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Markus Hoffmann

Bio: Markus Hoffmann is a academic researcher at German Primate Center who has co-authored 103 publication(s) receiving 14853 citation(s). The author has an hindex of 30. Previous affiliations of Markus Hoffmann include University of Veterinary Medicine Hanover & University of Veterinary Medicine Vienna. The author has done significant research in the topic(s): Coronavirus & Viral entry.

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Topics: Coronavirus, Viral entry, Virus ...read more
Papers
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Open accessJournal ArticleDOI: 10.1016/J.CELL.2020.02.052
16 Apr 2020-Cell
Abstract: The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency. Cell entry of coronaviruses depends on binding of the viral spike (S) proteins to cellular receptors and on S protein priming by host cell proteases. Unravelling which cellular factors are used by SARS-CoV-2 for entry might provide insights into viral transmission and reveal therapeutic targets. Here, we demonstrate that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. A TMPRSS2 inhibitor approved for clinical use blocked entry and might constitute a treatment option. Finally, we show that the sera from convalescent SARS patients cross-neutralized SARS-2-S-driven entry. Our results reveal important commonalities between SARS-CoV-2 and SARS-CoV infection and identify a potential target for antiviral intervention.

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Topics: Proteases (52%)

10,193 Citations


Open accessJournal ArticleDOI: 10.1016/J.MOLCEL.2020.04.022
21 May 2020-Molecular Cell
Abstract: The pandemic coronavirus SARS-CoV-2 threatens public health worldwide. The viral spike protein mediates SARS-CoV-2 entry into host cells and harbors a S1/S2 cleavage site containing multiple arginine residues (multibasic) not found in closely related animal coronaviruses. However, the role of this multibasic cleavage site in SARS-CoV-2 infection is unknown. Here, we report that the cellular protease furin cleaves the spike protein at the S1/S2 site and that cleavage is essential for S-protein-mediated cell-cell fusion and entry into human lung cells. Moreover, optimizing the S1/S2 site increased cell-cell, but not virus-cell, fusion, suggesting that the corresponding viral variants might exhibit increased cell-cell spread and potentially altered virulence. Our results suggest that acquisition of a S1/S2 multibasic cleavage site was essential for SARS-CoV-2 infection of humans and identify furin as a potential target for therapeutic intervention.

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Topics: Furin (56%), Entry into host (56%), Coronavirus (51%)

893 Citations


Open accessPosted ContentDOI: 10.1101/2020.01.31.929042
31 Jan 2020-bioRxiv
Abstract: The emergence of a novel, highly pathogenic coronavirus, 2019-nCoV, in China, and its rapid national and international spread pose a global health emergency. Coronaviruses use their spike proteins to select and enter target cells and insights into nCoV-2019 spike (S)-driven entry might facilitate assessment of pandemic potential and reveal therapeutic targets. Here, we demonstrate that 2019-nCoV-S uses the SARS-coronavirus receptor, ACE2, for entry and the cellular protease TMPRSS2 for 2019-nCoV-S priming. A TMPRSS2 inhibitor blocked entry and might constitute a treatment option. Finally, we show that the serum form a convalescent SARS patient neutralized 2019-nCoV-S-driven entry. Our results reveal important commonalities between 2019-nCoV and SARS-coronavirus infection, which might translate into similar transmissibility and disease pathogenesis. Moreover, they identify a target for antiviral intervention. One sentence summary The novel 2019 coronavirus and the SARS-coronavirus share central biological properties which can guide risk assessment and intervention.

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  • Fig. 1. 2019-nCoV-S and SARS-S facilitates entry into a similar panel of mammalian cell 333 lines. Analysis of 2019-nCoV-S expression (A) and pseudotype incorporation (B) by Western 334 blot. Representative blots from three experiments are shown. ß-Actin (cell lysates) and VSV-M 335 (particles) served as loading controls. (C) Cell lines of human and animal origin were inoculated 336 with pseudotyped VSV harboring VSV-G, SARS-S or 2019-nCoV-S. At 16 h postinoculation, 337 pseudotype entry was analyzed. Shown are the combined data of three experiments. Error bars 338 indicate SEM. 339 340 341 342
    Fig. 1. 2019-nCoV-S and SARS-S facilitates entry into a similar panel of mammalian cell 333 lines. Analysis of 2019-nCoV-S expression (A) and pseudotype incorporation (B) by Western 334 blot. Representative blots from three experiments are shown. ß-Actin (cell lysates) and VSV-M 335 (particles) served as loading controls. (C) Cell lines of human and animal origin were inoculated 336 with pseudotyped VSV harboring VSV-G, SARS-S or 2019-nCoV-S. At 16 h postinoculation, 337 pseudotype entry was analyzed. Shown are the combined data of three experiments. Error bars 338 indicate SEM. 339 340 341 342
  • Fig. 4. Serum from a convalescent SARS patient cross-neutralizes 2019-nCoV-S-driven 369 entry. Pseudotypes harboring the indicated viral surface proteins were incubated with different 370 dilutions of serum from a convalescent SARS patient and subsequently inoculated onto 293T 371 cells that transiently express ACE2 in order to evaluate cross-neutralization. The results from a 372 representative experiment with triplicate samples are shown and were confirmed in a separate 373 experiment. Error bars indicate SD. Statistical significance was tested by two-way ANOVA with 374 Dunnett posttest. 375
    Fig. 4. Serum from a convalescent SARS patient cross-neutralizes 2019-nCoV-S-driven 369 entry. Pseudotypes harboring the indicated viral surface proteins were incubated with different 370 dilutions of serum from a convalescent SARS patient and subsequently inoculated onto 293T 371 cells that transiently express ACE2 in order to evaluate cross-neutralization. The results from a 372 representative experiment with triplicate samples are shown and were confirmed in a separate 373 experiment. Error bars indicate SD. Statistical significance was tested by two-way ANOVA with 374 Dunnett posttest. 375
  • Fig. 2. 2019-nCoV-S utilizes ACE2 as cellular receptor. (A) The S protein of 2019-nCoV 344 clusters phylogenetically with S proteins of known bat-associated betacoronaviruses (see also SI 345 Figure 1 for more details). (B) Alignment of the receptor binding motif of SARS-S with 346 corresponding sequences of bat-associated betacoronavirus S proteins that are able or unable to 347 use ACE2 as cellular receptor reveals that 2019-nCoV possesses amino acid residues crucial for 348 ACE2 binding. (C) 293T cells transiently expressing ACE2 of human (dark blue) or bat (light 349 blue) origin, human APN (purple) or hDPP4 (green) were inoculated with pseudotyped VSV 350 harboring VSV-G, SARS-S, 2019-nCoV-S, MERS-S or 229E-S. At 16 h postinoculation, 351 pseudotype entry was analyzed. The average of three independent experiments is shown. Error 352 bars indicate SEM. 353 354
    Fig. 2. 2019-nCoV-S utilizes ACE2 as cellular receptor. (A) The S protein of 2019-nCoV 344 clusters phylogenetically with S proteins of known bat-associated betacoronaviruses (see also SI 345 Figure 1 for more details). (B) Alignment of the receptor binding motif of SARS-S with 346 corresponding sequences of bat-associated betacoronavirus S proteins that are able or unable to 347 use ACE2 as cellular receptor reveals that 2019-nCoV possesses amino acid residues crucial for 348 ACE2 binding. (C) 293T cells transiently expressing ACE2 of human (dark blue) or bat (light 349 blue) origin, human APN (purple) or hDPP4 (green) were inoculated with pseudotyped VSV 350 harboring VSV-G, SARS-S, 2019-nCoV-S, MERS-S or 229E-S. At 16 h postinoculation, 351 pseudotype entry was analyzed. The average of three independent experiments is shown. Error 352 bars indicate SEM. 353 354
Topics: Coronavirus (61%)

554 Citations


Open accessJournal ArticleDOI: 10.1016/J.CELL.2020.04.031
28 May 2020-Cell
Abstract: Coronaviruses make use of a large envelope protein called spike (S) to engage host cell receptors and catalyze membrane fusion. Because of the vital role that these S proteins play, they represent a vulnerable target for the development of therapeutics. Here, we describe the isolation of single-domain antibodies (VHHs) from a llama immunized with prefusion-stabilized coronavirus spikes. These VHHs neutralize MERS-CoV or SARS-CoV-1 S pseudotyped viruses, respectively. Crystal structures of these VHHs bound to their respective viral targets reveal two distinct epitopes, but both VHHs interfere with receptor binding. We also show cross-reactivity between the SARS-CoV-1 S-directed VHH and SARS-CoV-2 S and demonstrate that this cross-reactive VHH neutralizes SARS-CoV-2 S pseudotyped viruses as a bivalent human IgG Fc-fusion. These data provide a molecular basis for the neutralization of pathogenic betacoronaviruses by VHHs and suggest that these molecules may serve as useful therapeutics during coronavirus outbreaks.

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Topics: Coronavirus (51%)

370 Citations


Open accessJournal ArticleDOI: 10.1016/J.CELL.2021.03.036
Markus Hoffmann1, Markus Hoffmann2, Prerna Arora2, Prerna Arora1  +21 moreInstitutions (5)
29 Apr 2021-Cell
Abstract: The global spread of SARS-CoV-2/COVID-19 is devastating health systems and economies worldwide. Recombinant or vaccine-induced neutralizing antibodies are used to combat the COVID-19 pandemic. However, the recently emerged SARS-CoV-2 variants B.1.1.7 (UK), B.1.351 (South Africa), and P.1 (Brazil) harbor mutations in the viral spike (S) protein that may alter virus-host cell interactions and confer resistance to inhibitors and antibodies. Here, using pseudoparticles, we show that entry of all variants into human cells is susceptible to blockade by the entry inhibitors soluble ACE2, Camostat, EK-1, and EK-1-C4. In contrast, entry of the B.1.351 and P.1 variant was partially (Casirivimab) or fully (Bamlanivimab) resistant to antibodies used for COVID-19 treatment. Moreover, entry of these variants was less efficiently inhibited by plasma from convalescent COVID-19 patients and sera from BNT162b2-vaccinated individuals. These results suggest that SARS-CoV-2 may escape neutralizing antibody responses, which has important implications for efforts to contain the pandemic.

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Topics: Neutralizing antibody (59%)

338 Citations


Cited by
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Open accessJournal ArticleDOI: 10.1016/J.CELL.2020.02.052
16 Apr 2020-Cell
Abstract: The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency. Cell entry of coronaviruses depends on binding of the viral spike (S) proteins to cellular receptors and on S protein priming by host cell proteases. Unravelling which cellular factors are used by SARS-CoV-2 for entry might provide insights into viral transmission and reveal therapeutic targets. Here, we demonstrate that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. A TMPRSS2 inhibitor approved for clinical use blocked entry and might constitute a treatment option. Finally, we show that the sera from convalescent SARS patients cross-neutralized SARS-2-S-driven entry. Our results reveal important commonalities between SARS-CoV-2 and SARS-CoV infection and identify a potential target for antiviral intervention.

...read more

Topics: Proteases (52%)

10,193 Citations


Open accessJournal ArticleDOI: 10.1126/SCIENCE.ABB2507
13 Mar 2020-Science
Abstract: The outbreak of a novel coronavirus (2019-nCoV) represents a pandemic threat that has been declared a public health emergency of international concern. The CoV spike (S) glycoprotein is a key target for vaccines, therapeutic antibodies, and diagnostics. To facilitate medical countermeasure development, we determined a 3.5-angstrom-resolution cryo-electron microscopy structure of the 2019-nCoV S trimer in the prefusion conformation. The predominant state of the trimer has one of the three receptor-binding domains (RBDs) rotated up in a receptor-accessible conformation. We also provide biophysical and structural evidence that the 2019-nCoV S protein binds angiotensin-converting enzyme 2 (ACE2) with higher affinity than does severe acute respiratory syndrome (SARS)-CoV S. Additionally, we tested several published SARS-CoV RBD-specific monoclonal antibodies and found that they do not have appreciable binding to 2019-nCoV S, suggesting that antibody cross-reactivity may be limited between the two RBDs. The structure of 2019-nCoV S should enable the rapid development and evaluation of medical countermeasures to address the ongoing public health crisis.

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5,197 Citations


Open accessJournal ArticleDOI: 10.1016/J.CELL.2020.02.058
16 Apr 2020-Cell
Abstract: The emergence of SARS-CoV-2 has resulted in >90,000 infections and >3,000 deaths. Coronavirus spike (S) glycoproteins promote entry into cells and are the main target of antibodies. We show that SARS-CoV-2 S uses ACE2 to enter cells and that the receptor-binding domains of SARS-CoV-2 S and SARS-CoV S bind with similar affinities to human ACE2, correlating with the efficient spread of SARS-CoV-2 among humans. We found that the SARS-CoV-2 S glycoprotein harbors a furin cleavage site at the boundary between the S1/S2 subunits, which is processed during biogenesis and sets this virus apart from SARS-CoV and SARS-related CoVs. We determined cryo-EM structures of the SARS-CoV-2 S ectodomain trimer, providing a blueprint for the design of vaccines and inhibitors of viral entry. Finally, we demonstrate that SARS-CoV S murine polyclonal antibodies potently inhibited SARS-CoV-2 S mediated entry into cells, indicating that cross-neutralizing antibodies targeting conserved S epitopes can be elicited upon vaccination.

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Topics: Ectodomain (56%), Viral entry (55%), Epitope (53%) ...read more

4,968 Citations


Open accessJournal ArticleDOI: 10.1001/JAMAINTERNMED.2020.0994
Chaomin Wu1, Xiaoyan Chen1, Yanping Cai, Jia’an Xia2  +21 moreInstitutions (3)
Abstract: Importance Coronavirus disease 2019 (COVID-19) is an emerging infectious disease that was first reported in Wuhan, China, and has subsequently spread worldwide. Risk factors for the clinical outcomes of COVID-19 pneumonia have not yet been well delineated. Objective To describe the clinical characteristics and outcomes in patients with COVID-19 pneumonia who developed acute respiratory distress syndrome (ARDS) or died. Design, Setting, and Participants Retrospective cohort study of 201 patients with confirmed COVID-19 pneumonia admitted to Wuhan Jinyintan Hospital in China between December 25, 2019, and January 26, 2020. The final date of follow-up was February 13, 2020. Exposures Confirmed COVID-19 pneumonia. Main Outcomes and Measures The development of ARDS and death. Epidemiological, demographic, clinical, laboratory, management, treatment, and outcome data were also collected and analyzed. Results Of 201 patients, the median age was 51 years (interquartile range, 43-60 years), and 128 (63.7%) patients were men. Eighty-four patients (41.8%) developed ARDS, and of those 84 patients, 44 (52.4%) died. In those who developed ARDS, compared with those who did not, more patients presented with dyspnea (50 of 84 [59.5%] patients and 30 of 117 [25.6%] patients, respectively [difference, 33.9%; 95% CI, 19.7%-48.1%]) and had comorbidities such as hypertension (23 of 84 [27.4%] patients and 16 of 117 [13.7%] patients, respectively [difference, 13.7%; 95% CI, 1.3%-26.1%]) and diabetes (16 of 84 [19.0%] patients and 6 of 117 [5.1%] patients, respectively [difference, 13.9%; 95% CI, 3.6%-24.2%]). In bivariate Cox regression analysis, risk factors associated with the development of ARDS and progression from ARDS to death included older age (hazard ratio [HR], 3.26; 95% CI 2.08-5.11; and HR, 6.17; 95% CI, 3.26-11.67, respectively), neutrophilia (HR, 1.14; 95% CI, 1.09-1.19; and HR, 1.08; 95% CI, 1.01-1.17, respectively), and organ and coagulation dysfunction (eg, higher lactate dehydrogenase [HR, 1.61; 95% CI, 1.44-1.79; and HR, 1.30; 95% CI, 1.11-1.52, respectively] and D-dimer [HR, 1.03; 95% CI, 1.01-1.04; and HR, 1.02; 95% CI, 1.01-1.04, respectively]). High fever (≥39 °C) was associated with higher likelihood of ARDS development (HR, 1.77; 95% CI, 1.11-2.84) and lower likelihood of death (HR, 0.41; 95% CI, 0.21-0.82). Among patients with ARDS, treatment with methylprednisolone decreased the risk of death (HR, 0.38; 95% CI, 0.20-0.72). Conclusions and Relevance Older age was associated with greater risk of development of ARDS and death likely owing to less rigorous immune response. Although high fever was associated with the development of ARDS, it was also associated with better outcomes among patients with ARDS. Moreover, treatment with methylprednisolone may be beneficial for patients who develop ARDS.

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Topics: ARDS (52%)

4,614 Citations


Open accessJournal ArticleDOI: 10.1038/S41586-020-2196-X
Roman Wölfel1, Victor M. Corman2, Wolfgang Guggemos, M Seilmaier  +15 moreInstitutions (4)
01 Apr 2020-Nature
Abstract: Coronavirus disease 2019 (COVID-19) is an acute infection of the respiratory tract that emerged in late 20191,2. Initial outbreaks in China involved 13.8% of cases with severe courses, and 6.1% of cases with critical courses3. This severe presentation may result from the virus using a virus receptor that is expressed predominantly in the lung2,4; the same receptor tropism is thought to have determined the pathogenicity—but also aided in the control—of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of cases of COVID-19 in which the patient shows mild upper respiratory tract symptoms, which suggests the potential for pre- or oligosymptomatic transmission6–8. There is an urgent need for information on virus replication, immunity and infectivity in specific sites of the body. Here we report a detailed virological analysis of nine cases of COVID-19 that provides proof of active virus replication in tissues of the upper respiratory tract. Pharyngeal virus shedding was very high during the first week of symptoms, with a peak at 7.11 × 108 RNA copies per throat swab on day 4. Infectious virus was readily isolated from samples derived from the throat or lung, but not from stool samples—in spite of high concentrations of virus RNA. Blood and urine samples never yielded virus. Active replication in the throat was confirmed by the presence of viral replicative RNA intermediates in the throat samples. We consistently detected sequence-distinct virus populations in throat and lung samples from one patient, proving independent replication. The shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 7 days in 50% of patients (and by day 14 in all patients), but was not followed by a rapid decline in viral load. COVID-19 can present as a mild illness of the upper respiratory tract. The confirmation of active virus replication in the upper respiratory tract has implications for the containment of COVID-19. Detailed virological analysis of nine cases of coronavirus disease 2019 (COVID-19) provides proof of active replication of the SARS-CoV-2 virus in tissues of the upper respiratory tract.

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Topics: Virus receptor (62%), Coronavirus (60%), Viral shedding (59%) ...read more

4,325 Citations


Performance
Metrics

Author's H-index: 30

No. of papers from the Author in previous years
YearPapers
202146
202019
201911
20186
20175
20164

Top Attributes

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Author's top 5 most impactful journals

bioRxiv

16 papers, 834 citations

Journal of Virology

10 papers, 208 citations

PLOS ONE

7 papers, 253 citations

medRxiv

7 papers, 40 citations

Cell Reports

5 papers, 91 citations

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