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

de Neergaard, Therese (author): Lund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Molekylär patogenes,Forskargrupper vid Lunds universitet,Kvantitativ immunbiologi,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine,Molecular Pathogenesis,Lund University Research Groups,Quantitative immunobiology

Sundwall, Martin (author): Lund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Molekylär patogenes,Forskargrupper vid Lunds universitet,Kvantitativ immunbiologi,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine,Molecular Pathogenesis,Lund University Research Groups,Quantitative immunobiology

Wrighton, Sebastian (author): Lund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Kvantitativ immunbiologi,Forskargrupper vid Lunds universitet,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine,Quantitative immunobiology,Lund University Research Groups

Nordenfelt, Pontus (author): Lund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Molekylär patogenes,Forskargrupper vid Lunds universitet,Kvantitativ immunbiologi,SEBRA Sepsis and Bacterial Resistance Alliance,epIgG,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine,Molecular Pathogenesis,Lund University Research Groups,Quantitative immunobiology

(creator_code:org_t)

2021-01-01
2021
English 11 s.
In: Journal of immunology. - : The American Association of Immunologists. - 1550-6606 .- 0022-1767. ; 206:1, s. 214-224

Related links:: http://dx.doi.org/10...; show more...; https://www.jimmunol...; https://lup.lub.lu.s...; https://doi.org/10.4...; show less...

Abstract Subject headings

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.

By the author/editor: de Neergaard, Th ...; Sundwall, Martin; Wrighton, Sebast ...; Nordenfelt, Pont ...

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Basic Medicine; and Immunology in th ...

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