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Viruses. 2019 Mar 19;11(3). pii: E275. doi: 10.3390/v11030275.

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

Zapatero-Belinchón FJ1,2,3, Dietzel E4,5, Dolnik O6,7, Döhner K8, Costa R9,10,11,12, Hertel B13,14,15, Veselkova B16, Kirui J17, Klintworth A18,19,20, Manns MP21,22, Pöhlmann S23,24, Pietschmann T25, Krey T26,27, Ciesek S28, Gerold G29,30, Sodeik B31,32, Becker S33,34, von Hahn T35,36,37.

Author information

1
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. ZapateroBelinchon.Francisco@mh-hannover.de.
2
Institute for Molecular Biology, Hannover Medical School, 30625 Hannover, Germany. ZapateroBelinchon.Francisco@mh-hannover.de.
3
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. ZapateroBelinchon.Francisco@mh-hannover.de.
4
Institute of Virology, Philipps University Marburg, 35043 Marburg, Germany. erik.dietzel@staff.uni-marburg.de.
5
German Center for Infection Research (DZIF) Gießen-Marburg-Langen Site, Germany. erik.dietzel@staff.uni-marburg.de.
6
Institute of Virology, Philipps University Marburg, 35043 Marburg, Germany. olga.dolnik@staff.uni-marburg.de.
7
German Center for Infection Research (DZIF) Gießen-Marburg-Langen Site, Germany. olga.dolnik@staff.uni-marburg.de.
8
Institute of Virology, Hannover Medical School, 30625 Hannover, Germany. Doehner.Katinka@mh-hannover.de.
9
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. rui.costa@uk-essen.de.
10
Institute for Molecular Biology, Hannover Medical School, 30625 Hannover, Germany. rui.costa@uk-essen.de.
11
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. rui.costa@uk-essen.de.
12
Institute of Virology, Essen University Hospital, 45122 Essen, Germany. rui.costa@uk-essen.de.
13
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. bhertel@uni-potsdam.de.
14
Institute for Molecular Biology, Hannover Medical School, 30625 Hannover, Germany. bhertel@uni-potsdam.de.
15
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. bhertel@uni-potsdam.de.
16
Institute of Virology, Hannover Medical School, 30625 Hannover, Germany. Veselkova.Barbora@mh-hannover.de.
17
TWINCORE, Center for Experimental and Clinical Infection Research, Institute for Experimental Virology, 30625 Hannover, Germany. jared.kirui@twincore.de.
18
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. A.Klintworth@gmx.de.
19
Institute for Molecular Biology, Hannover Medical School, 30625 Hannover, Germany. A.Klintworth@gmx.de.
20
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. A.Klintworth@gmx.de.
21
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. Manns.Michael@mh-hannover.de.
22
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. Manns.Michael@mh-hannover.de.
23
Infection Biology Unit, German Primate Center ⁻ Leibniz Institute for Primate Research, 37077 Göttingen, Germany. SPoehlmann@dpz.eu.
24
Faculty of Biology and Psychology, University Göttingen, 37073 Göttingen, Germany. SPoehlmann@dpz.eu.
25
TWINCORE, Center for Experimental and Clinical Infection Research, Institute for Experimental Virology, 30625 Hannover, Germany. Pietschmann.Thomas@mh-hannover.de.
26
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. Krey.Thomas@mh-hannover.de.
27
Institute of Virology, Hannover Medical School, 30625 Hannover, Germany. Krey.Thomas@mh-hannover.de.
28
Institute of Virology, Essen University Hospital, 45122 Essen, Germany. Sandra.Ciesek@uk-essen.de.
29
TWINCORE, Center for Experimental and Clinical Infection Research, Institute for Experimental Virology, 30625 Hannover, Germany. gisa.gerold@twincore.de.
30
Department of Clinical Microbiology, Virology; Wallenberg Centre for Molecular Medicine (WCMM), Umeå University, 90187 Umeå, Sweden. gisa.gerold@twincore.de.
31
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. Sodeik.Beate@mh-hannover.de.
32
Institute of Virology, Hannover Medical School, 30625 Hannover, Germany. Sodeik.Beate@mh-hannover.de.
33
Institute of Virology, Philipps University Marburg, 35043 Marburg, Germany. becker@staff.uni-marburg.de.
34
German Center for Infection Research (DZIF) Gießen-Marburg-Langen Site, Germany. becker@staff.uni-marburg.de.
35
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 30625 Hannover, Germany. vonhahn.thomas@mh-hannover.de.
36
Institute for Molecular Biology, Hannover Medical School, 30625 Hannover, Germany. vonhahn.thomas@mh-hannover.de.
37
German Center for Infection Research (DZIF) Hannover-Braunschweig Site, Germany. vonhahn.thomas@mh-hannover.de.

Abstract

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.

KEYWORDS:

Filovirus cell entry; SH-SY5Y cell line; attachment factors redundancy; host–pathogen interactions

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