The release of inflammatory Cytokines from human peripheral blood mononuclear cells in vitro following exposure to adenovirus variants and capsid
The release of inflammatory Cytokines from human peripheral blood mononuclear cells in vitro following exposure to adenovirus variants and capsid
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DOI：
10.1089/10430340152712683
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发表时间：
2002-01-01
影响因子：
4.2
通讯作者：
Ramsey, WJ
中科院分区：
文献类型：
--
作者：
Higginbotham, JN;Seth, P;Ramsey, WJ
腺病毒载体的临床前和临床研究已经清楚地说明了这种基因转移系统的潜在优势。然而，许多研究也已经证明了针对载体和转导细胞的有效免疫应答。我们研究了在体外反应的人外周血单核细胞（PBMC）病毒暴露作为这种主机响应的模型。从正常供体中分离PBMC，并与野生型腺病毒（Ad 5）、缺失E1和/或E3片段的Ad 5变体和空病毒衣壳一起孵育。监测促炎细胞因子释放96小时。完整病毒体对TNF-α的诱导很低，尽管空衣壳的刺激产生了显著和持续的反应。完整病毒体对IL-6、GM-CSF和a组α-和β-趋化因子的诱导作用显著，通常接近用2.5 mg/ml脂多糖（LPS）获得的结果。反应一般是独立的病毒粒子的遗传组成，只有部分钝化时，紫外线灭活的病毒。剂量反应数据显示病毒体浓度增加100倍，细胞因子释放最多增加3倍，表明达到饱和。令人惊讶的是，在添加空衣壳后发生显著的刺激，其通常引起与LPS刺激所见的反应相当的反应。我们提出的论点，细胞信号转导机制激活结合病毒粒子/衣壳刺激转录的促炎细胞因子基因。
Preclinical and clinical studies with adenoviral vectors have clearly illustrated the potential advantages of this gene transfer system. However, many studies have also demonstrated potent immune responses directed at both vector and transduced cells. We examined in vitro responses of human peripheral blood mononuclear cells (PBMC) to virus exposure as a model for this host response. PBMC were isolated from normal donors and incubated with wild-type adenovirus (Ad5), Ad5 variants deleted for segments of E1 and/or E3, and empty viral capsids. Proinflammatory cytokine release was monitored for 96 hr. Induction of TNF-alpha by intact virions was low although stimulation by empty capsid gave a significant and sustained response. Induction of IL-6, GM-CSF, and a panel alpha- and beta-chemokines by intact virions was prominent, often approaching results obtained with 2.5 mg/ml of lipopolysaccharide (LPS). Responses were generally independent of virion genetic composition and were only partially blunted when UV-inactivated virus was used. Dose-response data showed 100-fold increases in virion concentration produced a maximum 3-fold increase in cytokine release, suggesting saturation. Surprisingly, prominent stimulation occurred after addition of empty capsid, which typically provoked responses equivalent to those seen with LPS stimulation. We present arguments that cellular signal transduction mechanisms activated by binding of virions/capsids stimulate transcription of proinflammatory cytokine genes.