Abnormal response to granulocyte colony-stimulating factor (G-CSF) in canine cyclic hematopoiesis is not caused by altered G-CSF receptor expression

Belinda R. Avalos, Virginia C. Broudy, Sarah K. Ceselski, Brian Druker, James D. Griffin, William P. Hammond

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

A decrease in responsiveness to granulocyte colony-stimulating factor (G- CSF) has been implicated in the pathophysiology of cyclic hematopoiesis. Using the canine model of cyclic neutropenia, we examined the response of neutrophil precursors to G-CSF in vitro and G-CSF receptor expression in neutrophils from grey collie dogs to determine whether the abnormal response observed to G-CSF in vivo in this disorder is present at the level of the progenitor cell and is caused by defective G-CSF receptor expression. Bone marrow mononuclear cells from grey collie dogs required sevenfold higher G- CSF concentrations than normal dog cells to achieve half-maximal colony growth (56 pmol/L v 8 pmol/L). Receptor binding assays with 125I-labeled G-CSF and Scatchard analyses of the equilibrium binding data were consistent with expression of a single class of high-affinity receptors for G-CSF on neutrophils from both normal dogs and grey collies with similar receptor numbers (56 to 446 sites/cell v 78 to 199 sites/cell) and binding affinities (28 to 206 pmol/L v 84 to 195 pmol/L). Chemical cross-linking studies identified a G-CSF binding protein of approximately 120 kD on neutrophils from grey collies, similar in size to that on normal dog neutrophils. No abnormal G-CSF receptor mRNA transcripts were detected in neutrophils from grey collie dogs by Northern blot analysis. Treatment of both normal and grey collie neutrophils with G-CSF rapidly induced tyrosine phosphorylation of an 80-kD protein that behaved like canine c-rel. These results demonstrate that the abnormal responsiveness to G-CSF in canine cyclic hematopoiesis is present in neutrophil precursors and is not associated with demonstrable alterations in the number, binding affinity, or overall size of the G-CSF receptor in neutrophils, or with defective tyrosine phosphorylation of p80. These data suggest that cyclic hematopoiesis is caused by a defect in the G- CSF signal transduction pathway at a point distal to G-CSF receptor binding that does not involve the early biochemical events leading to p80 tyrosine phosphorylation.

Original languageEnglish (US)
Pages (from-to)789-794
Number of pages6
JournalBlood
Volume84
Issue number3
StatePublished - Aug 1 1994
Externally publishedYes

Fingerprint

Granulocyte Colony-Stimulating Factor Receptors
Granulocyte Colony-Stimulating Factor
Canidae
Neutrophils
Dogs
Phosphorylation
Tyrosine
Cyclic neutropenia
Signal transduction
Bone Marrow Cells
Northern Blotting
Statistical Factor Analysis
Assays
Signal Transduction
Carrier Proteins
Bone
Stem Cells
Binding Sites

ASJC Scopus subject areas

  • Hematology

Cite this

Abnormal response to granulocyte colony-stimulating factor (G-CSF) in canine cyclic hematopoiesis is not caused by altered G-CSF receptor expression. / Avalos, Belinda R.; Broudy, Virginia C.; Ceselski, Sarah K.; Druker, Brian; Griffin, James D.; Hammond, William P.

In: Blood, Vol. 84, No. 3, 01.08.1994, p. 789-794.

Research output: Contribution to journalArticle

Avalos, Belinda R. ; Broudy, Virginia C. ; Ceselski, Sarah K. ; Druker, Brian ; Griffin, James D. ; Hammond, William P. / Abnormal response to granulocyte colony-stimulating factor (G-CSF) in canine cyclic hematopoiesis is not caused by altered G-CSF receptor expression. In: Blood. 1994 ; Vol. 84, No. 3. pp. 789-794.
@article{4831a55f41164b079841c10437c79ed6,
title = "Abnormal response to granulocyte colony-stimulating factor (G-CSF) in canine cyclic hematopoiesis is not caused by altered G-CSF receptor expression",
abstract = "A decrease in responsiveness to granulocyte colony-stimulating factor (G- CSF) has been implicated in the pathophysiology of cyclic hematopoiesis. Using the canine model of cyclic neutropenia, we examined the response of neutrophil precursors to G-CSF in vitro and G-CSF receptor expression in neutrophils from grey collie dogs to determine whether the abnormal response observed to G-CSF in vivo in this disorder is present at the level of the progenitor cell and is caused by defective G-CSF receptor expression. Bone marrow mononuclear cells from grey collie dogs required sevenfold higher G- CSF concentrations than normal dog cells to achieve half-maximal colony growth (56 pmol/L v 8 pmol/L). Receptor binding assays with 125I-labeled G-CSF and Scatchard analyses of the equilibrium binding data were consistent with expression of a single class of high-affinity receptors for G-CSF on neutrophils from both normal dogs and grey collies with similar receptor numbers (56 to 446 sites/cell v 78 to 199 sites/cell) and binding affinities (28 to 206 pmol/L v 84 to 195 pmol/L). Chemical cross-linking studies identified a G-CSF binding protein of approximately 120 kD on neutrophils from grey collies, similar in size to that on normal dog neutrophils. No abnormal G-CSF receptor mRNA transcripts were detected in neutrophils from grey collie dogs by Northern blot analysis. Treatment of both normal and grey collie neutrophils with G-CSF rapidly induced tyrosine phosphorylation of an 80-kD protein that behaved like canine c-rel. These results demonstrate that the abnormal responsiveness to G-CSF in canine cyclic hematopoiesis is present in neutrophil precursors and is not associated with demonstrable alterations in the number, binding affinity, or overall size of the G-CSF receptor in neutrophils, or with defective tyrosine phosphorylation of p80. These data suggest that cyclic hematopoiesis is caused by a defect in the G- CSF signal transduction pathway at a point distal to G-CSF receptor binding that does not involve the early biochemical events leading to p80 tyrosine phosphorylation.",
author = "Avalos, {Belinda R.} and Broudy, {Virginia C.} and Ceselski, {Sarah K.} and Brian Druker and Griffin, {James D.} and Hammond, {William P.}",
year = "1994",
month = "8",
day = "1",
language = "English (US)",
volume = "84",
pages = "789--794",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "3",

}

TY - JOUR

T1 - Abnormal response to granulocyte colony-stimulating factor (G-CSF) in canine cyclic hematopoiesis is not caused by altered G-CSF receptor expression

AU - Avalos, Belinda R.

AU - Broudy, Virginia C.

AU - Ceselski, Sarah K.

AU - Druker, Brian

AU - Griffin, James D.

AU - Hammond, William P.

PY - 1994/8/1

Y1 - 1994/8/1

N2 - A decrease in responsiveness to granulocyte colony-stimulating factor (G- CSF) has been implicated in the pathophysiology of cyclic hematopoiesis. Using the canine model of cyclic neutropenia, we examined the response of neutrophil precursors to G-CSF in vitro and G-CSF receptor expression in neutrophils from grey collie dogs to determine whether the abnormal response observed to G-CSF in vivo in this disorder is present at the level of the progenitor cell and is caused by defective G-CSF receptor expression. Bone marrow mononuclear cells from grey collie dogs required sevenfold higher G- CSF concentrations than normal dog cells to achieve half-maximal colony growth (56 pmol/L v 8 pmol/L). Receptor binding assays with 125I-labeled G-CSF and Scatchard analyses of the equilibrium binding data were consistent with expression of a single class of high-affinity receptors for G-CSF on neutrophils from both normal dogs and grey collies with similar receptor numbers (56 to 446 sites/cell v 78 to 199 sites/cell) and binding affinities (28 to 206 pmol/L v 84 to 195 pmol/L). Chemical cross-linking studies identified a G-CSF binding protein of approximately 120 kD on neutrophils from grey collies, similar in size to that on normal dog neutrophils. No abnormal G-CSF receptor mRNA transcripts were detected in neutrophils from grey collie dogs by Northern blot analysis. Treatment of both normal and grey collie neutrophils with G-CSF rapidly induced tyrosine phosphorylation of an 80-kD protein that behaved like canine c-rel. These results demonstrate that the abnormal responsiveness to G-CSF in canine cyclic hematopoiesis is present in neutrophil precursors and is not associated with demonstrable alterations in the number, binding affinity, or overall size of the G-CSF receptor in neutrophils, or with defective tyrosine phosphorylation of p80. These data suggest that cyclic hematopoiesis is caused by a defect in the G- CSF signal transduction pathway at a point distal to G-CSF receptor binding that does not involve the early biochemical events leading to p80 tyrosine phosphorylation.

AB - A decrease in responsiveness to granulocyte colony-stimulating factor (G- CSF) has been implicated in the pathophysiology of cyclic hematopoiesis. Using the canine model of cyclic neutropenia, we examined the response of neutrophil precursors to G-CSF in vitro and G-CSF receptor expression in neutrophils from grey collie dogs to determine whether the abnormal response observed to G-CSF in vivo in this disorder is present at the level of the progenitor cell and is caused by defective G-CSF receptor expression. Bone marrow mononuclear cells from grey collie dogs required sevenfold higher G- CSF concentrations than normal dog cells to achieve half-maximal colony growth (56 pmol/L v 8 pmol/L). Receptor binding assays with 125I-labeled G-CSF and Scatchard analyses of the equilibrium binding data were consistent with expression of a single class of high-affinity receptors for G-CSF on neutrophils from both normal dogs and grey collies with similar receptor numbers (56 to 446 sites/cell v 78 to 199 sites/cell) and binding affinities (28 to 206 pmol/L v 84 to 195 pmol/L). Chemical cross-linking studies identified a G-CSF binding protein of approximately 120 kD on neutrophils from grey collies, similar in size to that on normal dog neutrophils. No abnormal G-CSF receptor mRNA transcripts were detected in neutrophils from grey collie dogs by Northern blot analysis. Treatment of both normal and grey collie neutrophils with G-CSF rapidly induced tyrosine phosphorylation of an 80-kD protein that behaved like canine c-rel. These results demonstrate that the abnormal responsiveness to G-CSF in canine cyclic hematopoiesis is present in neutrophil precursors and is not associated with demonstrable alterations in the number, binding affinity, or overall size of the G-CSF receptor in neutrophils, or with defective tyrosine phosphorylation of p80. These data suggest that cyclic hematopoiesis is caused by a defect in the G- CSF signal transduction pathway at a point distal to G-CSF receptor binding that does not involve the early biochemical events leading to p80 tyrosine phosphorylation.

UR - http://www.scopus.com/inward/record.url?scp=0028285845&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028285845&partnerID=8YFLogxK

M3 - Article

VL - 84

SP - 789

EP - 794

JO - Blood

JF - Blood

SN - 0006-4971

IS - 3

ER -