Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells

Area of Science:

• Immunology
• Cell Biology
• Genomics

Background:

• T cell-based assays are crucial for understanding immune responses.
• Existing methods like intracellular cytokine staining (ICS) struggle to characterize rare T cell subsets.
• Significant heterogeneity exists within antigen-specific T cell populations.

Purpose of the Study:

• To explore advanced single-cell technologies for T cell analysis.
• To characterize the heterogeneity of antigen-specific T cell responses.
• To elucidate T cell differentiation and function at a single-cell level.

Main Methods:

• Enhanced flow cytometry with imaging capabilities.
• Microfluidics and digital PCR for single-cell sorting and multiplexed gene detection.
• Single-cell RNA sequencing (RNA-seq) and TCR RNA-seq.

Main Results:

• Advanced techniques reveal previously unrecognized heterogeneity in T cell responses.
• Single-cell RNA-seq elucidates splice variants and allele-specific expression.
• TCR RNA-seq provides insights into memory T cell differentiation pathways.

Conclusions:

• State-of-the-art technologies enable definitive elucidation of T cell transcriptomics and genomics.
• These methods enhance the characterization of rare T cell subsets.
• Deeper understanding of T cell heterogeneity and function is achievable.