Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

High-Sensitivity Assessment of Phagocytosis by Persistent Association-Based Normalization

de Neergaard, Therese LU ; Sundwall, Martin LU orcid ; Wrighton, Sebastian LU orcid and Nordenfelt, Pontus LU orcid (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)
Please use this url to cite or link to this publication:
author
; ; and
organization
publishing date
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
  • pmid:33268484
  • scopus:85098691560
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
2024-06-27 03:58:07
@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}},
}