Oleksiewicz M B, Wolfinbarger J B, Bloom M E
Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, MT, USA.
Virus Res. 1998 Jul;56(1):41-51. doi: 10.1016/s0168-1702(98)00053-7.
We used three-color fluorescent labeling and confocal microscopy to compare the permissive and the antibody-mediated, restricted replication of Aleutian mink disease parvovirus (ADV). In both permissive (CRFK cells) and restricted (K562 cells) situations, both ADV non-structural proteins (NS1 and NS2) concentrated at focal sites in the nucleus, which also contained viral DNA. Bromodeoxyuridine labeling demonstrated that these sites also supported active ADV single-strand DNA synthesis, indicating that they were replication compartments. ADV capsid proteins were located in intranuclear shells surrounding the replication compartments. At later time points, NS2 was readily detected in the cytoplasm of permissively infected CRFK cells, whereas the cytoplasmic presence of NS2 was much less pronounced in the K562 cells. These results showed that both permissive and restricted ADV replication are associated with a tight nuclear subcompartmentalization of viral products. Furthermore, differences between the permissive and restricted virus-cell interactions were noted, suggesting that there may be a morphological basis for examining the outcome of ADV infection.
我们使用三色荧光标记和共聚焦显微镜来比较阿留申水貂病细小病毒(ADV)的允许性复制和抗体介导的限制性复制。在允许性(CRFK细胞)和限制性(K562细胞)两种情况下,ADV的非结构蛋白(NS1和NS2)都集中在细胞核中的焦点部位,这些部位也含有病毒DNA。溴脱氧尿苷标记表明,这些部位也支持活跃的ADV单链DNA合成,表明它们是复制区室。ADV衣壳蛋白位于围绕复制区室的核内壳中。在后期时间点,在允许性感染的CRFK细胞的细胞质中很容易检测到NS2,而在K562细胞中NS2在细胞质中的存在则不太明显。这些结果表明,允许性和限制性ADV复制都与病毒产物紧密的核亚区室化有关。此外,还注意到允许性和限制性病毒-细胞相互作用之间的差异,这表明可能存在一个形态学基础来研究ADV感染的结果。