Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors

• Published in: Viruses Vol. 11; no. 3; p. 275
• Main Authors: Zapatero-Belinchón, Francisco J, Dietzel, Erik, Dolnik, Olga, Döhner, Katinka, Costa, Rui, Hertel, Barbara, Veselkova, Barbora, Kirui, Jared, Klintworth, Anneke, Manns, Michael P, Pöhlmann, Stefan, Pietschmann, Thomas, Krey, Thomas, Ciesek, Sandra, Gerold, Gisa, Sodeik, Beate, Becker, Stephan, von Hahn, Thomas
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 19.03.2019; MDPI
• Subjects: A549 Cells; attachment factors redundancy; Axl protein; Carrier Proteins - genetics; Cathepsin B; Cell adhesion; Cell Line; Cell lines; Cell Membrane - virology; Cell surface; DC-SIGN protein; Dendritic cells; Ebola virus; Ebolavirus - genetics; Ebolavirus - physiology; Epidemics; Filoviridae - genetics; Filoviridae - physiology; Filovirus cell entry; Gene expression; Gene Expression Profiling; HEK293 Cells; Heterokaryons; Host-Pathogen Interactions; host-pathogen teractions; Humans; Infections; Intracellular; Kinases; Lymphatic system; Lymphocytes; Lymphocytes T; Medical schools; Membrane Glycoproteins - genetics; Membrane proteins; Mucin; Niemann-Pick disease; Npc1 protein; Plasma; Plasmids; Proteins; Receptors, Virus - genetics; SH-SY5Y cell line; Viral Proteins - genetics; Virology; Virus Internalization; Viruses; United States > US; Germany; attachment factors redundancy; Filovirus cell entry; SH-SY5Y cell line; host–pathogen interactions
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v11030275

Filoviruses infect a wide range of cell types with the exception of lymphocytes. The intracellular proteins cathepsin B and L, two-pore channel 1 and 2, and bona fide receptor Niemann⁻Pick Disease C1 (NPC1) are essential for the endosomal phase of cell entry. However, earlier steps of filoviral infection remain poorly characterized. Numerous plasma membrane proteins have been implicated in attachment but it is still unclear which ones are sufficient for productive entry. To define a minimal set of host factors required for filoviral glycoprotein-driven cell entry, we screened twelve cell lines and identified the nonlymphocytic cell line SH-SY5Y to be specifically resistant to filovirus infection. Heterokaryons of SH-SY5Y cells fused to susceptible cells were susceptible to filoviruses, indicating that SH-SY5Y cells do not express a restriction factor but lack an enabling factor critical for filovirus entry. However, all tested cell lines expressed functional intracellular factors. Global gene expression profiling of known cell surface entry factors and protein expression levels of analyzed attachment factors did not reveal any correlation between susceptibility and expression of a specific host factor. Using binding assays with recombinant filovirus glycoprotein, we identified cell attachment as the step impaired in filovirus entry in SH-SY5Y cells. Individual overexpression of attachment factors T-cell immunoglobulin and mucin domain 1 (TIM-1), Axl, Mer, or dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) rendered SH-SY5Y cells susceptible to filovirus glycoprotein-driven transduction. Our study reveals that a lack of attachment factors limits filovirus entry and provides direct experimental support for a model of filoviral cell attachment where host factor usage at the cell surface is highly promiscuous.