High generation of reactive oxygen species from neutrophils in patients with severe COVID-19

Tonny Veenith, Helena Martin, Martin Le Breuilly, Tony Whitehouse, Fang Gao-Smith, Niharika Duggal, Janet M Lord, Rubina Mian, David Sarphie, Paul Moss, Tonny Veenith, Helena Martin, Martin Le Breuilly, Tony Whitehouse, Fang Gao-Smith, Niharika Duggal, Janet M Lord, Rubina Mian, David Sarphie, Paul Moss

Abstract

Neutrophilia and an elevated neutrophil:lymphocyte ratio are both characteristic features of severe COVID-19 infection. However, functional neutrophil responses have been poorly investigated in this setting. We utilised a novel PMA-based stimulation assay to determine neutrophil-derived reactive oxygen species (ROS) generation in patients with severe COVID-19 infection, non-COVID related sepsis and healthy study participants. ROS production was markedly elevated in COVID-19 patients with median values ninefold higher than in healthy controls and was particularly high in patients on mechanical ventilation. ROS generation correlated strongly with neutrophil count and elevated levels were also seen in patients with non-COVID related sepsis. Relative values, adjusted for neutrophil count, were high in both groups but extreme low or high values were seen in two patients who died shortly after testing, potentially indicating a predictive value for neutrophil function. Our results show that the high levels of neutrophils observed in patients with COVID-19 and sepsis exhibit functional capacity for ROS generation. This may contribute to the clinical features of acute disease and represents a potential novel target for therapeutic intervention.

Conflict of interest statement

Authors Mian and Sarphie declare that they are directors and principal shareholders of Seroxo Ltd, a private company developing clinical applications of the LIT™ system. The other authors declare no competing interests.

© 2022. The Author(s).

Figures

Figure 1
Figure 1
Dot plot (L) and violin plot (R) of LIT scores for patients with COVID-19 or healthy donors. Median LIT score for patients with COVID-19 was 1378 (IQR: 1276) and was ninefold higher than values within the control group (Median 153; IQR: 112, (Mann–Whitney U test, p 

Figure 2

Box plots showing distribution of…

Figure 2

Box plots showing distribution of LIT scores in patients with COVID-19 in relation…

Figure 2
Box plots showing distribution of LIT scores in patients with COVID-19 in relation to invasive ventilation status. Horizontal lines in each box represent the median LIT scores for each cohort. Median LIT score for patients on invasive ventilation was 2020 and compared with 916 for those not on invasive ventilation (Mann Whitney U test: p = 0.02).

Figure 3

Scatterplot to show correlation between…

Figure 3

Scatterplot to show correlation between LIT and neutrophil count in patients with COVID-19.…

Figure 3
Scatterplot to show correlation between LIT and neutrophil count in patients with COVID-19. LIT scores and neutrophil counts are positively correlated (Spearman’s rho = 0.80 [0.52, 0.92] (p = 0.0004)).

Figure 4

Dot plot (L) and violin…

Figure 4

Dot plot (L) and violin plot (R) of LIT scores in patients with…

Figure 4
Dot plot (L) and violin plot (R) of LIT scores in patients with sepsis compared to healthy volunteers. Median LIT score for patients with sepsis was 2120 (IQR: 2024) and was > 13-fold higher than values within the control group (median: 153; IQR: 112, Mann–Whitney U test, p 
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Figure 2
Figure 2
Box plots showing distribution of LIT scores in patients with COVID-19 in relation to invasive ventilation status. Horizontal lines in each box represent the median LIT scores for each cohort. Median LIT score for patients on invasive ventilation was 2020 and compared with 916 for those not on invasive ventilation (Mann Whitney U test: p = 0.02).
Figure 3
Figure 3
Scatterplot to show correlation between LIT and neutrophil count in patients with COVID-19. LIT scores and neutrophil counts are positively correlated (Spearman’s rho = 0.80 [0.52, 0.92] (p = 0.0004)).
Figure 4
Figure 4
Dot plot (L) and violin plot (R) of LIT scores in patients with sepsis compared to healthy volunteers. Median LIT score for patients with sepsis was 2120 (IQR: 2024) and was > 13-fold higher than values within the control group (median: 153; IQR: 112, Mann–Whitney U test, p 

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