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Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages

Identifying pro-inflammatory macrophages (M1) is of immense importance to diagnose, monitor, and treat various pathologies. In addition, adoptive cell therapies, where harvested cells are isolated, modified to express an M1-like phenotype, then re-implanted to the patient, are also becoming more pre...

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Autores principales: Hernandez, Derek S., Schunk, Hattie C., Shankar, Karan M., Rosales, Adrianne M., Suggs, Laura J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: RSC 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9416964/
https://www.ncbi.nlm.nih.gov/pubmed/36132778
http://dx.doi.org/10.1039/d0na00373e
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author Hernandez, Derek S.
Schunk, Hattie C.
Shankar, Karan M.
Rosales, Adrianne M.
Suggs, Laura J.
author_facet Hernandez, Derek S.
Schunk, Hattie C.
Shankar, Karan M.
Rosales, Adrianne M.
Suggs, Laura J.
author_sort Hernandez, Derek S.
collection PubMed
description Identifying pro-inflammatory macrophages (M1) is of immense importance to diagnose, monitor, and treat various pathologies. In addition, adoptive cell therapies, where harvested cells are isolated, modified to express an M1-like phenotype, then re-implanted to the patient, are also becoming more prevalent to treat diseases such as cancer. In a step toward identifying, labeling, and monitoring macrophage phenotype for adoptive cell therapies, we developed a reactive oxygen species (ROS)-sensitive, gold nanoparticle (AuNP) that fluorescently labels M1 macrophages. AuNPs are electrostatically coated with a proteolysis resistant, fluorescein isothiocyanate-conjugated, poly-d-lysine (PDL-FITC) that is susceptible to backbone cleavage by ROS. When PDL-FITC is bound to AuNPs, fluorescence is quenched via a combination of nanoparticle surface (NSET) and Forster resonance (FRET) energy transfer mechanisms. Upon ROS-induced cleavage of PDL-FITC, up to a 7-fold change in fluorescence is demonstrated. PDL-FITC AuNPs were loaded into RAW 264.7 macrophages (RAWs) and primary bone marrow- derived macrophages (BMDMs) prior to in vitro polarization. For both cell types, detectable differences in intracellular fluorescence were observed between M1 polarized and non-stimulated (M0) control groups after 24 h using both confocal imaging and flow cytometry. PDL-FITC AuNPs can potentially be useful in identifying M1 macrophages within diverse cell populations and provide longitudinal macrophage response data to external cues.
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spelling pubmed-94169642022-09-20 Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages Hernandez, Derek S. Schunk, Hattie C. Shankar, Karan M. Rosales, Adrianne M. Suggs, Laura J. Nanoscale Adv Chemistry Identifying pro-inflammatory macrophages (M1) is of immense importance to diagnose, monitor, and treat various pathologies. In addition, adoptive cell therapies, where harvested cells are isolated, modified to express an M1-like phenotype, then re-implanted to the patient, are also becoming more prevalent to treat diseases such as cancer. In a step toward identifying, labeling, and monitoring macrophage phenotype for adoptive cell therapies, we developed a reactive oxygen species (ROS)-sensitive, gold nanoparticle (AuNP) that fluorescently labels M1 macrophages. AuNPs are electrostatically coated with a proteolysis resistant, fluorescein isothiocyanate-conjugated, poly-d-lysine (PDL-FITC) that is susceptible to backbone cleavage by ROS. When PDL-FITC is bound to AuNPs, fluorescence is quenched via a combination of nanoparticle surface (NSET) and Forster resonance (FRET) energy transfer mechanisms. Upon ROS-induced cleavage of PDL-FITC, up to a 7-fold change in fluorescence is demonstrated. PDL-FITC AuNPs were loaded into RAW 264.7 macrophages (RAWs) and primary bone marrow- derived macrophages (BMDMs) prior to in vitro polarization. For both cell types, detectable differences in intracellular fluorescence were observed between M1 polarized and non-stimulated (M0) control groups after 24 h using both confocal imaging and flow cytometry. PDL-FITC AuNPs can potentially be useful in identifying M1 macrophages within diverse cell populations and provide longitudinal macrophage response data to external cues. RSC 2020-07-13 /pmc/articles/PMC9416964/ /pubmed/36132778 http://dx.doi.org/10.1039/d0na00373e Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Hernandez, Derek S.
Schunk, Hattie C.
Shankar, Karan M.
Rosales, Adrianne M.
Suggs, Laura J.
Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title_full Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title_fullStr Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title_full_unstemmed Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title_short Poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
title_sort poly-d-lysine coated nanoparticles to identify pro-inflammatory macrophages
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9416964/
https://www.ncbi.nlm.nih.gov/pubmed/36132778
http://dx.doi.org/10.1039/d0na00373e
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