Alexandersen S, Bloom M E, Wolfinbarger J
National Institute of Allergy and Infectious Diseases, Rocky Mountain Laboratories, Hamilton, Montana 59840.
J Virol. 1988 May;62(5):1495-507. doi: 10.1128/JVI.62.5.1495-1507.1988.
Strand-specific hybridization probes were used in in situ molecular hybridization specifically to localize cells containing replicative intermediates of Aleutian disease of mink parvovirus (ADV). When adult mink of Aleutian genotype were infected with ADV Utah I, the largest number of cells positive for viral replication (i.e., containing replicative-form DNA and RNA) were found in the mesenteric lymph nodes and spleens at 10 days after infection. The localization of positive cells in the middle of germinal centers suggested that they were B lymphoblasts. Circulating leukocytes and bone marrow cells also contained viral RNA, but the levels of replicative-form DNA were below detectability. The levels of viral DNA and RNA in adult mink cells replicating ADV were decreased compared with those in permissively infected cell cultures or neonatal mink, suggesting that the replication of ADV in adult mink might be semipermissive or restricted at some early stage of viral gene expression. The low level of viral replication and transcription in lymphoid cells might provide a mechanism for the development of immune disorders and for the maintenance of persistent infection. Single-stranded virion DNA was found in other organs, but the strand-specific probes made it possible to show that this DNA represented virus sequestration. In addition, glomerular immune complexes containing virion DNA were detected, suggesting that ADV virions, or perhaps free DNA, may have a role in the development of ADV-induced glomerulonephritis.
链特异性杂交探针用于原位分子杂交,专门定位含有水貂阿留申病细小病毒(ADV)复制中间体的细胞。当阿留申基因型的成年水貂感染ADV Utah I时,在感染后10天,在肠系膜淋巴结和脾脏中发现了最多的病毒复制阳性细胞(即含有复制型DNA和RNA的细胞)。生发中心中部阳性细胞的定位表明它们是B淋巴母细胞。循环白细胞和骨髓细胞也含有病毒RNA,但复制型DNA水平低于可检测范围。与允许感染的细胞培养物或新生水貂相比,成年水貂细胞中复制ADV的病毒DNA和RNA水平降低,这表明ADV在成年水貂中的复制可能在病毒基因表达的某些早期阶段是半允许的或受限的。淋巴细胞中病毒复制和转录水平较低可能为免疫紊乱的发展和持续感染的维持提供了一种机制。在其他器官中发现了单链病毒粒子DNA,但链特异性探针能够表明这种DNA代表病毒隔离。此外,检测到含有病毒粒子DNA的肾小球免疫复合物,这表明ADV病毒粒子或可能是游离DNA可能在ADV诱导的肾小球肾炎的发展中起作用。