Controlling T cells spreading, mechanics and activation by micropatterning

Area of Science:

• Immunology
• Biophysics
• Cell Biology

Background:

• T cell activation is crucial for adaptive immunity.
• Controlling T cell adhesion and mechanics is vital for understanding immune responses.
• Existing methods lack precise control over T cell biophysical properties during activation.

Purpose of the Study:

• To develop a microstamping strategy for precise control over T cell adhesion, spreading, and mechanical properties.
• To investigate the impact of controlled adhesion on T cell activation.
• To establish a standardized platform for dissecting T cell activation mechanisms.

Main Methods:

• Protein microstamping to create patterned substrates for T cell adhesion.
• Atomic Force Microscopy (AFM) for measuring T cell mechanics (Young's modulus).
• Flow cytometry to assess T cell calcium fluxes and activation markers.
• Expression of MEGF10 as a non-immune adhesion receptor in T cells.

Main Results:

• Microstamping enabled precise control over T cell position, shape, and spreading.
• Adhesion to anti-CD45 antibody-decorated surfaces did not affect T cell calcium fluxes.
• T cells exhibited similar mechanical properties (Young's modulus) and spreading areas regardless of patterned adhesion.
• Engineered T cells retained activation capabilities through soluble anti-CD3 or model APC contacts.

Conclusions:

• The developed microstamping system provides a robust platform for standardized T cell activation assays.
• This system allows for the study of T cell activation and anergy with defined adhesion and mechanical characteristics.
• It facilitates the dissection of fine details in T cell activation mechanisms by observing homogenized cell populations.