Using Spectral Flow Cytometry to Characterize Anti-Tumor Immunity in Orthotopic and Subcutaneous Mouse Models of...

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Mouse models are crucial for immuno-oncology research, but existing flow cytometry panels often lack the scope to capture complex anti-cancer immune responses.
• Current panels frequently focus on limited leukocyte subsets and analyze systemic samples (blood, spleen), neglecting the critical tumor microenvironment (TME).

Purpose of the Study:

• To develop and optimize a comprehensive spectral flow cytometry panel for identifying key immune cell types involved in anti-cancer immunity.
• To validate the panel's utility in analyzing immune cells from both the tumor microenvironment and peripheral immune sites in mouse models.

Main Methods:

• Optimization of a spectral flow cytometry panel using immune cells derived from pancreatic tumors (orthotopic and subcutaneous) in mouse models.
• Validation of the panel in peripheral immune districts including blood, spleen, and liver of tumor-bearing mice.
• Establishment of protocols for tumor induction, single-cell suspension preparation, and cell staining.

Main Results:

• The optimized panel effectively identifies major immune cell types within the tumor microenvironment, offering a more holistic view of anti-cancer immunity.
• The panel demonstrated flexibility and suitability across different mouse cancer models and validated for use with various immune tissues.
• Comparative analysis between local tumor immunity and systemic immunity is facilitated by the panel's validation in peripheral immune districts.

Conclusions:

• This optimized spectral flow cytometry panel provides a powerful tool for in-depth analysis of anti-cancer immunity in mouse models.
• The panel's ability to analyze both local (TME) and systemic immunity enhances our understanding of immune responses to cancer and aids in the development of novel immuno-oncology strategies.