Efficient Tonsillar T Follicular Helper Cell Processing and Functional Analysis through High-dimensional Flow...

Area of Science:

• Immunology
• Cell Biology

Background:

• T follicular helper (Tfh) cells are critical for adaptive humoral immunity, influencing B cell responses in infection, autoimmunity, and vaccination.
• Understanding Tfh cell development and function is key for vaccine design and targeted therapies for immune-related diseases.

Purpose of the Study:

• To provide a comprehensive protocol for the collection, isolation, preservation, and culture of human tonsil cells.
• To introduce optimized spectral flow cytometry panels for detailed Tfh cell characterization.
• To demonstrate the utility of high-dimensional data analysis for uncovering treatment-induced immune cell differences.

Main Methods:

• Human tonsil tissue collection, cell isolation, and in vitro culture.
• Development and optimization of two spectral flow cytometry panels for Tfh cell phenotyping.
• Application of unsupervised analysis on high-dimensional flow cytometry data.

Main Results:

• A standardized protocol for human tonsil cell processing was established.
• Optimized flow cytometry panels enable in-depth characterization of Tfh cells and related immune populations.
• Unsupervised analysis successfully identified subtle, treatment-induced changes in high-dimensional immune cell data.

Conclusions:

• Human tonsil cells provide a valuable model for studying Tfh cell biology and immune responses.
• The presented protocol and flow cytometry panels facilitate robust Tfh cell research.
• High-dimensional data analysis is a powerful tool for discovering immune cell alterations in response to stimuli or treatments.