FLT LIGAND PERIPHERAL BLOOD STEM CELL MOBILIZATION FOR STEM CELL GENE THERAPY
用于干细胞基因治疗的 FLT 配体外周血干细胞动员
基本信息
- 批准号:6591324
- 负责人:
- 金额:$ 11.11万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2002
- 资助国家:美国
- 起止时间:2002-05-01 至 2003-04-30
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Optimization of the harvesting and transduction of hematopoietic
stem cells is critical to successful stem cell gene therapy We
evaluated the utility of a novel protocol involving flt-3 and G-CSF
mobilization of peripheral blood stem cells and retroviral
transduction using hematopoietic growth factors to introduce a
reporter gene, murine CD24, into hematopoietic stem cells in nonhuman
primates Rhesus macaques were treated with flt-3 ligand (200 mg/kg)
and G-CSF (20 mg/kg) for 7 days and autologous CD34+ peripheral blood
stem cells harvested by leukapheresis CD34+ cells were transduced with
an MFG-based retroviral vector encoding murine CD24 using 4 daily
transductions with centrifugations in the presence of flt-3 ligand
(100 ng/ml), SCF (100 ng/ml) and PIXY (100 ng/ml) in serum free medium
Animals were sublethally irradiated (400 cGy) on the day of
transplantation Engraftment was monitored sequentially in the bone
marrow and blood using both multiparameter flo w cytometry and
semi-quantitative DNA PCR Our data demonstrate successful and
persistent engraftment in multiple hematopoietic lineages, including
granulocytes, monocytes and B and T lymphocytes At 46 and 117 days
post transplantation 75% and 6% of granulocytes, respectively,
expressed mCD24 antigen at the cell surface Peak in vivo levels of
genetically-modified peripheral blood lymphocytes (PBL) approached 60%
as assessed both by flow cytometry and PCR 18 days post
transplantation In addition, naive (CD45RA+ and CD62L+) CD4+ and
CD3+8+ cells were the predominant phenotype of the T cells detected at
this time Engraftment has subsequently plateaued but persisted at
between 10 and 15% of PBLs for 133 days Acytotoxic T cell response
against murine CD24 was detected in only one animal This degree of
persistent long-lived, high level gene marking of multiple
hematopoietic lineages, including naive T cells, has important
implications for the field of stem cell gene therapy
造血干细胞的收获和转导的优化
干细胞是成功的干细胞基因治疗的关键,
评估了一种新的方案,包括flt-3和G-CSF的效用
外周血干细胞动员和逆转录病毒
使用造血生长因子进行转导,
报告基因,鼠CD 24,进入非人造血干细胞
灵长类恒河猴用flt-3配体(200 mg/kg)处理
和G-CSF(20 mg/kg)7天,以及自体CD 34+外周血
通过白细胞去除术收获的干细胞CD 34+细胞用
基于MFG的编码鼠CD 24的逆转录病毒载体,每天使用4次
在FLT-3配体存在下离心转导
(100无血清培养基中的SCF(100 ng/ml)和PIXY(100 ng/ml
在给药当天对动物进行亚致死剂量辐照(400 cGy)。
在骨中连续监测移植物植入
使用多参数流式细胞术和
半定量DNA PCR我们的数据表明成功的,
在多种造血谱系中持续植入,包括
46天和117天时的粒细胞、单核细胞以及B和T淋巴细胞
移植后分别有75%和6%的粒细胞,
在细胞表面表达mCD 24抗原,体内峰值水平为
转基因外周血淋巴细胞(PBL)接近60%
如在18天后通过流式细胞术和PCR两者评估的,
此外,移植初始(CD 45 RA+和CD 62 L+)CD 4+和
CD 3 +8+细胞是T细胞的主要表型,
这一次,植入随后达到稳定状态,但仍持续,
10 - 15%的PBL持续133天细胞毒性T细胞应答
仅在一只动物中检测到针对鼠CD 24的抗体
持久的长寿命,高水平的基因标记的多种
造血谱系,包括幼稚T细胞,具有重要的
干细胞基因治疗领域的意义
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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M ROSENZWEIG其他文献
M ROSENZWEIG的其他文献
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{{ truncateString('M ROSENZWEIG', 18)}}的其他基金
STEM CELL GENE THERAPY USING LONG TERM BONE MARROW CULTURE TRANSDUCTION PROTOCOL
使用长期骨髓培养转导方案的干细胞基因治疗
- 批准号:
6591288 - 财政年份:2002
- 资助金额:
$ 11.11万 - 项目类别:
IN VIVO ANALYSIS OF T CELL TURNOVER IN RHESUS MACAQUES
恒河猴 T 细胞更新的体内分析
- 批准号:
6591317 - 财政年份:2002
- 资助金额:
$ 11.11万 - 项目类别:
EXPRESSION OF BCL 2 IN SIV INFECTED THYMIC TISSUE
BCL 2 在 SIV 感染的胸腺组织中的表达
- 批准号:
6591316 - 财政年份:2002
- 资助金额:
$ 11.11万 - 项目类别:
XENOGENEIC THYMIC TRANSPLANTATION AS AN ADJUNCT TO TREATMENT OF AIDS
异种胸腺移植作为艾滋病治疗的辅助手段
- 批准号:
6591306 - 财政年份:2002
- 资助金额:
$ 11.11万 - 项目类别:
EXPRESSION OF BCL 2 IN SIV INFECTED THYMIC TISSUE
BCL 2 在 SIV 感染的胸腺组织中的表达
- 批准号:
6453762 - 财政年份:2001
- 资助金额:
$ 11.11万 - 项目类别:
IN VIVO ANALYSIS OF T CELL TURNOVER IN RHESUS MACAQUES
恒河猴 T 细胞更新的体内分析
- 批准号:
6453763 - 财政年份:2001
- 资助金额:
$ 11.11万 - 项目类别:
STEM CELL GENE THERAPY USING LONG TERM BONE MARROW CULTURE TRANSDUCTION PROTOCOL
使用长期骨髓培养转导方案的干细胞基因治疗
- 批准号:
6453734 - 财政年份:2001
- 资助金额:
$ 11.11万 - 项目类别:
FLT LIGAND PERIPHERAL BLOOD STEM CELL MOBILIZATION FOR STEM CELL GENE THERAPY
用于干细胞基因治疗的 FLT 配体外周血干细胞动员
- 批准号:
6453770 - 财政年份:2001
- 资助金额:
$ 11.11万 - 项目类别:
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