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腺相关病毒载体对马关节组织外植体和培养的单层细胞表现出血清型特异性转导。

Adeno-associated viral vectors show serotype specific transduction of equine joint tissue explants and cultured monolayers.

作者信息

Hemphill Daniel D, McIlwraith C Wayne, Samulski R Jude, Goodrich Laurie R

机构信息

Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.

University of North Carolina Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.

出版信息

Sci Rep. 2014 Jul 29;4:5861. doi: 10.1038/srep05861.

Abstract

Adeno-associated virus (AAV) receptors range from heparan sulfate proteoglycan to sialic acid moieties present on cell surfaces. Abundance of the glycan profiles is greatly influenced by animal species, cell type, and culture conditions. The objective of this study was to determine whether AAV serotypes' transduction efficiencies specifically in the equine monolayer culture model are an accurate representation of transduction efficiencies in tissue explants, a model more closely related to in vivo transduction. It was found that AAV 2 and 2.5 transduced cells more efficiently in explants than in monolayers. Through experiments involving assessing enzyme degradation of cell surface proteoglycans, this change could not be attributed to differences in the extra cellular matrix (ECM), but a similar change in AAV 5 transduction efficiency could be readily explained by differences in cell surface sialylated glycan. Unexpectedly it was found that in a small but diverse sample of horses evidence for serum neutralizing antibodies was only found to AAV 5. This suggests a unique relationship between this capsid and the equine host or an unresolved relationship between similar bovine AAV and the AAV 5 capsid immune response.

摘要

腺相关病毒(AAV)的受体范围从细胞表面存在的硫酸乙酰肝素蛋白聚糖到唾液酸部分。聚糖谱的丰度受动物物种、细胞类型和培养条件的影响很大。本研究的目的是确定AAV血清型在马单层培养模型中的转导效率是否能准确代表组织外植体中的转导效率,组织外植体模型与体内转导的关系更为密切。研究发现,AAV 2和2.5在外植体中转导细胞的效率高于单层培养。通过评估细胞表面蛋白聚糖酶降解的实验,这种变化不能归因于细胞外基质(ECM)的差异,但AAV 5转导效率的类似变化可以很容易地用细胞表面唾液酸化聚糖的差异来解释。出乎意料的是,在一小部分但种类多样的马匹样本中,仅发现针对AAV 5的血清中和抗体证据。这表明该衣壳与马宿主之间存在独特关系,或者类似牛AAV与AAV 5衣壳免疫反应之间存在未解决的关系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0acf/4894424/c51d8ea709fa/srep05861-f1.jpg

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