Leon R P, Hedlund T, Meech S J, Li S, Schaack J, Hunger S P, Duke R C, DeGregori J
Department of Biochemistry and Molecular Genetics, University of Colorado Cancer Center, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA.
Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):13159-64. doi: 10.1073/pnas.95.22.13159.
Although adenovirus can infect a wide range of cell types, lymphocytes are not generally susceptible to adenovirus infection, in part because of the absence of the expression of the cellular receptor for the adenoviral fiber protein. The cellular receptor for adenovirus and coxsackievirus (CAR) recently was cloned and shown to mediate adenoviral entry by interaction with the viral fiber protein. We show that the ectopic expression of CAR in various lymphocyte cell lines, which are almost completely resistant to adenovirus infection, is sufficient to facilitate the efficient transduction of these cells by recombinant adenoviruses. Furthermore, this property of CAR does not require its cytoplasmic domain, consistent with the idea that CAR primarily serves as a high affinity binding site for the adenoviral fiber protein, and that viral entry is mediated by interaction of the viral penton base proteins with cellular integrins. As a demonstration of their functional utility, we used CAR-expressing lymphocytes transduced with an adenovirus expressing Fas ligand to efficiently kill Fas receptor-expressing tumor cells. The ability to efficiently manipulate gene expression in lymphocyte cells by using adenovirus vectors should facilitate the functional characterization of pathways affecting lymphocyte physiology.
尽管腺病毒能够感染多种细胞类型,但淋巴细胞通常不易受到腺病毒感染,部分原因是缺乏腺病毒纤维蛋白细胞受体的表达。腺病毒和柯萨奇病毒的细胞受体(CAR)最近被克隆出来,并显示通过与病毒纤维蛋白相互作用介导腺病毒进入细胞。我们发现,在几乎对腺病毒感染完全抗性的各种淋巴细胞系中异位表达CAR,足以促进重组腺病毒对这些细胞的有效转导。此外,CAR的这一特性并不需要其胞质结构域,这与CAR主要作为腺病毒纤维蛋白的高亲和力结合位点,以及病毒进入是由病毒五聚体基质蛋白与细胞整合素相互作用介导的观点一致。作为其功能效用的一个例证,我们使用表达Fas配体的腺病毒转导的表达CAR的淋巴细胞,有效杀伤表达Fas受体的肿瘤细胞。利用腺病毒载体有效调控淋巴细胞基因表达的能力,应有助于影响淋巴细胞生理学途径的功能特性研究。