A mass cytometry method pairing T cell receptor and differentiation state analysis.

Area of Science:

• Immunology
• Cellular Biology
• Mass Cytometry (CyTOF)

Background:

• Adaptive immune responses rely on T cell antigen receptor (TCR) recognition and clonal expansion.
• Understanding T cell dynamics during infection and disease is crucial for immunology.
• Current methods may not fully capture TCR diversity alongside T cell activation states.

Purpose of the Study:

• To develop and validate a mass cytometry (CyTOF) approach for tracking T cell responses.
• To link specific T cell receptor (TCR) Vα and Vβ chains to antigen recognition.
• To analyze T cell differentiation states and TCR diversity in response to various stimuli.

Main Methods:

• Utilized mass cytometry (CyTOF) with antibodies targeting TCR Vα/Vβ chains.
• Combined TCR staining with antibodies for T cell activation and differentiation markers.
• Applied the method to mouse models of Listeria monocytogenes infection, tumors, and influenza.

Main Results:

• Identified expansions of CD8+ and CD4+ T cells with specific TCR Vα/Vβ chains during immune responses.
• Linked expanded T cell populations to the recognition of specific antigens from Listeria, tumors, and influenza.
• Observed alterations in TCR diversity and T cell differentiation due to vaccination and serum transfer in influenza infection.

Conclusions:

• The developed CyTOF method enables comprehensive monitoring of TCR usage and T cell phenotypes.
• This approach allows direct correlation of specific TCRs with antigen recognition in vivo.
• Therapeutic interventions can significantly modulate TCR repertoire and T cell differentiation dynamics.