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Quantum sensing to monitor changes in free radical generation by intracellular vesicles of polarized macrophages
Aldona Mzyk1, Claudia Reyes-San-Martin2, Yasemin Doğan3
1Groningen University, University Medical Center Groningen, Antonius Deusinglaan 1, 9713AV Groningen, the Netherlands; Technical University of Denmark, Ørsteds Plads, 349, 2800 Kgs, Lyngby, Denmark.
Abstract:
Macrophages are immune cells crucial in clearing our tissues from bacteria, viruses, dying cells, cell debris and other waste products. They also regulate inflammation by differentiating from non-activated (M0) cells into macrophages that initiate inflammation (pro-inflammatory macrophages, M1), or resolve inflammation (anti-inflammatory macrophages, M2). One of their key functions is to ingest pathogens within vesicles where they are degraded. The production of free radical (FR) plays an important role in this degradation process but also in macrophage differentiation and signaling. Here we used diamond-based quantum sensing to track free radical changes in vesicles with nanoscale resolution. We further followed the oxidative stress status, through free radical measurement during the macrophage activation process. We found that the three macrophage subtypes differed significantly in free radical generation in their vesicles. Additionally, we showed that the FR generation evolves over time in the different subtypes. We observed a 50 % increase in radical production in M0 after 24 h compared to the T1 values measured after 4 h of cell culture, a decrease in M1 and constant radical levels in M2 macrophages. STATEMENT OF SIGNIFICANCE: Here we use quantum sensing for the first time to investigate the role that free radicals play in immune cells when they differentiate to fulfill their functions in the immune system. We were able to measure free radical generation specifically in vesicles while the macrophages differentiated.
Insights
Quantum sensing precisely measured free radical (FR) changes in immune cell vesicles during differentiation. Different macrophage subtypes (M0, M1, M2) showed distinct FR generation patterns, revealing insights into immune cell function and oxidative stress.
Area of Science:
- Immunology
- Cell Biology
- Quantum Technology
Background:
- Macrophages are vital immune cells involved in pathogen clearance and inflammation regulation.
- Free radical (FR) production is critical for macrophage functions, including pathogen degradation, differentiation, and signaling.
- Understanding FR dynamics during macrophage activation is essential for immune system research.
Purpose of the Study:
- To utilize diamond-based quantum sensing for nanoscale measurement of free radical changes within macrophage vesicles.
- To investigate the role of free radicals in macrophage differentiation and oxidative stress status.
- To quantify and compare free radical generation across different macrophage subtypes (M0, M1, M2) and over time.
Main Methods:
- Employing diamond-based quantum sensing for high-resolution free radical detection.
- Tracking oxidative stress levels through FR measurements during macrophage activation.
- Analyzing FR generation specifically within cellular vesicles of differentiating macrophages.
Main Results:
- Significant differences in vesicle free radical generation were observed among M0, M1, and M2 macrophage subtypes.
- Free radical production dynamics varied over time across the macrophage subtypes.
- M0 macrophages showed a 50% increase in radical production after 24h, M1 decreased, and M2 remained constant.
Conclusions:
- Quantum sensing provides unprecedented nanoscale resolution for studying free radicals in immune cell vesicles.
- This study demonstrates distinct free radical profiles associated with macrophage differentiation and activation states.
- Findings offer new insights into the role of oxidative stress in immune cell function and regulation.
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