Thrombopoietin enhances neutrophil production by bone marrow hematopoieticprogenitors with the aid of stem cell factor in congenital neutropenia

We examined the effects of granulocyte colony-stimulating factor (G-CSF), s tent cell factor (SCF), and thrombopoietin (TPO), alone or in combination, on the generation of neutrophils by bone marrow (BM) cells from three patie nts with severe congenital neutropenia (SCN) through the use of a serum-dep rived Liquid culture system. Synergistic effects of G-CSF and SCF on the ne utrophil 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 th e two groups, Single-cell culture experiments revealed that the three-facto r combination caused increases in both. the number and size of neutrophil c olonies compared with G-CSF + SCF in normal BM cells, whereas only a signif icant increment in the colony size was observed in SCN patients. Even in th e presence of SCF or SCF + TPO, the concentrations of G-CSF necessary for t he substantial production of neutrophils by CD39(+)CD38(+)c-kit(+) cells we re higher in two patients compared with the levels obtained by normal contr ol cells. In addition, TPO did not accelerate the maturation of neutrophili c 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.