Brown D R, Fan L, Jones J, Bryan J
Department of Medicine, Indiana University School of Medicine, Indianapolis 46202.
Virology. 1994 May 15;201(1):46-54. doi: 10.1006/viro.1994.1264.
The most abundant viral mRNA species in tissues infected with HPV 11 consists of two exons, joining a short segment of open reading frame (ORF) E1 to ORF E4, potentially encoding an protein of 10 kDa. E4 gene products have previously been identified by immunohistochemistry in human tissues infected with HPV 1 and HPV 16, and in HPV 11-infected raft cultures. The E1[symbol: see text]E4 mRNA is produced in abundance in HPV 11-infected human foreskin implants grown in athymic mice. In contrast, the L1 mRNA is present at low levels and appears late in the course of infection. To characterize the relationship of these proteins, polyclonal rabbit antisera were produced against bacterially expressed HPV 11 trpE/E1[symbol: see text]E4 and trpE/L1 fusion proteins and tested in an immunohistochemical assay of paraffin-embedded sections of HPV 11-infected human foreskin tissue fixed with 10% buffered formalin phosphate or zinc formalin. In sections fixed with either fixative, the anti-L1 serum stained nuclei of cells in the upper spinous and granular layers. In contrast, the anti-E1[symbol: see text]E4 serum stained the cell membrane and, to a lesser degree, the cytoplasm of cells in the upper spinous and granular layers of tissue fixed with zinc formalin, but not 10% buffered formalin phosphate. In sections treated with both the E1[symbol: see text]E4 and L1 antisera, cell membrane staining occurred in the same cells that exhibited nuclear staining. The HPV 11 E1[symbol: see text]E4 protein appears to be a cell membrane-associated protein. Expression of the HPV 11 E1[symbol: see text]E4 and L1 proteins may be influenced by similar factors in differentiating cells.
在感染人乳头瘤病毒11型（HPV 11）的组织中，最丰富的病毒信使核糖核酸（mRNA）种类由两个外显子组成，它将开放阅读框（ORF）E1的一小段与ORF E4连接起来，可能编码一种10千道尔顿的蛋白质。先前已通过免疫组织化学在感染HPV 1和HPV 16的人体组织以及感染HPV 11的筏式培养物中鉴定出E4基因产物。E1[符号：见正文]E4 mRNA在无胸腺小鼠体内生长的感染HPV 11的人包皮植入物中大量产生。相比之下，L1 mRNA水平较低，且在感染过程后期出现。为了表征这些蛋白质之间的关系，制备了针对细菌表达的HPV 11 trpE/E1[符号：见正文]E4和trpE/L1融合蛋白的兔多克隆抗血清，并在对用10%磷酸缓冲福尔马林或锌福尔马林固定的感染HPV 11的人包皮组织石蜡包埋切片进行的免疫组织化学检测中进行了测试。在用任何一种固定剂固定的切片中，抗L1血清对棘层上部和颗粒层细胞的细胞核进行了染色。相比之下，抗E1[符号：见正文]E4血清对用锌福尔马林固定而非10%磷酸缓冲福尔马林固定的组织的棘层上部和颗粒层细胞的细胞膜进行了染色，对细胞质的染色程度较轻。在用E1[符号：见正文]E4和L1抗血清处理的切片中，细胞膜染色出现在显示细胞核染色的相同细胞中。HPV 11 E1[符号：见正文]E4蛋白似乎是一种与细胞膜相关的蛋白。HPV 11 E1[符号：见正文]E4和L1蛋白的表达可能在分化细胞中受到相似因素的影响。
