Diphtheria toxin A gene-mediated HIV-1 protection of cord blood-derived T cells in the SCID-hu mouse model.
Diphtheria toxin A gene-mediated HIV-1 protection of cord blood-derived T cells in the SCID-hu mouse model.
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DOI：
10.1089/scd.1.1998.7.319
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发表时间：
1998-08
期刊：
影响因子：
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通讯作者：
N. Banda;R. Akkina;Kristina L. Terrell;Elizabeth J. Shpall;J. Tomczak;Julie Campain;Henry N. Claman;Linda Cagle;Gail S. Harrison
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文献类型：
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作者：
N. Banda;R. Akkina;Kristina L. Terrell;Elizabeth J. Shpall;J. Tomczak;Julie Campain;Henry N. Claman;Linda Cagle;Gail S. Harrison
CD34+来源的脐带血（CB）细胞被调节白喉毒素a （DT-A）链基因转导，在含有人类胸腺和肝脏组成的联合器官（thy/liv）的SCID-hu小鼠中检测了其重建潜力。将经dt - a转导的细胞直接注射到活体器官中，显示出与用非dt - a亲本载体转导的对照CB细胞相同的移植潜力。通过HLA标记B7与thy/ live细胞区分开来的CB细胞在离体培养中被优先保留。在肝/活器官中，移植的CB细胞占细胞总数的80%。大多数细胞（约70%）在thy/liv器官也是CD4+CD8+，正如预期的成熟胸腺细胞。注射CB细胞后44天（与30天和80天相比）双阳性细胞的发生率最高。这表明，细胞在活体器官中实现最佳增殖需要最短的时间，但在后来的时间里，所有早期细胞都成熟了。因此，用于移植的群体包含早期细胞，但不包含自我更新细胞。双阳性细胞在离体培养中迅速成熟为单阳性细胞（CD4+或CD8+）。在thy/ live -derived细胞中可以很容易地检测到标记细胞（neo+）。经DT-A转导的细胞在离体培养中对HIV T淋巴细胞NL4-3具有长期的保护作用。本研究表明，dt - a转导细胞在移植活体器官时没有明显的缺陷，在体内也没有任何毒性作用。这些细胞即使在转导后超过2个月和在thy/ live小鼠中植入44天后仍能抵抗HIV感染。这些数据支持毒素基因治疗作为HIV感染策略的可行性。
The reconstitutive potential of CD34+-derived cord blood (CB) cells, transduced with a regulated diphtheria toxin A (DT-A) chain gene, was examined in SCID-hu mice harboring a conjoint organ composed of human thymus and liver (thy/liv). The DT-A-transduced cells, injected directly into the thy/liv organ, showed the same engraftment potential as control CB cells transduced with the non-DT-A parental vector. CB cells, distinguishable from the thy/liv cells by the HLA marker B7, were preferentially maintained in ex vivo culture. In the thy/liv organ, the engrafted CB cells represented >80% of the total cells. A majority of cells (>70%) in the thy/liv organ were also CD4+CD8+, as would be expected of maturing thymocytes. The incidence of double-positive cells was highest at 44 days (compared with 30 days and 80 days) after injection of CB cells. This suggested that a minimum time was required to achieve optimal proliferation of cells in the thy/liv organ but that, at later times, all of the early cells had matured. Thus, the population used for engraftment contained early cells but not self-renewing cells. The double-positive cells matured rapidly into single-positive cells (either CD4+ or CD8+) when placed in ex vivo culture. Marked cells (neo+) could readily be detected in the thy/liv-derived cells. The cells transduced with DT-A showed long-term protection in ex vivo culture against HIV T lymphotropic isolate NL4-3. This study shows that DT-A-transduced cells had no apparent disadvantage in engraftment of the thy/liv organ and did not have any toxic effects in vivo. Such cells were protected against HIV infection even when challenged more than 2 months after transduction and after a 44-day engraftment period in the thy/liv mice. These data support the feasibility of toxin gene therapy as a strategy for HIV infection.