Neutrophil elastase-processing defect in cyclic hematopoietic dogs

Ronghua Meng, Roger Bridgman, Maria Toivio-Kinnucan, Glenn P. Niemeyer, William Vernau, Tommy Hock, Clinton D. Lothrop

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Objective: Canine cyclic hematopoiesis (CH), a model of human cyclic neutropenia and severe congenital neutropenia, is characterized by a periodic reduced neutrophil count and decreased neutrophil elastase (NE) enzymatic activity. Canine CH is caused by a mutation of AP3B1 encoding the β3A subunit of adaptor protein complex-3 (AP-3). It has been proposed that trafficking of elastase is affected by AP-3. The aim of this study was to study intracellular sorting/trafficking of NE in CH dogs using antibodies specific to canine NE. Materials and Methods: Polyclonal and monoclonal antibodies were generated to immunogenic epitopes in the middle (aa85-98) and C-terminal (aa269-282) regions of NE. The antibodies to canine NE were characterized by Western immunoblotting and immunocytochemistry. Results: Antibody ELA85 (antibody to canine NE aa 85-98) specifically recognized mature 28-kD NE. Immunocytochemical analysis using ELA85 and an antibody to myeloperoxidase demonstrated colocalizaton of NE and myeloperoxidase in primary granules of normal dogs. Antibody ELA269 (antibody to canine NE aa 269-282) reacted exclusively with the 33-kD NE presumptive precursor form. Immunocytochemical analysis demonstrated that the NE precursor was not colocalized with myeloperoxidase in the primary granules of normal or CH dogs. Western immunoblotting using these antibodies demonstrated that CH dogs contained reduced mature NE, but accumulated a large amount of the NE precursor protein that was not enzymatically active. Conclusion: Antibodies ELA85 and ELA269 were found to be useful reagents for studying the biosynthesis, processing, and trafficking of NE during normal myelopoiesis. Neutrophils from CH dogs accumulated large amounts of higher molecular weight elastase precursors compared to normal dogs.

Original languageEnglish (US)
Pages (from-to)104-115
Number of pages12
JournalExperimental Hematology
Volume38
Issue number2
DOIs
StatePublished - Feb 2010

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Leukocyte Elastase
Dogs
Canidae
Antibodies
Adaptor Protein Complex 3
Peroxidase
Pancreatic Elastase
Neutrophils
Western Blotting
Myelopoiesis
Protein Precursors
Cyclic neutropenia

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Hematology

Cite this

Meng, R., Bridgman, R., Toivio-Kinnucan, M., Niemeyer, G. P., Vernau, W., Hock, T., & Lothrop, C. D. (2010). Neutrophil elastase-processing defect in cyclic hematopoietic dogs. Experimental Hematology, 38(2), 104-115. https://doi.org/10.1016/j.exphem.2009.09.010

Neutrophil elastase-processing defect in cyclic hematopoietic dogs. / Meng, Ronghua; Bridgman, Roger; Toivio-Kinnucan, Maria; Niemeyer, Glenn P.; Vernau, William; Hock, Tommy; Lothrop, Clinton D.

In: Experimental Hematology, Vol. 38, No. 2, 02.2010, p. 104-115.

Research output: Contribution to journalArticle

Meng, R, Bridgman, R, Toivio-Kinnucan, M, Niemeyer, GP, Vernau, W, Hock, T & Lothrop, CD 2010, 'Neutrophil elastase-processing defect in cyclic hematopoietic dogs', Experimental Hematology, vol. 38, no. 2, pp. 104-115. https://doi.org/10.1016/j.exphem.2009.09.010
Meng, Ronghua ; Bridgman, Roger ; Toivio-Kinnucan, Maria ; Niemeyer, Glenn P. ; Vernau, William ; Hock, Tommy ; Lothrop, Clinton D. / Neutrophil elastase-processing defect in cyclic hematopoietic dogs. In: Experimental Hematology. 2010 ; Vol. 38, No. 2. pp. 104-115.
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