Linking antigen specific T-cell dynamics in a microfluidic chip to single cell transcription patterns

Area of Science:

• Immunology
• Biotechnology
• Microfluidics

Background:

• Adoptive immunotherapy requires precisely activated T-cells for effective cancer recognition and elimination.
• Current methods for assessing T-cell activation can be insufficient for single-cell analysis and therapeutic optimization.

Purpose of the Study:

• To develop a non-invasive screening profile for determining T-cell activation state at the single-cell level.
• To enhance the efficiency and specificity of antigen-specific T-cell selection for adoptive immunotherapy.

Main Methods:

• Utilizing a microfluidic chip to analyze intracellular Ca2+ intensity and T-cell motility during interaction with Antigen Presenting Cells (APCs).
• Testing the system with OVA257-264 peptide-pulsed APCs and modified affinities, using T-cells from OT-1 mice.
• Employing single-cell RNA sequencing to correlate cellular profiles with T-cell activation states.

Main Results:

• A novel screening profile accurately determines T-cell activation states at the single-cell level.
• The microfluidic system demonstrated high efficiency in screening antigen-specific T-cells with stable proliferation.
• Single-cell RNA sequencing data corroborated the activation states identified by the screening profile.

Conclusions:

• The developed non-invasive screening system is versatile for advancing T-cell-based immunotherapy.
• This method enables efficient and high-proliferation screening of antigen-specific T-cells, crucial for therapeutic applications.
• The integration of Ca2+ intensity, motility, and transcriptomic data provides a comprehensive assessment of T-cell activation.