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Establishment of a scalable microfluidic assay for characterization of population‐based neutrophil chemotaxis
Author(s) -
Grigolato Fulvio,
Egholm Cecilie,
Impellizzieri Daniela,
Arosio Paolo,
Boyman Onur
Publication year - 2020
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1111/all.14195
Subject(s) - chemotaxis , cxcl2 , chemokine , interleukin 8 , microbiology and biotechnology , chemotaxis assay , biology , immunology , inflammation , microfluidics , chemokine receptor , receptor , materials science , nanotechnology , biochemistry
Background Regulation of neutrophil chemotaxis and activation plays crucial roles in immunity, and dysregulated neutrophil responses can lead to pathology as seen in neutrophilic asthma. Neutrophil recruitment is key for initiating immune defense and inflammation, and its modulation is a promising therapeutic target. Microfluidic technology is an attractive tool for characterization of neutrophil migration. Compared to transwell assays, microfluidic approaches could offer several advantages, including precis e control of defined chemokine gradients in space and time, automated quantitative analysis of chemotaxis, and high throughput. Methods We established a microfluidic device for fully automated, quantitative assessment of neutrophil chemotaxis. Freshly isolated mouse neutrophils from bone marrow or human neutrophils from peripheral blood were assessed in real time using an epifluorescence microscope for their migration toward the potent chemoattractants C‐X‐C‐motif ligand 2 (CXCL2) and CXCL8, without or with interleukin‐4 (IL‐4) pre‐incubation. Results Our microfluidic device allowed the precise and reproducible determination of the optimal CXCL2 and CXCL8 concentrations for mouse and human neutrophil chemotaxis, respectively. Furthermore, our microfluidic assay was able to measure the equilibrium and real‐time dynamic effects of specific modulators of neutrophil chemotaxis. We demonstrated this concept by showing that IL‐4 receptor signaling in mouse and human neutrophils inhibited their migration toward CXCL2 and CXCL8, respectively, and this inhibition was time‐dependent. Conclusion Collectively, our microfluidic device shows several advantages over traditional transwell migration assays and its design is amenable to future integration into multiplexed high‐throughput platforms for screening of molecules that modulate the chemotaxis of different immune cells.