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Molecular cloning and in vivo evaluation of canine granulocyte- macrophage
colony-stimulating factor
RA Nash, F Schuening, F Appelbaum, WP Hammond, T Boone, CF Morris, SJ Slichter and R Storb
Division of Clinical Research, Fred Hutchinson Cancer Research Center,
Seattle, WA 98104.
Canine granulocyte-macrophage colony-stimulating factor (caGM-CSF) was
cloned and expressed to allow further investigation of GM-CSF in a large
animal model. The cDNA is 850 base pairs (bp) long and encodes a peptide of
144 amino acids. The nucleotide and amino acid sequence homology between
caGM-CSF and human GM-CSF (hGM-CSF) is 80% and 70%, respectively. A
mammalian expression vector pCMV/CAGM was constructed and used to transfect
COS cells for expression of caGM-CSF. Supernatant from transfected COS
cells enriched with caGM-CSF was shown to have significant stimulating
activity in granulocyte-macrophage colony forming unit (CFU-GM) assays of
canine marrow. caGM-CSF, expressed from bacteria, was used to treat seven
dogs at varying doses twice daily subcutaneously (sc) for 14 to 16 days.
Circulating blood neutrophils and monocytes increased significantly. The
increase in circulating eosinophils was variable. Thrombocytopenia
developed during administration of caGM-CSF but corrected rapidly after
cessation of treatment. Evaluation of survival times of 51Cr-labeled
autologous platelets suggested increased consumption as the primary reason
for thrombocytopenia. A species-specific GM-CSF will be a useful tool for
hematologic or immunologic studies in dogs.
Volume 78,
Issue 4,
pp. 930-937,
08/15/1991
Copyright © 1991 by The American Society of Hematology
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