Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan.
Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University, Hiroshima, Japan.
Methods Mol Biol. 2021;2274:141-154. doi: 10.1007/978-1-0716-1258-3_13.
Visualization of virus-infected cells is usually performed by immunostaining with an antiviral antibody. On the other hand, we established an easy method for fluorescence (FL) imaging of cells infected with influenza A and B viruses and some paramyxoviruses without the need for cell fixation and an antiviral antibody. These viruses and the cells they have infected express the viral surface enzyme "neuraminidase" or "hemagglutinin-neuraminidase" that shows sialidase activity. Sialidase activity is fluorescently visualized by using a sialidase fluorogenic probe developed in our previous study. The probe enables histochemical FL imaging of the virus-infected cells and is applicable to virus isolation and detection of an influenza virus resistant to antiinfluenza drugs of sialidase inhibitors.
病毒感染细胞的可视化通常通过用抗病毒抗体进行免疫染色来实现。另一方面，我们建立了一种无需细胞固定和抗病毒抗体即可对甲型和乙型流感病毒以及一些副粘病毒感染的细胞进行荧光（FL）成像的简便方法。这些病毒及其感染的细胞表达具有唾液酸酶活性的病毒表面酶“神经氨酸酶”或“血凝素-神经氨酸酶”。我们之前的研究中开发的唾液酸酶荧光探针可用于检测该酶的活性，从而实现对具有荧光活性的病毒感染细胞的荧光成像，该探针可用于病毒分离和检测对流感病毒抑制剂唾液酸酶抑制剂具有耐药性的流感病毒。
