Okimoto M A, Fan H
Department of Molecular Biology and Biochemistry and Cancer Research Institute, University of California, Irvine, California 92697-3900, USA.
J Virol. 1999 Mar;73(3):2509-16. doi: 10.1128/JVI.73.3.2509-2516.1999.
The nature of Moloney murine leukemia virus (M-MuLV) infection after a subcutaneous (s.c.) inoculation was studied. We have previously shown that an enhancer variant of M-MuLV, Mo+PyF101 M-MuLV, is poorly leukemogenic when used to inoculate mice s.c., but not when inoculated intraperitoneally. This attenuation of leukemogenesis correlated with an inability of Mo+PyF101 M-MuLV to establish infection in the bone marrow of mice at early times postinfection. These results suggested that a cell type(s) is infected in the skin by wild-type but not Mo+PyF101 M-MuLV after s.c. inoculation and that this infection is important for the delivery of infection to the bone marrow, as well as for efficient leukemogenesis. To determine the nature of the cell types infected by M-MuLV and Mo+PyF101 M-MuLV in the skin after a s.c. inoculation, immunohistochemistry with an anti-M-MuLV CA antibody was performed. Cells of developing hair follicles, specifically cells of the outer root sheath (ORS), were extensively infected by M-MuLV after s.c. inoculation. The Mo+PyF101 M-MuLV variant also infected cells of the ORS but the level of infection was lower. By Western blot analysis, the level of infection in skin by Mo+PyF101 M-MuLV was approximately 4- to 10-fold less than that of wild-type M-MuLV. Similar results were seen when a mouse keratinocyte line was infected in vitro with both viruses. Cells of the ORS are a primary target of infection in vivo, since a replication defective M-MuLV-based vector expressing beta-galactosidase also infected these cells after a s.c. inoculation.
研究了皮下接种后莫洛尼鼠白血病病毒(M-MuLV)感染的性质。我们之前已经表明,M-MuLV的一种增强子变体Mo+PyF101 M-MuLV用于皮下接种小鼠时致白血病性较差,但腹腔接种时并非如此。这种白血病发生的减弱与Mo+PyF101 M-MuLV在感染后早期无法在小鼠骨髓中建立感染有关。这些结果表明,皮下接种后野生型而非Mo+PyF101 M-MuLV在皮肤中感染了某种细胞类型,并且这种感染对于将感染传递至骨髓以及高效的白血病发生很重要。为了确定皮下接种后皮肤中被M-MuLV和Mo+PyF101 M-MuLV感染的细胞类型的性质,使用抗M-MuLV CA抗体进行了免疫组织化学分析。皮下接种后,发育中的毛囊细胞,特别是外根鞘(ORS)细胞被M-MuLV广泛感染。Mo+PyF101 M-MuLV变体也感染了ORS细胞,但感染水平较低。通过蛋白质免疫印迹分析,Mo+PyF101 M-MuLV在皮肤中的感染水平比野生型M-MuLV低约4至10倍。当用这两种病毒体外感染小鼠角质形成细胞系时,也观察到了类似的结果。ORS细胞是体内感染的主要靶标,因为一种表达β-半乳糖苷酶的基于M-MuLV的复制缺陷型载体在皮下接种后也感染了这些细胞。