High-Sensitivity Assessment of Phagocytosis by Persistent Association-Based Normalization
(2021) In Journal of immunology 206(1). p.214-224- Abstract
Phagocytosis is measured as a functional outcome in many research fields, but accurate quantification can be challenging, with no robust method available for cross-laboratory reproducibility. In this study, we identified a simple, measurable parameter, persistent prey-phagocyte association, to use for normalization and dose-response analysis. We apply this in a straightforward analytical method, persistent association-based normalization, in which the multiplicity of prey (MOP) ratio needed to elicit half of the phagocytes to associate persistently (MOP50) is determined first. MOP50 is then applied to normalize for experimental factors, separately analyzing association and internalization. We use reference human phagocyte THP-1 cells... (More)
Phagocytosis is measured as a functional outcome in many research fields, but accurate quantification can be challenging, with no robust method available for cross-laboratory reproducibility. In this study, we identified a simple, measurable parameter, persistent prey-phagocyte association, to use for normalization and dose-response analysis. We apply this in a straightforward analytical method, persistent association-based normalization, in which the multiplicity of prey (MOP) ratio needed to elicit half of the phagocytes to associate persistently (MOP50) is determined first. MOP50 is then applied to normalize for experimental factors, separately analyzing association and internalization. We use reference human phagocyte THP-1 cells with different prey and opsonization conditions to compare the persistent association-based normalization method to standard ways of assessing phagocytosis and find it to perform better, exhibiting increased robustness, sensitivity, and reproducibility. The approach is easily incorporated into most existing phagocytosis assays and allows for reproducible results with high sensitivity.
(Less)
- author
- de Neergaard, Therese LU ; Sundwall, Martin LU ; Wrighton, Sebastian LU and Nordenfelt, Pontus LU
- organization
- publishing date
- 2021-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of immunology
- volume
- 206
- issue
- 1
- pages
- 11 pages
- publisher
- American Association of Immunologists
- external identifiers
-
- scopus:85098691560
- pmid:33268484
- ISSN
- 1550-6606
- DOI
- 10.4049/jimmunol.2000032
- project
- The Molecular Basis of Phagocytosis
- language
- English
- LU publication?
- yes
- additional info
- Copyright © 2020 by The American Association of Immunologists, Inc.
- id
- 144de961-a3ae-404b-a401-bc41ca5a0c88
- date added to LUP
- 2020-12-17 15:43:05
- date last changed
- 2024-09-19 11:41:44
@article{144de961-a3ae-404b-a401-bc41ca5a0c88, abstract = {{<p>Phagocytosis is measured as a functional outcome in many research fields, but accurate quantification can be challenging, with no robust method available for cross-laboratory reproducibility. In this study, we identified a simple, measurable parameter, persistent prey-phagocyte association, to use for normalization and dose-response analysis. We apply this in a straightforward analytical method, persistent association-based normalization, in which the multiplicity of prey (MOP) ratio needed to elicit half of the phagocytes to associate persistently (MOP50) is determined first. MOP50 is then applied to normalize for experimental factors, separately analyzing association and internalization. We use reference human phagocyte THP-1 cells with different prey and opsonization conditions to compare the persistent association-based normalization method to standard ways of assessing phagocytosis and find it to perform better, exhibiting increased robustness, sensitivity, and reproducibility. The approach is easily incorporated into most existing phagocytosis assays and allows for reproducible results with high sensitivity.</p>}}, author = {{de Neergaard, Therese and Sundwall, Martin and Wrighton, Sebastian and Nordenfelt, Pontus}}, issn = {{1550-6606}}, language = {{eng}}, month = {{01}}, number = {{1}}, pages = {{214--224}}, publisher = {{American Association of Immunologists}}, series = {{Journal of immunology}}, title = {{High-Sensitivity Assessment of Phagocytosis by Persistent Association-Based Normalization}}, url = {{http://dx.doi.org/10.4049/jimmunol.2000032}}, doi = {{10.4049/jimmunol.2000032}}, volume = {{206}}, year = {{2021}}, }