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A new generation of sensitive T cell-based assays facilitates the direct quantitation and characterization of antigen-specific T cell responses. Single-cell analyses have focused on measuring the quality and breadth of a response. Accumulating data from these studies demonstrate that there is considerable, previously-unrecognized, heterogeneity. Standard assays, such as the ICS, are often insufficient for characterization of rare subsets of cells. Enhanced flow cytometry with imaging capabilities enables the determination of cell morphology, as well as the spatial localization of the protein molecules within a single cell. Advances in both microfluidics and digital PCR have improved the efficiency of single-cell sorting and allowed multiplexed gene detection at the single-cell level. Delving further into the transcriptome of single-cells using RNA-seq is likely to reveal the fine-specificity of cellular events such as alternative splicing (i.e., splice variants) and allele-specific expression, and will also define the roles of new genes. Finally, detailed analysis of clonally related antigen-specific T cells using single-cell TCR RNA-seq will provide information on pathways of differentiation of memory T cells. With these state of the art technologies the transcriptomics and genomics of Ag-specific T cells can be more definitively elucidated.

Original publication

DOI

10.3390/ijms160818878

Type

Journal article

Journal

Int J Mol Sci

Publication Date

12/08/2015

Volume

16

Pages

18878 - 18893

Keywords

antigen-specific T cells, digital PCR, microfluidics, mingle-cell RNA-seq, Antigens, Flow Cytometry, Genomics, High-Throughput Nucleotide Sequencing, Humans, Lymphocyte Activation, Microfluidic Analytical Techniques, Polymerase Chain Reaction, Single-Cell Analysis, T-Cell Antigen Receptor Specificity, T-Lymphocytes