Luo et al. developed ATLAS-seq (Aptamer-based T-Lymphocyte Activity Screening and Sequencing), a microfluidic-based high-throughput platform for isolating and profiling antigen-reactive T cells. The system utilizes cholesterol-modified aptamer beacons to detect cytokine secretion, specifically IFNγ, from single activated T cells upon antigen stimulation, and ensures precise screening and TCR clonotype profiling. Compared to MHC multimer staining, ATLAS-seq identified distinct TCR clonotype populations with superior activation potential for cytomegalovirus (including superior cytotoxicity), and PSA-specific T cells.
Contributed by Shishir Pant
ABSTRACT: Discovering antigen-reactive T cell receptors (TCRs) is central to developing effective engineered T cell immunotherapies. However, the conventional technologies for isolating antigen-reactive TCRs (i.e., major histocompatibility complex (MHC) multimer staining) focus on high-affinity interactions between the TCR and MHC-antigen complex, and may fail to identify TCRs with high efficacy for activating T cells. Here, we develop a microfluidic single-cell screening method for antigen-reactive T cells named ATLAS-seq (Aptamer-based T Lymphocyte Activity Screening and SEQuencing). This technology isolates and characterizes activated T cells via an aptamer-based fluorescent molecular sensor, which monitors the cytotoxic cytokine IFN_ secretion from single T cells upon antigen stimulation, followed by single-cell RNA and single-cell TCR sequencing. We use ATLAS-seq to screen TCRs reactive to cytomegalovirus (CMV) or prostate specific antigen (PSA) from peripheral blood mononuclear cells (PBMCs). ATLAS-seq identifies distinct TCR clonotype populations with higher T cell activation levels compared to TCRs recovered by MHC multimer staining. Select TCR clonotypes from ATLAS-seq are more efficient in target cell killing than those from MHC multimer staining. Collectively, ATLAS-seq provides an efficient and broadly applicable technology to screen antigen-reactive TCRs for engineered T cell immunotherapy.
Author Info: (1) Raymond G. Perelman Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. Center for Computational and Genomic Medicine, C
Author Info: (1) Raymond G. Perelman Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. Center for Computational and Genomic Medicine, Children's Hospital of Philadelphia, Philadelphia, PA, USA. (2) Raymond G. Perelman Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. Center for Computational and Genomic Medicine, Children's Hospital of Philadelphia, Philadelphia, PA, USA. (3) Raymond G. Perelman Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. linlan@chop.edu. Center for Computational and Genomic Medicine, Children's Hospital of Philadelphia, Philadelphia, PA, USA. linlan@chop.edu. Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA. linlan@chop.edu.
