Background: Different asthma phenotypes are driven by molecular endotypes. A Th1-high phenotype is linked to severe, therapy-refractory asthma, subclinical infections and neutrophil inflammation. Previously, we found neutrophil granulocytes (NGs) from asthmatics exhibit decreased chemotaxis towards leukotriene B4 (LTB(4)), a chemoattractant involved in inflammation response. We hypothesized that this pattern is driven by asthma in general and aggravated in a Th1-high phenotype. Methods: NGs from asthmatic nd healthy children were stimulated with 10 nM LTB(4)/100 nM N-formylmethionine-leucyl-phenylalanine and neutrophil migration was documented following our prior SiMA (simplified migration assay) workflow, capturing morphologic and dynamic parameters from single-cell tracking in the images. Demographic, clinical and serum cytokine data were determined in the ALLIANCE cohort. Results: A reduced chemotactic response towards LTB(4) was confirmed in asthmatic donors regardless of inhaled corticosteroid (ICS) treatment. By contrast, only NGs from ICS-treated asthmatic children migrate similarly to controls with the exception of Th1-high donors, whose NGs presented a reduced and less directed migration towards the chemokines. ICS-treated and Th1-high asthmatic donors present an altered surface receptor profile, which partly correlates with migration. Conclusions: Neutrophil migration in vitro may be affected by ICS-therapy or a Th1-high phenotype. This may be explained by alteration of receptor expression and could be used as a tool to monitor asthma treatment.
- Lemmel, S.
- Weckmann, M.
- Wohlers, A.
- Jirmo, A. C.
- Grychtol, R.
- Ricklefs, I.
- Nissen, G.
- Bachmann, A.
- Singh, S.
- Caicedo, J.
- Bahmer, T.
- Hansen, G.
- Von Mutius, E.
- Rabe, K. F.
- Fuchs, O.
- Dittrich, A. M.
- Schaub, B.
- Happle, C.
- Carpenter, A. E.
- Kopp, M. V.
- Becker, T.
Keywords
- Ltb4
- fMLP
- high-content image analysis
- migration
- neutrophil granulocytes
- single-cell analysis
