Dendritic cells transduced with an adenovirus vector encoding Epstein-Barr virus latent membrane protein 2B: A new modality for vaccination
Dendritic cells transduced with an adenovirus vector encoding Epstein-Barr virus latent membrane protein 2B: A new modality for vaccination
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DOI：
10.1128/jvi.73.12.10416-10425.1999
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发表时间：
1999-12-01
影响因子：
5.4
通讯作者：
Storkus, WJ
中科院分区：
文献类型：
--
作者：
Ranieri, E;Herr, W;Storkus, WJ
爱泼斯坦-巴尔病毒(EBV)是一种疱疹病毒，通常与多种恶性肿瘤有关，尤其是在免疫功能低下的宿主中。作为一种刺激抗EBV免疫的策略用于治疗EBV相关肿瘤，我们利用重组腺病毒载体，通过基因工程的树突状细胞(DC)表达EBV抗原，如潜伏膜蛋白2B(LMP2B)。用携带增强型绿色荧光蛋白(EGFP)或AdLMP2B的重组腺病毒载体AdEGFP感染的自体DC培养CD8(+)T淋巴细胞，感染倍数为250。48h后，荧光激活细胞分选分析显示，95%的DC远端EGFP表达阳性，表明基因转移有效。用AdLMP2转导的DC刺激CD8(+)T细胞。用酶联斑点(ELISPOT)法检测应答CD8(+)T细胞的干扰素(IFN-γ)释放和细胞毒活性。在体外刺激之前，通过干扰素-γ斑点形成(T细胞频率，
Epstein-Barr virus (EBV) is a herpesvirus commonly associated with several malignancies, particularly in immunocompromised hosts. As a strategy for stimulating immunity against EBV for the treatment of EBV-associated tumors, we have genetically engineered dendritic cells (DC) to express EBV antigens, such as latent membrane protein 2B (LMP2B), using recombinant adenovirus vectors. CD8(+) T lymphocytes from HLA-A2.1(+), EBV-seropositive healthy donors were cultured with autologous DC infected with recombinant adenovirus vector AdEGFP, encoding an enhanced green fluorescent protein (EGFP), or AdLMP2B at a multiplicity of infection of 250. After 48 h, >95% of the DC were positive far EGFP expression as assessed by fluorescence-activated cell sorting analysis, indicating efficient gene transfer. AdLMP2-transduced DC were used to stimulate CD8(+) T cells. Responder CD8(+) T cells were tested for gamma interferon (IFN-gamma) release by enzyme-linked spot (ELISPOT) assay and cytotoxic activity. Prior to in vitro stimulation, the frequencies of T-cells directed against two HLA-A2-presented LMP2 peptides (LMP2 329-337 and LMP2 426-434) were very low as assessed by IFN-gamma spot formation (T-cell frequency,