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Prepublished online as a Blood First Edition Paper on August 22, 2002; DOI 10.1182/blood-2002-03-0756.
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Blood, 15 January 2003, Vol. 101, No. 2, pp. 492-497
HEMATOPOIESIS
Fetal hemoglobin modulation during human erythropoiesis: stem
cell factor has "late" effects related to the expression
pattern of CD117
Urszula Wojda,
Kristina R. Leigh,
Joyce M. Njoroge,
Kaedrea A. Jackson,
Bhanu Natarajan,
Michael Stitely, and
Jeffery L. Miller
From the Laboratory of Chemical Biology, National
Institute of Diabetes and Digestive and Kidney Diseases, National
Institutes of Health, Bethesda, MD.
A cytokine-screening assay of cultured peripheral blood cells
obtained using immune rosetting and separation of progenitors was
developed to identify determinants of fetal hemoglobin (HbF) modulation
during adult erythropoiesis. Among the 12 erythroid growth-promoting
cytokines tested, stem cell factor (SCF) at a concentration of 50 ng/mL
resulted in the most significant increase in cell proliferation
and HbF content. The average HbF/hemoglobin A (HbA) ratio was
30.9% ± 18.7% in cultures containing SCF compared with
4.1% ± 2.2% in those grown with erythropoietin (EPO) alone (P = 8.5E-8). To further investigate the
hemoglobin-modulating effects of SCF, we examined the surface
expression pattern of the SCF receptor, CD117, among maturing
erythroblasts. CD117 expression increased during the first week of
culture and peaked on culture days 7 to 9. After culture day 9, the
level of CD117 declined to lower levels. The rise in CD117 expression
to high levels mirrored that of the transferrin receptor (CD71), and
the subsequent reduction in CD117 was inversely related to increases in
expression of glycophorin A. SCF-related increases in the HbF/HbA ratio
correlated with the expression pattern of CD117. SCF added during days
7 to 14 resulted in a more pancellular distribution of HbF on day 14 compared with the heterocellular distribution present in cultures
supplemented with SCF on days 0 to 7. A significant SCF-mediated
increase in HbF was also measured using progenitors derived from cord
blood. These results suggest that the HbF response to SCF is greatest at the late progenitor stage as a function of surface CD117 expression.
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