Influence of Cell Quality on Inflammatory Biomarkers in COPD Sputum Supernatant

Frauke Pedersen; Frederik Trinkmann; Mustafa Abdo; Anne-Marie Kirsten; Klaus F Rabe; Henrik Watz; Simonetta Baraldo; Marina Saetta; Jens M Hohlfeld; Olaf Holz

doi:10.2147/COPD.S284938

Influence of Cell Quality on Inflammatory Biomarkers in COPD Sputum Supernatant

Int J Chron Obstruct Pulmon Dis. 2021 Mar 1:16:487-493. doi: 10.2147/COPD.S284938. eCollection 2021.

Authors

Frauke Pedersen^{1

2}, Frederik Trinkmann³, Mustafa Abdo², Anne-Marie Kirsten¹, Klaus F Rabe², Henrik Watz¹, Simonetta Baraldo⁴, Marina Saetta⁴, Jens M Hohlfeld^{5

6}, Olaf Holz⁵

Affiliations

¹ Pulmonary Research Institute at LungenClinic Grosshansdorf, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Grosshansdorf, Germany.
² LungenClinic Grosshansdorf, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Grosshansdorf, Germany.
³ Pneumology and Critical Care Medicine, Thoraxklinik at University Hospital Heidelberg, Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany.
⁴ Department of Cardiac, Thoracic, Vascular Sciences and Public Health, Respiratory Diseases Clinic, University of Padova, Padova, Italy.
⁵ Fraunhofer ITEM, Clinical Airway Research - Biomedical Research in End-Stage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover, Germany.
⁶ Department of Respiratory Medicine, Hannover Medical School (MHH), Biomedical Research in End-Stage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover, Germany.

Abstract

Purpose: We recently introduced a sputum cell quality score to rate how cell morphology, cellular debris and squamous cell contamination influence inflammatory cell identification during microscopic evaluation. However, sputum cell quality is generally not considered for the interpretation of sputum fluid phase biomarkers. Therefore, we compared the soluble protein concentrations between sputum samples with different cell quality. The impact of cell quality was compared to other factors potentially affecting soluble biomarker concentrations.

Methods: A comprehensive sputum dataset from 154 clinically stable COPD patients was used to analyse the differences and the variability of sputum supernatant concentrations for 23 proteins between low, medium, and high sputum cell quality samples. A model was developed and tested to compare the impact of different factors on sputum supernatant protein levels.

Results: Mean percentages of sputum macrophages, neutrophils, eosinophils, monocytes and lymphocytes showed no significant differences between low, medium and high cell quality levels. The mean percentage of squamous cells were lower, while total cell count/mL sputum and cell viability were significantly higher in sputum samples with higher cell quality. The concentrations of Interleukin-6, Interleukin-8 and Tumor Necrosis Factor Receptor 2 were significantly increased in sputum samples of higher cell quality. The variability of most protein concentrations declined with increasing cell quality levels. Sixteen proteins showed significantly negative correlations with the percentage of squamous cells. For 14 proteins we observed a positive correlation with cell number/mL sputum. Multiple regression analysis shows that generally less than 30% of the protein variability can be explained by the included factors.

Conclusion: Sputum cell quality has a significant impact on some soluble biomarker concentrations in sputum supernatant. Sputum samples with low sputum cell quality show a higher variability of fluid phase proteins in comparison to medium and high sputum cell quality levels.

Keywords: soluble biomarkers; sputum cell quality score; sputum supernatant.

MeSH terms

Biomarkers
Eosinophils
Humans
Leukocyte Count
Neutrophils
Pulmonary Disease, Chronic Obstructive* / diagnosis
Sputum*

Substances

Biomarkers