Differential engraftment of genetically modified CD34+ and CD34- hematopoietic cell subsets in lethally irradiated baboons
Differential engraftment of genetically modified CD34+ and CD34- hematopoietic cell subsets in lethally irradiated baboons
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DOI：
10.1016/s0301-472x(00)00137-5
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发表时间：
2000-05-01
影响因子：
2.6
通讯作者：
Kiem, HP
中科院分区：
文献类型：
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作者：
Andrews, RG;Peterson, LJ;Kiem, HP
Objective.为了测试骨髓亚群中有助于体内造血重建的猩猩白血病病毒（GALV）假型载体转导。将分别通过与PG 13/LN、PG 13/LNX和PG 13/LNY载体产生细胞共培养转导的自体CD 34（+）Lin（-）、CD 34 + tint和CD 34 Lin-骨髓细胞移植到三只雌性狒狒中。还向两只雌性狒狒移植了来自MHC匹配的雄性同胞的新鲜同种异体CD 34（+）Lin（-）骨髓细胞，并为确保存活，分别用PG 13/LN和PG 13/LNX转导的自体CD 34（+）Lin（-）和CD 34（+）Lin（+）骨髓细胞。LN、LNX和LNY载体除了细菌新霉素磷酸转移酶（neo）基因的3'末端的长度序列不同之外是相同的。移植后所有动物的血液和骨髓中分别检测到来自CD 34（+）Lin（-）和CD 34（+）Lin（+）细胞的LN+和LNX+细胞，但未检测到来自CD 34（-）Lin（-）细胞的LNY+细胞。LN+、CD 34（+）Lin（-）细胞参与T、B和髓系的重建，LNX+、CD 34（+）Lin（+）细胞仅参与B和髓系的重建。通过聚合酶链反应在两只同种异体移植动物的血液和骨髓中检测到雄性细胞CD 34（-）Lin（-），估计频率在两只动物移植后1个月均小于或等于0.001%。雄性细胞在一只动物中变得无法检测到，并且在另一只动物中保持可检测到，频率下降，超过15个月。在该动物中，未检测到雄性CD 34（+）或集落形成细胞。CD 34（+）Lin（-）和CD 34（+）Lin（+）骨髓细胞可作为GALV假型逆转录病毒介导的基因转移的靶细胞。CD 34（+）Lin（-）细胞有助于所有造血谱系的重建，自体CD 34（+）Lin（-）细胞使用当前方法不被GALV假型逆转录病毒载体转导，或者不显著有助于重建，如同种异体移植所表明的。(C)2000年国际实验血液学学会，Elsevier Science Inc.出版。
Objective. To test gibbon ape leukemia virus (GALV) pseudotype vector transduction of marrow subpopulations that contribute to hematopoietic reconstitution in vivo.Materials and Methods. Autologous CD34(+) Lin(-), CD34+ tint, and CD34 Lin- marrow cells, transduced by coculture with PG13/LN, PG13/LNX, and PG13/LNY vector-producing cells, respectively, were transplanted in three female baboons. Two female baboons also were transplanted with fresh allogeneic CD34(+)Lin(-) marrow cells from MHC-matched male siblings and, to ensure survival, with autologous CD34(+)Lin(-) and CD34(+)Lin(+) marrow cells transduced with PG13/LN and PG13/LNX, respectively. The LN, LNX, and LNY vectors are identical except for different length sequences at the 3' end of the bacterial neomycin phosphotransferase (neo) gene.Results. LN+ and LNX+ cells from CD34(+)Lin(-) and CD34(+)Lin(+) cells, respectively, but no LNY+ from CD34(-)Lin(-) cells were detectable in blood and marrow of all animals after transplant. LN+, CD34(+)Lin(-) cells contributed to reconstitution of the T, B, and myeloid lineages, LNX+, CD34(+)Lin(+) cells contributed only to B and myeloid lineages. Male cells, CD34(-)Lin(-), were detected by polymerase chain reaction in blood and marrow of the two allogeneic transplanted animals at estimated frequencies of less than or equal to 0.001% 1 month after transplant in both animals. Male cells became undetectable in one animal and have remained detectable, with declining frequency, in the other for more than 15 months. In this animal, no male CD34(+) or colony-forming cells have been detected.Conclusions. CD34(+)Lin(-) and CD34(+)Lin(+) marrow cells can serve as targets for GALV pseudotype retrovirus-mediated gene transfer. CD34(+)Lin(-) cells contribute to reconstitution of all hematopoietic lineages, Autologous CD34(+)Lin(-) cells were either not transduced by GALV pseudotype retrovirus vectors using current approaches or did not contribute significantly to reconstitution, as suggested by allogeneic transplants. (C) 2000 International Society for Experimental Hematology, Published by Elsevier Science Inc.