Synergistic protecting effect of cord blood CD34+ cells over-expressing both interleukin-3 and Flt3 ligand on lethally irradiated mice.
The objective of the present work was to study the effect of interleukin-3 (IL-3) and Flt3 ligand (FL) over-expression in human cord blood CD34(+) cells on rescuing lethally irradiated mice by transplantation. CD34(+) cells were isolated from human umbilical cord blood (UCB) and infected with recombinant retrovirus expressing FL, IL-3 or FL/IL-3 genes, respectively. Stably transduced CD34(+) cells were transplanted i.v. into NOD/SCID/IL2rgamma(null) mice that had been conditioned with 8.0 Gy of irradiation. At 6 weeks post-irradiation, chimerism in the animal bone marrow (BM) and the spleen were investigated. Recovery of peripheral blood cell and animal survival were recorded 2, 4, and 6 weeks post-irradiation. The chimerism was further investigated by serial transplantation assay. At 6 weeks post-transplantation, each of the three transduced human UCB CD34(+) cell types could be observed in all examined BM of chimeric mice; however, more human cells could be found in BM, spleen and peripheral blood of those mice transplanted with CD34(+) cells over-expressing IL-3/FL cells. Over-expression of IL-3 and FL at mRNA and protein levels could be detected in the BM cells of chimeric mice. Accordingly, hIL-3/FL co-overexpressing mice showed greater recovery of peripheral blood counts as early as 2 weeks after irradiation, and a much higher survival rate (11/12) than other groups at 6 weeks post-irradiation. Serial transplantation assay indicated that human cord blood CD34(+) cells could recover potentials of proliferation and self-renewal after being transplanted into mice. Our results suggested that transplant of CD34(+) cord blood cells over-expressing IL-3/FL genes improved rescue of radiation-injured mice, which probably results from the synergistic effect between hIL-3 and hFL causing the rapid reconstitution of hematopoiesis.
Zhang
Yong
Y
Department of Hematology, Southwest Hospital, Third Military Medical University, 400038, Chongqing, China. nibingxi@yahoo.com.
Guo
Chaohua
C
Institute of Combined Injury of PLA, Third Military Medical University, 400038, Chongqing, China.
Zhang
Hongbin
H
Department of Hematology, Southwest Hospital, Third Military Medical University, 400038, Chongqing, China.
Dong
Shiwu
S
Institute of Combined Injury of PLA, Third Military Medical University, 400038, Chongqing, China.
D000818:Animals; D018952:Antigens, CD34; D036101:Cord Blood Stem Cell Transplantation; D006410:Hematopoiesis / Q000528:radiation effects; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D007377:Interleukin-3 / Q000096:biosynthesis*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011833:Radiation Injuries, Experimental / Q000378:metabolism / Q000628:therapy*; D013997:Time Factors; D018183:Transplantation Chimera / Q000378:metabolism; D014183:Transplantation, Heterologous; D014916:Whole-Body Irradiation; D051941:fms-Like Tyrosine Kinase 3 / Q000096:biosynthesis*
D018952:Antigens, CD34; C508596:IL3 protein, human; D007377:Interleukin-3; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3