Transgene expression by a large fraction of dendritic cells following autologous transplantation of retrovirally transduced CD34 cells.
Transgene expression by a large fraction of dendritic cells following autologous transplantation of retrovirally transduced CD34 cells.
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逆转录病毒转导的 CD34+ 细胞自体移植后，大部分树突状细胞进行转基因表达。
DOI：
10.1089/scd.2006.15.619
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发表时间：
2006
影响因子：
4
通讯作者：
Kiem,Hans-Peter
中科院分区：
文献类型：
--
作者：
Storek,Jan;Kiem,Hans-Peter
树突状细胞（DC）在对感染性微生物和癌细胞的免疫应答中起着至关重要的作用（1，2）。它们对于维持外周对自身抗原的耐受性也是必不可少的（3，4）。这两种功能都是通过抗原捕获和加工以及抗原片段（与主要组织相容性复合体结合）的表面表达来介导的。如果DC表面上的抗原片段与共刺激分子如CD 40或CD 86共表达，则T细胞被刺激。如果不是，或者如果DC分泌色氨酸代谢物，则T细胞可能变得无反应性或缺失（5，6）。基因工程改造的DC将来可用于治疗，以刺激免疫（如果工程改造以表达微生物或癌症抗原和共刺激分子）（7）或抑制自身免疫（如果工程改造以表达自身抗原并且不表达共刺激分子或产生色氨酸代谢物）。在这里，我们提出了一个观察，工程化的DC（表达转基因）是有效地产生在体内从携带转基因的CD 34细胞。对6只幼年狒狒进行辐照（10.2戈伊），并按照所述输注自体CD 34细胞（8，9）。如所述（10），用携带MNDMFGeYFP转基因（MNDMFG启动子和增强型黄色荧光蛋白基因）的凤凰巨猿白血病病毒假型肿瘤逆转录病毒转导CD 34细胞。输注的CD 34细胞中有39 - 70%表达YFP（因此携带转基因）。移植后，通过流式细胞术测定循环粒细胞（侧向散射-高细胞）、CD 4 T细胞[通过前向和侧向散射测定的CD 3 + CD 4 + CD 8+单核细胞（MNC）]、CD 8 T细胞（CD 3 + CD 4 + CD 8 + MNC）、B细胞（CD 20 + MNC）、单核细胞（CD 14 + MNC）、自然杀伤（NK）细胞中YFP+细胞的百分比
DENDRITIC CELLS (DCs) play a crucial role in immune responses to infectious microorganisms and cancer cells (1, 2). They are also essential for maintaining peripheral tolerance to self-antigens (3, 4). Both functions are mediated through antigen capture and processing and surface expression of fragments of the antigen (bound to the major histocompatibility complex). If the antigen fragments on the DC surface are coexpressed with costimulatory molecules like CD40 or CD86, T cells become stimulated. If not or if the DC secretes tryptophan metabolites, T cells may be rendered anergic or deleted (5, 6). Genetically engineered DCs may be used therapeutically in the future either to stimulate immunity (if engineered to express microbial or cancer antigens and costimulatory molecules)(7) or inhibit autoimmunity (if engineered to express autoantigens and to either not express costimulatory molecules or produce tryptophan metabolites). Here we present an observation that engineered DCs (expressing a transgene) are efficiently generated in vivo from CD34 cells carrying the transgene. Six juvenile baboons were irradiated (10.2 Gy) and infused with autologous CD34 cells as described (8, 9). The CD34 cells were transduced with a Phoenix Gibbon Ape Leukemia Virus-pseudotyped oncoretrovirus carrying the MNDMFGeYFP transgene (MNDMFG promoter and enhanced yellow fluorescent protein gene) as described (10). Thirty-nine to 70% of the infused CD34 cells expressed YFP (thus carried the transgene). Posttransplant, the percentage of YFP+ cells was determined by flow cytometry among circulating granulocytes (side scatter-high cells), CD4 T cells [CD3+ CD4+ CD8 J mononuclear cells (MNCs) by forward and side scatter], CD8 T cells (CD3+ CD4 J CD8+ MNCs), B cells (CD20+ MNCs), monocytes (CD14+ MNCs), natural killer (NK) cells