TY - JOUR
T1 - Donor chimerism and stem cell function in a murine congenic transplantation model after low-dose radiation conditioning
T2 - Effects of a retroviral-mediated gene transfer protocol and implications for gene therapy
AU - Goebel, W. Scott
AU - Yoder, Mervin C.
AU - Pech, Nancy K.
AU - Dinauer, Mary C.
N1 - Funding Information:
This work was supported by National Heart, Lung, and Blood Institute Grants P01 HL53586, (M.C.D.), and R01 HL63169 (M.C.Y.) and a Clinical Research Award from the March of Dimes (No. 1FY97; M.C.D.). The Wells Center for Pediatric Research is a Center for Excellence in Molecular Hematology funded by the National Institute of Diabetes and Digestive and Kidney Diseases (P50 DK 49218). W.S.G. is a William Kennedy Fellow of the National Childhood Cancer Foundation. The authors thank Laura Haneline for helpful discussions regarding the competitive repopulation assays, and Shari Upchurch for assistance with preparing the manuscript.
PY - 2002/11/1
Y1 - 2002/11/1
N2 - Objective. We investigated low-dose radiation conditioning for the transplantation of retrovirus-transduced cells in a C57Bl6/J murine model. Materials and Methods. The effect of low-dose radiation on stem cell function was investigated using a competitive repopulation assay. Stem cell function of marrow cells that underwent a retroviral-mediated gene transfer (RMGT) protocol was examined by this assay, and donor chimerism of these cells when transplanted into 160-cGy conditioned syngeneic hosts was compared to fresh marrow. Results. Irradiation with 300 or 160 cGy substantially decreased stem cell function as measured by competitive repopulation. Animals conditioned with 160 cGy and transplanted with 20 × 106 fresh marrow cells permitted donor cell engraftment of 53.6% ± 11.4% 6 months after transplant compared to 100% donor cell engraftment after 1100 cGy irradiation. Lymphoid and myeloid engraftment did not significantly differ from total engraftment in submyeloablated hosts. When transplanted into lethally irradiated hosts, the competitive repopulating activity of marrow treated with a single dose of 5-fluorouracil followed by ex vivo culture according to a standard RMGT protocol was equal to 5-fluorouracil-only treated marrow. However, cells treated with 5-fluorouracil or 5-fluorouracil plus ex vivo culture for RMGT repopulated less well than fresh marrow cells in 160 cGy conditioned hosts. Conclusions. Low-dose irradiation decreases host stem cell function, allowing engraftment of both fresh and RMGT protocol-treated marrow, although the engraftment of 5-fluorouracil-treated cells was reduced at least two-fold, and 5-fluorouracil plus RMGT protocol-treated cells at least three-fold, compared to fresh marrow. Modification of current RMGT protocols may be important for optimizing engraftment under these conditions.
AB - Objective. We investigated low-dose radiation conditioning for the transplantation of retrovirus-transduced cells in a C57Bl6/J murine model. Materials and Methods. The effect of low-dose radiation on stem cell function was investigated using a competitive repopulation assay. Stem cell function of marrow cells that underwent a retroviral-mediated gene transfer (RMGT) protocol was examined by this assay, and donor chimerism of these cells when transplanted into 160-cGy conditioned syngeneic hosts was compared to fresh marrow. Results. Irradiation with 300 or 160 cGy substantially decreased stem cell function as measured by competitive repopulation. Animals conditioned with 160 cGy and transplanted with 20 × 106 fresh marrow cells permitted donor cell engraftment of 53.6% ± 11.4% 6 months after transplant compared to 100% donor cell engraftment after 1100 cGy irradiation. Lymphoid and myeloid engraftment did not significantly differ from total engraftment in submyeloablated hosts. When transplanted into lethally irradiated hosts, the competitive repopulating activity of marrow treated with a single dose of 5-fluorouracil followed by ex vivo culture according to a standard RMGT protocol was equal to 5-fluorouracil-only treated marrow. However, cells treated with 5-fluorouracil or 5-fluorouracil plus ex vivo culture for RMGT repopulated less well than fresh marrow cells in 160 cGy conditioned hosts. Conclusions. Low-dose irradiation decreases host stem cell function, allowing engraftment of both fresh and RMGT protocol-treated marrow, although the engraftment of 5-fluorouracil-treated cells was reduced at least two-fold, and 5-fluorouracil plus RMGT protocol-treated cells at least three-fold, compared to fresh marrow. Modification of current RMGT protocols may be important for optimizing engraftment under these conditions.
UR - https://www.scopus.com/pages/publications/0036840102
U2 - 10.1016/S0301-472X(02)00927-X
DO - 10.1016/S0301-472X(02)00927-X
M3 - Article
C2 - 12423686
AN - SCOPUS:0036840102
SN - 0301-472X
VL - 30
SP - 1324
EP - 1332
JO - Experimental Hematology
JF - Experimental Hematology
IS - 11
ER -