Engraftment of NOD/SCID mice with human CD34+ cells transduced by concentrated oncoretroviral vector particles pseudotyped with the feline endogenous retrovirus (RD 114) envelope protein

被引:39
作者
Gatlin, J
Melkus, MW
Padgett, A
Kelly, PF
Garcia, JV [1 ]
机构
[1] Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9206, Dallas, TX 75390 USA
[2] St Jude Childrens Res Hosp, Dept Hematol, Memphis, TN 38105 USA
关键词
D O I
10.1128/JVI.75.20.9995-9999.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Oncoretrovirus vectors pseudotyped with the feline endogenous retrovirus (RD114) envelope protein produced by the FLYRD18 packaging cell line have previously been shown to transduce human hematopoietic progenitor cells with a greater efficiency than similar amphotropic envelope-pseudotyped vectors. In this report, we describe the production and efficient concentration of RD114-pseudotyped murine leukemia virus (MLV)-based vectors. Following a single round of centrifugation, vector supernatants were concentrated approximately 200-fold with a 50 to 70% yield. Concentrated vector stocks transduced prestimulated human CD34(+) (hCD34(+)) cells with approximately 69% efficiency (n = 7, standard deviation = 4.4%) using a single addition of vector at a low multiplicity of infection (MOI = 5). Introduction of transduced hCD34(+) cells into irradiated NOD/SCID recipients resulted in multilineage engraftment with long-term transgene expression. These data demonstrate that RD114-pseudotyped MLV-based vectors can be efficiently concentrated to high titers and that hCD34(+) cells transduced with concentrated vector stocks retain in vivo repopulating potential. These results highlight the potential of RD114-pseudotyped oncoretrovirus vectors for future clinical implementation in hematopoietic stem cell gene transfer.
引用
收藏
页码:9995 / 9999
页数:5
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