Department of Applied Biochemistry, Institute of Biotechnology, Technische Universität Berlin, Gustav-Meyer-Allee 25, Berlin, Germany.
Department of Vetrinary Medicine, Institute of Veterinary Pathology, Freie Universität Berlin, Robert-von-Ostertag-Strasse 15, Berlin, Germany.
Gene Ther. 2015 Jun;22(6):458-66. doi: 10.1038/gt.2015.19. Epub 2015 Mar 19.
Immunosuppressed (IS) patients, such as recipients of hematopoietic stem cell transplantation, occasionally develop severe and fatal adenovirus (Ad) infections. Here, we analyzed the potential of a virus receptor trap based on a soluble coxsackievirus and Ad receptor (sCAR) for inhibition of Ad infection. In vitro, a dimeric fusion protein, sCAR-Fc, consisting of the extracellular domain of CAR and the Fc portion of human IgG1 and a monomeric sCAR lacking the Fc domain, were expressed in cell culture. More sCAR was secreted into the cell culture supernatant than sCAR-Fc, but it had lower Ad neutralization activity than sCAR-Fc. Further investigations showed that sCAR-Fc reduced the Ad infection by a 100-fold and Ad-induced cytotoxicity by ~20-fold. Not only was Ad infection inhibited by sCAR-Fc applied prior to infection, it also inhibited infection when used to treat ongoing Ad infection. In vivo, sCAR-Fc was delivered to IS mice by an AAV9 vector, resulting in persistent and high (>40 μg ml(-1)) sCAR-Fc serum levels. The sCAR-Fc serum concentration was sufficient to significantly inhibit hepatic and cardiac wild-type Ad5 infection. Treatment with sCAR-Fc did not induce side effects. Thus, sCAR-Fc virus receptor trap may be a promising novel therapeutic for treatment of Ad infections.
免疫抑制(IS)患者,如接受造血干细胞移植的患者,偶尔会发生严重和致命的腺病毒(Ad)感染。在这里,我们分析了基于可溶性柯萨奇病毒和 Ad 受体(sCAR)的病毒受体陷阱抑制 Ad 感染的潜力。在体外,二聚体融合蛋白 sCAR-Fc 由 CAR 的细胞外结构域和人 IgG1 的 Fc 部分组成,单体 sCAR 缺乏 Fc 结构域,在细胞培养中表达。与 sCAR-Fc 相比,更多的 sCAR 分泌到细胞培养上清中,但它的 Ad 中和活性低于 sCAR-Fc。进一步的研究表明,sCAR-Fc 将 Ad 感染减少了 100 倍,Ad 诱导的细胞毒性减少了约 20 倍。sCAR-Fc 不仅在感染前抑制 Ad 感染,而且在治疗正在进行的 Ad 感染时也抑制感染。在体内,sCAR-Fc 通过 AAV9 载体递送至 IS 小鼠,导致持续和高(>40μg/ml(-1))的 sCAR-Fc 血清水平。sCAR-Fc 的血清浓度足以显著抑制肝和心脏野生型 Ad5 感染。sCAR-Fc 的治疗没有引起副作用。因此,sCAR-Fc 病毒受体陷阱可能是治疗 Ad 感染的一种很有前途的新型治疗方法。