Abstract
Eosinophils (Eos) play an important role in the immune system’s response releasing several inflammatory factors and contributing to allergic rhinitis, asthma, or atopic dermatitis. Since Eos have a relatively short lifetime after isolation from blood, usually eosinophilic cell line (EoL-1) is used to study mechanisms of their activation and to test therapies. In particular, EoL-1 cells are examined in terms of signalling pathways of the inflammatory response manifested by the presence of lipid bodies (LBs). Here we examined the differences in response to inflammation modelled by various factors, between isolated human eosinophils and EoL-1 cells, as manifested in the number and chemical composition of LBs. The analysis was performed using fluorescence, Raman, and coherent anti-Stokes Raman scattering (CARS) microscopy, which recognised the inflammatory process in the cells, but it is manifested slightly differently depending on the method used. We showed that unstimulated EoL-1 cells, compared to isolated eosinophils, contained more LBs, displayed different nucleus morphology and did not have eosinophilic peroxidase (EPO). In EoL-1 cells stimulated with various proinflammatory agents, including butyric acid (BA), liposaccharide (LPS), or cytokines (IL-1β, TNF-α), an increased production of LBs with a various degree of lipid unsaturation was observed in spontaneous Raman spectra. Furthermore, stimulation of EoL-1 cells resulted in alterations of the LBs morphology. In conclusion, a level of lipid unsaturation and eosinophilic peroxidase as well as LBs distribution among cell population mainly accounted for the biochemistry of eosinophils upon inflammation.
Highlights
Eosinophils (Eos) belong to granulocytes, the type of white blood cells in the innate immune system, characterized by the presence of granules in their cytoplasm
Conventional fluorescence microscopy assesses the morphological characteristic of stained nuclei and lipid bodies (LBs) in these blood cells and estimates an averaged number of LBs present in control and induced by proinflammatory agents
Label-free coherent anti-Stokes Raman scattering (CARS) imaging with similar time of data collection as for fluorescence microscopy, indicates the detailed distribution of lipid bodies within cells and its results can be semi-automatically used for the quantification of LBs in each cell individually
Summary
Eosinophils (Eos) belong to granulocytes, the type of white blood cells in the innate immune system, characterized by the presence of granules in their cytoplasm. They participate in the regulation of inflammatory conditions by releasing several inflammatory mediators such as cytokines and reactive oxygen species. Eos are involved in the communication between the immune system cells [1] They play an important role in allergic rhinitis, asthma, atopic dermatitis, and the immune system’s responses, including fighting parasites. Eos are considered to play two opposite roles They inactivate histamine—a slow reactive substance of anaphylaxis and platelet-activating factor (PAF), which is responsible for attenuation of the allergic inflammation products. Eosinophil peroxidase (EPO) and eosinophil cationic protein present in granules worsen and prolong
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