|
|
 Previous Article | Table of Contents | Next Article 
c-kit ligand reactivates fetal hemoglobin synthesis in serum-free culture
of stringently purified normal adult burst-forming unit- erythroid
C Peschle, M Gabbianelli, U Testa, E Pelosi, T Barberi, C Fossati, M Valtieri and L Leone
Department of Hematology-Oncology, Istituto Superiore di Sanita, Rome,
Italy.
We have analyzed the reactivation of fetal hemoglobin (HbF) synthesis under
rigorous in vitro conditions, ie, in mature erythroblasts generated by
erythroid burst-forming units (BFU-E) stringently purified from normal
adult peripheral blood and grown in fetal calf serum(FCS)- free semisolid
or liquid phase culture. In clonogenetic dishes, graded amounts of c-kit
ligand (KL) were added together with saturating levels of erythropoietin
(Ep) and variable amounts of interleukin-3 and granulocyte-macrophage
colony stimulating factor (IL-3/GM-CSF), ie, high or low level, or no
IL-3/GM-CSF addition. In all conditions, KL induced a sharp, dose-dependent
increase in the percentage of F cells and HbF content from nearly normal
levels (< 10% and < 2.5%, respectively, at 0.1 and 1 ng/mL) up to 40%
to 50% and 10% to 15% at 100 to 200 ng/mL. This increase was not associated
with significant differences of burst number or stage of maturation at the
time of analysis (as evaluated on the basis of percent mature erythroblasts
and Hb content per cell). However, the KL-induced reactivation of HbF
synthesis was strictly and directly correlated with a sharp increase of
colony size, ie, cell number per burst. Addition of large amounts of IL- 3
and GM-CSF (10 to 100 U and 1 to 10 ng/mL, respectively) significantly
potentiated the KL-induced reactivation of HbF, as compared with low levels
(0.1 U and 0.01 to 0.1 ng) or no addition of these growth factors: this
increase was highly significant at low KL doses (ie, 1 to 10 ng/mL).
Single-burst analysis showed that the KL- induced HbF reactivation occurs
homogeneously in the erythroid colonies within each of these culture
conditions. We have analyzed the effect of KL in liquid phase BFU-E culture
treated with the IL-3/GM-CSF/Ep combination at sequential times until
terminal erythroid maturation: KL causes a sharp increase in the percentage
of F cells and HbF content in all stages of maturation, whereas the
IL-3/GM-CSF/Ep combination alone has a markedly lower effect. These results
suggest that KL plays a key role in the reactivation of HbF synthesis in
adult life, whereas IL- 3/GM-CSF potentiate this effect at low KL levels.
The KL-induced HbF reactivation is seemingly related to an enhanced
proliferation of erythroid progenitors in the erythropoietic
differentiation pathway.
Volume 81,
Issue 2,
pp. 328-336,
01/15/1993
Copyright © 1993 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
W. Aerbajinai, J. Zhu, C. Kumkhaek, K. Chin, and G. P. Rodgers
SCF induces {gamma}-globin gene expression by regulating downstream transcription factor COUP-TFII
Blood,
July 2, 2009;
114(1):
187 - 194.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Gabbianelli, O. Morsilli, A. Massa, L. Pasquini, P. Cianciulli, U. Testa, and C. Peschle
Effective erythropoiesis and HbF reactivation induced by kit ligand in -thalassemia
Blood,
January 1, 2008;
111(1):
421 - 429.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Amoyal, E. Prus, and E. Fibach
Vanadate Elevates Fetal Hemoglobin in Human Erythroid Precursors by Inhibiting Cell Maturation
Experimental Biology and Medicine,
May 1, 2007;
232(5):
654 - 661.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. V. Bhanu, T. A. Trice, Y. T. Lee, N. M. Gantt, P. Oneal, J. D. Schwartz, P. Noel, and J. L. Miller
A sustained and pancellular reversal of gamma-globin gene silencing in adult human erythroid precursor cells
Blood,
January 1, 2005;
105(1):
387 - 393.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. V. Bhanu, T. A. Trice, Y. T. Lee, and J. L. Miller
A signaling mechanism for growth-related expression of fetal hemoglobin
Blood,
March 1, 2004;
103(5):
1929 - 1933.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Zeuner, F. Pedini, M. Signore, U. Testa, E. Pelosi, C. Peschle, and R. De Maria
Stem cell factor protects erythroid precursor cells from chemotherapeutic agents via up-regulation of BCL-2 family proteins
Blood,
July 1, 2003;
102(1):
87 - 93.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Gabbianelli, U. Testa, A. Massa, O. Morsilli, E. Saulle, N. M. Sposi, E. Petrucci, G. Mariani, and C. Peschle
HbF reactivation in sibling BFU-E colonies: synergistic interaction of kit ligand with low-dose dexamethasone
Blood,
April 1, 2003;
101(7):
2826 - 2832.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
U. Wojda, K. R. Leigh, J. M. Njoroge, K. A. Jackson, B. Natarajan, M. Stitely, and J. L. Miller
Fetal hemoglobin modulation during human erythropoiesis: stem cell factor has "late" effects related to the expression pattern of CD117
Blood,
January 15, 2003;
101(2):
492 - 497.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Gabbianelli, U. Testa, A. Massa, E. Pelosi, N. M. Sposi, R. Riccioni, L. Luchetti, and C. Peschle
Hemoglobin switching in unicellular erythroid culture of sibling erythroid burst-forming units: kit ligand induces a dose-dependent fetal hemoglobin reactivation potentiated by sodium butyrate
Blood,
June 1, 2000;
95(11):
3555 - 3561.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Malik, T. C. Fisher, L. L.W. Barsky, L. Zeng, P. Izadi, A. L. Hiti, K. I. Weinberg, T. D. Coates, H. J. Meiselman, and D. B. Kohn
An In Vitro Model of Human Red Blood Cell Production From Hematopoietic Progenitor Cells
Blood,
April 15, 1998;
91(8):
2664 - 2671.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. C. Broudy
Stem Cell Factor and Hematopoiesis
Blood,
August 15, 1997;
90(4):
1345 - 1364.
[Full Text]
[PDF]
|
|
|
|
|
