Journal: Viruses

Article Title: Infection of Human Tracheal Epithelial Cells by H5 Avian Influenza Virus Is Regulated by the Acid Stability of Hemagglutinin and the pH of Target Cell Endosomes

doi: 10.3390/v12010082

Figure Lengend Snippet: Expression of virus receptors by human tracheal epithelial cell clones. The expression of sialic acid (SA) receptors on the surface of the HTEpC-T clones was analyzed by flow cytometry. α2,3SAGal and α2,6SAGal receptors were detected by Maackia amurensis (MAA) and Sambucus nigra (SNA) lectins, respectively. α2,3SAGal residues are shown in red (when detected by MAA-I) or blue (when detected by MAA-II). α2,6SAGal is shown as a black line. Control cells (without lectin) are represented by the black bold line. SAEC-T clones (1A5 and 21E5), which were derived from primary human bronchiolar epithelial cells, were also examined, and receptor expression patterns were compared with those of HTEpC-T clones. Madin–Darby canine kidney (MDCK) cells (virus-susceptible controls) express both α2,3SA and α2,6SA receptors. MDCK cells were first treated with neuraminidase to confirm the reliability of lectin staining.

Article Snippet: After washing once with PBS (−), the cells were incubated for 1 h at 4 °C with 2.5 μg/mL Sambucus nigra (SNA)-FITC (Vector Laboratories) in PBS (−) containing 0.1% bovine serum albumin to detect sialic acid α2,6-galactose (SAα2,6Gal) moieties or with 2.5 μg/mL Maackia amurensis (MAA)-I-FITC (Vector Laboratories) in PBS (−) containing 0.1% bovine serum albumin to detect SAα2,3Gal (Siaα2-3Galβ(1-4) GlcNAc) moieties.

Techniques: Expressing, Clone Assay, Flow Cytometry, Derivative Assay, Staining