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麻疹病毒RNA在麻疹、亚急性硬化性全脑炎及自身免疫性疾病患者外周血单个核细胞中的表达。

Expression of measles virus RNA in peripheral blood mononuclear cells of patients with measles, SSPE, and autoimmune diseases.

作者信息

Schneider-Schaulies S, Kreth H W, Hofmann G, Billeter M, Ter Meulen V

机构信息

Institut für Virologie und Immunbiologie, Universität Würzburg, Germany.

出版信息

Virology. 1991 Jun;182(2):703-11. doi: 10.1016/0042-6822(91)90611-e.

Abstract

In order to characterize measles virus (MV) infection in peripheral blood mononuclear cells (PBMCs), RNA was isolated from PBMCs after PHA-stimulation for 72 hr of 9 patients with acute measles, 16 patients with subacute sclerosing panencephalitis (SSPE), 13 patients with various autoimmune diseases, and 16 healthy control donors. The RNA obtained was screened for the presence of MV N (nucleocapsid) gene specific transcripts of either positive or negative orientation in a S1 nuclease protection assay. The sensitivity of this assay allowed us to detect one infected cell in 20,000 PBMCs or 0.1 to 0.05 copies of MV-specific RNA per cell. Using single-stranded DNA or RNA probes expression of MV genomic RNA of negative polarity could be detected in only one case of acute measles and one healthy control donor. Conversely, N-specific transcripts of positive polarity, indicating active transcription, could only be detected in patients with acute measles. In addition, in infected PBMCs and in a persistently MV-infected B cell line positive stranded N-specific transcripts containing leader usually present at very low frequency have been found in relatively increased amounts in comparison with transcripts lacking leader. Whereas the ratio of these RNA species during lytic infection with MV in Vero cells is about 1:50, the ratio found here ranges from 1:3 to 1:10. This altered ratio indicates a specific regulation of MV specific transcription in cells of lymphoid origin that has not been found in any other cell system analyzed.

摘要

为了表征外周血单个核细胞(PBMC)中的麻疹病毒(MV)感染情况,从9例急性麻疹患者、16例亚急性硬化性全脑炎(SSPE)患者、13例各种自身免疫性疾病患者以及16名健康对照供体的PBMC中提取RNA,这些PBMC在PHA刺激72小时后进行提取。在S1核酸酶保护试验中,对获得的RNA进行筛选,以检测MV N(核衣壳)基因特定转录本的正向或负向存在情况。该试验的灵敏度使我们能够在20,000个PBMC中检测到一个感染细胞,或每个细胞检测到0.1至0.05拷贝的MV特异性RNA。使用单链DNA或RNA探针,仅在1例急性麻疹患者和1名健康对照供体中检测到了负极性MV基因组RNA的表达。相反,仅在急性麻疹患者中检测到了指示活跃转录的正极性N特异性转录本。此外,在感染的PBMC和持续感染MV的B细胞系中,与缺乏前导序列的转录本相比,含有前导序列的正极性N特异性转录本的相对含量有所增加,而这些前导序列通常以非常低的频率出现。在Vero细胞中,MV裂解感染期间这些RNA种类的比例约为1:50,而这里发现的比例范围为1:3至1:10。这种改变的比例表明在淋巴起源的细胞中MV特异性转录存在特定调节,这在任何其他分析的细胞系统中均未发现。

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