Severe congenital neutropenia: abnormal growth and differentiation of myeloid progenitors to granulocyte colony-stimulating factor (G-CSF) but normal response to G-CSF plus stem cell factor

Several mechanisms have been proposed to explain the pathogenesis of severe congenital neutropenia (SCN); however, the mechanism(s) still remains unknown. In particular, clinical observations suggest that abnormal responsiveness of myeloid progenitors to hematopoietic growth factors (HGFs) is a possible mechanism. Therefore, to better define the status of hematopoietic progenitors in the bone marrow (BM) of patients with SCN, the responsiveness of myeloid progenitors to HGFs from two SCN patients was compared with the responsiveness of progenitors from healthy individuals. BM cells (BMCs) from the first SCN patient required higher (10- to 100-fold) concentrations of granulocyte colony- stimulating factor (G-CSF) to achieve maximal and half-maximal colony growth in vitro compared with BMCs from controls. In contrast, the dose- response of interleukin-3 (IL-3) and granulocyte-macrophage-CSF (GM- CSF) in colony formation was normal. Interestingly, IL-3, GM-CSF, and G- CSF at optimal doses showed reduced ability to induce neutrophil differentiation of BMCs from a SCN patient compared with BMCs from controls. Despite an abnormal responsiveness of mature myeloid progenitors to G-CSF in this SCN patient, myeloid progenitors responsive to the combination of stem cell factor (SCF) and G-CSF showed normal dose-response. In contrast to G-CSF alone, the combination of G-CSF and SCF induced the formation of neutrophils almost to the same extent compared with cultures of normal BMCs. Furthermore, also on BM progenitor cells obtained from the second patient with SCN, SCF highly synergized with G-CSF to promote neutrophil progenitor cell growth and differentiation in vitro. Thus, these results indicate that one mechanism of the pathogenesis in SCN patients is reduced responsiveness of neutrophil progenitor cells to G- CSF and that SCF can enhance the responsiveness of these cells to G-CSF.     

Theophylline potentiates glucagon-induced hepatic glucose production in man but does not prevent hepatic downregulation to glucagon

Prolonged hyperglucagonemia causes only a transient increase in hepatic glucose production. To determine whether activation of hepatic phosphodiesterase by glucagon is responsible for the transient nature of this response, the effect of infusion of the phosphodiestrase inhibitor theophylline alone, glucagon alone, and glucagon plus theophylline on isotopically determined glucose production was examined in normal human subjects. Infusion of theophylline alone did not alter rates of glucose production or utilization. Infusion of glucagon alone increased glucose production transiently from a basal rate of 1.9 ± 0.1 mg/kg/min to a maximum at min 30 of 2.8 ± 0.3 mg/kg/min followed by a return to rates no different from basal by min 60; plasma glucose increased from 89 ± 3 mg/dl to a maximum of 114 ± 5 mg/dl. Infusion of glucagon in the presence of theophylline resulted in greater increases in both plasma glucose (maximum at min 60 of 134 ± 9 mg/dl) and glucose production (maximum at min 30 of 3.5 ± 0.3 mg/kg/min) than had occurred during infusion of glucagon alone; the increase in glucose production, however, was not sustained. Thus theophylline potentiated glucagon-induced stimulation of hepatic glucose production, but it did not prevent the evanescent hepatic response to sustained hyperglucagonemia. Therefore, the present studies indicate that glucagon activation of hepatic phosphodiesterase does not appear to be responsible for the transient nature of the increase in hepatic glucose production observed during prolonged hyperglucagonemia.     

Dose-response characteristics for arginine-stimulated insulin secretion in man and influence of hyperglycemia

To test the hypothesis that glucose only affects the responsiveness (maximum velocity) of the beta-cell to arginine without changing the sensitivity (ED50) of the beta-cell to arginine, we investigated the influence of hyperglycemia on the responsiveness and sensitivity of arginine-induced insulin secretion in eight healthy male volunteers. Plasma C-peptide and insulin levels achieved during infusions of five doses of arginine (30 min) with and without a 60-min hyperglycemic clamp (17 mmol/L) were analyzed using a modified Michaelis-Menten equation. At euglycemia, the ED50 (half-maximally stimulating serum arginine concentration) was significantly less for first phase than for second phase plasma C-peptide secretion (0.7 +/- 0.1 vs. 2.7 +/- 0.4 mmol/L; P less than 0.002). Hyperglycemia significantly increased arginine-induced insulin secretion at all arginine infusion rates (P less than 0.01) without significantly altering the ED50 for either phase. We conclude 1) that the regulation of arginine-induced insulin secretion differs between both phases of insulin secretion, and 2) that a 1-h infusion with glucose significantly potentiates arginine-induced insulin secretion without influencing the difference in regulation of both phases of arginine-induced insulin secretion, supporting the validity of the use of arginine as a secretagogue in studies involving hyperglycemia.