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In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy
Published on: August 24, 2018
Neutrophil elastase-processing defect in cyclic hematopoietic dogs
Ronghua Meng1, Roger Bridgman, Maria Toivio-Kinnucan
1Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University, Auburn, Ala., USA.
Experimental Hematology
|November 28, 2009
Summary
Canine cyclic hematopoiesis (CH) involves abnormal neutrophil elastase (NE) processing. CH dogs accumulate inactive NE precursors, unlike normal dogs, impacting neutrophil function.
Area of Science:
- Hematology
- Molecular Biology
- Immunology
Background:
- Canine cyclic hematopoiesis (CH) is a model for human neutropenias.
- CH is linked to AP3B1 mutations affecting protein trafficking.
- Neutrophil elastase (NE) activity is reduced in CH.
Purpose of the Study:
- Investigate intracellular sorting and trafficking of NE in CH dogs.
- Utilize specific antibodies against canine NE.
Main Methods:
- Generated polyclonal and monoclonal antibodies to canine NE.
- Characterized antibodies using Western immunoblotting and immunocytochemistry.
- Analyzed NE precursor and mature NE localization and quantity.
Main Results:
- Developed antibodies recognizing mature NE (ELA85) and NE precursor (ELA269).
- Normal dogs show colocalization of mature NE and myeloperoxidase in primary granules.
- CH dogs accumulate inactive, higher molecular weight NE precursors, not colocalized with myeloperoxidase.
Conclusions:
- ELA85 and ELA269 are valuable tools for studying NE biosynthesis and trafficking.
- CH neutrophils exhibit abnormal accumulation of NE precursors, impacting enzyme activity.
