A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

Area of Science:

• Immunology
• Drug Discovery
• Biotechnology

Background:

• High-throughput screening (HTS) is crucial for identifying bioactive small molecules in drug discovery.
• Despite advances, novel immunomodulatory compounds for transplantation and autoimmune diseases remain elusive.
• Oncology dominates drug screening, leaving a gap in immunomodulatory research.

Purpose of the Study:

• To develop and validate a novel in vitro fluorescent-based lymphocyte assay for identifying immunomodulatory compounds.
• To establish a screening tool for T- and B-cell receptor-mediated responses.
• To facilitate drug discovery programs targeting immune system modulation.

Main Methods:

• Utilized a transgenic murine model with GFP reporter expression under the Nur77 promoter.
• Developed an in vitro assay using T or B lymphocytes stimulated via their respective receptors.
• Quantified GFP intensity as a measure of cellular activation and transcriptional activity.

Main Results:

• Successfully established a fluorescent-based lymphocyte assay for screening compounds.
• Identified 160 potential immunomodulatory hits from a library of 4,398 compounds in a primary screen.
• Demonstrated the assay's utility in identifying compounds with immunomodulatory effects without a predefined target hypothesis.

Conclusions:

• The novel fluorescent assay is a valuable tool for drug discovery programs seeking immunomodulatory agents.
• This method enables efficient screening of large chemical libraries for potential therapeutic compounds.
• The assay supports further in vitro and in vivo validation studies for identified hits.