CoChat
Sign Up
Cited by 462SARS-CoV-2 and COVID-19 ResearchCOVID-19 Clinical Research Studiesinterferon and immune responses

Syncytia formation by SARS‐CoV‐2‐infected cells

Open access
Buchrieser, Julian, Dufloo, Jérémy, Hubert, Mathieu, Monel, Blandine, Planas, Delphine, Rajah, Maaran Michael, Planchais, Cyril, Porrot, Françoise +6 more·2020·The EMBO Journal
DOI 10.15252/embj.2020106267PMC PMC7646020

Published version of a preprint

Open this paper in CoChat — free accountOpen PDF

Abstract

Severe cases of COVID-19 are associated with extensive lung damage and the presence of infected multinucleated syncytial pneumocytes. The viral and cellular mechanisms regulating the formation of these syncytia are not well understood. Here, we show that SARS-CoV-2-infected cells express the Spike protein (S) at their surface and fuse with ACE2-positive neighboring cells. Expression of S without any other viral proteins triggers syncytia formation. Interferon-induced transmembrane proteins (IFITMs), a family of restriction factors that block the entry of many viruses, inhibit S-mediated fusion, with IFITM1 being more active than IFITM2 and IFITM3. On the contrary, the TMPRSS2 serine protease, which is known to enhance infectivity of cell-free virions, processes both S and ACE2 and increases syncytia formation by accelerating the fusion process. TMPRSS2 thwarts the antiviral effect of IFITMs. Our results show that SARS-CoV-2 pathological effects are modulated by cellular proteins that either inhibit or facilitate syncytia formation.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.15252/embj.2020106267

Cites

7 of 7 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • U2OS-ACE2 GFP1-10 or GFP11 cells (S-Fuse cells) become GFP+ when productively infected by SARS-CoV-2.

    “U2OS-ACE2 GFP1-10 or GFP 11 cells, also termed S-Fuse cells, become GFP + when they are productively infected by SARS-CoV-2 10,11 .”

    From Reduced sensitivity of SARS-CoV-2 variant Delta to antibody neutralization (2021) · cites this paper as [10]

    The Results section identifies the U2OS-ACE2 split-GFP cells as “S-Fuse” cells and reports GFP-positive syncytia after SARS-CoV-2 exposure. But it says GFP appears only upon cell fusion and notes that some infected single cells have not yet fused. Productive infection alone is therefore not shown to make each cell GFP-positive. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 53

  1. Abdel Motal UM, Zhou X, Joki A, Siddiqi AR, Srinivasa BR, Stenvall K, Dahmen J, Jondal M (1993) Major Histocompatibility Complex class I‐binding peptides are recycled to the cell surface after internalization. Eur J Immunol 23: 3224– 3229 8258337
  2. Bertram S, Dijkman R, Habjan M, Heurich A, Gierer S, Glowacka I, Welsch K, Winkler M, Schneider H, Hofmann‐Winkler H et al (2013) TMPRSS2 activates the human coronavirus 229E for cathepsin‐independent host cell entry and is expressed in viral target cells in the respiratory epithelium. J Virol 87: 6150– 6160 23536651 PMC3648130
  3. Blanco‐Melo D, Nilsson‐Payant BE, Liu WC, Uhl S, Hoagland D, Moller R, Jordan TX, Oishi K, Panis M, Sachs D et al (2020) Imbalanced host response to SARS‐CoV‐2 drives development of COVID‐19. Cell 181: 1036– 1045.e9 32416070 PMC7227586
  4. Bost P, Giladi A, Liu Y, Bendjelal Y, Xu G, David E, Blecher‐Gonen R, Cohen M, Medaglia C, Li H et al (2020) Host‐viral infection maps reveal signatures of severe COVID‐19 patients. Cell 181: 1475– 1488 e12 32479746 PMC7205692
  5. Bozzo C, Nchioua R, Volcic M, Wettstein L, Weil T, Krüger J, Heller S, Conzelmann C, Müller J, Gross R et al (2020) IFITM proteins promote SARS‐CoV‐2 infection of human lung cells. bioRxiv 10.1101/2020.08.18.255935 [PREPRINT] · DOI
  6. Broggi A, Ghosh S, Sposito B, Spreafico R, Balzarini F, Lo Cascio A, Clementi N, De Santis M, Mancini N, Granucci F et al (2020) Type III interferons disrupt the lung epithelial barrier upon viral recognition. Science 369: 706– 712 32527925 PMC7292499
  7. Buchrieser J, Degrelle SA, Couderc T, Nevers Q, Disson O, Manet C, Donahue DA, Porrot F, Hillion KH, Perthame E et al (2019) IFITM proteins inhibit placental syncytiotrophoblast formation and promote fetal demise. Science 365: 176– 180 31296770
  8. Chan JF, Chan KH, Choi GK, To KK, Tse H, Cai JP, Yeung ML, Cheng VC, Chen H, Che XY et al (2013) Differential cell line susceptibility to the emerging novel human betacoronavirus 2c EMC/2012: implications for disease pathogenesis and clinical manifestation. J Infect Dis 207: 1743– 1752 23532101 PMC7107374
  9. Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF, McCallion AS, Katsanis N, Reeves RH (2018) Survey of human chromosome 21 gene expression effects on early development in Danio rerio. G3 (Bethesda) 8: 2215– 2223 29760202 PMC6027891
  10. Franks TJ, Chong PY, Chui P, Galvin JR, Lourens RM, Reid AH, Selbs E, McEvoy CPL, Hayden CDL, Fukuoka J et al (2003) Lung pathology of severe acute respiratory syndrome (SARS): a study of 8 autopsy cases from Singapore. Hum Pathol 34: 743– 748 14506633 PMC7119137
  11. Freundt EC, Yu L, Goldsmith CS, Welsh S, Cheng A, Yount B, Liu W, Frieman MB, Buchholz UJ, Screaton GR et al (2010) The open reading frame 3a protein of severe acute respiratory syndrome‐associated coronavirus promotes membrane rearrangement and cell death. J Virol 84: 1097– 1109 19889773 PMC2798367
  12. Giacca M, Bussani R, Schneider E, Zentilin L, Collesi C, Ali H, Braga L, Secco I, Volpe MC, Colliva A et al(2020) Persistence of viral RNA, widespread thrombosis and abnormal cellular syncytia are hallmarks of COVID‐19 lung pathology. medRxiv 10.1101/2020.06.22.20136358 [PREPRINT] PMC7677597 33158808 · DOI
  13. Glowacka I, Bertram S, Müller MA, Allen P, Soilleux E, Pfefferle S, Steffen I, Tsegaye TS, He Y, Gnirss K et al (2011) Evidence that TMPRSS2 activates the severe acute respiratory syndrome coronavirus spike protein for membrane fusion and reduces viral control by the humoral immune response. J Virol 85: 4122– 4134 21325420 PMC3126222
  14. Gorshkov K, Chen CZ, Bostwick R, Rasmussen L, Xu M, Pradhan M, Tran BN, Zhu W, Shamim K, Huang W et al(2020) The SARS‐CoV‐2 cytopathic effect is blocked with autophagy modulators. bioRxiv 10.1101/2020.05.16.091520 [PREPRINT] · DOI
  15. Hadjadj J, Yatim N, Barnabei L, Corneau A, Boussier J, Pere H, Charbit B, Bondet V, Chenevier‐Gobeaux C, Breillat P et al (2020) Impaired type I interferon activity and exacerbated inflammatory responses in severe Covid‐19 patients. medRxiv 10.1101/2020.04.19.20068015 [PREPRINT] PMC7402632 32661059 · DOI
  16. Haga S, Yamamoto N, Nakai‐Murakami C, Osawa Y, Tokunaga K, Sata T, Yamamoto N, Sasazuki T, Ishizaka Y (2008) Modulation of TNF‐α‐converting enzyme by the spike protein of SARS‐CoV and ACE2 induces TNF‐α production and facilitates viral entry. Proc Natl Acad Sci USA 105: 7809 18490652 PMC2409424
  17. Heurich A, Hofmann‐Winkler H, Gierer S, Liepold T, Jahn O, Pöhlmann S (2014) TMPRSS2 and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. J Virol 88: 1293– 1307 24227843 PMC3911672
  18. Hoffmann M, Kleine‐Weber H, Schroeder S, Kruger N, Herrler T, Erichsen S, Schiergens TS, Herrler G, Wu NH, Nitsche A et al (2020b) SARS‐CoV‐2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell 181: 271– 280 e8 32142651 PMC7102627
  19. Hou YJ, Okuda K, Edwards CE, Martinez DR, Asakura T, Dinnon 3rd KH, Kato T, Lee RE, Yount BL, Mascenik TM et al (2020) SARS‐CoV‐2 reverse genetics reveals a variable infection gradient in the respiratory tract. Cell 182: 429– 446.e14 32526206 PMC7250779
  20. Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, Zhang L, Fan G, Xu J, Gu X et al (2020) Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 395: 497– 506 31986264 PMC7159299
  21. Huang IC, Bailey CC, Weyer JL, Radoshitzky SR, Becker MM, Chiang JJ, Brass AL, Ahmed AA, Chi X, Dong L et al (2011) Distinct patterns of IFITM‐mediated restriction of filoviruses, SARS coronavirus, and influenza A virus. PLoS Pathog 7: e1001258 21253575 PMC3017121
  22. Kleine‐Weber H, Elzayat MT, Hoffmann M, Pohlmann S (2018) Functional analysis of potential cleavage sites in the MERS‐coronavirus spike protein. Sci Rep 8: 16597 30413791 PMC6226446
  23. Kodaka M, Yang Z, Nakagawa K, Maruyama J, Xu X, Sarkar A, Ichimura A, Nasu Y, Ozawa T, Iwasa H et al (2015) A new cell‐based assay to evaluate myogenesis in mouse myoblast C2C12 cells. Exp Cell Res 336: 171– 181 26116467
  24. Lan J, Ge J, Yu J, Shan S, Zhou H, Fan S, Zhang Q, Shi X, Wang Q, Zhang L et al (2020) Structure of the SARS‐CoV‐2 spike receptor‐binding domain bound to the ACE2 receptor. Nature 581: 215– 220 32225176
  25. Li W, Moore MJ, Vasilieva N, Sui J, Wong SK, Berne MA, Somasundaran M, Sullivan JL, Luzuriaga K, Greenough TC et al (2003) Angiotensin‐converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426: 450– 454 14647384 PMC7095016
  26. Major J, Crotta S, Llorian M, McCabe TM, Gad HH, Priestnall SL, Hartmann R, Wack A (2020) Type I and III interferons disrupt lung epithelial repair during recovery from viral infection. Science 369: 712– 717 32527928 PMC7292500
  27. Matsuyama S, Nao N, Shirato K, Kawase M, Saito S, Takayama I, Nagata N, Sekizuka T, Katoh H, Kato F et al (2020) Enhanced isolation of SARS‐CoV‐2 by TMPRSS2‐expressing cells. Proc Natl Acad Sci USA 117: 7001– 7003 32165541 PMC7132130
  28. Ogando NS, Dalebout TJ, Zevenhoven‐Dobbe JC, Limpens RWAL, van der Meer Y, Caly L, Druce J, de Vries JJC, Kikkert M, Bárcena M et al (2020) SARS‐coronavirus‐2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology. J Gen Virol 101: 925– 940 32568027 PMC7654748
  29. Ong EZ, Chan YFZ, Leong WY, Lee NMY, Kalimuddin S, Haja Mohideen SM, Chan KS, Tan AT, Bertoletti A, Ooi EE et al (2020) A dynamic immune response shapes COVID‐19 progression. Cell Host Microbe 27: 879– 882 e2 32359396 PMC7192089
  30. Park A, Iwasaki A (2020) Type I and Type III interferons – induction, signaling, evasion, and application to combat COVID‐19. Cell Host Microbe 27: 870– 878 32464097 PMC7255347
  31. Pemble H, Kumar P, van Haren J, Wittmann T (2017) GSK3‐mediated CLASP2 phosphorylation modulates kinetochore dynamics. J Cell Sci 130: 1404– 1412 28232523 PMC5399784
  32. Pfaender S, Mar KB, Michailidis E, Kratzel A, Boys IN, V'Kovski P, Fan W, Kelly JN, Hirt D, Ebert N et al (2020) LY6E impairs coronavirus fusion and confers immune control of viral disease. Nat Microbiol 5: 1330– 1339 32704094 PMC7916999
  33. Qian Z, Dominguez SR, Holmes KV (2013) Role of the spike glycoprotein of human Middle East respiratory syndrome coronavirus (MERS‐CoV) in virus entry and syncytia formation. PLoS One 8: e76469 24098509 PMC3789674
  34. Reinke LM, Spiegel M, Plegge T, Hartleib A, Nehlmeier I, Gierer S, Hoffmann M, Hofmann‐Winkler H, Winkler M, Pöhlmann S (2017) Different residues in the SARS‐CoV spike protein determine cleavage and activation by the host cell protease TMPRSS2. PLoS One 12: e0179177 28636671 PMC5479546
  35. Ren Y, Shu T, Wu D, Mu J, Wang C, Huang M, Han Y, Zhang X‐Y, Zhou W, Qiu Y et al (2020) The ORF3a protein of SARS‐CoV‐2 induces apoptosis in cells. Cell Mol Immunol 17: 881– 883 32555321 PMC7301057
  36. Shi G, Kenney AD, Kudryashova E, Zhang L, Hall‐Stoodley L, Robinson RT, Kudryashov DS, Compton AA, Yount JS (2020) Opposing activities of IFITM proteins in SARS‐CoV‐2 infection. bioRxiv 10.1101/2020.08.11.246678 [PREPRINT] PMC7744865 33270927 · DOI
  37. Shi G, Schwartz O, Compton AA (2017) More than meets the I: the diverse antiviral and cellular functions of interferon‐induced transmembrane proteins. Retrovirology 14: 53 29162141 PMC5697417
  38. Shirato K, Kawase M, Matsuyama S (2013) Middle east respiratory syndrome coronavirus infection mediated by the transmembrane serine protease TMPRSS2. J Virol 87: 12552– 12561 24027332 PMC3838146
  39. Shulla A, Heald‐Sargent T, Subramanya G, Zhao J, Perlman S, Gallagher T (2011) A transmembrane serine protease is linked to the severe acute respiratory syndrome coronavirus receptor and activates virus entry. J Virol 85: 873– 882 21068237 PMC3020023
  40. Siu YL, Teoh KT, Lo J, Chan CM, Kien F, Escriou N, Tsao SW, Nicholls JM, Altmeyer R, Peiris JSM et al (2008) The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus‐like particles. J Virol 82: 11318– 11330 18753196 PMC2573274
  41. Subbarao K, Mahanty S (2020) Respiratory virus infections: understanding COVID‐19. Immunity 52: 905– 909 32497522 PMC7237932
  42. Tang D, Comish P, Kang R (2020) The hallmarks of COVID‐19 disease. PLoS Pathog 16: e1008536 32442210 PMC7244094
  43. Tian S, Hu W, Niu L, Liu H, Xu H, Xiao SY (2020) Pulmonary pathology of early‐phase 2019 novel coronavirus (COVID‐19) pneumonia in two patients with lung cancer. J Thorac Oncol 15: 700– 704 32114094 PMC7128866
  44. Walls AC, Park YJ, Tortorici MA, Wall A, McGuire AT, Veesler D (2020) Structure, function, and antigenicity of the SARS‐CoV‐2 spike glycoprotein. Cell 181: 281– 292.e6 32155444 PMC7102599
  45. Warren CJ, Griffin LM, Little AS, Huang IC, Farzan M, Pyeon D (2014) The antiviral restriction factors IFITM1, 2 and 3 do not inhibit infection of human papillomavirus, cytomegalovirus and adenovirus. PLoS One 9: e96579 24827144 PMC4020762
  46. Wrensch F, Winkler M, Pohlmann S (2014) IFITM proteins inhibit entry driven by the MERS‐coronavirus spike protein: evidence for cholesterol‐independent mechanisms. Viruses 6: 3683– 3698 25256397 PMC4189045
  47. Zang R, Case JB, Gomez Castro MF, Liu Z, Zeng Q, Zhao H, Son J, Rothlauf PW, Hou G, Bose S, Wang X, Vahey MD, Kirchhausen T, Fremont DH, Diamond MS, Whelan SPJ, Ding S (2020) Cholesterol 25‐hydroxylase suppresses SARS‐CoV‐2 replication by blocking membrane fusion. bioRxiv 10.1101/2020.06.08.141077 [PREPRINT] PMC7749331 33239446 · DOI
  48. Zani A, Yount JS (2018) Antiviral protection by IFITM3 in vivo. Curr Clin Microbiol Rep 5: 229– 237 30662816 PMC6334760
  49. Zhao X, Guo F, Liu F, Cuconati A, Chang J, Block TM, Guo JT (2014) Interferon induction of IFITM proteins promotes infection by human coronavirus OC43. Proc Natl Acad Sci USA 111: 6756– 6761 24753610 PMC4020042
  50. Zhao X, Sehgal M, Hou Z, Cheng J, Shu S, Wu S, Guo F, Le Marchand SJ, Lin H, Chang J et al (2018) Identification of residues controlling restriction versus enhancing activities of IFITM proteins on entry of human coronaviruses. J Virol 92: e01535‐17 29263263 PMC5827390
  51. Zhao X, Zheng S, Chen D, Zheng M, Li X, Li G, Lin H, Chang J, Zeng H, Guo JT (2020) LY6E restricts entry of human coronaviruses, including currently pandemic SARS‐CoV‐2. J Virol 94: e00562‐20 32641482 PMC7459569
  52. Zheng M, Zhao X, Zheng S, Chen D, Du P, Li X, Jiang D, Guo J‐T, Zeng H, Lin H (2020) Bat SARS‐Like WIV1 coronavirus uses the ACE2 of multiple animal species as receptor and evades IFITM3 restriction via TMPRSS2 activation of membrane fusion. Emerg Microbes Infect 9: 1567– 1579 32602823 PMC7473123
  53. Zhou Z, Ren L, Zhang L, Zhong J, Xiao Y, Jia Z, Guo L, Yang J, Wang C, Jiang S et al (2020) Heightened Innate Immune Responses in the Respiratory Tract of COVID‐19 Patients. Cell Host Microbe 27: 883– 890 e2 32407669 PMC7196896

Cited by

Sorted by citations

Authors
Buchrieser, J., Dufloo, J., Hubert, M., Monel, B., Planas, D., Rajah, M. M., Planchais, C., Porrot, F., Guivel‐Benhassine, F., Van der Werf, S., Casartelli, N., Mouquet, H., Bruel, T., & Schwartz, O.
Affiliations
  • Centre National de la Recherche Scientifique (FR)
  • Inserm (FR)
Corresponding author
Schwartz, Olivier
ORCID
Venue
The EMBO Journal
Publisher
Springer Science and Business Media LLC
Published
2020-11-04
Volume
39
Issue
23
Pages
e106267-e106267
ISSN
  • 0261-4189 (ISSN-L)
  • 1460-2075
DOI
10.15252/embj.2020106267
PMC
PMC7646020
OpenAlex
W4206142017
Cited by
462
References
7
Citations by year
2026: 35 · 2025: 45 · 2024: 60 · 2023: 87 · 2022: 113 · 2021: 109 · 2020: 13
FWCI
10.56
Type
journal article
Language
en
Update notices
Open access
Licence
Locations
  • The EMBO Journal (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • HAL (submittedVersion)
Funders
  • Institut Pasteur
  • Agence Nationale de Recherches sur le Sida et les Hépatites Virales
  • The Vaccine Research Institute (ANR‐10‐LABX‐77)
  • LABex IBEID (ANR‐10‐LABX‐62‐IBEID)
  • TIMTAMDEM (ANR‐14‐CE14‐0029)
  • CHIKV‐Viro‐Immuno (ANR‐14‐CE14‐0015‐01)
  • Gilead HIV cure Program
  • ANR/FRM Flash COVID Proeto‐SARS‐COv‐2
  • IDISCOVR
  • Agence Nationale de la Recherche (ANR‐10‐INSB‐04‐01)
  • Region Ile‐de‐France program DIM1 Health
  • Milieu Interier Program (ANR‐10‐LABX‐69‐01)
  • Reacting and EU recover grants
  • Centre National de la Recherche Scientifique
  • Sante Public France
  • Université de paris
  • Pasteur Paris Univeristy International Doctoral Program
  • Institut National de la Santé et de la Recherche Médicale
Topics
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Health Sciences › Medicine › Infectious Diseases › COVID-19 Clinical Research Studies
  • Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
Keywords
SARS-CoV-2, syncytium formation, COVID-19 pandemic, ACE2, spike protein, cell fusion, interferon-induced transmembrane proteins, IFITM1, IFITM2, IFITM3, TMPRSS2, host restriction factors, viral entry, alveolar epithelial cells, lung injury
MeSH terms
Angiotensin-Converting Enzyme 2, COVID-19, SARS-CoV-2, Animals, Antigens, Differentiation, Cell Fusion, Cell Line, Chlorocebus aethiops, Humans, Membrane Proteins, Serine Endopeptidases, Vero Cells, Giant Cells, RNA-Binding Proteins, Host-Pathogen Interactions, HEK293 Cells, Spike Glycoprotein, Coronavirus

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review