Influence of Cell Quality on Inflammatory Biomarkers in COPD Sputum Supernatant

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:

• sputum cell quality score
• soluble biomarkers
• sputum supernatant

Data Sharing Statement

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to restrictions eg their containing information that could compromise the privacy of research participants.

Disclosure

Dr Frederik Trinkmann reports personal fees from Actelion, Berlin Chemie, Boehringer Ingelheim, Chiesi, Novartis, Mundipharma, TEVA, AstraZeneca, Bristol-Myers Squibb, GlaxoSmithKline, Roche, and Sanofi Aventis, outside the submitted work. Prof. Dr. Klaus F Rabe reports personal fees from Boehringer Ingelheim, AstraZeneca, Novartis, Chiesi, Teva, Sanofi & Regeneron, Berlin Chemie, GSK, and Orion Menarini, outside the submitted work. Prof. Dr. Marina Saetta reports grants from Chiesi Farmaceutici, outside the submitted work. Prof. Dr. Jens M Hohlfeld reports personal fees from Boehringer Ingelheim for consultancy and HAL for lecture fee, grants to my institution from AstraZeneca AB, Novartis, Janssen Pharmaceutica NV, ALK, Boehringer Ingelheim, LETI, GlaxoSmithKline, GSK, Astellas Pharma, Allergopharma, and Sanofi-Aventis, personal fees for consultancy from Merck & Co, Inc., lecture fee from Novartis, outside the submitted work. The authors report no other conflicts of interest in this work.