Immunogenicity of a p210(BCR-ABL) fusion domain candidate DNA vaccine targeted to dendritic cells by a recombinant adeno-associated virus vector in vitro.
Immunogenicity of a p210(BCR-ABL) fusion domain candidate DNA vaccine targeted to dendritic cells by a recombinant adeno-associated virus vector in vitro.
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发表时间：
2002-06
期刊：
影响因子：
11.2
通讯作者：
Ji-Yao Sun;R. Krouse;S. Forman;D. Senitzer;I. Sniecinski;S. Chatterjee;K. Wong
中科院分区：
文献类型：
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作者：
Ji-Yao Sun;R. Krouse;S. Forman;D. Senitzer;I. Sniecinski;S. Chatterjee;K. Wong
慢性髓性白血病（CML）的特点是t（9;22）易位，导致嵌合BCR-ABL融合癌蛋白的表达，这是肿瘤发生所必需的，是白血病克隆所特有的，是免疫治疗的诱人靶点。作为CML的免疫治疗策略，我们构建了一个重组腺相关病毒载体，编码p210(BCR-ABL) b3a2变异融合区和侧翼序列（CWRBA），并利用它在原代人树突状细胞（dc）中表达BCR-ABL融合区，dc是目前已知的最有效的抗原呈递细胞。来自健康供者的外周血单核细胞在体外被纯化的CWRBA、CWRAP（阴性对照）转导的自体dc引物和再刺激（阳性对照），或用与融合结构域对应的肽脉冲（阳性对照）。用CWRAP转导的dc未产生特异性反应。相比之下，cwrba转导的dc以抗原特异性、mhc限制的方式启动自体T细胞，其水平与阳性对照相当。cwrba转导的dc引起细胞毒性CD4+/Th1和CD8+反应，尽管前者在该系统中更容易检测到。对内源性表达p210(BCR-ABL) b3a2变异融合区的肿瘤细胞系的细胞毒性也被证实。此外，HLA-DRB5(*)0101+DRA （DR2a）被鉴定为能够呈递b3a2 BCR-ABL融合区表位的新限制性元件。因此，本文开发的构建物可作为CML基因抗原特异性免疫治疗的候选疫苗，并可作为使用重组腺相关病毒载体编码多表位免疫原转导的dc用于疫苗开发的范例。
Chronic myelogenous leukemia (CML) is characterized by a t(9;22) translocation, which results in the expression of chimeric BCR-ABL fusion oncoproteins that are necessary for oncogenesis, unique to the leukemic clones, and represent enticing targets for immunotherapy. As a strategy for the immunotherapy of CML, we constructed a recombinant adeno-associated virus vector encoding the p210(BCR-ABL) b3a2 variant fusion region with flanking sequences (CWRBA) and used it to express the BCR-ABL fusion region within primary human dendritic cells (DCs), the most potent antigen-presenting cells currently known. Peripheral blood mononuclear cells from healthy donors were primed and restimulated in vitro with autologous DCs transduced with purified CWRBA, CWRAP (negative control), or pulsed with a peptide corresponding to the fusion domain (positive control). No specific responses were generated using DCs transduced with CWRAP. In contrast, CWRBA-transduced DCs primed autologous T cells in an antigen-specific, MHC-restricted fashion to levels comparable with the positive control. CWRBA-transduced DCs elicited both cytotoxic CD4+/Th1 and CD8+ responses, although the former were more readily detected in this system. Cytotoxicity against a tumor cell line endogenously expressing the p210(BCR-ABL) b3a2 variant fusion region was also demonstrable. In addition, HLA-DRB5(*)0101+DRA (DR2a) was identified as a new restriction element capable of presenting the b3a2 BCR-ABL fusion region epitope. Thus, the construct developed herein may serve as a candidate vaccine for gene-based antigen-specific immunotherapy of CML and may serve as a paradigm for the use of DCs transduced with recombinant adeno-associated virus vectors encoding multiepitope immunogens for vaccine development.