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Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
Derek S Hernandez1, Hattie C Schunk1,2, Karan M Shankar1
1Department of Biomedical Engineering, University of Texas at Austin Austin TX 78712 USA suggs@utexas.edu.
Nanoscale Advances
|September 22, 2022
Summary
Researchers developed a novel gold nanoparticle (AuNP) probe to detect pro-inflammatory M1 macrophages. This ROS-sensitive probe offers a new method for monitoring macrophage phenotypes in adoptive cell therapies and disease diagnostics.
Area of Science:
- Nanotechnology
- Immunology
- Biomedical Engineering
Background:
- Identifying pro-inflammatory M1 macrophages is crucial for diagnosing and treating various pathologies.
- Adoptive cell therapies, including those targeting cancer, increasingly utilize M1-like phenotype cells.
- Accurate labeling and monitoring of macrophage phenotypes are essential for these applications.
Purpose of the Study:
- To develop a reactive oxygen species (ROS)-sensitive gold nanoparticle (AuNP) for fluorescently labeling M1 macrophages.
- To enable identification, labeling, and monitoring of macrophage phenotypes for adoptive cell therapies.
- To assess the utility of the developed probe for distinguishing M1 macrophages from other cell types.
Main Methods:
- Developed ROS-sensitive AuNPs coated with poly-d-lysine conjugated to fluorescein isothiocyanate (PDL-FITC).
- Utilized nanoparticle surface (NSET) and Forster resonance (FRET) energy transfer mechanisms for fluorescence quenching.
- Loaded PDL-FITC AuNPs into RAW 264.7 macrophages and primary bone marrow-derived macrophages (BMDMs) for in vitro polarization studies.
Main Results:
- ROS-induced cleavage of PDL-FITC resulted in up to a 7-fold increase in fluorescence.
- Confocal imaging and flow cytometry revealed detectable differences in intracellular fluorescence between M1 polarized and M0 control macrophages after 24 hours.
- The probe successfully distinguished between M1 and M0 macrophage phenotypes in both cell types.
Conclusions:
- PDL-FITC AuNPs provide a sensitive and specific method for identifying M1 macrophages.
- This probe can be valuable for monitoring macrophage responses in diverse cell populations and in vivo.
- The developed nanoparticle offers potential for improved diagnostics and longitudinal tracking in adoptive cell therapies.
