The DOI matches its Crossref record; 15 of the 16 references it cites were found in a registry or an index.
A novel method to determine the engulfment of apoptotic cells by macrophages using pHrodo succinimidyl ester
Open accessAbstract
Apoptotic cell phagocytosis has recently raised considerable interest, particularly due to its intricate molecular mechanisms and negative immunologic impact of incompetent clearance of apoptotic cells. There is a need for simple and reliable methods to clearly determine the internalization of apoptotic cells. Labeling with pHrodo succinimidyl ester (SE), a pH-sensitive fluorescent dye, makes engulfed apoptotic cells detectable due to the increased post-phagocytic light emission. This is a valuable tool for phagocytosis studies via FACS. We designed an ex vivo assay, using apoptotic pHrodo-labeled lymphocytes as prey and anti-CD11b-labeled tissue macrophages. To demonstrate its validity of detecting internalized apoptotic lymphocytes, we used MFGE8(-/-) macrophages, known to have impaired phagocytic ability. Uptake of apoptotic lymphocytes was accelerated and enhanced in splenic macrophages after stimulation with recombinant MFGE8, while peritoneal macrophages were able to compensate for the delayed uptake. This novel assay is a quick and reliable method to evaluate the internalization of apoptotic cells.
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“Because pHrodo only fluoresces once it enters acidic compartments of lysosomes, any pHrodo-positive fluorescence within a microglia confirms phagocytosis of RGC inputs.”
From Microglia Sculpt Postnatal Neural Circuits in an Activity and Complement-Dependent Manner (2012) · cites this paper as Miksa et al., 2009
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“pHrodo only fluoresces significantly when in the low pH environment of lysosomes (39).”
From LRPAP1 is released from activated microglia and inhibits microglial phagocytosis and amyloid beta aggregation (2023) · cites this paper as [39]
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A blast from the past: clearance of apoptotic cells regulates immune responses
Savill, J., Dransfield, I., Gregory, C. D., & Haslett, C. (2002) · Nature reviews. Immunology · doi:10.1038/nri957 · Cited by 1,574
Identification of a factor that links apoptotic cells to phagocytes
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Regulation of Phagosome Maturation by Signals from Toll-Like Receptors
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Engulfment of apoptotic cells: signals for a good meal
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Bacterial strategies for overcoming host innate and adaptive immune responses
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Lactadherin Deficiency Leads to Apoptotic Cell Accumulation and Accelerated Atherosclerosis in Mice
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100 years of the phagocytosis doctrine (Ilya Ilyich Mechnikov).
Nn, K. (1983) · PubMed · Cited by 1
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Microglia Sculpt Postnatal Neural Circuits in an Activity and Complement-Dependent Manner
Schafer, D. P., Lehrman, E. K., Kautzman, A. G., Koyama, R., Mardinly, A. R., Yamasaki, R., Ransohoff, R. M., Greenberg, M. E., Barres, B. A., & Stevens, B. (2012) · Neuron · doi:10.1016/j.neuron.2012.03.026 · Cited by 4,089
“To further validate that microglia phagocytose RGC inputs, pHrodo-dextran, an anterograde tracer and pH-sensitive dye, was used to label RGC inputs (Figures S1A and S1B; Deriy et al., 2009; Miksa et al., 2009).”
Fluorescent Indicators for Intracellular pH
Han, J., & Burgess, K. (2009) · Chemical Reviews · doi:10.1021/cr900249z · Cited by 1,623
CD24 signalling through macrophage Siglec-10 is a target for cancer immunotherapy
Barkal, A. A., Brewer, R. E., Markovic, M., Kowarsky, M., Barkal, S. A., Zaro, B. W., Krishnan, V., Hatakeyama, J., Dorigo, O., Barkal, L. J., et al. (2019) · Nature · doi:10.1038/s41586-019-1456-0 · Cited by 1,409
Apoptosis and Clearance of Apoptotic Cells
Nagata, S. (2018) · Annual Review of Immunology · doi:10.1146/annurev-immunol-042617-053010 · Cited by 1,135
Sellgren, C. M., Gracias, J., Watmuff, B., Biag, J. D., Thanos, J. M., Whittredge, P. B., Fu, T., Worringer, K. A., Brown, H. E., Wang, J. P., et al. (2019) · Nature Neuroscience · doi:10.1038/s41593-018-0334-7 · Cited by 810
Metadata
- Authors
- Miksa, M., Komura, H., Wu, R., Shah, K. G., & Wang, P.
- Affiliations
- Feinstein Institute for Medical Research (US)
- Corresponding author
- Wang, Ping
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 2009-03
- Volume
- 342
- Issue
- 1-2
- Pages
- 71-77
- ISSN
- 0022-1759 (ISSN-L)
- PMC
- PMC2675277
- OpenAlex
- W2100326315
- Cited by
- 320
- References
- 16
- Citations by year
- 2026: 11 · 2025: 14 · 2024: 23 · 2023: 12 · 2022: 17 · 2021: 39 · 2020: 15 · 2019: 19 · 2018: 26 · 2017: 25 · 2016: 23 · 2015: 32 · 2014: 19 · 2013: 15 · 2012: 7
- FWCI
- 4.31
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute on Aging (R01AG028352)
- National Institute of General Medical Sciences (R01GM053008, R01GM057468)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Phagocytosis and Immune Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Autophagy in Disease and Therapy
- Keywords
- phagocytosis of apoptotic cells, macrophages, pH-sensitive fluorescent dye, efferocytosis, MFG-E8, splenic macrophages, peritoneal macrophages, lymphocytes
- MeSH terms
- Animals, Flow Cytometry, Fluorescent Dyes, Male, Mice, Inbred C57BL, Microscopy, Fluorescence, Phagocytosis, Spleen, Succinimides, Thymus Gland, Rats, Sprague-Dawley, Apoptosis, Macrophages, Peritoneal, Mice, Rats
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