Optimizing protocols for human regulatory T isolation, expansion, and characterization

• Published in: Methods in cell biology Vol. 191; pp. 59 - 77
• Main Authors: Inés, Sánchez-Moreno, Celia, Martín-Otal, Lasarte, Juan José, Teresa, Lozano
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 2025
• Subjects: Human regulatory T cells; In vitro Treg suppression assay; Human regulatory T cells; In vitro Treg suppression assay
• ISSN: 0091-679X
• DOI: 10.1016/bs.mcb.2024.10.005

Regulatory T cells (Tregs) are a subset of CD4+ T lymphocytes known for their immunosuppressive function, playing a vital role in maintaining peripheral tolerance and controlling autoimmune responses. Despite their beneficial roles, Tregs can hinder specific anti-tumor immune responses, contributing to tumor growth and progression. Elevated Treg levels within the tumor microenvironment are associated with poorer prognosis and reduced survival rates in various human cancers. Conversely, the therapeutic potential of Treg cells in managing autoimmune diseases, transplant rejection, graft versus host disease (GvHD) and allergic has gained increasing attention. This interest has prompted the investigation of novel treatments focused on inhibiting or eliminating Tregs to enhance anti-tumor immunity, while also considering their potential application as adoptive cell therapy for autoimmune diseases. Establishing standardized protocols for obtaining regulatory T cells is crucial for efficiently evaluating and implementing targeted therapies. The development of a simple protocol for the large-scale generation of human regulatory T cells, along with methods to assess the efficacy of new drugs that inhibit Treg activity, is detailed herein.