof. Luca Gattinoni. His doctoral research provided the first in-depth immunomonitoring of patients treated with donor-derived CD19 CAR-modified TSCM, a landmark study recently published in Cell. In parallel, Gabriele is conducting the first comprehensive study to extensively characterize alloreactive CAR T cell dynamics directly in patients following allogeneic CAR T cell therapy, unveiling the mechanisms that limit graft-versus-host disease. This work was selected for an oral presentation at the American Society of Hematology Annual Meeting, where it received a Best Abstract Award, and was further recognized with a Beacon Discovery Global Grant from Bruker, as the sole European awardee.

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