Inflammatory cytokines can be monitored in exhaled breath particles following segmental and inhalation endotoxin challenge in healthy volunteers

Olaf Holz, Meike Müller, Saskia Carstensen, Anna-Carin Olin, Jens M Hohlfeld, Olaf Holz, Meike Müller, Saskia Carstensen, Anna-Carin Olin, Jens M Hohlfeld

Abstract

Particles in exhaled air (PEx) are generated when collapsed small airways reopen during breathing. PEx can be noninvasively collected by particle impaction, allowing the analysis of undiluted epithelial lining fluid (ELF). We used the endotoxin (LPS) challenge model to proof the concept that PEx can be used to monitor inflammatory changes in the lung. In this pilot study PEx were collected from ten healthy nonsmoking subjects using the PExA® instrument twice before and twice after a segmental LPS challenge (5, 21 h). Following a 4-week washout period, PEx were collected during the week before and 5 h after a whole lung LPS inhalation challenge. PEx biomarkers were compared to blood, bronchoalveolar lavage (BAL) following segmental challenge and induced sputum (ISP) following inhalation challenge. A clear LPS-induced inflammatory response was detectable in BAL fluid, ISP and blood. Albumin and surfactant-protein D were detectable in all PEx samples. While most baseline cytokines were close to or below the detection limit, the median (IQR) IL-6 and IL-8 concentrations in PEx increased significantly after segmental (0.04 (0.03; 0.06) fg/ng PEx; 0.10 (0.08; 0.17) fg/ng PEx) and inhalation LPS challenge (0.19 (0.15; 0.23) fg/ng PEx; 0.32 (0.23; 0.42) fg/ng PEx). Using a highly sensitive analysis platform, we were able to detect a cytokine response in PEx during the early phase of LPS-induced inflammation. This will broaden the spectrum of applications for this noninvasive method to monitor inflammatory processes in the lung, including its use in clinical trials for respiratory drug development.Trial registration: The study has been registered on 07.02.2017 at Clinicaltrials.gov (NCT03044327).

Conflict of interest statement

OH, MM, SC and JMH have no competing interest. ACO is a chairholder of PExA AB.

© 2022. The Author(s).

Figures

Figure 1
Figure 1
Study design. The study was designed to test the ability of novel noninvasive methods to detect airway inflammation. Here we focus on exhaled breath particles (PEx). BAL bronchoalveolar lavage, V visit (index pre: refers to samples taken before a challenge, post: refers to samples taken after a challenge), NaCl isotonic saline solution, LPS endotoxin, w week, h hours. PEx samples were collected on V3 in the afternoon 5 h after the segmental challenge. On V4 PEx were sampled prior to the BAL procedure 21 h after the LPS challenge. On visit 5 and 6 PEx were collected before the sputum induction. On V6 this was done 5 h following the inhalation LPS challenge.
Figure 2
Figure 2
Individual and median concentrations of interleukin-6 and interleukin-8 per ng PEx. Two hundred microliters of elution buffer was used for a membrane with 120 ng of exhaled breath particles (PEx).*p 

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