Related Experiment Video
Updated: Sep 13, 2026
Advanced Imaging of Lung Homing Human Lymphocytes in an Experimental In Vivo Model of Allergic Inflammation Based on Light-sheet Microscopy
Published on: April 16, 2019
IL-5-stimulated eosinophils adherent to periostin undergo stereotypic morphological changes and ADAM8-dependent
M W Johansson1, M Khanna1, V Bortnov1
1Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI, USA.
Background:
IL-5 causes suspended eosinophils to polarize with filamentous (F)-actin and granules at one pole and the nucleus in a specialized uropod, the "nucleopod," which is capped with P-selectin glycoprotein ligand-1 (PSGL-1). IL-5 enhances eosinophil adhesion and migration on periostin, an extracellular matrix protein upregulated in asthma by type 2 immunity mediators.
Objective:
Determine how the polarized morphology evolves to foster migration of IL-5-stimulated eosinophils on a surface coated with periostin.
Methods:
Blood eosinophils adhering to adsorbed periostin were imaged at different time points by fluorescent microscopy, and migration of eosinophils on periostin was assayed.
Results:
After 10 minutes in the presence of IL-5, adherent eosinophils were polarized with PSGL-1 at the nucleopod tip and F-actin distributed diffusely at the opposite end. After 30-60 minutes, the nucleopod had dissipated such that PSGL-1 was localized in a crescent or ring away from the cell periphery, and F-actin was found in podosome-like structures. The periostin layer, detected with monoclonal antibody Stiny-1, shown here to recognize the FAS1 4 module, was cleared in wide areas around adherent eosinophils. Clearance was attenuated by metalloproteinase inhibitors or antibodies to disintegrin metalloproteinase 8 (ADAM8), a major eosinophil metalloproteinase previously implicated in asthma pathogenesis. ADAM8 was not found in podosome-like structures, which are associated with proteolytic activity in other cell types. Instead, immunoblotting demonstrated proteoforms of ADAM8 that lack the cytoplasmic tail in the supernatant. Anti-ADAM8 inhibited migration of IL-5-stimulated eosinophils on periostin.
Conclusions And Clinical Relevance:
Migrating IL-5-activated eosinophils on periostin exhibit loss of nucleopodal features and appearance of prominent podosomes along with clearance of the Stiny-1 periostin epitope. Migration and epitope clearance are both attenuated by inhibitors of ADAM8. We propose, therefore, that eosinophils remodel and migrate on periostin-rich extracellular matrix in the asthmatic airway in an ADAM8-dependent manner, making ADAM8 a possible therapeutic target.
Insights
Interleukin-5 (IL-5) activated eosinophils migrate on periostin by losing their polarized structure and clearing the matrix via ADAM8. This ADAM8-dependent process suggests a therapeutic target for asthma.
Area of Science:
- Immunology
- Cell Biology
- Extracellular Matrix Biology
Background:
- Interleukin-5 (IL-5) induces eosinophil polarization, forming a specialized
- nucleopod
- capped with P-selectin glycoprotein ligand-1 (PSGL-1).
- IL-5 also enhances eosinophil adhesion and migration on periostin, an extracellular matrix protein upregulated in asthma.
Purpose of the Study:
- To investigate the morphological changes of IL-5-stimulated eosinophils during migration on periostin.
- To elucidate the role of ADAM8 in eosinophil migration and periostin remodeling.
Main Methods:
- Fluorescent microscopy of adhering eosinophils on periostin at various time points.
- Assaying eosinophil migration on periostin.
- Inhibition studies using metalloproteinase inhibitors and anti-ADAM8 antibodies.
Main Results:
- IL-5-stimulated eosinophils initially polarized with PSGL-1 at the nucleopod tip, later losing this structure.
- F-actin reorganized into podosome-like structures, and periostin was cleared around migrating eosinophils.
- Periostin clearance and eosinophil migration were inhibited by metalloproteinase inhibitors and anti-ADAM8 antibodies, with ADAM8 proteoforms detected in the supernatant.
Conclusions:
- Migrating eosinophils on periostin lose nucleopodal features and develop podosomes, clearing the matrix in an ADAM8-dependent manner.
- ADAM8 plays a crucial role in eosinophil migration and extracellular matrix remodeling.
- ADAM8 represents a potential therapeutic target for asthma.
Related Concept Videos
01:09Cell Migration
01:19Cell Migration
01:39Mechanism of Filopodia Formation
Their main function is to guide migrating cells during normal tissue morphogenesis or cancer metastasis by recognizing and making initial contacts with the extracellular matrix. However, they can also act as stationary cell anchors or help to establish communication...
01:31Mechanism of Lamellipodia Formation
01:16Cell Motility through Blebbing
Blebbing Through the Matrix
In multicellular...
01:22Role of Ephrin-Eph Signalling in Intestinal Stem Cell Renewal