Document Detail


Human eosinophils express functional CCR7.
MedLine Citation:
PMID:  23449735     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human eosinophils display directed chemotactic activity toward an array of soluble chemokines. Eosinophils have been observed to migrate to draining lymph nodes in experimental models of allergic inflammation, yet it is unknown whether eosinophils express CCR7, a key chemokine receptor in coordinating leukocyte trafficking to lymph nodes. The purpose of this study is to demonstrate expression of CCR7 by human eosinophils and functional responses to CCL19 and CCL21, the known ligands of CCR7. Human eosinophils were purified by negative selection from healthy donors. CCR7 expression of freshly purified, unstimulated eosinophils and of IL-5-primed eosinophils was determined by flow cytometry and Western blot. Chemotaxis to CCL19 and CCL21 was measured in transwell assays. Shape changes to CCL19 and CCL21 were analyzed by flow cytometry and microscopy. Calcium fluxes of fluo-4 AM-loaded eosinophils were recorded by flow cytometry after chemokine stimulation. ERK phosphorylation of CCL19- and CCL21-stimulated eosinophils was measured by Western blot and Luminex assay. Human eosinophils expressed CCR7 as demonstrated by flow cytometry and Western blots. Eosinophils exhibited detectable cell surface expression of CCR7. IL-5-primed eosinophils exhibited chemotaxis toward CCL19 and CCL21 in a dose-dependent fashion. Upon stimulation with CCL19 or CCL21, IL-5-primed eosinophils demonstrated dose-dependent shape changes with polarization of F-actin and exhibited calcium influxes. Finally, primed eosinophils stimulated with CCL19 or CCL21 exhibited increased phosphorylation of ERK in response to both CCR7 ligands. We demonstrate that human eosinophils express CCR7 and have multipotent responses to the known ligands of CCR7.
Authors:
Praveen Akuthota; Shigeharu Ueki; Jessica Estanislau; Peter F Weller
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  American journal of respiratory cell and molecular biology     Volume:  48     ISSN:  1535-4989     ISO Abbreviation:  Am. J. Respir. Cell Mol. Biol.     Publication Date:  2013 Jun 
Date Detail:
Created Date:  2013-06-04     Completed Date:  2013-08-05     Revised Date:  2014-03-19    
Medline Journal Info:
Nlm Unique ID:  8917225     Medline TA:  Am J Respir Cell Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  758-64     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Aniline Compounds / metabolism
Blotting, Western
Calcium / metabolism
Cell Shape
Chemokine CCL19 / immunology,  pharmacology
Chemokine CCL21 / immunology,  pharmacology
Chemotaxis
Dose-Response Relationship, Drug
Eosinophils / drug effects,  immunology,  metabolism*
Extracellular Signal-Regulated MAP Kinases / metabolism*
Flow Cytometry
Humans
Interleukin-5 / metabolism
Phosphorylation
Receptors, CCR7 / genetics,  metabolism*
Recombinant Proteins / metabolism
Xanthenes / metabolism
Grant Support
ID/Acronym/Agency:
F32AI081513/AI/NIAID NIH HHS; R01 AI051645/AI/NIAID NIH HHS; R01/R37AI020241/AI/NIAID NIH HHS; R01AI051645/AI/NIAID NIH HHS; R37 AI020241/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Actins; 0/Aniline Compounds; 0/CCL19 protein, human; 0/CCL21 protein, human; 0/CCR7 protein, human; 0/Chemokine CCL19; 0/Chemokine CCL21; 0/Fluo 4; 0/IL5 protein, human; 0/Interleukin-5; 0/Receptors, CCR7; 0/Recombinant Proteins; 0/Xanthenes; EC 2.7.11.24/Extracellular Signal-Regulated MAP Kinases; SY7Q814VUP/Calcium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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