Perreau Matthieu, Mennechet Franck, Serratrice Nicolas, Glasgow Joel N, Curiel David T, Wodrich Harald, Kremer Eric J
Institut de Génétique Moléculaire de Montpellier, CNRS 5535, 1919 Route de Mende, 34293 Montpellier, France.
J Virol. 2007 Apr;81(7):3272-84. doi: 10.1128/JVI.01530-06. Epub 2007 Jan 17.
Antipathogen immune responses create a balance between immunity, tolerance, and immune evasion. However, during gene therapy most viral vectors are delivered in substantial doses and are incapable of expressing gene products that reduce the host's ability to detect transduced cells. Gene transfer efficacy is also modified by the in vivo transduction of dendritic cells (DC), which notably increases the immunogenicity of virions and vector-encoded genes. In this study, we evaluated parameters that are relevant to the use of canine adenovirus serotype 2 (CAV-2) vectors in the clinical setting by assaying their effect on human monocyte-derived DC (hMoDC). We compared CAV-2 to human adenovirus (HAd) vectors containing the wild-type virion, functional deletions in the penton base RGD motif, and the CAV-2 fiber knob. In contrast to the HAd type 5 (HAd5)-based vectors, CAV-2 poorly transduced hMoDC, provoked minimal upregulation of major histocompatibility complex class I/II and costimulatory molecules (CD40, CD80, and CD86), and induced negligible morphological changes indicative of DC maturation. Functional maturation assay results (e.g., reduced antigen uptake; tumor necrosis factor alpha, interleukin-1beta [IL-1beta], gamma interferon [IFN-gamma], IL-10, IL-12, and IFN-alpha/beta secretion; and stimulation of heterologous T-cell proliferation) were also significantly lower for CAV-2. Our data suggested that this was due, in part, to the use of an alternative receptor and a block in vesicular escape. Additionally, HAd5 vector-induced hMoDC maturation was independent of the aforementioned cytokines. Paradoxically, an HAd5/CAV-2 hybrid vector induced the greatest phenotypical and functional maturation of hMoDC. Our data suggest that CAV-2 and the HAd5/CAV-2 vector may be the antithesis of Adenoviridae immunogenicity and that each may have specific clinical advantages.
抗病原体免疫反应在免疫、耐受和免疫逃逸之间建立了一种平衡。然而，在基因治疗期间，大多数病毒载体是以高剂量递送的，并且无法表达能够降低宿主检测转导细胞能力的基因产物。树突状细胞（DC）的体内转导也会改变基因转移效率，这显著增加了病毒粒子和载体编码基因的免疫原性。在本研究中，我们通过检测犬腺病毒2型（CAV-2）载体对人单核细胞来源的DC（hMoDC）的影响，评估了与在临床环境中使用CAV-2载体相关的参数。我们将CAV-2与含有野生型病毒粒子、五聚体基质RGD基序功能性缺失以及CAV-2纤维钮的人腺病毒（HAd）载体进行了比较。与基于5型人腺病毒（HAd5）的载体相比，CAV-2对hMoDC的转导效率较低，引发主要组织相容性复合体I/II类分子和共刺激分子（CD40、CD80和CD86）的上调最少，并诱导出可忽略不计的形态学变化，表明DC成熟。CAV-2的功能成熟检测结果（例如，抗原摄取减少；肿瘤坏死因子α、白细胞介素-1β[IL-1β]、γ干扰素[IFN-γ]、IL-10、IL-12和IFN-α/β分泌减少；以及对异源T细胞增殖的刺激减少）也显著较低。我们的数据表明，这部分是由于使用了替代受体以及囊泡逃逸受阻。此外，HAd5载体诱导的hMoDC成熟与上述细胞因子无关。矛盾的是，一种HAd5/CAV-2杂交载体诱导了hMoDC最大程度的表型和功能成熟。我们的数据表明，CAV-2和HAd5/CAV-2载体可能是腺病毒科免疫原性的对立面，并且每种载体可能都有特定的临床优势。
