Multi-dimensional flow cytometry analysis reveals increasing changes in the systemic neutrophil compartment during seven consecutive days of endurance exercise

Selma van Staveren, Twan Ten Haaf, Margot Klöpping, Bart Hilvering, Gerjen H. Tinnevelt, Karin de Ruiter, Maria F. Piacentini, Bart Roelands, Romain Meeusen, Jos J. de Koning, Jeroen J. Jansen, Nienke Vrisekoop, Leo Koenderman

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Endurance exercise is associated with a transient increase in neutrophil counts in the peripheral blood. Here we investigate the impact of intensified endurance exercise on the neutrophil compartment. We hypothesized that intensified endurance exercise leads to mobilization of neutrophil subsets, which are normally absent in the blood. Furthermore, we followed the potential build-up of neutrophil activation and the impact on overnight recovery of the neutrophil compartment during a seven-day cycling tour. The neutrophil compartment was studied in 28 healthy amateur cyclists participating in an eight-day strenuous cycling tour. Blood samples were taken at baseline, after 4 days and after 7 days of cycling. The neutrophil compartment was analyzed in terms of numbers and its phenotype by deep phenotyping of flow cytometry data with the multi-dimensional analysis method FLOOD. Repeated endurance exercise led to a gradual increase in total neutrophil counts over the days leading to a 1.26 fold-increase (95%CI 1.01-1.51 p = 0.0431) in the morning of day 8. Flow cytometric measurements revealed the appearance of 2 additional neutrophil subsets: CD16brightCD62Ldim and CD16dimCD62Lbright. A complex change in neutrophil phenotypes was present characterized by decreased expression of both CD11b and CD62L and marked increased expression of LAIR-1, VLA-4 and CBRM1/5. The changes in expression were found on all neutrophils present in the blood. Strikingly, in strong contrast to our findings during acute inflammation evoked by LPS challenge, these neutrophils did not upregulate classical degranulation markers. In fact, our FLOOD analysis revealed that the exercise induced neutrophil phenotype did not overlap with the neutrophil subsets arising upon acute inflammation. In conclusion, during multiple days of endurance exercise the neutrophil compartment does not regain homeostasis overnight. Thereby our study supports the concept of a build-up of inflammatory cues during repeated endurance exercise training, causing a prolonged change of the systemic neutrophil compartment.

Original languageEnglish
Article numbere0206175
Pages (from-to)1-23
Number of pages23
JournalPLoS ONE
Volume13
Issue number10
DOIs
Publication statusPublished - 30 Oct 2018

Funding

This research received funding from the Netherlands Organization for Scientific Research (NWO) in the framework of the Technology Area COAST. We are very grateful to all participants who voluntarily took part in all measurements. Also we would like to thank Tour for Life organizer Emolife, especially Nienke Wynia and Kelian Lankester, for their collaboration. We thank Julia Drylewicz for her help with statistics and André van den Doel for his early work for the development of the FLOOD analysis method. Not the least thanks to Danilo Ian-netta, Erik Oudenhoven and Hans de Koning for their enthusiastic help with sample collection.

FundersFunder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
Tekniikan Akatemia

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