Signal through gp130 activated by soluble interleukin (IL)-6 receptor (R) and IL-6 or IL-6R/IL-6 fusion protein enhances ex vivo expansion of human peripheral blood-derived hematopoietic progenitors.
This study was designed to investigate the effects of a combination of soluble interleukin (sIL)-6 receptor (R) and IL-6 on the ex vivo expansion of human peripheral blood (PB)-derived hematopoietic progenitor cells in a short-term serum-free liquid suspension culture system, using PB-derived CD34(+)IL-6R(+/-) cells as a target. In combination with stem cell factor (SCF), IL-3, and sIL-6R/IL-6, the expansion efficiency (EE) for granulocyte/macrophage colony-forming unit (CFU-GM) reached a peak level on day 10 of incubation. On the other hand, the EE for erythroid burst (BFU-E) and mixed colony-forming unit (CFU-Mix) reached a peak level on day 7 of incubation. Among the cytokine combinations tested, SCF + IL-3 + sIL-6R/IL-6 + flt3 ligand (FL) most effectively expanded CFU-GM and CFU-Mix. The maximum EEs for CFU-GM and CFU-Mix were 208-fold and 42-fold, respectively. While the EE for BFU-E was 70-90-fold in the presence of SCF + IL-3 + sIL-6R/IL-6, FL significantly augmented the EE for CFU-GM and CFU-Mix. In contrast, thrombopoietin (TPO) significantly augmented the EE for CFU-Mix. Interestingly, in combination with IL-3 and SCF, newly generated IL-6R/IL-6 fusion protein (FP) expanded PB-derived BFU-E and CFU-Mix twice more effectively than a combination of sIL-6R and IL-6. These results demonstrated that human PB-derived committed progenitors were effectively expanded in vitro using sIL-6R/IL-6 or FP, in combination with IL-3, SCF and/or FL or TPO, and that FP may transduce a stronger intracellular signal than a combination of sIL-6R and IL-6.
Kimura
T
T
Department of Hygiene, the Department of Gastroenterological Surgery, and the Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Wang
J
J
Minamiguchi
H
H
Fujiki
H
H
Harada
S
S
Okuda
K
K
Kaneko
H
H
Yokota
S
S
Yasukawa
K
K
Abe
T
T
Sonoda
Y
Y
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D050822:Cytokine Receptor gp130; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism* / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism / Q000494:pharmacology; D019947:Receptors, Interleukin-6 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology; D015398:Signal Transduction; D012995:Solubility; D019089:Stem Cell Factor / Q000378:metabolism / Q000494:pharmacology; D013926:Thrombopoietin / Q000378:metabolism / Q000494:pharmacology
D015703:Antigens, CD; C494199:IL6ST protein, human; D007377:Interleukin-3; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019947:Receptors, Interleukin-6; D011993:Recombinant Fusion Proteins; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D050822:Cytokine Receptor gp130; D013926:Thrombopoietin