A high-throughput chemotaxis detection method for CCR4⁺ T cell migration inhibition using image cytometry

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Chemotaxis, or cell movement, is vital in the tumor microenvironment (TME), influencing immune responses.
• Regulatory T cells (Tregs) migrate via chemotaxis, suppressing anti-tumor immunity and hindering immunotherapy.
• Current methods for studying chemotaxis are time-consuming, limiting high-throughput screening.

Purpose of the Study:

• To develop a high-throughput method for measuring immune cell chemotaxis in the TME.
• To screen for therapeutic antibodies that modulate immune cell recruitment.
• To assess the efficacy of anti-chemokine antibodies in inhibiting Treg migration.

Main Methods:

• Utilized the Celigo Image Cytometer for a novel image-based chemotaxis assay.
• Developed a method to directly count CCR4+ T cells migrating towards CCL17 and CCL22 chemokines in Transwell plates.
• Applied the assay to evaluate dose-dependent inhibition by anti-CCL17 and anti-CCL22 antibodies.

Main Results:

• The image cytometry method successfully quantified T cell migration induced by CCL17 and CCL22.
• Commercially available anti-CCL17 and anti-CCL22 antibodies demonstrated dose-dependent inhibition of T cell chemotaxis.
• The method enabled simultaneous screening of multiple antibodies at various concentrations.

Conclusions:

• The developed image cytometry assay provides an efficient, high-throughput platform for studying chemotaxis.
• This method accelerates the discovery of therapeutic antibodies targeting immune cell recruitment in the TME.
• The assay facilitates the identification of agents that can enhance or block immune cell migration for cancer immunotherapy.