193 results on '"Mothes W"'
Search Results
2. Cell-to-cell transmission of retroviruses along filopodial bridges: C3-L1
- Author
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Mothes, W. and Sherer, N.
- Published
- 2007
3. Native structure of a retroviral envelope protein and its conformational change upon interaction with the target cell
- Author
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Riedel, C, Vasishtan, D, Siebert, CA, Whittle, CJ, Lehmann, MJ, Mothes, W, and Grünewald, K
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Env ,Binding Sites ,Protein Conformation ,cET, electron cryo tomography ,viruses ,aa, amino acid ,Cryoelectron Microscopy ,cEM, electron cryo microscopy ,FrMLV, Friends MLV ,SU, Env surface unit ,mCAT, murine cationic amino acid transporter (SLC7a1, Gene ID: 11987) ,Article ,YFP, yellow fluorescent protein ,Sub-volume averaging ,MoMLV, Moloney MLV ,Leukemia Virus, Murine ,TM, Env transmembrane Unit ,Virus entry ,Env, envelope glycoprotein ,Retroviridae ,Electron cryo tomography ,Viral Envelope Proteins ,Structural Biology ,Murine leukemia virus ,MLV, murine leukemia virus - Abstract
Enveloped viruses enter their host cells by membrane fusion. The process of attachment and fusion in retroviruses is mediated by a single viral envelope glycoprotein (Env). Conformational changes of Env in the course of fusion are a focus of intense studies. Here we provide further insight into the changes occurring in retroviral Env during its initial interaction with the cell, employing murine leukemia virus (MLV) as model system. We first determined the structure of both natively membrane anchored MLV Env and MLV Env tagged with YFP in the proline rich region (PRR) by electron cryo tomography (cET) and sub-volume averaging. At a resolution of ∼20Å, native MLV Env presents as a hollow trimer (height ∼85Å, diameter ∼120Å) composed of step-shaped protomers. The major difference to the YFP-tagged protein was in regions outside of the central trimer. Next, we focused on elucidating the changes in MLV Env upon interaction with a host cell. Virus interaction with the plasma membrane occurred over a large surface and Env clustering on the binding site was observed. Sub-volume averaging did yield a low-resolution structure of Env interacting with the cell, which had lost its threefold symmetry and was elongated by ∼35Å in comparison to the unbound protein. This indicates a major rearrangement of Env upon host cell binding. At the site of virus interaction, the otherwise clearly defined bilayer structure of the host cell plasma membrane was much less evident, indicative of integral membrane protein accumulation and/or a change in membrane lipid composition.
- Published
- 2016
4. Colostomien im Säuglings- und Kindesalter
- Author
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Joppich, I., Mothes, W., and Merkatz, G. F. v.
- Published
- 1968
- Full Text
- View/download PDF
5. Murine Leukemia Virus Gag Localizes to the Uropod of Migrating Primary Lymphocytes
- Author
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Li, F., primary, Sewald, X., additional, Jin, J., additional, Sherer, N. M., additional, and Mothes, W., additional
- Published
- 2014
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6. Time-Resolved Imaging of Single Retrovirus-Cell Fusion
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Melikyan, G, primary, Barnard, R, additional, Abrahamyan, L, additional, Mothes, W, additional, and Young, J, additional
- Published
- 2005
- Full Text
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7. Hemmstoffe und Aktivatoren des Herzsuccinatoxydasesystems in Erythrozyten und Retikulozyten
- Author
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Raporort, S., Gerischer-Mothes, W., and Nieradt, Ch.
- Published
- 1955
- Full Text
- View/download PDF
8. Systematic probing of the environment of a translocating secretory protein during translocation through the ER membrane.
- Author
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Mothes, W., primary, Prehn, S., additional, and Rapoport, T.A., additional
- Published
- 1994
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9. Über die Wirkung von Al und Ca auf das Succinatoxydase-System des Herzmuskels
- Author
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Rapoport, S., Nieradt, Ch., and Gerischer-Mothes, W.
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- 1955
- Full Text
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10. Germanium Incorporation in Ga.
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Müller, K., Mothes, W., Jacobs, B., and Butter, E.
- Published
- 1982
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11. Follow-up studies in 50 totally colectomised children
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Rode H, H. Kraeft, Menardi G, Knutrud O, Stirrat A, Schickedanz H, Hecker Wc, A. M. Holschneider, Mothes W, and Bürger D
- Subjects
Adult ,Pediatrics ,medicine.medical_specialty ,Adolescent ,Colon ,medicine.medical_treatment ,Frequent stools ,Enteritis ,Colonic Diseases ,Postoperative Complications ,Consistency (statistics) ,medicine ,Humans ,Hirschsprung Disease ,Child ,Colectomy ,Enterocolitis, Pseudomembranous ,business.industry ,Follow up studies ,Infant ,Intestinal Polyps ,medicine.disease ,Colitis ,Child, Preschool ,Pediatrics, Perinatology and Child Health ,Colonic Neoplasms ,Dysentery, Amebic ,Surgery ,business ,Follow-Up Studies - Abstract
We report on 50 totally colectomised children, most of whom suffered from Hirschsprung's disease. Of the 50, one child died postoperatively of enteritis. On an average, the children were re-examined 5 1/2 years after the colectomy. The findings were as follows: With the exception of four, the size and weight of the patients were within the norm; 20 passed frequent stools of pulpy consistency; 16 suffered from disturbances of continence; 14 developed severe, partially recurrent enteritis. We did not find any advantage of a single method of operation, e.g. Martin's operation.
- Published
- 1985
12. Kinetic and thermodynamic analysis of the control of C3b by the complement regulatory proteins factors H and I
- Author
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Gerischer-Mothes W, Rapoport S, and Wagenknecht C
- Subjects
Sheep ,Chemistry ,Thermodynamics ,Biochemistry ,Complement (complexity) ,Molecular Weight ,Kinetics ,Spectrometry, Fluorescence ,Complement Factor I ,Complement Factor H ,Complement C3b ,Endopeptidases ,Complement C3b Inactivator Proteins ,Animals - Published
- 1983
13. TRIM5 is an innate immune sensor for the retrovirus capsid lattice
- Author
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Pertel, T, Hausmann, S, Morger, D, Züger, S, Guerra, J, Lascano, J, Reinhard, C, Santoni, F A, Uchil, P D, Chatel, L, Bisiaux, A, Albert, M L, Strambio-De-Castillia, C, Mothes, W, Pizzato, M, Grütter, M G, and Luban, J
- Subjects
3. Good health
14. AAS-Untersuchungen über Verdampfungs- und plasmaprozesse in der Graphitrohrküvette Zur Optimierung der Bestimmungen von Cd-, Zn- und Mn-Spuren in AIIIBV-Halbleiter-Matrices
- Author
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Dittrich, K., primary, Mothes, W., additional, and Weber, P., additional
- Published
- 1978
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15. Follow-Up Studies in 50 Totally Colectomised Children
- Author
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Kraeft, H., primary, Holschneider, A., additional, Hecker, W., additional, Bürger, D., additional, Knutrud, O., additional, Menardi, G., additional, Mothes, W., additional, Rode, H., additional, Schickedanz, H., additional, and Stirrat, A., additional
- Published
- 1985
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16. Phosphatide als Inaktivatoren des Reticulocyten-Hemmstoffs der Mitochondrien-Atmung
- Author
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Rapoport, S., primary and Gerischer-Mothes, W., additional
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- 1959
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17. Häminkatalysierte Oxydation ungesättigter Fettsäuren im Blut anämischer Kaninchen
- Author
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Rapoport, S., primary, Gerischer-Mothes, W., additional, and Nieradt, C., additional
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- 1955
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18. Biochemische Vorgänge bei der Reticulocyten-Reifung: Auftreten und Abklingen des Reticulocyten-Hemmstoffes
- Author
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Rapoport, S., primary and Gerischer-Mothes, W., additional
- Published
- 1956
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- View/download PDF
19. �ber die Wirkung von Al und Ca auf das Succinatoxydase-System des Herzmuskels
- Author
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Rapoport, S., primary, Nieradt, Ch., additional, and Gerischer-Mothes, W., additional
- Published
- 1955
- Full Text
- View/download PDF
20. Biochemische Vorgänge bei der Reticulocyten-Reifung: Über die Inaktivierung des Reticulocyten-Hemmstoffes der Atmungskette
- Author
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Rapoport, S., primary, Gerischer-Mothes, W., additional, and Nieradt-Hiebsch, Ch., additional
- Published
- 1956
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21. Biochemische Vorgänge bei der Erythrocytenreifung: Über einen Hemmstoff des Succinatoxydase-Systems in Reticulocyten
- Author
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Rapoport, S., primary and Gerischer-Mothes, W., additional
- Published
- 1955
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22. TRIM5alpha contributes to the anti-viral state
- Author
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Zufferey Madeleine, Hausman Stephane, Neagu Martha R, Pertel Thomas, Uchil Pradeep, Mothes Walther, and Luban Jeremy
- Subjects
Immunologic diseases. Allergy ,RC581-607 - Published
- 2009
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23. Follow-Up Studies in 50 Totally Colectomised Children.
- Author
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Kraeft, H., Holschneider, A. M., Hecker, W. Ch., B�rger, D., Knutrud, O., Menardi, G., Mothes, W., Rode, H., Schickedanz, H., and Stirrat, A.
- Published
- 1985
- Full Text
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24. Investigations of trace analysis of A IIIB V semiconductor microsamples by atomic spectroscopy—VII : Investigation of trace and thin-layer analysis of doping elements (Ag, Au, Bi, Cd, Sn, Tl) in InAs by atomic-absorption with electrothermal evaporation
- Author
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Dittrich, K., Mothes, W., Yudelevich, I.G., and Papina, T.S.
- Published
- 1985
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25. Untersuchungen zur atomspektroskopischen Spurenanalyse in A III B V-Halbleitermikroproben—VI Untersuchungen zur Schichtabtrennung, Profil- und Spurenanalyse an InSb-Materialien
- Author
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Schelpakowa, I.R., Schtscherbakowa, O.I., Judelewitsch, I.G., Beisel, N.F., Dittrich, K., and Mothes, W.
- Published
- 1982
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26. TRIM22 inhibits HIV-1 transcription independently of its E3 ubiquitin ligase activity, Tat, and NF-kappaB-responsive long terminal repeat elements
- Author
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Elisa Vicenzi, Pradeep D. Uchil, Nadir Mechti, Walther Mothes, Guido Poli, Thomas Pertel, Cinzia Pultrone, Jeremy Luban, Sara Marelli, Anna Kajaste-Rudnitski, Chair of Risk, Safety and Uncertainty Quantification [ETH Zurich], Institute of Structural Engineering [ETH Zürich] (IBK), Department of Civil, Environmental and Geomatic Engineering [ETH Zürich] (D-BAUG), Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)- Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)-Department of Civil, Environmental and Geomatic Engineering [ETH Zürich] (D-BAUG), Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)- Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Kajaste Rudnitski, A, Marelli, S. S., Pultrone, C, Pertel, T, Uchil, P. D., Mechti, N, Mothes, W, Poli, Guido, Luban, J, and Vicenzi, E.
- Subjects
Transcription, Genetic ,T-Lymphocytes ,[SDV]Life Sciences [q-bio] ,T cell ,Immunology ,Cellular Response to Infection ,Biology ,Virus Replication ,TRIM22 ,Microbiology ,Monocytes ,Cell Line ,Minor Histocompatibility Antigens ,Tripartite Motif Proteins ,03 medical and health sciences ,Transactivation ,Transcription (biology) ,Virology ,medicine ,Humans ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,0303 health sciences ,030302 biochemistry & molecular biology ,HEK 293 cells ,Molecular biology ,Long terminal repeat ,3. Good health ,Repressor Proteins ,medicine.anatomical_structure ,Viral replication ,Cell culture ,Insect Science ,HIV-1 - Abstract
Previous studies identified clones of the U937 promonocytic cell line that were either permissive or nonpermissive for human immunodeficiency virus type 1 (HIV-1) replication. These clones were investigated further in the search for host restriction factors that could explain their differential capacity to support HIV-1 replication. Among known HIV-1 restriction factors screened, tri partite m otif-containing protein 22 (TRIM22) was the only factor constitutively expressed in nonpermissive and absent in permissive U937 cells. Stable TRIM22 knockdown (KD) rescued HIV-1 long-terminal-repeat (LTR)-driven transcription in KD-nonpermissive cells to the levels observed in permissive cells. Conversely, transduction-mediated expression of TRIM22 in permissive cells reduced LTR-driven luciferase expression by ∼7-fold, supporting a negative role of TRIM22 in HIV-1 transcription. This finding was further confirmed in the human T cell line A3.01 expressing TRIM22. Moreover, overexpression of TRIM22 in 293T cells significantly impaired basal and phorbol myristate acetate-ionomycin-induced HIV-1 LTR-driven gene expression, whereas inhibition of tumor necrosis factor alpha-induced viral transcription was a consequence of lower basal expression. In agreement, TRIM22 equally inhibited an LTR construct lacking the tandem NF-κB binding sites. In addition, TRIM22 did not affect Tat-mediated LTR transactivation. Finally, these effects were independent of TRIM22 E3 ubiquitin-ligase activity. In the context of replication-competent virus, significantly higher levels of HIV-1 production were observed in KD-nonpermissive versus control nonpermissive U937 cells after infection. In contrast, lower peak levels of HIV-1 replication characterized U937 and A3.01 cells expressing TRIM22 versus their control transduced counterpart. Thus, nuclear TRIM22 significantly impairs HIV-1 replication, likely by interfering with Tat- and NF-κB-independent LTR-driven transcription.
- Published
- 2011
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27. The asymmetric opening of HIV-1 Env by a potent CD4 mimetic enables anti-coreceptor binding site antibodies to mediate ADCC.
- Author
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Richard J, Grunst MW, Niu L, Díaz-Salinas MA, Tolbert WD, Marchitto L, Zhou F, Bourassa C, Yang D, Chiu TJ, Chen HC, Benlarbi M, Gottumukkala S, Li W, Dionne K, Bélanger É, Chatterjee D, Medjahed H, Hendrickson WA, Sodroski J, Lang ZC, Morton AJ, Huang RK, Matthies D, Smith AB 3rd, Mothes W, Munro JB, Pazgier M, and Finzi A
- Abstract
HIV-1 envelope glycoproteins (Env) from primary HIV-1 isolates typically adopt a pretriggered "closed" conformation that resists to CD4-induced (CD4i) non-neutralizing antibodies (nnAbs) mediating antibody-dependent cellular cytotoxicity (ADCC). CD4-mimetic compounds (CD4mcs) "open-up" Env allowing binding of CD4i nnAbs, thereby sensitizing HIV-1-infected cells to ADCC. Two families of CD4i nnAbs, the anti-cluster A and anti-coreceptor binding site (CoRBS) Abs, are required to mediate ADCC in combination with the indane CD4mc BNM-III-170. Recently, new indoline CD4mcs with improved potency and breadth have been described. Here, we show that the lead indoline CD4mc, CJF-III-288, sensitizes HIV-1-infected cells to ADCC mediated by anti-CoRBS Abs alone, contributing to improved ADCC activity. Structural and conformational analyses reveal that CJF-III-288, in combination with anti-CoRBS Abs, potently stabilizes an asymmetric "open" State-3 Env conformation, This Env conformation orients the anti-CoRBS Ab to improve ADCC activity and therapeutic potential.
- Published
- 2024
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28. Design of soluble HIV-1 envelope trimers free of covalent gp120-gp41 bonds with prevalent native-like conformation.
- Author
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Zhang P, Gorman J, Tsybovsky Y, Lu M, Liu Q, Gopan V, Singh M, Lin Y, Miao H, Seo Y, Kwon A, Olia AS, Chuang GY, Geng H, Lai YT, Zhou T, Mascola JR, Mothes W, Kwong PD, and Lusso P
- Subjects
- Humans, Antibodies, Neutralizing immunology, CD4 Antigens metabolism, Protein Conformation, Protein Binding, HIV Envelope Protein gp120 metabolism, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 immunology, HIV Envelope Protein gp41 metabolism, HIV Envelope Protein gp41 chemistry, HIV Envelope Protein gp41 immunology, HIV-1 metabolism, HIV-1 immunology, Protein Multimerization, Solubility
- Abstract
Soluble HIV-1 envelope (Env) trimers may serve as effective vaccine immunogens. The widely utilized SOSIP trimers have been paramount for structural studies, but the disulfide bond they feature between gp120 and gp41 constrains intersubunit mobility and may alter antigenicity. Here, we report an alternative strategy to generate stabilized soluble Env trimers free of covalent gp120-gp41 bonds. Stabilization was achieved by introducing an intrasubunit disulfide bond between the inner and outer domains of gp120, defined as interdomain lock (IDL). Correctly folded IDL trimers displaying a native-like antigenic profile were produced for HIV-1 Envs of different clades. Importantly, the IDL design abrogated CD4 binding while not affecting recognition by potent neutralizing antibodies to the CD4-binding site. By cryoelectron microscopy, IDL trimers were shown to adopt a closed prefusion configuration, while single-molecule fluorescence resonance energy transfer documented a high prevalence of native-like conformation. Thus, IDL trimers may be promising candidates as vaccine immunogens., Competing Interests: Declaration of interests P.L. and P.Z. applied for a patent describing HIV-1 envelope trimers stabilized in a native-like, non-CD4-binding conformation (US Provisional Patent Application No. 62/306,006, filed on March 9, 2016, entitled “Recombinant HIV-1 envelope proteins and their use”)., (Published by Elsevier Inc.)
- Published
- 2024
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29. Structure and inhibition of SARS-CoV-2 spike refolding in membranes.
- Author
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Grunst MW, Qin Z, Dodero-Rojas E, Ding S, Prévost J, Chen Y, Hu Y, Pazgier M, Wu S, Xie X, Finzi A, Onuchic JN, Whitford PC, Mothes W, and Li W
- Subjects
- Humans, Virus Internalization, Protein Refolding, Electron Microscope Tomography, Protein Multimerization, Betacoronavirus immunology, Betacoronavirus chemistry, Cell Membrane metabolism, COVID-19 virology, COVID-19 immunology, Peptidyl-Dipeptidase A chemistry, Peptidyl-Dipeptidase A metabolism, Spike Glycoprotein, Coronavirus chemistry, Spike Glycoprotein, Coronavirus metabolism, Spike Glycoprotein, Coronavirus immunology, Spike Glycoprotein, Coronavirus genetics, SARS-CoV-2 immunology, SARS-CoV-2 genetics, Angiotensin-Converting Enzyme 2 metabolism, Angiotensin-Converting Enzyme 2 chemistry, Molecular Dynamics Simulation, Antibodies, Neutralizing immunology, Antibodies, Neutralizing chemistry, Cryoelectron Microscopy, Antibodies, Viral immunology, Antibodies, Viral chemistry, Protein Domains
- Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds the receptor angiotensin converting enzyme 2 (ACE2) and drives virus-host membrane fusion through refolding of its S2 domain. Whereas the S1 domain contains high sequence variability, the S2 domain is conserved and is a promising pan-betacoronavirus vaccine target. We applied cryo-electron tomography to capture intermediates of S2 refolding and understand inhibition by antibodies to the S2 stem-helix. Subtomogram averaging revealed ACE2 dimers cross-linking spikes before transitioning into S2 intermediates, which were captured at various stages of refolding. Pan-betacoronavirus neutralizing antibodies targeting the S2 stem-helix bound to and inhibited refolding of spike prehairpin intermediates. Combined with molecular dynamics simulations, these structures elucidate the process of SARS-CoV-2 entry and reveal how pan-betacoronavirus S2-targeting antibodies neutralize infectivity by arresting prehairpin intermediates.
- Published
- 2024
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30. Proof-of-concept studies with a computationally designed M pro inhibitor as a synergistic combination regimen alternative to Paxlovid.
- Author
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Papini C, Ullah I, Ranjan AP, Zhang S, Wu Q, Spasov KA, Zhang C, Mothes W, Crawford JM, Lindenbach BD, Uchil PD, Kumar P, Jorgensen WL, and Anderson KS
- Subjects
- Humans, Animals, Mice, Clinical Protocols, Drug Combinations, Antiviral Agents pharmacology, Hepatitis C, Chronic, Ritonavir, Hydroxylamines, Cytidine analogs & derivatives, Nitriles, Leucine, Proline, Lactams
- Abstract
As the SARS-CoV-2 virus continues to spread and mutate, it remains important to focus not only on preventing spread through vaccination but also on treating infection with direct-acting antivirals (DAA). The approval of Paxlovid, a SARS-CoV-2 main protease (M
pro ) DAA, has been significant for treatment of patients. A limitation of this DAA, however, is that the antiviral component, nirmatrelvir, is rapidly metabolized and requires inclusion of a CYP450 3A4 metabolic inhibitor, ritonavir, to boost levels of the active drug. Serious drug-drug interactions can occur with Paxlovid for patients who are also taking other medications metabolized by CYP4503A4, particularly transplant or otherwise immunocompromised patients who are most at risk for SARS-CoV-2 infection and the development of severe symptoms. Developing an alternative antiviral with improved pharmacological properties is critical for treatment of these patients. By using a computational and structure-guided approach, we were able to optimize a 100 to 250 μM screening hit to a potent nanomolar inhibitor and lead compound, Mpro61. In this study, we further evaluate Mpro61 as a lead compound, starting with examination of its mode of binding to SARS-CoV-2 Mpro . In vitro pharmacological profiling established a lack of off-target effects, particularly CYP450 3A4 inhibition, as well as potential for synergy with the currently approved alternate antiviral, molnupiravir. Development and subsequent testing of a capsule formulation for oral dosing of Mpro61 in B6-K18-hACE2 mice demonstrated favorable pharmacological properties, efficacy, and synergy with molnupiravir, and complete recovery from subsequent challenge by SARS-CoV-2, establishing Mpro61 as a promising potential preclinical candidate., Competing Interests: Competing interests statement:Yale University has filed a patent application covering Mpro61.- Published
- 2024
- Full Text
- View/download PDF
31. Bioorthogonal click labeling of an amber-free HIV-1 provirus for in-virus single molecule imaging.
- Author
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Ao Y, Grover JR, Gifford L, Han Y, Zhong G, Katte R, Li W, Bhattacharjee R, Zhang B, Sauve S, Qin W, Ghimire D, Haque MA, Arthos J, Moradi M, Mothes W, Lemke EA, Kwong PD, Melikyan GB, and Lu M
- Subjects
- Humans, Single Molecule Imaging, Proteins metabolism, Peptides metabolism, Proviruses, HIV-1
- Abstract
Structural dynamics of human immunodeficiency virus 1 (HIV-1) envelope (Env) glycoprotein mediate cell entry and facilitate immune evasion. Single-molecule FRET using peptides for Env labeling revealed structural dynamics of Env, but peptide use risks potential effects on structural integrity/dynamics. While incorporating noncanonical amino acids (ncAAs) into Env by amber stop-codon suppression, followed by click chemistry, offers a minimally invasive approach, this has proved to be technically challenging for HIV-1. Here, we develope an intact amber-free HIV-1 system that overcomes hurdles of preexisting viral amber codons. We achieved dual-ncAA incorporation into Env on amber-free virions, enabling single-molecule Förster resonance energy transfer (smFRET) studies of click-labeled Env that validated the previous peptide-based labeling approaches by confirming the intrinsic propensity of Env to dynamically sample multiple conformational states. Amber-free click-labeled Env also enabled real-time tracking of single virion internalization and trafficking in cells. Our system thus permits in-virus bioorthogonal labeling of proteins, compatible with studies of virus entry, trafficking, and egress from cells., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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- View/download PDF
32. Plasma Human Immunodeficiency Virus 1 Soluble Glycoprotein 120 Association With Correlates of Immune Dysfunction and Inflammation in Antiretroviral Therapy-Treated Individuals With Undetectable Viremia.
- Author
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Benlarbi M, Richard J, Bourassa C, Tolbert WD, Chartrand-Lefebvre C, Gendron-Lepage G, Sylla M, El-Far M, Messier-Peet M, Guertin C, Turcotte I, Fromentin R, Verly MM, Prévost J, Clark A, Mothes W, Kaufmann DE, Maldarelli F, Chomont N, Bégin P, Tremblay C, Baril JG, Trottier B, Trottier S, Duerr R, Pazgier M, Durand M, and Finzi A
- Subjects
- Humans, Viremia, Cohort Studies, Cross-Sectional Studies, Canada, HIV Antibodies, Glycoproteins, HIV Envelope Protein gp120, HIV-1, HIV Infections drug therapy
- Abstract
Background: Chronic inflammation persists in some people living with human immunodeficiency virus (HIV) during antiretroviral therapy and is associated with premature aging. The glycoprotein 120 (gp120) subunit of HIV-1 envelope sheds and can be detected in plasma, showing immunomodulatory properties even in the absence of detectable viremia. We evaluated whether plasma soluble gp120 (sgp120) and a family of gp120-specific anti-cluster A antibodies, linked to CD4 depletion in vitro, contribute to chronic inflammation, immune dysfunction, and subclinical cardiovascular disease in participants of the Canadian HIV and Aging Cohort Study with undetectable viremia., Methods: Cross-sectional assessment of sgp120 and anti-cluster A antibodies was performed in 386 individuals from the cohort. Their association with proinflammatory cytokines and subclinical coronary artery disease was assessed using linear regression models., Results: High levels of sgp120 and anti-cluster A antibodies were inversely correlated with CD4+ T cell count and CD4/CD8 ratio. The presence of sgp120 was associated with increased levels of interleukin 6. In participants with detectable atherosclerotic plaque and detectable sgp120, anti-cluster A antibodies and their combination with sgp120 levels correlated positively with the total volume of atherosclerotic plaques., Conclusions: This study showed that sgp120 may act as a pan toxin causing immune dysfunction and sustained inflammation in a subset of people living with HIV, contributing to the development of premature comorbid conditions., Competing Interests: Potential conflicts of interest. W. M., C. T., M. P., and A. F. received funding from ViiV Healthcare. A. C. is a full-time employee of ViiV Healthcare. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2023. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)
- Published
- 2024
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33. Epistatic pathways can drive HIV-1 escape from integrase strand transfer inhibitors.
- Author
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Hikichi Y, Grover JR, Schäfer A, Mothes W, and Freed EO
- Subjects
- Humans, Raltegravir Potassium pharmacology, Mutation, HIV Integrase Inhibitors pharmacology, HIV-1 genetics, HIV-1 metabolism, HIV Integrase genetics, HIV Integrase metabolism
- Abstract
People living with human immunodeficiency virus (HIV) receiving integrase strand transfer inhibitors (INSTIs) have been reported to experience virological failure in the absence of resistance mutations in integrase. To elucidate INSTI resistance mechanisms, we propagated HIV-1 in the presence of escalating concentrations of the INSTI dolutegravir. HIV-1 became resistant to dolutegravir by sequentially acquiring mutations in the envelope glycoprotein (Env) and the nucleocapsid protein. The selected Env mutations enhance the ability of the virus to spread via cell-cell transfer, thereby increasing the multiplicity of infection (MOI). While the selected Env mutations confer broad resistance to multiple classes of antiretrovirals, the fold resistance is ~2 logs higher for INSTIs than for other classes of drugs. We demonstrate that INSTIs are more readily overwhelmed by high MOI than other classes of antiretrovirals. Our findings advance the understanding of how HIV-1 can evolve resistance to antiretrovirals, including the potent INSTIs, in the absence of drug-target gene mutations.
- Published
- 2024
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34. Bioluminescence imaging reveals enhanced SARS-CoV-2 clearance in mice with combinatorial regimens.
- Author
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Ullah I, Escudie F, Scandale I, Gilani Z, Gendron-Lepage G, Gaudette F, Mowbray C, Fraisse L, Bazin R, Finzi A, Mothes W, Kumar P, Chatelain E, and Uchil PD
- Abstract
Direct acting antivirals (DAAs) represent critical tools for combating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) that have escaped vaccine-elicited spike-based immunity and future coronaviruses with pandemic potential. Here, we used bioluminescence imaging to evaluate therapeutic efficacy of DAAs that target SARS-CoV-2 RNA-dependent RNA polymerase (favipiravir, molnupiravir) or main protease (nirmatrelvir) against Delta or Omicron VOCs in K18-hACE2 mice. Nirmatrelvir displayed the best efficacy followed by molnupiravir and favipiravir in suppressing viral loads in the lung. Unlike neutralizing antibody treatment, DAA monotherapy regimens did not eradicate SARS-CoV-2 in mice, but combining molnupiravir with nirmatrelvir exhibited superior additive efficacy and led to virus clearance. Furthermore, combining molnupiravir with caspase-1/4 inhibitor mitigated inflammation and lung pathology whereas combining molnupiravir with COVID-19 convalescent plasma demonstrated synergy, rapid virus clearance, and 100% survival. Thus, our study provides insights into in vivo treatment efficacies of DAAs and other effective combinations to bolster COVID-19 therapeutic arsenal., Competing Interests: The authors declare no competing interests., (© 2024 The Author(s).)
- Published
- 2024
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35. HIV-1 Env trimers asymmetrically engage CD4 receptors in membranes.
- Author
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Li W, Qin Z, Nand E, Grunst MW, Grover JR, Bess JW Jr, Lifson JD, Zwick MB, Tagare HD, Uchil PD, and Mothes W
- Subjects
- Humans, AIDS Vaccines chemistry, AIDS Vaccines immunology, Capsid chemistry, Capsid metabolism, Capsid ultrastructure, Cryoelectron Microscopy, Electron Microscope Tomography, HIV Antibodies immunology, HIV Infections virology, Virion chemistry, Virion metabolism, Virion ultrastructure, CD4 Antigens chemistry, CD4 Antigens metabolism, CD4 Antigens ultrastructure, Cell Membrane chemistry, Cell Membrane metabolism, Cell Membrane ultrastructure, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 metabolism, HIV Envelope Protein gp120 ultrastructure, HIV-1 chemistry, HIV-1 ultrastructure, Protein Multimerization
- Abstract
Human immunodeficiency virus 1 (HIV-1) infection is initiated by binding of the viral envelope glycoprotein (Env) to the cell-surface receptor CD4
1-4 . Although high-resolution structures of Env in a complex with the soluble domains of CD4 have been determined, the binding process is less understood in native membranes5-13 . Here we used cryo-electron tomography to monitor Env-CD4 interactions at the membrane-membrane interfaces formed between HIV-1 and CD4-presenting virus-like particles. Env-CD4 complexes organized into clusters and rings, bringing the opposing membranes closer together. Env-CD4 clustering was dependent on capsid maturation. Subtomogram averaging and classification revealed that Env bound to one, two and finally three CD4 molecules, after which Env adopted an open state. Our data indicate that asymmetric HIV-1 Env trimers bound to one and two CD4 molecules are detectable intermediates during virus binding to host cell membranes, which probably has consequences for antibody-mediated immune responses and vaccine immunogen design., (© 2023. The Author(s).)- Published
- 2023
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36. Immunogenicity and Pre-Clinical Efficacy of an OMV-Based SARS-CoV-2 Vaccine.
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Grandi A, Tomasi M, Ullah I, Bertelli C, Vanzo T, Accordini S, Gagliardi A, Zanella I, Benedet M, Corbellari R, Di Lascio G, Tamburini S, Caproni E, Croia L, Ravà M, Fumagalli V, Di Lucia P, Marotta D, Sala E, Iannacone M, Kumar P, Mothes W, Uchil PD, Cherepanov P, Bolognesi M, Pizzato M, and Grandi G
- Abstract
The vaccination campaign against SARS-CoV-2 relies on the world-wide availability of effective vaccines, with a potential need of 20 billion vaccine doses to fully vaccinate the world population. To reach this goal, the manufacturing and logistic processes should be affordable to all countries, irrespective of economical and climatic conditions. Outer membrane vesicles (OMVs) are bacterial-derived vesicles that can be engineered to incorporate heterologous antigens. Given the inherent adjuvanticity, such modified OMVs can be used as vaccines to induce potent immune responses against the associated proteins. Here, we show that OMVs engineered to incorporate peptides derived from the receptor binding motif (RBM) of the spike protein from SARS-CoV-2 elicit an effective immune response in vaccinated mice, resulting in the production of neutralizing antibodies (nAbs) with a titre higher than 1:300. The immunity induced by the vaccine is sufficient to protect the animals from intranasal challenge with SARS-CoV-2, preventing both virus replication in the lungs and the pathology associated with virus infection. Furthermore, we show that OMVs can be effectively decorated with the RBM of the Omicron BA.1 variant and that such engineered OMVs induce nAbs against Omicron BA.1 and BA.5, as measured using the pseudovirus neutralization infectivity assay. Importantly, we show that the RBM
438-509 ancestral-OMVs elicited antibodies which efficiently neutralize in vitro both the homologous ancestral strain, the Omicron BA.1 and BA.5 variants with a neutralization titre ranging from 1:100 to 1:1500, suggesting its potential use as a vaccine targeting diverse SARS-CoV-2 variants. Altogether, given the convenience associated with the ease of engineering, production and distribution, our results demonstrate that OMV-based SARS-CoV-2 vaccines can be a crucial addition to the vaccines currently available.- Published
- 2023
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37. Plasmatic HIV-1 soluble gp120 is associated with immune dysfunction and inflammation in ART-treated individuals with undetectable viremia.
- Author
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Benlarbi M, Richard J, Bourassa C, Tolbert WD, Chartrand-Lefebvre C, Gendron-Lepage G, Sylla M, El-Far M, Messier-Peet M, Guertin C, Turcotte I, Fromentin R, Verly MM, Prévost J, Clark A, Mothes W, Kaufmann DE, Maldarelli F, Chomont N, Bégin P, Tremblay C, Baril JG, Trottier B, Trottier S, Duerr R, Pazgier M, Durand M, and Finzi A
- Abstract
Background: Chronic inflammation persists in some people living with HIV (PLWH), even during antiretroviral therapy (ART) and is associated with premature aging. The gp120 subunit of the HIV-1 envelope glycoprotein can shed from viral and cellular membranes and can be detected in plasma and tissues, showing immunomodulatory properties even in the absence of detectable viremia. We evaluated whether plasmatic soluble gp120 (sgp120) and a family of gp120-specific anti-cluster A antibodies, which were previously linked to CD4 depletion in vitro , could contribute to chronic inflammation, immune dysfunction, and sub-clinical cardiovascular disease in participants of the Canadian HIV and Aging cohort (CHACS) with undetectable viremia., Methods: Cross-sectional assessment of plasmatic sgp120 and anti-cluster A antibodies was performed in 386 individuals from CHACS. Their association with pro-inflammatory cytokines, as well as subclinical coronary artery disease measured by computed tomography coronary angiography was assessed using linear regression models., Results: In individuals with high levels of sgp120, anti-cluster A antibodies inversely correlated with CD4 count (p=0.042) and CD4:CD8 ratio (p=0.004). The presence of sgp120 was associated with increased plasma levels of IL-6. In participants with detectable atherosclerotic plaque and detectable sgp120, sgp120 levels, anti-cluster A antibodies and their combination correlated positively with the total volume of atherosclerotic plaques (p=0.01, 0.018 and 0.006, respectively)., Conclusion: Soluble gp120 may act as a pan toxin causing immune dysfunction and sustained inflammation in a subset of PLWH, contributing to the development of premature comorbidities. Whether drugs targeting sgp120 could mitigate HIV-associated comorbidities in PLWH with suppressed viremia warrants further studies., Key Points: Soluble gp120 is detected in the plasma of people living with HIV-1 with undetectable viremia. The presence of soluble gp120 and anti-cluster A antibodies is associated with immune dysfunction, chronic inflammation, and sub-clinical cardiovascular disease.
- Published
- 2023
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38. Antiviral HIV-1 SERINC restriction factors disrupt virus membrane asymmetry.
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Leonhardt SA, Purdy MD, Grover JR, Yang Z, Poulos S, McIntire WE, Tatham EA, Erramilli SK, Nosol K, Lai KK, Ding S, Lu M, Uchil PD, Finzi A, Rein A, Kossiakoff AA, Mothes W, and Yeager M
- Subjects
- Humans, Membrane Proteins metabolism, Antiviral Restriction Factors, Membrane Glycoproteins, Antiviral Agents, HIV-1 metabolism, HIV Infections
- Abstract
The host proteins SERINC3 and SERINC5 are HIV-1 restriction factors that reduce infectivity when incorporated into the viral envelope. The HIV-1 accessory protein Nef abrogates incorporation of SERINCs via binding to intracellular loop 4 (ICL4). Here, we determine cryoEM maps of full-length human SERINC3 and an ICL4 deletion construct, which reveal that hSERINC3 is comprised of two α-helical bundles connected by a ~ 40-residue, highly tilted, "crossmember" helix. The design resembles non-ATP-dependent lipid transporters. Consistently, purified hSERINCs reconstituted into proteoliposomes induce flipping of phosphatidylserine (PS), phosphatidylethanolamine and phosphatidylcholine. Furthermore, SERINC3, SERINC5 and the scramblase TMEM16F expose PS on the surface of HIV-1 and reduce infectivity, with similar results in MLV. SERINC effects in HIV-1 and MLV are counteracted by Nef and GlycoGag, respectively. Our results demonstrate that SERINCs are membrane transporters that flip lipids, resulting in a loss of membrane asymmetry that is strongly correlated with changes in Env conformation and loss of infectivity., (© 2023. The Author(s).)
- Published
- 2023
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39. PLSCR1 is a cell-autonomous defence factor against SARS-CoV-2 infection.
- Author
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Xu D, Jiang W, Wu L, Gaudet RG, Park ES, Su M, Cheppali SK, Cheemarla NR, Kumar P, Uchil PD, Grover JR, Foxman EF, Brown CM, Stansfeld PJ, Bewersdorf J, Mothes W, Karatekin E, Wilen CB, and MacMicking JD
- Subjects
- Animals, Humans, Mice, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Chiroptera, Exome Sequencing, Hepatocytes immunology, Hepatocytes metabolism, Interferon-gamma immunology, Lung immunology, Lung metabolism, Membrane Fusion, Virus Internalization, COVID-19 immunology, COVID-19 metabolism, COVID-19 prevention & control, COVID-19 virology, Phospholipid Transfer Proteins chemistry, Phospholipid Transfer Proteins genetics, Phospholipid Transfer Proteins immunology, Phospholipid Transfer Proteins metabolism, SARS-CoV-2 classification, SARS-CoV-2 immunology, SARS-CoV-2 metabolism, SARS-CoV-2 pathogenicity
- Abstract
Understanding protective immunity to COVID-19 facilitates preparedness for future pandemics and combats new SARS-CoV-2 variants emerging in the human population. Neutralizing antibodies have been widely studied; however, on the basis of large-scale exome sequencing of protected versus severely ill patients with COVID-19, local cell-autonomous defence is also crucial
1-4 . Here we identify phospholipid scramblase 1 (PLSCR1) as a potent cell-autonomous restriction factor against live SARS-CoV-2 infection in parallel genome-wide CRISPR-Cas9 screens of human lung epithelia and hepatocytes before and after stimulation with interferon-γ (IFNγ). IFNγ-induced PLSCR1 not only restricted SARS-CoV-2 USA-WA1/2020, but was also effective against the Delta B.1.617.2 and Omicron BA.1 lineages. Its robust activity extended to other highly pathogenic coronaviruses, was functionally conserved in bats and mice, and interfered with the uptake of SARS-CoV-2 in both the endocytic and the TMPRSS2-dependent fusion routes. Whole-cell 4Pi single-molecule switching nanoscopy together with bipartite nano-reporter assays found that PLSCR1 directly targeted SARS-CoV-2-containing vesicles to prevent spike-mediated fusion and viral escape. A PLSCR1 C-terminal β-barrel domain-but not lipid scramblase activity-was essential for this fusogenic blockade. Our mechanistic studies, together with reports that COVID-associated PLSCR1 mutations are found in some susceptible people3,4 , identify an anti-coronavirus protein that interferes at a late entry step before viral RNA is released into the host-cell cytosol., (© 2023. The Author(s).)- Published
- 2023
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40. Combinatorial Regimens Augment Drug Monotherapy for SARS-CoV-2 Clearance in Mice.
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Ullah I, Escudie F, Scandale I, Gilani Z, Gendron-Lepage G, Gaudette F, Mowbray C, Fraisse L, Bazin R, Finzi A, Mothes W, Kumar P, Chatelain E, and Uchil PD
- Abstract
Direct acting antivirals (DAAs) represent critical tools for combating SARS-CoV-2 variants of concern (VOCs) that evolve to escape spike-based immunity and future coronaviruses with pandemic potential. Here, we used bioluminescence imaging to evaluate therapeutic efficacy of DAAs that target SARS-CoV-2 RNA-dependent RNA polymerase (favipiravir, molnupiravir) or Main protease (nirmatrelvir) against Delta or Omicron VOCs in K18-hACE2 mice. Nirmatrelvir displayed the best efficacy followed by molnupiravir and favipiravir in suppressing viral loads in the lung. Unlike neutralizing antibody treatment, DAA monotherapy did not eliminate SARS-CoV-2 in mice. However, targeting two viral enzymes by combining molnupiravir with nirmatrelvir resulted in superior efficacy and virus clearance. Furthermore, combining molnupiravir with Caspase-1/4 inhibitor mitigated inflammation and lung pathology whereas combining molnupiravir with COVID-19 convalescent plasma yielded rapid virus clearance and 100% survival. Thus, our study provides insights into treatment efficacies of DAAs and other effective combinations to bolster COVID-19 therapeutic arsenal.
- Published
- 2023
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41. Vaccinia Virus Strain MVA Expressing a Prefusion-Stabilized SARS-CoV-2 Spike Glycoprotein Induces Robust Protection and Prevents Brain Infection in Mouse and Hamster Models.
- Author
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Lorenzo MM, Marín-López A, Chiem K, Jimenez-Cabello L, Ullah I, Utrilla-Trigo S, Calvo-Pinilla E, Lorenzo G, Moreno S, Ye C, Park JG, Matía A, Brun A, Sánchez-Puig JM, Nogales A, Mothes W, Uchil PD, Kumar P, Ortego J, Fikrig E, Martinez-Sobrido L, and Blasco R
- Abstract
The COVID-19 pandemic has underscored the importance of swift responses and the necessity of dependable technologies for vaccine development. Our team previously developed a fast cloning system for the modified vaccinia virus Ankara (MVA) vaccine platform. In this study, we reported on the construction and preclinical testing of a recombinant MVA vaccine obtained using this system. We obtained recombinant MVA expressing the unmodified full-length SARS-CoV-2 spike (S) protein containing the D614G amino-acid substitution (MVA-Sdg) and a version expressing a modified S protein containing amino-acid substitutions designed to stabilize the protein a in a pre-fusion conformation (MVA-Spf). S protein expressed by MVA-Sdg was found to be expressed and was correctly processed and transported to the cell surface, where it efficiently produced cell-cell fusion. Version Spf, however, was not proteolytically processed, and despite being transported to the plasma membrane, it failed to induce cell-cell fusion. We assessed both vaccine candidates in prime-boost regimens in the susceptible transgenic K18-human angiotensin-converting enzyme 2 (K18-hACE2) in mice and in golden Syrian hamsters. Robust immunity and protection from disease was induced with either vaccine in both animal models. Remarkably, the MVA-Spf vaccine candidate produced higher levels of antibodies, a stronger T cell response, and a higher degree of protection from challenge. In addition, the level of SARS-CoV-2 in the brain of MVA-Spf inoculated mice was decreased to undetectable levels. Those results add to our current experience and range of vaccine vectors and technologies for developing a safe and effective COVID-19 vaccine.
- Published
- 2023
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42. Temsavir blocks the immunomodulatory activities of HIV-1 soluble gp120.
- Author
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Richard J, Prévost J, Bourassa C, Brassard N, Boutin M, Benlarbi M, Goyette G, Medjahed H, Gendron-Lepage G, Gaudette F, Chen HC, Tolbert WD, Smith AB 3rd, Pazgier M, Dubé M, Clark A, Mothes W, Kaufmann DE, and Finzi A
- Subjects
- CD4-Positive T-Lymphocytes metabolism, Down-Regulation, HIV Envelope Protein gp120, Cytokines metabolism, HIV-1
- Abstract
While HIV-1-mediated CD4 downregulation protects infected cells from antibody-dependent cellular cytotoxicity (ADCC), shed gp120 binds to CD4 on uninfected bystander CD4
+ T cells, sensitizing them to ADCC mediated by HIV+ plasma. Soluble gp120-CD4 interaction on multiple immune cells also triggers a cytokine burst. The small molecule temsavir acts as an HIV-1 attachment inhibitor by preventing envelope glycoprotein (Env)-CD4 interaction and alters the overall antigenicity of Env by affecting its processing and glycosylation. Here we show that temsavir also blocks the immunomodulatory activities of shed gp120. Temsavir prevents shed gp120 from interacting with uninfected bystander CD4+ cells, protecting them from ADCC responses and preventing a cytokine burst. Mechanistically, this depends on temsavir's capacity to prevent soluble gp120-CD4 interaction, to reduce gp120 shedding, and to alter gp120 antigenicity. This suggests that the clinical benefits provided by temsavir could extend beyond blocking viral entry., Competing Interests: Declaration of interests W.M. and A.F. received funding from ViiV Healthcare. A.C. is a full-time employee of ViiV Healthcare., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)- Published
- 2023
- Full Text
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43. Molecular basis for antiviral activity of two pediatric neutralizing antibodies targeting SARS-CoV-2 Spike RBD.
- Author
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Chen Y, Prévost J, Ullah I, Romero H, Lisi V, Tolbert WD, Grover JR, Ding S, Gong SY, Beaudoin-Bussières G, Gasser R, Benlarbi M, Vézina D, Anand SP, Chatterjee D, Goyette G, Grunst MW, Yang Z, Bo Y, Zhou F, Béland K, Bai X, Zeher AR, Huang RK, Nguyen DN, Sherburn R, Wu D, Piszczek G, Paré B, Matthies D, Xia D, Richard J, Kumar P, Mothes W, Côté M, Uchil PD, Lavallée VP, Smith MA, Pazgier M, Haddad E, and Finzi A
- Abstract
Neutralizing antibodies (NAbs) hold great promise for clinical interventions against SARS-CoV-2 variants of concern (VOCs). Understanding NAb epitope-dependent antiviral mechanisms is crucial for developing vaccines and therapeutics against VOCs. Here we characterized two potent NAbs, EH3 and EH8, isolated from an unvaccinated pediatric patient with exceptional plasma neutralization activity. EH3 and EH8 cross-neutralize the early VOCs and mediate strong Fc-dependent effector activity in vitro . Structural analyses of EH3 and EH8 in complex with the receptor-binding domain (RBD) revealed the molecular determinants of the epitope-driven protection and VOC evasion. While EH3 represents the prevalent IGHV3-53 NAb whose epitope substantially overlaps with the ACE2 binding site, EH8 recognizes a narrow epitope exposed in both RBD-up and RBD-down conformations. When tested in vivo, a single-dose prophylactic administration of EH3 fully protected stringent K18-hACE2 mice from lethal challenge with Delta VOC. Our study demonstrates that protective NAbs responses converge in pediatric and adult SARS-CoV-2 patients., Competing Interests: A.F. has filed a provisional patent application on the following monoclonal antibodies: CV3-1 and CV3-25. A.F., J.P., V.L. M.A.S., V.-P.L., and E.H. have filed a provisional patent application on the following monoclonal antibodies: EH3 and EH8., (© 2022 The Author(s).)
- Published
- 2023
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44. The Fc-effector function of COVID-19 convalescent plasma contributes to SARS-CoV-2 treatment efficacy in mice.
- Author
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Ullah I, Beaudoin-Bussières G, Symmes K, Cloutier M, Ducas E, Tauzin A, Laumaea A, Grunst MW, Dionne K, Richard J, Bégin P, Mothes W, Kumar P, Bazin R, Finzi A, and Uchil PD
- Subjects
- Animals, Mice, COVID-19 Serotherapy, Treatment Outcome, Immunoglobulin G, SARS-CoV-2, COVID-19 therapy
- Abstract
COVID-19 convalescent plasmas (CCPs) are chosen for plasma therapy based on neutralizing titers and anti-Spike immunoglobulin levels. However, CCP characteristics that promote SARS-CoV-2 control are complex and incompletely defined. Using an in vivo imaging approach, we demonstrate that CCPs with low neutralizing (ID
50 ≤ 1:250), but moderate to high Fc-effector activity, in contrast to those with poor Fc function, delay mortality and/or improve survival of SARS-CoV-2-challenged K18-hACE2 mice. The impact of innate immune cells on CCP efficacy depended on their residual neutralizing activity. Fractionation of a selected CCP revealed that IgG and Ig(M + A) were required during therapy, but the IgG fraction alone sufficed during prophylaxis. Finally, despite reduced neutralization, ancestral SARS-CoV-2-elicited CCPs significantly delayed Delta and Beta-induced mortality suggesting that Fc-effector functions contribute to immunity against VOCs. Thus, Fc activity of CCPs provide a second line of defense when neutralization is compromised and can serve as an important criterion for CCP selection., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2023
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45. VE607 stabilizes SARS-CoV-2 Spike in the "RBD-up" conformation and inhibits viral entry.
- Author
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Ding S, Ullah I, Gong SY, Grover JR, Mohammadi M, Chen Y, Vézina D, Beaudoin-Bussières G, Verma VT, Goyette G, Gaudette F, Richard J, Yang D, Smith AB 3rd, Pazgier M, Côté M, Abrams C, Kumar P, Mothes W, Uchil PD, Finzi A, and Baron C
- Abstract
SARS-CoV-2 infection of host cells starts by binding the Spike glycoprotein (S) to the ACE2 receptor. The S-ACE2 interaction is a potential target for therapies against COVID-19 as demonstrated by the development of immunotherapies blocking this interaction. VE607 - a commercially available compound composed of three stereoisomers - was described as an inhibitor of SARS-CoV-1. Here, we show that VE607 broadly inhibits pseudoviral particles bearing the Spike from major VOCs (D614G, Alpha, Beta, Gamma, Delta, Omicron - BA.1, and BA.2) as well as authentic SARS-CoV-2 at low micromolar concentrations. In silico docking, mutational analysis, and smFRET revealed that VE607 binds to the receptor binding domain (RBD)-ACE2 interface and stabilizes RBD in its "up" conformation. Prophylactic treatment with VE607 did not prevent SARS-CoV-2-induced mortality in K18-hACE2 mice, but it did reduce viral replication in the lungs by 37-fold. Thus, VE607 is an interesting lead for drug development for the treatment of SARS-CoV-2 infection., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)
- Published
- 2022
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46. Engineered ACE2-Fc counters murine lethal SARS-CoV-2 infection through direct neutralization and Fc-effector activities.
- Author
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Chen Y, Sun L, Ullah I, Beaudoin-Bussières G, Anand SP, Hederman AP, Tolbert WD, Sherburn R, Nguyen DN, Marchitto L, Ding S, Wu D, Luo Y, Gottumukkala S, Moran S, Kumar P, Piszczek G, Mothes W, Ackerman ME, Finzi A, Uchil PD, Gonzalez FJ, and Pazgier M
- Abstract
Soluble angiotensin-converting enzyme 2 (ACE2) constitutes an attractive antiviral capable of targeting a wide range of coronaviruses using ACE2 as their receptor. Using structure-guided approaches, we developed a series of bivalent ACE2-Fcs harboring functionally and structurally validated mutations that enhance severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor binding domain recognition by up to ~12-fold and remove angiotensin enzymatic activity. The lead variant M81 potently cross-neutralized SARS-CoV-2 variants of concern (VOCs), including Omicron, at subnanomolar half-maximal inhibitory concentration and was capable of robust Fc-effector functions, including antibody-dependent cellular cytotoxicity, phagocytosis, and complement deposition. When tested in a stringent K18-hACE2 mouse model, Fc-enhanced ACE2-Fc delayed death by 3 to 5 days or effectively resolved lethal SARS-CoV-2 infection in both prophylactic and therapeutic settings via the combined effects of neutralization and Fc-effector functions. These data add to the demonstrated utility of soluble ACE2 as a valuable SARS-CoV-2 antiviral and indicate that Fc-effector functions may constitute an important component of ACE2-Fc therapeutic activity.
- Published
- 2022
- Full Text
- View/download PDF
47. Cancer Microbiology.
- Author
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DiMaio D, Emu B, Goodman AL, Mothes W, and Justice A
- Subjects
- Anti-Bacterial Agents, Carcinogens, Delivery of Health Care, Humans, Microbiota, Neoplasms prevention & control
- Abstract
Microbes play important roles in cancer from direct carcinogenic effects to their use in treatment. Cancers caused by microorganisms account for approximately 15% of cancers, primarily in low- and middle-income countries. Unique features of infectious carcinogens include their transmissibility, mutability, and specific immune interactions, which provide challenges and opportunities for cancer prevention and treatment. For these agents, infection control through exposure reduction, antivirals, antibiotics, and vaccines is cancer control. In addition, developing evidence suggests that microorganisms including the human microbiome can indirectly modulate cancer formation and influence the effectiveness and toxicity of cancer treatments. Finally, microorganisms themselves can be used to prevent or treat cancer. The convergence of these factors signals the emergence of a new field, cancer microbiology. Recognition of cancer microbiology will spur research, stimulate cross-disciplinary training, inform drug development, and improve public health., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2022
- Full Text
- View/download PDF
48. Antigenic analysis of the HIV-1 envelope trimer implies small differences between structural states 1 and 2.
- Author
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Cale EM, Driscoll JI, Lee M, Gorman J, Zhou T, Lu M, Geng H, Lai YT, Chuang GY, Doria-Rose NA, Mothes W, Kwong PD, and Mascola JR
- Subjects
- Broadly Neutralizing Antibodies chemistry, Broadly Neutralizing Antibodies metabolism, HIV Antibodies, Humans, Protein Conformation, HIV Infections, HIV-1 metabolism, env Gene Products, Human Immunodeficiency Virus chemistry, env Gene Products, Human Immunodeficiency Virus metabolism
- Abstract
The conformationally dynamic HIV-1 envelope trimer (Env) is the target of broadly neutralizing antibodies (bnAbs) that block viral entry. Single-molecule Förster resonance energy transfer (smFRET) has revealed that HIV-1 Env exists in at least three conformational states on the virion. Prior to complete host-receptor engagement (State 3), Env resides most prevalently in the smFRET-defined State 1, which is preferentially recognized by most bnAbs that are elicited by natural infection. smFRET has also revealed that soluble trimers containing prefusion-stabilizing disulfide and isoleucine-to-proline substitutions reside primarily in State 2, which is a required intermediate between States 1 and 3. While high-resolution Env structures have been determined for States 2 and 3, the structure of these trimers in State 1 is unknown. To provide insight into the State 1 structure, here we characterized antigenic differences between smFRET-defined states and then correlated these differences with known structural differences between States 2 and 3. We found that cell surface-expressed Env was enriched in each state using state-enriching antibody fragments or small-molecule virus entry inhibitors and then assessed binding to HIV-1 bnAbs preferentially binding different states. We observed small but consistent differences in binding between Env enriched in States 1 and 2, and a more than 10-fold difference in binding to Env enriched in these states versus Env enriched in State 3. We conclude that structural differences between HIV-1 Env States 1 and 3 are likely more than 10-fold greater than those between States 1 and 2, providing important insight into State 1., Competing Interests: Conflict of interest The authors declare no competing interests with the contents of this article., (Published by Elsevier Inc.)
- Published
- 2022
- Full Text
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49. A Fc-enhanced NTD-binding non-neutralizing antibody delays virus spread and synergizes with a nAb to protect mice from lethal SARS-CoV-2 infection.
- Author
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Beaudoin-Bussières G, Chen Y, Ullah I, Prévost J, Tolbert WD, Symmes K, Ding S, Benlarbi M, Gong SY, Tauzin A, Gasser R, Chatterjee D, Vézina D, Goyette G, Richard J, Zhou F, Stamatatos L, McGuire AT, Charest H, Roger M, Pozharski E, Kumar P, Mothes W, Uchil PD, Pazgier M, and Finzi A
- Subjects
- Animals, Antibodies, Viral chemistry, Antibody-Dependent Cell Cytotoxicity, COVID-19 mortality, COVID-19 prevention & control, COVID-19 transmission, Disease Models, Animal, Epitopes, Humans, Immunization, Passive mortality, Immunoglobulin Fab Fragments chemistry, Immunoglobulin Fab Fragments metabolism, Immunoglobulin Fc Fragments genetics, Immunoglobulin Fc Fragments immunology, Mice, Protein Binding, Protein Conformation, SARS-CoV-2 immunology, SARS-CoV-2 pathogenicity, Spike Glycoprotein, Coronavirus chemistry, Spike Glycoprotein, Coronavirus immunology, Spike Glycoprotein, Coronavirus metabolism, COVID-19 Serotherapy, Antibodies, Neutralizing therapeutic use, Antibodies, Viral immunology, Antibodies, Viral therapeutic use, COVID-19 therapy
- Abstract
Emerging evidence indicates that both neutralizing and Fc-mediated effector functions of antibodies contribute to protection against SARS-CoV-2. It is unclear whether Fc-effector functions alone can protect against SARS-CoV-2. Here, we isolated CV3-13, a non-neutralizing antibody, from a convalescent individual with potent Fc-mediated effector functions. The cryoelectron microscopy structure of CV3-13 in complex with the SARS-CoV-2 spike reveals that the antibody binds from a distinct angle of approach to an N-terminal domain (NTD) epitope that only partially overlaps with the NTD supersite recognized by neutralizing antibodies. CV3-13 does not alter the replication dynamics of SARS-CoV-2 in K18-hACE2 mice, but its Fc-enhanced version significantly delays virus spread, neuroinvasion, and death in prophylactic settings. Interestingly, the combination of Fc-enhanced non-neutralizing CV3-13 with Fc-compromised neutralizing CV3-25 completely protects mice from lethal SARS-CoV-2 infection. Altogether, our data demonstrate that efficient Fc-mediated effector functions can potently contribute to the in vivo efficacy of anti-SARS-CoV-2 antibodies., Competing Interests: Declaration of interests L.S., A.T.M., and A.F. have filed a provisional patent application on the following monoclonal antibodies: CV3-1, CV3-13, and CV3-25., (Copyright © 2022. Published by Elsevier Inc.)
- Published
- 2022
- Full Text
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50. Structural basis and mode of action for two broadly neutralizing antibodies against SARS-CoV-2 emerging variants of concern.
- Author
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Li W, Chen Y, Prévost J, Ullah I, Lu M, Gong SY, Tauzin A, Gasser R, Vézina D, Anand SP, Goyette G, Chaterjee D, Ding S, Tolbert WD, Grunst MW, Bo Y, Zhang S, Richard J, Zhou F, Huang RK, Esser L, Zeher A, Côté M, Kumar P, Sodroski J, Xia D, Uchil PD, Pazgier M, Finzi A, and Mothes W
- Abstract
Emerging variants of concern for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can transmit more efficiently and partially evade protective immune responses, thus necessitating continued refinement of antibody therapies and immunogen design. Here, we elucidate the structural basis and mode of action for two potent SARS-CoV-2 spike (S)-neutralizing monoclonal antibodies, CV3-1 and CV3-25, which remain effective against emerging variants of concern in vitro and in vivo. CV3-1 binds to the (485-GFN-487) loop within the receptor-binding domain (RBD) in the "RBD-up" position and triggers potent shedding of the S1 subunit. In contrast, CV3-25 inhibits membrane fusion by binding to an epitope in the stem helix region of the S2 subunit that is highly conserved among β-coronaviruses. Thus, vaccine immunogen designs that incorporate the conserved regions in the RBD and stem helix region are candidates to elicit pan-coronavirus protective immune responses., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021. Published by Elsevier Inc.)
- Published
- 2022
- Full Text
- View/download PDF
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