Immunohistochemical and in situ hybridization studies of influenza A virus infection in human lungs
Immunohistochemical and in situ hybridization studies of influenza A virus infection in human lungs
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DOI：
10.1309/hv74-n24t-2k2c-3e8q
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发表时间：
2000-08-01
影响因子：
3.5
通讯作者：
Zaki, SR
中科院分区：
文献类型：
--
作者：
Guarner, J;Shieh, WJ;Zaki, SR
流感病毒是引起急性发热性呼吸道疾病的原因。当发生死亡时，明确诊断需要病毒分离，因为没有发现特征性的病毒包涵体。我们使用2种免疫组化检测（核蛋白[NP]和血凝素[HA]的单克隆抗体）和2种原位杂交（ISH）检测（地高辛标记的HA和NP基因杂交探针）检测了8例致命感染患者组织中甲型流感病毒的分布。5例患者有明显的支气管炎;通过免疫组织化学检测，甲型流感病毒染色存在于较大支气管上皮（完整和分离的坏死细胞）和罕见的间质细胞中。抗NP抗体主要染色细胞核，抗HA抗体主要染色细胞质。在其中4例病例中，在相同区域鉴定出核酸（ISH）。3例患者有淋巴组织细胞性肺泡炎，未显示免疫组化或ISH染色。这两种技术都是有用的检测流感病毒抗原和核酸在福尔马林固定石蜡包埋的组织，可以使进一步了解致命的甲型流感病毒感染的人。
Influenza viruses are responsible for acute febrile respiratory disease. When deaths occur, definitive diagnosis requires viral isolation because no characteristic viral inclusions are seen. We examined the distribution of influenza A virus in tissues from 8 patients with fatal infection using 2 immunohistochemical assays (monoclonal antibodies to nucleoprotein [NP] and hemagglutinin [HA]) and 2 in situ hybridization (ISH) assays (digoxigenin-labeled probes that hybridized to HA and NP genes). Five patients had prominent bronchitis; by immunohistochemical assay, influenza A staining was present focally in the epithelium of larger bronchi (intact and detached necrotic cells) and in rare interstitial cells. The anti-NP antibody stained primarily cell nuclei, and the anti-HA antibody stained mainly the cytoplasm. In 4 of these cases, nucleic acids (ISH) were identified in the same areas. Three patients had lymphohistiocytic alveolitis and showed no immunohistochemical or ISH staining. Both techniques were useful for detection of influenza virus antigens and nucleic acids in formalin-fixed paraffin-embedded tissues and can enable further understanding of fatal influenza A virus infections in humans.