Cross-reactive antigenicity of nucleoproteins of lyssaviruses recognized by a monospecific antirabies virus nucleoprotein antiserum on paraffin sections of formalin-fixed tissues
Cross-reactive antigenicity of nucleoproteins of lyssaviruses recognized by a monospecific antirabies virus nucleoprotein antiserum on paraffin sections of formalin-fixed tissues
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DOI：
10.1046/j.1440-1827.2003.01511.x
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发表时间：
2003-08-01
影响因子：
2.2
通讯作者：
Iwasaki, T
中科院分区：
文献类型：
--
作者：
Inoue, S;Sato, Y;Iwasaki, T
狂犬病的诊断通常通过丙酮固定的冷冻脑组织或印迹涂片中的狂犬病病毒抗原的检测来确认，该检测采用免疫荧光法，使用市售的抗狂犬病病毒抗体。由于最近的分子分析揭示了狂犬病病毒株和相关狂犬病病毒的基因组序列的广泛异质性，因此有必要确认这些狂犬病病毒中存在共同表位。在这项研究中，我们证实了各种狂犬病病毒的交叉反应抗原的存在下，在石蜡切片的福尔马林固定的组织使用单特异性兔抗血清制备的狂犬病病毒的重组核蛋白免疫。通过免疫组织化学应用，抗原主要在感染狂犬病病毒、Duvenhage病毒、Mokola病毒和欧洲蝙蝠狂犬病病毒-1的小鼠脑中神经元的细胞质中检测到，而在未感染的人和动物（包括狗、蝙蝠和浣熊）中未观察到交叉反应。此外，我们检查了一例在狂犬病流行国家感染并于1970年返回日本后出现狂犬病临床表现的尸检病例，发现抗原在福尔马林固定组织的石蜡切片中保存良好。因此，这表明狂犬病病毒核蛋白的单特异性抗体识别狂犬病病毒特异性抗原，并且通过免疫组织化学分析在福尔马林固定的石蜡包埋组织上可检测到这种交叉反应抗原。
Diagnosis of rabies is routinely confirmed by detection of rabies virus antigens in acetone-fixed frozen brain tissues or imprint smears using an immunofluorescence method with commercial antirabies virus antibodies. Since recent molecular analyses disclosed wide heterogeneity in the genome sequences of rabies virus strains and related lyssaviruses, it is necessary to confirm the presence of common epitopes in these lyssaviruses. In this study we confirmed the presence of cross-reactive antigens of various lyssaviruses in paraffin sections of formalin-fixed tissue using a monospecific rabbit antiserum prepared by immunization with a recombinant nucleoprotein of rabies virus. By immunohistochemical application, the antigen was detected predominantly in the cytoplasm of neurons in the brains of mice infected with rabies virus, Duvenhage virus, Mokola virus and European bat lyssavirus-1, while no cross-reaction was observed in uninfected humans and animals including dogs, bats, and raccoons. In addition, we examined one autopsy case that was infected in a rabies-endemic nation and developed the clinical manifestation of rabies after returning to Japan in 1970, and found that the antigen was well preserved in paraffin sections of formalin-fixed tissues. Thus, this suggests that the lyssavirus-specific antigen is recognized by the monospecific antibody against rabies virus nucleoprotein, and that this cross-reactive antigen is detectable on formalin-fixed paraffin-embedded tissues by immunohistochemical analysis.