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Related Experiment Video

Updated: Jul 23, 2025

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

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

Published on: March 10, 2017

A protocol for high-throughput screening for immunomodulatory compounds using human primary cells.

Katherine Chew¹, Branden Lee¹, Al Ozonoff²

¹Precision Vaccines Program, Division of Infectious Diseases, Boston Children's Hospital, Boston, MA, USA.

|July 15, 2023

Summary

This summary is machine-generated.

This study presents a 72-hour protocol using human peripheral blood mononuclear cells (PBMCs) to screen compounds for immune effects. It measures cytokine release and cell activation markers, aiding immunomodulator discovery.

Keywords:

Biotechnology and bioengineering Cell-based Assays Flow Cytometry/Mass Cytometry High-Throughput Screening Immunology

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Last Updated: Jul 23, 2025

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

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

Published on: March 10, 2017

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

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Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

Published on: July 22, 2011

Area of Science:

Immunology and Pharmacology
High-throughput screening methodologies

Background:

High-throughput screening (HTS) enables rapid assessment of compound libraries for cytotoxic, immunological, and phenotypical effects.
Human peripheral blood mononuclear cells (PBMCs) offer a relevant model for studying human immune responses in vitro.
Developing efficient protocols is crucial for leveraging HTS in drug discovery and immunomodulation research.

Purpose of the Study:

To establish and validate a 72-hour protocol for stimulating PBMCs and analyzing immune responses.
To enable simultaneous measurement of cytokine secretion and cell surface activation markers.
To facilitate the discovery of novel immunomodulators and vaccine adjuvant candidates.

Main Methods:

Cryopreserved PBMCs were stimulated for 72 hours with small molecule libraries.
Cytokine secretion (TNF-α, IFN-γ, IL-10) was quantified using AlphaLISA assays.
Cell surface activation markers (CD80, CD86, HLA-DR, OX40) were analyzed by flow cytometry.

Main Results:

The protocol successfully measured key cytokine secretion levels.
Simultaneous analysis of cell surface activation markers was achieved.
A multiplexed readout workflow was developed for comprehensive immune profiling.

Conclusions:

This validated protocol provides a robust platform for immunomodulatory compound screening.
The multiplexed approach aids in the phenotypic identification of potential therapeutic agents.
The method supports the discovery of novel vaccine adjuvants and immunomodulators.