Neutrophil Phagocytic Capacity and Priming Assessment

Neutrophil phagocytic capacity was assessed using two independent methods: a light microscopic analysis of isolated neutrophils using zymosan particles pre-opsonized using autologous serum [10 (link), 11 (link)], and a flow cytometric analysis of whole blood using pHrodo^™ Green Staphylococcus aureus Bioparticles^® (Thermofisher, MA, USA) [12 (link)]. Neutrophil priming was assessed by quantifying expression of the cell surface markers CD11b and CD62L using a whole blood flow cytometric method as previously described [4 (link), 13 (link)].

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Funston W., Ruchaud-Sparagano M.H., Scott J., Powell J., Cooles F.A., Shelmerdine L., McDowell C., O’Leary D., Booth K.L., Clark S.C., Ledingham S.J., Rostron A.J., Dark J.H, & Simpson A.J. (2022). A human model of bilateral pulmonary vein sampling to assess the effects of one-lung ventilation on neutrophil function. PLoS ONE, 17(7), e0271958.

Publication 2022

pHrodo™ Green Staphylococcus aureus Bioparticles

Blood analysis Blood flow Cd11b Cd62l Cell Flow cytometric Microscopic light Neutrophil Phagocytic Serum Staphylococcus aureus Zymosan

Corresponding Organization : Newcastle University

Other organizations : Health Education North West, Royal Victoria Hospital, University of Ulster, Newcastle upon Tyne Hospitals NHS Foundation Trust, Northumbria University

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Variable analysis

independent variables

Zymosan particles pre-opsonized using autologous serum
PHrodo™ Green Staphylococcus aureus Bioparticles®

dependent variables

Neutrophil phagocytic capacity
Neutrophil priming (expression of cell surface markers CD11b and CD62L)

control variables

Whole blood flow cytometric method for assessing neutrophil priming

controls

Autologous serum used for opsonization of zymosan particles (positive control)
No controls explicitly mentioned for the flow cytometric analysis of neutrophil phagocytic capacity using pHrodo™ Green Staphylococcus aureus Bioparticles®

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