The whole blood phagocytosis assay: a clinically relevant test of neutrophil function and dysfunction in community-acquired pneumonia

Objective: To refine and validate a neutrophil function assay with clinical relevance for patients with communityacquired

pneumonia (CAP).

Design: Two phase cross-sectional study to standardise and refine the assay in blood from healthy volunteers and

test neutrophil phagocytic function in hospital patients with CAP.

Participants: Phase one: Healthy adult volunteers (n = 30). Phase two: Critical care patients with severe CAP (n = 16),

ward-level patients with moderate CAP (n = 15) and respiratory outpatients (no acute disease, n = 15).

Results: Our full standard operating procedure for the assay is provided. Patients with severe CAP had significantly

decreased neutrophil function compared to moderate severity disease (median phagocytic index 2.8 vs. 18.0,

p = 0.014). Moderate severity pneumonia neutrophil function was significantly higher than control samples (median

18.0 vs. 1.6, p = 0.015). There was no significant difference between critical care and control neutrophil function

(median 2.8 vs. 1.6, p = 0.752).

Conclusions: Our whole blood neutrophil assay is simple, reproducible and clinically relevant. Changes in neutrophil

function measured in this pneumonia cohort is in agreement with previous studies. The assay has potential to be

used to identify individuals for clinical trials of immunomodulatory therapies, to risk-stratify patients with pneumonia,

and to refine our understanding of ‘normal’ neutrophil function in infection.

Keywords: Neutrophils, Sepsis, Flow cytometry, Phagocytosis, Pneumonia