Defer C, Belin M T, Caillet-Boudin M L, Boulanger P
Unité de Virologie Moléculaire, Institut National de la Santé et de la Recherche Médicale, (U-233), Lille, France.
J Virol. 1990 Aug;64(8):3661-73. doi: 10.1128/JVI.64.8.3661-3673.1990.
Host cell interactions of human adenovirus serotypes belonging to subgroups B (adenovirus type 3 [Ad3] and Ad7) and C (Ad2 and Ad5) were comparatively analyzed at three levels: (i) binding of virus particles with host cell receptors; (ii) cointernalization of macromolecules with adenovirions; and (iii) adenovirus-induced cytoskeletal alterations. The association constants with human cell receptors were found to be similar for Ad2 and Ad3 (8 x 10(9) to 9 x 10(9) M-1), and the number of receptor sites per cell ranged from 5,000 (Ad2) to 7,000 (Ad3). Affinity blottings, competition experiments, and immunofluorescence stainings suggested that the receptor sites for adenovirus were distinct for members of subgroups B and C. Adenovirions increased the permeability of cells to macromolecules. We showed that this global effect could be divided into two distinct events: (i) cointernalization of macromolecules and virions into endocytotic vesicles, a phenomenon that occurred in a serotype-independent way, and (ii) release of macromolecules into the cytoplasm upon adenovirus-induced lysis of endosomal membranes. The latter process was found to be type specific and to require unaltered and infectious virus particles of serotype 2 or 5. Perinuclear condensation of the vimentin filament network was observed at early stages of infection with Ad2 or Ad5 but not with Ad3, Ad7, and noninfectious particles of Ad2 or Ad5, obtained by heat inactivation of wild-type virions or with the H2 ts1 mutant. This phenomenon appeared to be a cytological marker for cytoplasmic transit of infectious virions within adenovirus-infected cells. It could be experimentally dissociated from vimentin proteolysis, which was found to be serotype dependent, occurring only with members of subgroup C, regardless of the infectivity of the input virus.
对属于B亚组（腺病毒3型[Ad3]和Ad7）和C亚组（Ad2和Ad5）的人腺病毒血清型的宿主细胞相互作用进行了三个层面的比较分析：（i）病毒颗粒与宿主细胞受体的结合；（ii）大分子与腺病毒粒子的共内化；（iii）腺病毒诱导的细胞骨架改变。发现Ad2和Ad3与人细胞受体的结合常数相似（8×10⁹至9×10⁹ M⁻¹），每个细胞的受体位点数量从5000（Ad2）到7000（Ad3）不等。亲和印迹、竞争实验和免疫荧光染色表明，B亚组和C亚组成员的腺病毒受体位点不同。腺病毒粒子增加了细胞对大分子的通透性。我们表明，这种整体效应可分为两个不同的事件：（i）大分子和病毒粒子共内化到内吞小泡中，这一现象以血清型无关的方式发生，以及（ii）腺病毒诱导内体膜裂解后大分子释放到细胞质中。发现后一过程具有类型特异性，并且需要血清型2或5的未改变的感染性病毒粒子。在感染Ad2或Ad5的早期阶段观察到波形蛋白丝网络的核周凝聚，但在感染Ad3、Ad7以及通过野生型病毒粒子热灭活或H2 ts1突变体获得的Ad2或Ad5的非感染性粒子时未观察到。这种现象似乎是腺病毒感染细胞内感染性病毒粒子细胞质转运的细胞学标记。它可以通过实验与波形蛋白的蛋白水解分离，波形蛋白的蛋白水解发现是血清型依赖性的，仅在C亚组成员中发生，与输入病毒的感染性无关。
