Retroviral vector backbone immunogenicity: identification of cytotoxic T-cell epitopes in retroviral vector-packaging sequences
Retroviral vector backbone immunogenicity: identification of cytotoxic T-cell epitopes in retroviral vector-packaging sequences
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DOI：
10.1038/sj.gt.3302406
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发表时间：
2005-02-01
期刊：
影响因子：
5.1
通讯作者：
Takahashi, T
中科院分区：
文献类型：
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作者：
Kondo, E;Akatsuka, Y;Takahashi, T
逆转录病毒载体是临床基因治疗中常用的基因传递载体，但其对插入基因编码蛋白的特异性免疫原性是一个需要克服的问题。在这里，我们描述了人类细胞毒性t淋巴细胞（CTL）克隆识别来自逆转录病毒载体主干编码的蛋白质的表位，这是在我们尝试在体外产生抗巨细胞病毒（CMV）或人乳头瘤病毒（HPV）的CTL过程中建立的。在健康cmv血清阴性供者的情况下，通过用CMV-pp65逆转录病毒转导的cd40 -活化B （CD40-B）细胞刺激CD8 T细胞，在8个供者中有4个产生了针对逆转录病毒转导细胞的CTL系。从其中一条CTL系衍生的两个CTL克隆分别识别HLA-B* 4403和-B*4601的gag表位。同样，来自宫颈癌患者的HLA-B* 3501限制性CTL克隆识别出位于gag和pol序列连接区域的表位。这些结果表明，由逆转录病毒载体主干成分编码的多肽实际上具有免疫原性，在体外的人类细胞中产生ctl。因此，在临床环境中也应考虑对逆转录病毒产品的潜在CTL反应。
Retroviral vectors are the frequently applied gene delivery vehicles for clinical gene therapy, but specificity of the immunogenicity to the protein encoded by the inserted gene of interest is a problem which needs to be overcome. Here, we describe human cytotoxic T-lymphocyte (CTL) clones recognizing epitopes derived from the protein encoded by the retroviral vector backbone, which were established during the course of our attempts to generate CTLs against cytomegalovirus (CMV) or human papilloma virus (HPV) in vitro. In the case of healthy CMV-seronegative donors, CTL lines specific for retrovirally transduced cells were generated in four out of eight donors by stimulating CD8 T cells with CD40-activated B (CD40-B) cells retrovirally transduced with CMV-pp65. Two CTL clones derived from one of the CTL lines were found to recognize epitopes from gag in the context of HLA-B* 4403 and -B*4601, respectively. Similarly, an HLA-B* 3501-restricted CTL clone from a cervical cancer patient recognized an epitope located in the junctional regions of the gag and pol sequences. These results show that polypeptides encoded by components of the retroviral vector backbone are in fact immunogenic, generating CTLs in vitro in human cells. Thus, potential CTL responses to retroviral products should also be considered in clinical settings.