Defects of granulopoiesis in patients with severe congenital neutropenia

To confirm the abnormalities of primitive myeloid progenitor cells in patients with severe congenital neutropenia (SCN), we studied their responsiveness to hematopoietic factors including granulocyte colony-stimulating factor (G-CSF). In all SCN patients studied no abnormalities of granulocyte colony-stimulating factor receptor (G-CSFR) gene were detected by polymerase chain reaction-single-strand conformation polymorphism analysis and sequence analysis. A flow cytometric analysis of bone marrow cells based on the expression of CD34, Kit receptor, and G-CSFR demonstrated a reduced frequency of CD34+/Kit+/G-CSFR+ cells in patients with SCN. The granulocyte/macrophage (GM)-colony formation of CD34+/Kit+/G-CSFR+ cells in patients was markedly decreased at all concentrations of G-CSF in serum-deprived semisolid culture. The responsiveness of CD34+/Kit+/G-CSFR+ cells in patients showed a reduced response to the combination of stem cell factor, the ligand for flk2/flt3, and interleukin-3 with or without G-CSF in serum-deprived semisolid and liquid suspension cultures. In contrast, no difference in the responsiveness of CD34+/Kit+/G-CSFR- cells was noted between SCN patients and normal subjects. The bone marrow cells from a patient who underwent bone marrow transplantation showed a restoration of both the reduced frequency and the decreased level of GM-colony formation of CD34+/Kit+/G-CSFR+ cells. These results demonstrate that the presence of qualitative and quantitative abnormalities of primitive myeloid progenitor cells expressing G-CSFR may play an important role in the impairment of granulopoiesis in patients with SCN.