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Updated: Mar 20, 2026
Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
Published on: January 30, 2019
Direct measurement of T cell receptor affinity and sequence from naïve antiviral T cells
Shu-Qi Zhang1, Patricia Parker2, Ke-Yue Ma3
1McKetta Department of Chemical Engineering, University of Texas at Austin, Austin, TX 78712, USA.
Abstract:
T cells recognize and kill a myriad of pathogen-infected or cancer cells using a diverse set of T cell receptors (TCRs). The affinity of TCR to cognate antigen is of high interest in adoptive T cell transfer immunotherapy and antigen-specific T cell repertoire immune profiling because it is widely known to correlate with downstream T cell responses. We introduce the in situ TCR affinity and sequence test (iTAST) for simultaneous measurement of TCR affinity and sequence from single primary CD8(+) T cells in human blood. We demonstrate that the repertoire of primary antigen-specific T cells from pathogen-inexperienced individuals has a surprisingly broad affinity range of 1000-fold composed of diverse TCR sequences. Within this range, samples from older individuals contained a reduced frequency of high-affinity T cells compared to young individuals, demonstrating an age-related effect of T cell attrition that could cause holes in the repertoire. iTAST should enable the rapid selection of high-affinity TCRs ex vivo for adoptive immunotherapy and measurement of T cell response for immune monitoring applications.
Insights
We developed a new test (iTAST) to measure T cell receptor (TCR) affinity and sequence simultaneously. Older individuals have fewer high-affinity T cells, suggesting age impacts immune response and potentially adoptive T cell therapy effectiveness.
Area of Science:
- Immunology
- Cellular Biology
- Biotechnology
Background:
- T cells utilize diverse T cell receptors (TCRs) to identify and eliminate infected or cancerous cells.
- TCR affinity to cognate antigen is crucial for T cell responses, impacting immunotherapy and immune profiling.
- Current methods lack simultaneous measurement of TCR affinity and sequence from single primary cells.
Purpose of the Study:
- To introduce a novel method, the in situ TCR affinity and sequence test (iTAST), for simultaneous measurement of TCR affinity and sequence.
- To analyze the affinity range and sequence diversity of antigen-specific T cells in primary human blood.
- To investigate age-related differences in T cell repertoire and high-affinity T cell frequency.
Main Methods:
- Development and application of the in situ TCR affinity and sequence test (iTAST).
- Simultaneous measurement of TCR affinity and sequence from single primary CD8(+) T cells in human blood.
- Analysis of T cell repertoires in pathogen-inexperienced individuals, comparing young and older cohorts.
Main Results:
- Primary antigen-specific T cells exhibit a broad, 1000-fold affinity range with diverse TCR sequences.
- Older individuals show a reduced frequency of high-affinity T cells compared to younger individuals.
- Evidence of age-related T cell attrition potentially creating gaps in the immune repertoire.
Conclusions:
- iTAST enables simultaneous measurement of TCR affinity and sequence from single primary T cells.
- The T cell repertoire shows significant age-related decline in high-affinity cells.
- iTAST can facilitate ex vivo selection of high-affinity TCRs for adoptive immunotherapy and immune monitoring.
