T cells play many critical roles in immune surveillance and defense against infections and cancer. Over the past few decades, improved understanding of T cell functionality has led to considerable breakthroughs in fields such as cancer immunotherapy, autoimmune diseases, and infectious diseases. Recent breakthroughs, including the first FDA approved TCR-T therapy and advanced designs of CAR-Ts to target solid tumors and autoimmune disease signal a promising future for T cell research.
Despite recent technological advances, researchers have only begun to scratch the surface of our potential to harness T cells for therapeutics. As the ambition for T cell centric therapy evolves, so to do the tools and techniques researchers need to fully understand T cells. In this blog, we’ll explore emerging trends in technologies used for T cell research and dive into how these trends will shape the future of T cell research.
Single-Cell Technologies: Unlocking Insights into T Cell Heterogeneity
Ensemble and bulk measurements provide cheap and easily accessible information on T cells but population averages have been shown to oftentimes mask the presence of rare cellular subpopulations with unique roles in disease and therapy.[1, 2] As such, recent research is trending towards incorporating single-cell platforms to garner improved biological resolution and capture critical details about individual cells. With PubMed entries including “single-cell” as a keyword increasing by over 500% in the past decade, it’s clear that single-cell technologies such as scRNA-seq, mass cytometry, and a variety of technologies looking at single-cell functions such as cytokine secretion, cytotoxicity, and metabolomics are pushing research forward at a rapid pace.
Functional Behavior: Moving Beyond Transcriptomics and Genomics
While genomic and transcriptomic data have been invaluable for understanding T cell biology, it is becoming increasingly clear that they cannot provide the full story about T cell behavior alone. In particular, the correlation between mRNA and protein levels has been shown to vary dramatically depending on a wide variety of factors such as cell subsets and functional status.[3] Given some of the limitations of sequencing, research is now shifting toward incorporating more functional data to directly characterize T cell activity. Functional assays, which measure a wide variety of data such as cytokine secretion, cytotoxic activity, and metabolic states, offer a more comprehensive view of T cell performance and can be used to directly assess the behaviors of T cells that directly contribute to immune responses.
Real Time Data: Monitoring T Cell Dynamics
T cells function dynamically with functional behaviors such as cytokine secretion, cytotoxicity, and surface marker expression intimately linked with outcomes such as activation, persistence, and exhaustion. These behaviors occur on a temporal spectrum that cannot be picked up using technologies reliant on single timepoint data such as flow cytometry and scRNA-seq. Critically, studies have shown that potency of T cells and engineered T cells can be linked to fast or slow cytotoxicity which highlights that designing effective cellular therapeutics may rely on dynamic measurements to optimize therapeutic potential.[4]
T cell research promises to revolutionize the future of immune therapeutics and more advanced tools are needed to fully understand the behavior of individual cells and harness their potential. One technology designed to simultaneously take advantage of all these trends simultaneously is the Beacon® Optofluidic Platform. The Beacon platform is a single-cell technology focused on providing real time functional data on individual cells. The Beacon instrument automatically isolates single T cells into 0.74 nL NanoPens and can perform multiple assays to interrogate single-cell function. For example, co-culturing a single T cell with a target cell can showcase the cytotoxicity, cytokine secretion, and surface marker expression capabilities of thousands of individual cells simultaneously. After performing assays, the Beacon instrument can also retrieve cells of interest to perform multiomic sequencing and reveal the transcriptomic drivers of functional behaviors.

References
- [1] Altschuler, S.J. and L.F. Wu, Cellular heterogeneity: do differences make a difference? Cell, 2010. 141(4): p. 559-563.
- [2] Colina, A.S., et al., Current advances in experimental and computational approaches to enhance CAR T cell manufacturing protocols and improve clinical efficacy. Front Mol Med, 2024. 4: p. 1310002.
- [3] Weerakoon, H., et al., Integrative temporal multi-omics reveals uncoupling of transcriptome and proteome during human T cell activation. npj Systems Biology and Applications, 2024. 10(1): p. 21.
- [4] Liadi, I., et al., Defining potency of CAR(+) T cells: Fast and furious or slow and steady. Oncoimmunology, 2019. 8(10): p. e1051298.