Thrombopoietin enhances neutrophil production by bone marrow hematopoietic progenitors with the aid of stem cell factor in congenital neutropenia.
We examined the effects of granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF), and thrombopoietin (TPO), alone or in combination, on the generation of neutrophils by bone marrow (BM) cells from three patients with severe congenital neutropenia (SCN) through the use of a serum-deprived liquid culture system. Synergistic effects of G-CSF and SCF on the neutrophil production by BM CD34+CD38+c-kit+ cells were observed in SCN patients as well as in normal controls. The addition of TPO to the culture containing G-CSF and SCF further augmented the growth of neutrophils in the two groups. Single-cell culture experiments revealed that the three-factor combination caused increases in both the number and size of neutrophil colonies compared with G-CSF + SCF in normal BM cells, whereas only a significant increment in the colony size was observed in SCN patients. Even in the presence of SCF or SCF + TPO, the concentrations of G-CSF necessary for the substantial production of neutrophils by CD34+CD38+c-kit+ cells were higher in two patients compared with the levels obtained by normal control cells. In addition, TPO did not accelerate the maturation of neutrophilic cells supported by G-CSF + SCF. When BM CD34+CD38-c-kit+ cells were targeted, the addition of TPO to the culture containing G-CSF and SCF was required for significant neutrophil colony growth in the two groups. These results suggest that TPO enhances the G-CSF-dependent neutrophil production with the aid of SCF in this disorder.
Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan.
N
N
Sawai
K
K
Koike
H H
HH
Mwamtemi
S
S
Ito
Y
Y
Kurokawa
K
K
Sakashita
T
T
Kinoshita
T
T
Higuchi
K
K
Takeuchi
M
M
Shiohara
T
T
Kamijo
Y
Y
Higuchi
H
H
Miyazaki
T
T
Kato
M
M
Kobayashi
M
M
Miyake
K
K
Yasui
A
A
Komiyama
D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D017209:Apoptosis; D001853:Bone Marrow / Q000187:drug effects* / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D002675:Child, Preschool; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D007223:Infant; D008297:Male; D008562:Membrane Glycoproteins; D009244:NAD+ Nucleosidase / Q000032:analysis; D009503:Neutropenia / Q000188:drug therapy / Q000473:pathology*; D009504:Neutrophils / Q000473:pathology*; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis / Q000187:drug effects; D019089:Stem Cell Factor / Q000008:administration & dosage / Q000494:pharmacology*; D013926:Thrombopoietin / Q000008:administration & dosage / Q000494:pharmacology*
D015703:Antigens, CD; D018952:Antigens, CD34; D000943:Antigens, Differentiation; D016895:Culture Media, Serum-Free; D008562:Membrane Glycoproteins; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D013926:Thrombopoietin; D019009:Proto-Oncogene Proteins c-kit; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1