Abstract | BACKGROUND: METHODS: Human eosinophils and other cells were purified from peripheral blood by density gradient sedimentation and negative/positive selections using immunomagnetic beads. Protease/ proteinase expression in eosinophils and release into the supernatant were evaluated by microarray analysis, qPCR, ELISA, flow cytometry and immunofluorescence staining before and after stimulation with eosinophil-activating cytokines and secretagogues. mRNAs for extracellular matrix proteins in human normal fibroblasts were measured by qPCR after exposure to recombinant protease serine 33 (PRSS33) protein (rPRSS33), created with a baculovirus system. RESULTS: Human eosinophils expressed relatively high levels of mRNA for metalloproteinase 25 ( MMP25), a disintegrin and metalloprotease 8 (ADAM8), ADAM10, ADAM19 and PRSS33. Expression of PRSS33 was the highest and eosinophil-specific. PRSS33 mRNA expression was not affected by eosinophil-activating cytokines. Immunofluorescence staining showed that PRSS33 was co-localized with an eosinophil granule protein. PRSS33 was not detected in the culture supernatant of eosinophils even after stimulation with secretagogues, but its cell surface expression was increased. rPRSS33 stimulation of human fibroblasts increased expression of collagen and fibronectin mRNAs, at least in part via protease-activated receptor-2 activation. CONCLUSIONS:
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Authors | Sumika Toyama, Naoko Okada, Akio Matsuda, Hideaki Morita, Hirohisa Saito, Takao Fujisawa, Susumu Nakae, Hajime Karasuyama, Kenji Matsumoto |
Journal | Allergology international : official journal of the Japanese Society of Allergology
(Allergol Int)
Vol. 66
Issue 3
Pg. 463-471
(Jul 2017)
ISSN: 1440-1592 [Electronic] England |
PMID | 28216055
(Publication Type: Journal Article)
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Copyright | Copyright © 2017 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved. |
Chemical References |
- Biomarkers
- Serine Proteases
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Topics |
- Asthma
(immunology, metabolism, pathology)
- Biomarkers
- Cell Degranulation
(immunology)
- Cells, Cultured
- Eosinophils
(immunology, metabolism)
- Fibroblasts
(metabolism)
- Flow Cytometry
- Gene Expression
- Gene Expression Profiling
- Humans
- Leukocytes
(immunology, metabolism)
- Organ Specificity
(genetics)
- Protein Transport
- Serine Proteases
(genetics, metabolism)
- Transcriptome
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