Bone marrow stromal cell line co-transfected with IL-2 and IL-3 genes can accelerate restoration of T-cell immunity in allo-BMT mice.
BACKGROUND
After T-cell depleted allogeneic bone marrow transplantation, impaired immune reconstitution is a major cause of morbidity and mortality in the recipient. The purpose of this study was to observe the effects of the gene-engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 on the reconstitution of T-cell immunity in allo-BMT mice.


METHODS
The bone marrow stromal cell line QXMSC1 was co-transfected with IL-2 and IL-3 genes using a Tet-on gene expression system. T lymphocyte subset counts per spleen were analyzed by flow cytometry. Lymphocyte proliferation response to ConA was examined to evaluate T-cell function. CDR3 spectratyping techniques were performed to evaluate TCR repertoire diversity at various time points post-transplantation.


RESULTS
Gene engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 could express IL-2 and IL-3 [1,300 ng.day(-1).10(-6) cells and 1100 ng.day(-1).10(-6) cells, respectively] under the control of doxycycline. QXMSC1-IL-2 + IL-3 in combination with allogeneic bone marrow could significantly increase the counts of CD(4)(+) and CD(8)(+) T cell, 1.72 and 1.27-fold respectively at week 3 compared with TCD-BMT group (P < 0.01); make CD(4)(+)/CD(8)(+) ratio return to normal level at week 4; enhance splenocytes mitotic response to ConA (P < 0.01), and accelerate restoration of TCR repertoire diversity in the lethally irradiated mice (P < 0.05).


CONCLUSIONS
The gene transduced stromal cell line QXMSC1-IL-2 + IL-3 is able to accelerate T-cell immunity in allo-BMT mice.
Li
Ai-ling
AL
Department of Immunology, Peking University Health Science Center, Beijing 100083, China.
Jiang
Ji-yang
JY
Ma
Jian-bo
JB
Wang
Guang-ming
GM
Hao
Jie
J
Gao
Xiang
X
Xie
Shu-Sheng
SS
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D016026:Bone Marrow Transplantation / Q000276:immunology*; D002460:Cell Line; D022801:Complementarity Determining Regions; D004318:Doxycycline / Q000494:pharmacology; D005260:Female; D007376:Interleukin-2 / Q000235:genetics*; D007377:Interleukin-3 / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D017154:Stromal Cells / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection; D014184:Transplantation, Homologous
D022801:Complementarity Determining Regions; D007376:Interleukin-2; D007377:Interleukin-3; D004318:Doxycycline