Human eosinophils release matrix metalloproteinase-9 on stimulation with TNF-α☆,☆☆,★
Section snippets
Isolation of peripheral blood eosinophils
Peripheral blood eosinophils were isolated from atopic asthmatic volunteers who had given their informed consent, as described previously.20, 21 Briefly, heparinized blood (100 mL) was mixed with 20 mL of dextran (6%; Fluka, Buchs, Switzerland) in RPMI-1640 medium (Bio Whittaker, Walkersville, Md). Erythrocytes were then sedimented for 30 minutes at room temperature. The plasma was layered on a Ficoll (Pharmacia, Quebec, Canada) cushion (15 mL) and centrifuged for 25 minutes at 1000g at room
RT-PCR
Total RNA was isolated from 2 × 106 eosinophils by using the Quiagen RNeasy kit (Ontario, Canada). The average amount of RNA obtained from 2 × 106 eosinophils was 300 ng. One third of the RNA was reverse transcribed by using superscript II reverse transcriptase (Gibco BRL, Ontario, Canada) and random hexamers (50A 260 U; Boehringer Mannheim GmbH, Mannheim, Germany) as primers. Thereafter, PCR was performed in 20-μL reactions by using the following primers (25 μmol/L) for MMP-9
MMP-9 and TIMP expression in human eosinophils
Transcriptional expression of MMP-9, as well as that of TIMP-1 and TIMP-2, was studied in human peripheral blood eosinophils by means of RT-PCR. MMP-9 and TIMP-2 gene expression could be detected in both resting and TNF-α–stimulated eosinophils (Fig 1, A ), whereas TIMP-1 gene expression appeared to vary between different subjects (data not shown).
DISCUSSION
The recent description of the capacity of eosinophils to release MMP-9 has opened new vistas toward our understanding of the process of airway remodeling in asthma.25 Our novel data revealed that human eosinophil–derived MMP-9 expression and secretion is upregulated on TNF-α stimulation. Gelatin zymography provided an excellent method to show MMP-9 activity in these cells. NFκB was previously shown to be involved in MMP-9 activation in fibroblasts,26 transformed cells,27, 28 and renal cells.29
Acknowledgements
We thank Jolanta Sawicka for technical help, and Dr Harissios Vliagoftis and Dr Greg Sawicki for useful and ongoing discussions.
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Cited by (114)
Tumor-associated tissue eosinophilia promotes angiogenesis and metastasis in head and neck squamous cell carcinoma
2023, Neoplasia (United States)Citation Excerpt :Eosinophils, a component of immune cells involved in chronic inflammatory conditions such as asthma, had shown in previous studies its ability to mediate the degradation of extracellular matrix through the activation of matrix metalloproteinases (MMP), such as MMP-2, MMP-9, collagenase and stromelysin [32]. Evidence also showed MMP-9 production by eosinophils under activated T cell stimulation and under regulation of stem cell factor (SCF), TGF-β, and TNF-α [32–35]. This release of MMP-9 also facilitates degradation of the basement membrane [36].
Eosinophils, Mast Cells and Basophils
2022, Comprehensive PharmacologyInnate effector cells in angiogenesis and lymphangiogenesis
2018, Current Opinion in ImmunologyCitation Excerpt :Human eosinophils produce several angiogenic factors such as VEGF-A [80,81•], fibroblast growth factors (FGF-2) [82,83], CXCL8/IL-8 [84], and osteopontin [85]. Human eosinophils also produce MMP9 [86–88]. Eosinophils have been detected in metastatic lymph nodes of cancer patients, but the production of lymphangiogenic factors by these cells should be further addressed.
Eosinophil Cytokines in Allergy
2017, Cytokine Effector Functions in Tissues
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Supported by the Medical Research Council of Canada, the Alberta Heritage Foundation for Medical Research, and the Glaxo-Wellcome Heritage Award, University of Alberta. A. S. was also supported by Hoffman-La Roche, ESCF. R. M. and M. D. are Alberta Heritage Senior Medical Scholars.
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Reprint requests: Redwan Moqbel, PhD, FRCPath, Pulmonary Research Group, University of Alberta, 5-74 HMRC, Edmonton, Alberta T6G 2S2, Canada.
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