Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells.
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.
Kirby Institute, University of New South Wales, 2031 Sydney, Australia. c.phetsouphanh@gmail.com.
Chansavath
C
Phetsouphanh
Kirby Institute, University of New South Wales, 2031 Sydney, Australia. jzaunders@kirby.unsw.edu.au.|Centre for Applied Medical Research, St. Vincent's Hospital, 2010 Sydney, Australia. jzaunders@kirby.unsw.edu.au.
John James
JJ
Zaunders
Kirby Institute, University of New South Wales, 2031 Sydney, Australia. akelleher@kirby.unsw.edu.au.|Centre for Applied Medical Research, St. Vincent's Hospital, 2010 Sydney, Australia. akelleher@kirby.unsw.edu.au.
Anthony Dominic
AD
Kelleher
D000941:Antigens / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods; D023281:Genomics / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D046210:Microfluidic Analytical Techniques; D016133:Polymerase Chain Reaction; D059010:Single-Cell Analysis / Q000379:methods; D037182:T-Cell Antigen Receptor Specificity / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*
D000941:Antigens
antigen-specific T cells; digital PCR; microfluidics; mingle-cell RNA-seq