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Related Experiment Video

Updated: Jun 30, 2026

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Published on: October 30, 2017

A high-throughput chemotaxis assay for assessing immunotoxicity using multiple human immune cell types.

Drake W Phelps^1,2, Nadia Barbo^1,2, Stephanie N Caty^1,2

¹Center for Computational Toxicology and Exposure, Office of Research and Development, United States Environmental Protection Agency, Research Triangle Park, NC, US.

NAM Journal

|June 29, 2026

Summary

A new in vitro assay effectively measures immune cell chemotaxis, a key indicator of immunotoxicity. This new approach methodology (NAM) aids in identifying harmful chemicals and reduces animal testing.

Keywords:

Chemotaxis Immunotoxicity Neutrophil T cell

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Published on: May 24, 2024

Area of Science:

Toxicology
Immunology
Cell Biology

Background:

New Approach Methodologies (NAMs) are essential for assessing chemical immunotoxicity, addressing data gaps, and minimizing animal use.
Current immunotoxicity testing relies on low-throughput rodent models that do not fully assess immune cell functions like chemotaxis.
Chemotaxis alteration is a critical hallmark of immunotoxicant action, yet high-throughput methods for its measurement are lacking.

Purpose of the Study:

To develop and validate a novel in vitro assay for measuring immune cell chemotaxis.
To establish a high-throughput screening method for identifying immunotoxicants that affect chemotaxis.
To assess the suitability of Jurkat T cells and HL60 (nHL60) cells for immunotoxicity screening.

Main Methods:

Development of an in vitro chemotaxis assay using transwell plates and automated high-content imaging.
Testing of Jurkat T cells and nHL60 cells with specific chemoattractants (CXCL12 and fMLP).
Evaluation of the effects of inhibitors (Cytochalasin D, CXCR4 inhibitor, cyclosporin A, rapamycin) on cell chemotaxis.

Main Results:

Jurkat T cells and nHL60 cells exhibited biphasic chemotaxis responses to CXCL12 and fMLP, respectively.
Cytochalasin D inhibited chemotaxis in both cell types in a concentration-dependent manner.
Cyclosporin A and a CXCR4 inhibitor reduced Jurkat cell chemotaxis, while rapamycin and cyclosporin A did not affect nHL60 cell chemotaxis.

Conclusions:

The developed in vitro assay is a promising New Approach Methodology (NAM) for assessing immunotoxicity by measuring chemotaxis.
Jurkat T cells and nHL60 cells demonstrate potential suitability for screening immunotoxicants affecting chemotaxis.
Further studies are required to fully determine the sensitivity and specificity of this assay for broad immunotoxicity screening.