44 results on '"Doranz, Benjamin J."'
Search Results
2. Single B cell transcriptomics identifies multiple isotypes of broadly neutralizing antibodies against flaviviruses.
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Lubow, Jay, Levoir, Lisa M., Ralph, Duncan K., Belmont, Laura, Contreras, Maya, Cartwright-Acar, Catiana H., Kikawa, Caroline, Kannan, Shruthi, Davidson, Edgar, Duran, Veronica, Rebellon-Sanchez, David E., Sanz, Ana M., Rosso, Fernando, Doranz, Benjamin J., Einav, Shirit, Matsen IV, Frederick A., and Goo, Leslie
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B cells ,MONOCLONAL antibodies ,IMMUNOGLOBULINS ,ZIKA virus infections ,FLAVIVIRAL diseases ,FLAVIVIRUSES ,DENGUE viruses ,FOOT & mouth disease - Abstract
Sequential dengue virus (DENV) infections often generate neutralizing antibodies against all four DENV serotypes and sometimes, Zika virus. Characterizing cross-flavivirus broadly neutralizing antibody (bnAb) responses can inform countermeasures that avoid enhancement of infection associated with non-neutralizing antibodies. Here, we used single cell transcriptomics to mine the bnAb repertoire following repeated DENV infections. We identified several new bnAbs with comparable or superior breadth and potency to known bnAbs, and with distinct recognition determinants. Unlike all known flavivirus bnAbs, which are IgG1, one newly identified cross-flavivirus bnAb (F25.S02) was derived from IgA1. Both IgG1 and IgA1 versions of F25.S02 and known bnAbs displayed neutralizing activity, but only IgG1 enhanced infection in monocytes expressing IgG and IgA Fc receptors. Moreover, IgG-mediated enhancement of infection was inhibited by IgA1 versions of bnAbs. We demonstrate a role for IgA in flavivirus infection and immunity with implications for vaccine and therapeutic strategies. Author summary: A central challenge for developing clinical interventions for dengue virus or the closely related Zika virus is the ability of IgG antibodies to enhance, rather than neutralize infection under certain conditions. When present prior to infection, as in the case of vaccination, these antibodies can worsen disease outcome. In this study, we analyzed B cells of individuals who experienced dengue or Zika infection to identify those expressing antibodies that can potently neutralize these viruses with minimal potential to enhance infection. We used a method that captured a larger number and wider variety of antibodies than previous approaches. We discovered several potent antibodies that simultaneously neutralized dengue and Zika viruses, including those of IgG isotype, which are common, and one of IgA isotype, which had never been described against this group of viruses. Although IgG antibodies enhanced infection in certain cases, the IgA antibody did not. We further showed that modifying a region of IgG antibodies to convert them to IgA antibodies eliminated their ability to enhance infection. Moreover, the modified IgA versions inhibited the ability of IgG versions to enhance infection. These results suggest that inducing IgA antibodies may be an attractive goal for safe and effective vaccines. [ABSTRACT FROM AUTHOR]
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- 2023
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3. Breaking barriers in antibody discovery: harnessing divergent species for accessing difficult and conserved drug targets.
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Banik, Soma S.R., Kushnir, Natasha, Doranz, Benjamin J., and Chambers, Ross
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- 2023
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4. Rapid discovery of diverse neutralizing SARS-CoV-2 antibodies from large-scale synthetic phage libraries.
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Yuan, Tom Z., Garg, Pankaj, Wang, Linya, Willis, Jordan R., Kwan, Eric, Hernandez, Ana G Lujan, Tuscano, Emily, Sever, Emily N., Keane, Erica, Soto, Cinque, Mucker, Eric M., Fouch, Mallorie E., Davidson, Edgar, Doranz, Benjamin J., Kailasan, Shweta, Aman, M. Javad, Li, Haoyang, Hooper, Jay W., Saphire, Erica Ollmann, and Crowe, James E.
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- 2022
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5. Proteo-Genomic Analysis Identifies Two Major Sites of Vulnerability on Ebolavirus Glycoprotein for Neutralizing Antibodies in Convalescent Human Plasma.
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Gilchuk, Pavlo, Guthals, Adrian, Bonissone, Stefano R., Shaw, Jared B., Ilinykh, Philipp A., Huang, Kai, Bombardi, Robin G., Liang, Jenny, Grinyo, Ariadna, Davidson, Edgar, Chen, Elaine C., Gunn, Bronwyn M., Alter, Galit, Saphire, Erica Ollmann, Doranz, Benjamin J., Bukreyev, Alexander, Zeitlin, Larry, Castellana, Natalie, and Crowe Jr., James E.
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CONVALESCENT plasma ,EBOLA virus ,EBOLA virus disease ,BONE marrow cells ,IMMUNOGLOBULINS - Abstract
Three clinically relevant ebolaviruses – Ebola (EBOV), Bundibugyo (BDBV), and Sudan (SUDV) viruses, are responsible for severe disease and occasional deadly outbreaks in Africa. The largest Ebola virus disease (EVD) epidemic to date in 2013-2016 in West Africa highlighted the urgent need for countermeasures, leading to the development and FDA approval of the Ebola virus vaccine rVSV-ZEBOV (Ervebo
® ) in 2020 and two monoclonal antibody (mAb)-based therapeutics (Inmazeb® [atoltivimab, maftivimab, and odesivimab-ebgn] and Ebanga® (ansuvimab-zykl) in 2020. The humoral response plays an indispensable role in ebolavirus immunity, based on studies of mAbs isolated from the antibody genes in peripheral blood circulating ebolavirus-specific human memory B cells. However, antibodies in the body are not secreted by circulating memory B cells in the blood but rather principally by plasma cells in the bone marrow. Little is known about the protective polyclonal antibody responses in convalescent plasma. Here we exploited both single-cell antibody gene sequencing and proteomic sequencing approaches to assess the composition of the ebolavirus glycoprotein (GP)-reactive antibody repertoire in the plasma of an EVD survivor. We first identified 1,512 GP-specific mAb variable gene sequences from single cells in the memory B cell compartment. Using mass spectrometric analysis of the corresponding GP-specific plasma IgG, we found that only a portion of the large B cell antibody repertoire was represented in the plasma. Molecular and functional analysis of proteomics-identified mAbs revealed recognition of epitopes in three major antigenic sites - the GP head domain, the glycan cap, and the base region, with a high prevalence of neutralizing and protective mAb specificities that targeted the base and glycan cap regions on the GP. Polyclonal plasma antibodies from the survivor reacted broadly to EBOV, BDBV, and SUDV GP, while reactivity of the potently neutralizing mAbs we identified was limited mostly to the homologous EBOV GP. Together these results reveal a restricted diversity of neutralizing humoral response in which mAbs targeting two antigenic sites on GP – glycan cap and base – play a principal role in plasma-antibody-mediated protective immunity against EVD. [ABSTRACT FROM AUTHOR]- Published
- 2021
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6. An affinity-matured human monoclonal antibody targeting fusion loop epitope of dengue virus with in vivo therapeutic potency.
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Kotaki, Tomohiro, Kurosu, Takeshi, Grinyo-Escuer, Ariadna, Davidson, Edgar, Churrotin, Siti, Okabayashi, Tamaki, Puiprom, Orapim, Mulyatno, Kris Cahyo, Sucipto, Teguh Hari, Doranz, Benjamin J., Ono, Ken-ichiro, Soegijanto, Soegeng, and Kameoka, Masanori
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MONOCLONAL antibodies ,EPITOPES ,DENGUE viruses ,VACCINE development ,SOMATIC mutation - Abstract
Dengue virus (DENV), from the genus flavivirus of the family flaviviridae, causes serious health problems globally. Human monoclonal antibodies (HuMAb) can be used to elucidate the mechanisms of neutralization and antibody-dependent enhancement (ADE) of DENV infections, leading to the development of a vaccine or therapeutic antibodies. Here, we generated eight HuMAb clones from an Indonesian patient infected with DENV. These HuMAbs exhibited the typical characteristics of weak neutralizing antibodies including high cross-reactivity with other flaviviruses and targeting of the fusion loop epitope (FLE). However, one of the HuMAbs, 3G9, exhibited strong neutralization (NT
50 < 0.1 μg/ml) and possessed a high somatic hyper-mutation rate of the variable region, indicating affinity-maturation. Administration of this antibody significantly prolonged the survival of interferon-α/β/γ receptor knockout C57BL/6 mice after a lethal DENV challenge. Additionally, Fc-modified 3G9 that had lost their in vitro ADE activity showed enhanced therapeutic potency in vivo and competed strongly with an ADE-prone antibody in vitro. Taken together, the affinity-matured FLE-targeting antibody 3G9 exhibits promising features for therapeutic application including a low NT50 value, potential for treatment of various kinds of mosquito-borne flavivirus infection, and suppression of ADE. This study demonstrates the therapeutic potency of affinity-matured FLE-targeting antibodies. [ABSTRACT FROM AUTHOR]- Published
- 2021
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7. Molecular determinants and mechanism for antibody cocktail preventing SARS-CoV-2 escape.
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Ku, Zhiqiang, Xie, Xuping, Davidson, Edgar, Ye, Xiaohua, Su, Hang, Menachery, Vineet D., Li, Yize, Yuan, Zihao, Zhang, Xianwen, Muruato, Antonio E., i Escuer, Ariadna Grinyo, Tyrell, Breanna, Doolan, Kyle, Doranz, Benjamin J., Wrapp, Daniel, Bates, Paul F., McLellan, Jason S., Weiss, Susan R., Zhang, Ningyan, and Shi, Pei-Yong
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SARS-CoV-2 ,IMMUNOGLOBULINS ,PROTEIN folding ,COVID-19 ,VIRAL antibodies - Abstract
Antibody cocktails represent a promising approach to prevent SARS-CoV-2 escape. The determinants for selecting antibody combinations and the mechanism that antibody cocktails prevent viral escape remain unclear. We compared the critical residues in the receptor-binding domain (RBD) used by multiple neutralizing antibodies and cocktails and identified a combination of two antibodies CoV2-06 and CoV2-14 for preventing viral escape. The two antibodies simultaneously bind to non-overlapping epitopes and independently compete for receptor binding. SARS-CoV-2 rapidly escapes from individual antibodies by generating resistant mutations in vitro, but it doesn't escape from the cocktail due to stronger mutational constraints on RBD-ACE2 interaction and RBD protein folding requirements. We also identified a conserved neutralizing epitope shared between SARS-CoV-2 and SARS-CoV for antibody CoV2-12. Treatments with CoV2-06 and CoV2-14 individually and in combination confer protection in mice. These findings provide insights for rational selection and mechanistic understanding of antibody cocktails as candidates for treating COVID-19. Antibody cocktails represent a promising approach to prevent SARS-CoV-2 escape. Here, Ku et al., identify SARS-CoV-2 neutralizing antibodies from a phage library and identify an antibody combination that prevents viral escape and protects mice from viral challenge. [ABSTRACT FROM AUTHOR]
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- 2021
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8. Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications.
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Whitbeck, J. Charles, Thomas, Anu, Kadash-Edmondson, Kathryn, Grinyo-Escuer, Ariadna, Stafford, Lewis J., Cheng, Celine, Liao, Grant C., Holtsberg, Frederick W., Aman, M. Javad, Simmons, Graham, Davidson, Edgar, and Doranz, Benjamin J.
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ZIKA virus ,ZIKA virus infections ,VACCINE development ,NEUTRALIZATION tests ,DENGUE viruses - Abstract
The development of vaccines against flaviviruses, including Zika virus (ZIKV) and dengue virus (DENV), continues to be a major challenge, hindered by the lack of efficient and reliable methods for screening neutralizing activity of sera or antibodies. To address this need, we previously developed a plasmid-based, replication-incompetent DENV reporter virus particle (RVP) production system as an efficient and safe alternative to the Plaque Reduction Neutralization Test (PRNT). As part of the response to the 2015–2016 ZIKV outbreak, we developed pseudo-infectious ZIKV RVPs by modifying our DENV RVP system. The use of ZIKV RVPs as critical reagents in human clinical trials requires their further validation using stability and reproducibility metrics for large-scale applications. In the current study, we validated ZIKV RVPs using infectivity, neutralization, and enhancement assays with monoclonal antibodies (MAbs) and human ZIKV-positive patient serum. ZIKV RVPs are antigenically equivalent to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of live virus replication assays. We demonstrate reproducible neutralization titer data (NT
50 values) across different RVP production lots, volumes, time frames, and laboratories. We also show RVP stability across experimentally relevant time intervals and temperatures. Our results demonstrate that ZIKV RVPs provide a safe, high-throughput, and reproducible reagent for large-scale, long-term studies of neutralizing antibodies and sera, which can facilitate large-scale screening and epidemiological studies to help expedite ZIKV vaccine development. Author summary: ZIKV is a mosquito-borne virus that can cause severe birth defects and other disorders. Large outbreaks of ZIKV occurred in 2015 and 2016 and there are still no drugs or vaccines available to protect against ZIKV infection. Vaccine development has been hindered by the lack of safe and efficient methods to screen ZIKV neutralizing properties of patient sera or antibodies, especially in the context of large clinical trials. To address this unmet need, we developed and validated the use of ZIKV reporter virus particles (RVPs), a safe, high-throughput, and quantitative alternative to using live virus for neutralization studies. We show that ZIKV RVPs are stable, show lot-to-lot consistency, and provide reproducible neutralization data that is suitable for the large-scale studies needed for development of a ZIKV vaccine, epidemiologic surveillance, and high-throughput screening. [ABSTRACT FROM AUTHOR]- Published
- 2020
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9. Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics.
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Gilchuk, Pavlo, Bombardi, Robin G., Erasmus, Jesse H., Tan, Qing, Nargi, Rachel, Soto, Cinque, Abbink, Peter, Suscovich, Todd J., Durnell, Lorellin A., Khandhar, Amit, Archer, Jacob, Liang, Jenny, Fouch, Mallorie E., Davidson, Edgar, Doranz, Benjamin J., Jones, Taylor, Larson, Elise, Ertel, Stacey, Granger, Brian, and Fuerte-Stone, Jasmine
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- 2020
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10. Potent neutralizing antibodies elicited by dengue vaccine in rhesus macaque target diverse epitopes.
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Li, Leike, Meng, Weixu, Horton, Melanie, DiStefano, Daniel R., Thoryk, Elizabeth A., Pfaff, Jennifer M., Wang, Qihui, Salazar, Georgina T., Barnes, Trevor, Doranz, Benjamin J., Bett, Andrew J., Casimiro, Danilo R., Vora, Kalpit A., An, Zhiqiang, and Zhang, Ningyan
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DENGUE viruses ,RHESUS monkeys ,EPITOPES ,MONOCLONAL antibodies ,MEMBRANE proteins ,ENZYME-linked immunosorbent assay - Abstract
There is still no safe and effective vaccine against dengue virus infection. Epidemics of dengue virus infection are increasingly a threat to human health around the world. Antibodies generated in response to dengue infection have been shown to impact disease development and effectiveness of dengue vaccine. In this study, we investigated monoclonal antibody responses to an experimental dengue vaccine in rhesus macaques. Variable regions of both heavy chain (VH) and light chain (VL) were cloned from single antibody-secreting B cells. A total of 780 monoclonal antibodies (mAbs) composed of paired VH and VL were characterized. Results show that the vaccination induces mAbs with diverse germline sequences and a wide range of binding affinities. Six potent neutralizing mAbs were identified among 130 dengue envelope protein binders. Critical amino acids for each neutralizing antibody binding to the dengue envelope protein were identified by alanine scanning of mutant libraries. Diverse epitopes were identified, including epitopes on the lateral ridge of DIII, the I-III hinge, the bc loop adjacent to the fusion loop of DII, and the β-strands and loops of DI. Significantly, one of the neutralizing mAbs has a previously unknown epitope in DII at the interface of the envelope and membrane protein and is capable of neutralizing all four dengue serotypes. Taken together, the results of this study not only provide preclinical validation for the tested experimental vaccine, but also shed light on a potential application of the rhesus macaque model for better dengue vaccine evaluation and design of vaccines and immunization strategies. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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11. Optimal therapeutic activity of monoclonal antibodies against chikungunya virus requires Fc-FcγR interaction on monocytes.
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Fox, Julie M., Roy, Vicky, Gunn, Bronwyn M., Huang, Ling, Edeling, Melissa A., Mack, Matthias, Fremont, Daved H., Doranz, Benjamin J., Johnson, Syd, Alter, Galit, and Diamond, Michael S.
- Abstract
Engaging monocytes to battle chikungunya: Antibody-binding receptors, including Fc receptors and complement receptors, play a central role in mediating antibody-dependent immune activation. Here, Fox et al. have examined the role of Fcγ receptors and complement component 1q (C1q) in meditating the therapeutic effects of monoclonal IgG antibodies targeting chikungunya virus. Using antibody engineering in conjunction with mouse strains lacking C1q or Fcγ receptors, they report that the therapeutic effects of these antibodies are dependent on expression of Fcγ receptors. Furthermore, by depleting distinct immune cell types, they found that engagement of Fc receptors on monocytes is central in driving antibody-dependent clearance of chikungunya virus. Chikungunya virus (CHIKV) is an emerging mosquito-borne virus that has caused explosive outbreaks worldwide. Although neutralizing monoclonal antibodies (mAbs) against CHIKV inhibit infection in animals, the contribution of Fc effector functions to protection remains unknown. Here, we evaluated the activity of therapeutic mAbs that had or lacked the ability to engage complement and Fcγ receptors (FcγR). When administered as post-exposure therapy in mice, the Fc effector functions of mAbs promoted virus clearance from infected cells and reduced joint swelling—results that were corroborated in antibody-treated transgenic animals lacking activating FcγR. The control of CHIKV infection by antibody-FcγR engagement was associated with an accelerated influx of monocytes. A series of immune cell depletions revealed that therapeutic mAbs required monocytes for efficient clearance of CHIKV infection. Overall, our study suggests that in mice, FcγR expression on monocytes is required for optimal therapeutic activity of antibodies against CHIKV and likely other related viruses. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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12. Structural basis of a potent human monoclonal antibody against Zika virus targeting a quaternary epitope.
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Feng Long, Doyle, Michael, Fernandez, Estefania, Miller, Andrew S., Klose, Thomas, Sevvana, Madhumati, Bryan, Aubrey, Davidson, Edgar, Doranz, Benjamin J., Kuhn, Richard J., Diamond, Michael S., Crowe Jr., James E., and Rossmann, Michael G.
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ZIKA virus ,IMMUNOGLOBULINS ,FLAVIVIRUSES ,CONGENITAL disorders ,PROTEINS - Abstract
Zika virus (ZIKV) is a major human pathogen and member of the Flavivirus genus in the Flaviviridae family. In contrast to most other insect-transmitted flaviviruses, ZIKV also can be transmitted sexually and from mother to fetus in humans. During recent outbreaks, ZIKV infections have been linked to microcephaly, congenital disease, and Guillain-Barré syndrome. Neutralizing antibodies have potential as therapeutic agents. We report here a 4-Å-resolution cryo-electron microscopy structure of the ZIKV virion in complex with Fab fragments of the potently neutralizing human monoclonal antibody ZIKV-195. The footprint of the ZIKV-195 Fab fragment expands across two adjacent envelope (E) protein protomers. ZIKV neutralization by this antibody is presumably accomplished by cross-linking the E proteins, which likely prevents formation of E protein trimers required for fusion of the viral and cellular membranes. A single dose of ZIKV-195 administered 5 days after virus inoculation showed marked protection against lethality in a stringent mouse model of infection. [ABSTRACT FROM AUTHOR]
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- 2019
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13. Increased breadth of HIV-1 neutralization achieved by diverse antibody clones each with limited neutralization breadth.
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Chukwuma, Valentine U., Kose, Nurgun, Sather, D. Noah, Sapparapu, Gopal, Falk, Rachel, King, Hannah, Singh, Vidisha, Lampley, Rebecca, Malherbe, Delphine C., Ditto, Noah T., Sullivan, Jonathan T., Barnes, Trevor, Doranz, Benjamin J., Labranche, Celia C., Montefiori, David C., Kalams, Spyros A., Haigwood, Nancy L., and Jr.Crowe, James E.
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THERAPEUTIC use of monoclonal antibodies ,HIV infections ,MOLECULAR cloning ,EPITOPES ,CD4 antigen - Abstract
Broadly neutralizing antibodies (bNAbs) are rarely elicited by current human immunodeficiency virus type 1 (HIV-1) vaccine designs, but the presence of bNAbs in naturally infected individuals may be associated with high plasma viral loads, suggesting that the magnitude, duration, and diversity of viral exposure may contribute to the development of bNAbs. Here, we report the isolation and characterization of a panel of human monoclonal antibodies (mAbs) from two subjects who developed broadly neutralizing autologous antibody responses during HIV-1 infection. In both subjects, we identified collections of mAbs that exhibited specificity only to a few autologous envelopes (Envs), with some mAbs exhibiting specificity only to a subset of Envs within the quasispecies of a particular sample at one time point. Neutralizing antibodies (NAbs) isolated from these subjects mapped mostly to epitopes in the Env V3 loop region and the CD4 binding site. None of the individual neutralizing mAbs recovered exhibited the cumulative breadth of neutralization present in the serum of the subjects. Surprisingly, however, the activity of polyclonal mixtures comprising individual mAbs that each possessed limited neutralizing activity, could achieve increased breadth of neutralizing activity against autologous isolates. While a single broadly neutralizing antibody targeting one epitope can mediate neutralization breadth, the findings presented here suggest that a cooperative polyclonal process mediated by diverse antibodies with more limited breadth targeting multiple epitopes also can achieve neutralization breadth against HIV-1. [ABSTRACT FROM AUTHOR]
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- 2018
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14. Isolation of state-dependent monoclonal antibodies against the 12-transmembrane domain glucose transporter 4 using virus-like particles.
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Tucker, David F., Sullivan, Jonathan T., Mattia, Kimberly-Anne, Fisher, Christine R., Barnes, Trevor, Mabila, Manu N., Wilf, Rona, Sulli, Chidananda, Pitts, Meghan, Payne, Riley J., Hall, Moniquetta, Huston-Paterson, Duncan, Xiaoxiang Deng, Davidson, Edgar, Willis, Sharon H., Doranz, Benjamin J., Chambers, Ross, and Rucker, Joseph B.
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MONOCLONAL antibodies ,MEMBRANE potential ,CARRIER proteins ,ADIPOSE tissues ,PROTEOMICS - Abstract
The insulin-responsive 12-transmembrane transporter GLUT4 changes conformation between an inward-open state and an outward-open state to actively facilitate cellular glucose uptake. Because of the difficulties of generating conformational mAbs against complex and highly conserved membrane proteins, no reliable tools exist to measure GLUT4 at the cell surface, follow its trafficking, or detect the conformational state of the protein. Here we report the isolation and characterization of conformational mAbs that recognize the extracellular and intracellular domains of GLUT4, including mAbs that are specific for the inward-open and outward-open states of GLUT4. mAbs against GLUT4 were generated using virus-like particles to present this complex membrane protein in its native conformation and using a divergent host species (chicken) for immunization to overcome immune tolerance. As a result, the isolated mAbs recognize conformational epitopes on native GLUT4 in cells, with apparent affinities as high as 1 pM and with specificity for GLUT4 across the human membrane proteome. Epitope mapping using shotgun mutagenesis alanine scanning across the 509 amino acids of GLUT4 identified the binding epitopes for mAbs specific for the states of GLUT4 and allowed the comprehensive identification of the residues that functionally control the GLUT4 inward-open and outward-open states. The mAbs identified here will be valuable molecular tools for monitoring GLUT4 structure, function, and trafficking, for differentiating GLUT4 conformational states, and for the development of novel therapeutics for the treatment of diabetes. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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15. Mxra8 is a receptor for multiple arthritogenic alphaviruses.
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Zhang, Rong, Kim, Arthur S., Fox, Julie M., Nair, Sharmila, Basore, Katherine, Klimstra, William B., Rimkunas, Rebecca, Fong, Rachel H., Lin, Hueylie, Poddar, Subhajit, Crowe, James E., Doranz, Benjamin J., Fremont, Daved H., and Diamond, Michael S.
- Abstract
Arthritogenic alphaviruses comprise a group of enveloped RNA viruses that are transmitted to humans by mosquitoes and cause debilitating acute and chronic musculoskeletal disease [1]. The host factors required for alphavirus entry remain poorly characterized [2]. Here we use a genome-wide CRISPR-Cas9-based screen to identify the cell adhesion molecule Mxra8 as an entry mediator for multiple emerging arthritogenic alphaviruses, including chikungunya, Ross River, Mayaro and O’nyong nyong viruses. Gene editing of mouse Mxra8 or human MXRA8 resulted in reduced levels of viral infection of cells and, reciprocally, ectopic expression of these genes resulted in increased infection. Mxra8 bound directly to chikungunya virus particles and enhanced virus attachment and internalization into cells. Consistent with these findings, Mxra8-Fc fusion protein or anti-Mxra8 monoclonal antibodies blocked chikungunya virus infection in multiple cell types, including primary human synovial fibroblasts, osteoblasts, chondrocytes and skeletal muscle cells. Mutagenesis experiments suggest that Mxra8 binds to a surface-exposed region across the A and B domains of chikungunya virus E2 protein, which are a speculated site of attachment. Finally, administration of the Mxra8-Fc protein or anti-Mxra8 blocking antibodies to mice reduced chikungunya and O’nyong nyong virus infection as well as associated foot swelling. Pharmacological targeting of Mxra8 could form a strategy for mitigating infection and disease by multiple arthritogenic alphaviruses. The cell adhesion molecule Mxra8 is identified as a receptor for multiple arthritogenic alphaviruses such as chikungunya virus, and anti-Mxra8 monoclonal antibodies are shown to reduce rates of chikungunya virus infection in mice and a range of human cells. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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16. Human antibody recognition of antigenic site IV on Pneumovirus fusion proteins.
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Mousa, Jarrod J., Binshtein, Elad, Human, Stacey, Fong, Rachel H., Alvarado, Gabriela, Doranz, Benjamin J., Moore, Martin L., Ohi, Melanie D., and Jr.Crowe, James E.
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RESPIRATORY syncytial virus ,CHIMERIC proteins ,IMMUNOGLOBULINS ,ANTIGENIC variation ,ELECTRON microscopy - Abstract
Respiratory syncytial virus (RSV) is a major human pathogen that infects the majority of children by two years of age. The RSV fusion (F) protein is a primary target of human antibodies, and it has several antigenic regions capable of inducing neutralizing antibodies. Antigenic site IV is preserved in both the pre-fusion and post-fusion conformations of RSV F. Antibodies to antigenic site IV have been described that bind and neutralize both RSV and human metapneumovirus (hMPV). To explore the diversity of binding modes at antigenic site IV, we generated a panel of four new human monoclonal antibodies (mAbs) and competition-binding suggested the mAbs bind at antigenic site IV. Mutagenesis experiments revealed that binding and neutralization of two mAbs (3M3 and 6F18) depended on arginine (R) residue R429. We discovered two R429-independent mAbs (17E10 and 2N6) at this site that neutralized an RSV R429A mutant strain, and one of these mAbs (17E10) neutralized both RSV and hMPV. To determine the mechanism of cross-reactivity, we performed competition-binding, recombinant protein mutagenesis, peptide binding, and electron microscopy experiments. It was determined that the human cross-reactive mAb 17E10 binds to RSV F with a binding pose similar to 101F, which may be indicative of cross-reactivity with hMPV F. The data presented provide new concepts in RSV immune recognition and vaccine design, as we describe the novel idea that binding pose may influence mAb cross-reactivity between RSV and hMPV. Characterization of the site IV epitope bound by human antibodies may inform the design of a pan-Pneumovirus vaccine. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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17. Enhancing antibody patent protection using epitope mapping information.
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Deng, Xiaoxiang, Storz, Ulrich, and Doranz, Benjamin J.
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- 2018
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18. Probing the antigenicity of hepatitis C virus envelope glycoprotein complex by high-throughput mutagenesis.
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Gopal, Radhika, Jackson, Kelli, Tzarum, Netanel, Kong, Leopold, Ettenger, Andrew, Guest, Johnathan, Pfaff, Jennifer M., Barnes, Trevor, Honda, Andrew, Giang, Erick, Davidson, Edgar, Wilson, Ian A., Doranz, Benjamin J., and Law, Mansun
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HEPATITIS C virus ,GLYCOPROTEINS ,PROTEIN precursors ,MEMBRANE proteins ,MUTAGENESIS - Abstract
The hepatitis C virus (HCV) envelope glycoproteins E1 and E2 form a non-covalently linked heterodimer on the viral surface that mediates viral entry. E1, E2 and the heterodimer complex E1E2 are candidate vaccine antigens, but are technically challenging to study because of difficulties in producing natively folded proteins by standard protein expression and purification methods. To better comprehend the antigenicity of these proteins, a library of alanine scanning mutants comprising the entirety of E1E2 (555 residues) was created for evaluating the role of each residue in the glycoproteins. The mutant library was probed, by a high-throughput flow cytometry-based assay, for binding with the co-receptor CD81, and a panel of 13 human and mouse monoclonal antibodies (mAbs) that target continuous and discontinuous epitopes of E1, E2, and the E1E2 complex. Together with the recently determined crystal structure of E2 core domain (E2c), we found that several residues in the E2 back layer region indirectly impact binding of CD81 and mAbs that target the conserved neutralizing face of E2. These findings highlight an unexpected role for the E2 back layer in interacting with the E2 front layer for its biological function. We also identified regions of E1 and E2 that likely located at or near the interface of the E1E2 complex, and determined that the E2 back layer also plays an important role in E1E2 complex formation. The conformation-dependent reactivity of CD81 and the antibody panel to the E1E2 mutant library provides a global view of the influence of each amino acid (aa) on E1E2 expression and folding. This information is valuable for guiding protein engineering efforts to enhance the antigenic properties and stability of E1E2 for vaccine antigen development and structural studies. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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19. Systematic analysis of human antibody response to ebolavirus glycoprotein shows high prevalence of neutralizing public clonotypes.
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Chen, Elaine C., Gilchuk, Pavlo, Zost, Seth J., Ilinykh, Philipp A., Binshtein, Elad, Huang, Kai, Myers, Luke, Bonissone, Stefano, Day, Samuel, Kona, Chandrahaas R., Trivette, Andrew, Reidy, Joseph X., Sutton, Rachel E., Gainza, Christopher, Diaz, Summer, Williams, Jazmean K., Selverian, Christopher N., Davidson, Edgar, Saphire, Erica Ollmann, and Doranz, Benjamin J.
- Abstract
Understanding the human antibody response to emerging viral pathogens is key to epidemic preparedness. As the size of the B cell response to a pathogenic-virus-protective antigen is poorly defined, we perform deep paired heavy- and light-chain sequencing in Ebola virus glycoprotein (EBOV-GP)-specific memory B cells, allowing analysis of the ebolavirus-specific antibody repertoire both genetically and functionally. This approach facilitates investigation of the molecular and genetic basis for the evolution of cross-reactive antibodies by elucidating germline-encoded properties of antibodies to EBOV and identification of the overlap between antibodies in the memory B cell and serum repertoire. We identify 73 public clonotypes of EBOV, 20% of which encode antibodies with neutralization activity and capacity to protect mice in vivo. This comprehensive analysis of the public and private antibody repertoire provides insight into the molecular basis of the humoral immune response to EBOV GP, which informs the design of vaccines and improved therapeutics. [Display omitted] • The estimated size of an EBOV-GP-specific repertoire is about 20,000 clonal families • The clonally expanded repertoire in response to EBOV is functionally diverse • EBOV mAbs have germline-encoded cross-reactivity and neutralization characteristics • A high prevalence of public clonotypes with neutralizing properties is identified Chen et al. investigate the complexity of the human antibody response to a single viral protein, the EBOV GP, using paired single-cell sequencing. The study shows an immense diversity in the clonally expanded B cell repertoire and a high prevalence of public clonotypes. [ABSTRACT FROM AUTHOR]
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- 2023
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20. Cooperativity Enables Non-neutralizing Antibodies to Neutralize Ebolavirus.
- Author
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Howell, Katie A., Brannan, Jennifer M., Bryan, Christopher, McNeal, Andrew, Davidson, Edgar, Turner, Hannah L., Vu, Hong, Shulenin, Sergey, He, Shihua, Kuehne, Ana, Herbert, Andrew S., Qiu, Xiangguo, Doranz, Benjamin J., Holtsberg, Frederick W., Ward, Andrew B., Dye, John M., and Aman, M. Javad
- Abstract
Summary Drug combinations are synergistic when their combined efficacy exceeds the sum of the individual actions, but they rarely include ineffective drugs that become effective only in combination. We identified several “enabling pairs” of neutralizing and non-neutralizing anti-ebolavirus monoclonal antibodies, whose combination exhibited new functional profiles, including transforming a non-neutralizing antibody to a neutralizer. Sub-neutralizing concentrations of antibodies 2G4 or m8C4 enabled non-neutralizing antibody FVM09 (IC 50 >1 μM) to exhibit potent neutralization (IC 50 1–10 nM). While FVM09 or m8C4 alone failed to protect Ebola-virus-infected mice, a combination of the two antibodies provided 100% protection. Furthermore, non-neutralizers FVM09 and FVM02 exponentially enhanced the potency of two neutralizing antibodies against both Ebola and Sudan viruses. We identified a hotspot for the binding of these enabling antibody pairs near the interface of the glycan cap and GP2. Enabling cooperativity may be an underappreciated phenomenon for viruses, with implications for the design and development of immunotherapeutics and vaccines. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
21. Neutralizing human antibodies prevent Zika virus replication and fetal disease in mice.
- Author
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Sapparapu, Gopal, Fernandez, Estefania, Kose, Nurgun, Bin Cao, Fox, Julie M., Bombardi, Robin G., Zhao, Haiyan, Nelson, Christopher A., Bryan, Aubrey L., Barnes, Trevor, Davidson, Edgar, Mysorekar, Indira U., Fremont, Daved H., Doranz, Benjamin J., Diamond, Michael S., and Crowe, James E.
- Abstract
Zika virus (ZIKV) is an emerging mosquito-transmitted flavivirus that can cause severe disease, including congenital birth defects during pregnancy. To develop candidate therapeutic agents against ZIKV, we isolated a panel of human monoclonal antibodies from subjects that were previously infected with ZIKV. We show that a subset of antibodies recognize diverse epitopes on the envelope (E) protein and exhibit potent neutralizing activity. One of the most inhibitory antibodies, ZIKV-117, broadly neutralized infection of ZIKV strains corresponding to African and Asian-American lineages. Epitope mapping studies revealed that ZIKV-117 recognized a unique quaternary epitope on the E protein dimer-dimer interface. We evaluated the therapeutic efficacy of ZIKV-117 in pregnant and non-pregnant mice. Monoclonal antibody treatment markedly reduced tissue pathology, placental and fetal infection, and mortality in mice. Thus, neutralizing human antibodies can protect against maternal-fetal transmission, infection and disease, and reveal important determinants for structure-based rational vaccine design efforts. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
22. Structural basis for nonneutralizing antibody competition at antigenic site II of the respiratory syncytial virus fusion protein.
- Author
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Mousa, Jarrod J., Sauer, Marion F., Sevy, Alexander M., Finn, Jessica A., Bates, John T., Alvarado, Gabriela, King, Hannah G., Loerin, Leah B., Fong, Rachel H., Doranz, Benjamin J., Correi, Bruno E., Kalyuzhniy, Oleksandr, Xiaolin Wen, Jardetzky, Theodore S., Schief, William R., Ohi, Melanie D., Meiler, Jens, and Crowe Jr., James E.
- Subjects
RESPIRATORY syncytial virus ,CHIMERIC proteins ,MONOCLONAL antibodies ,PALIVIZUMAB ,PREVENTIVE medicine - Abstract
Palivizumab was the first antiviral monoclonal antibody (mAb) approved for therapeutic use in humans, and remains a prophylactic treatment for infants at risk for severe disease because of respiratory syncytial virus (RSV). Palivizumab is an engineered humanized version of a murine mAb targeting antigenic site II of the RSV fusion (F) protein, a key target in vaccine development. There are limited reported naturally occurring human mAbs to site II; therefore, the structural basis for human antibody recognition of this major antigenic site is poorly understood. Here, we describe a nonneutralizing class of site II-specific mAbs that competed for binding with palivizumab to postfusion RSV F protein. We also describe two classes of site II-specific neutralizing mAbs, one of which escaped competition with nonneutralizing mAbs. An X-ray crystal structure of the neutralizing mAb 14N4 in complex with F protein showed that the binding angle at which human neutralizing mAbs interact with antigenic site II determines whether or not nonneutralizing antibodies compete with their binding. Fine-mapping studies determined that nonneutralizing mAbs that interfere with binding of neutralizing mAbs recognize site II with a pose that facilitates binding to an epitope containing F surface residues on a neighboring protomer. Neutralizing antibodies, like motavizumab and a new mAb designated 3J20 that escape interference by the inhibiting mAbs, avoid such contact by binding at an angle that is shifted away from the nonneutralizing site. Furthermore, binding to rationally and computationally designed site II helix- loop-helix epitope-scaffold vaccines distinguished neutralizing from nonneutralizing site II antibodies. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
23. Signal transmission through the CXC chemokine receptor 4 (CXCR4) transmembrane helices.
- Author
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Wescott, Melanie P., Kufareva, Irina, Paes, Cheryl, Goodman, Jason R., Thaker, Yana, Puffer, Bridget A., Berdougo, Eli, Rucker, Joseph B., Handel, Tracy M., and Doranz, Benjamin J.
- Subjects
G protein-coupled receptor kinases ,LIGAND binding (Biochemistry) ,G proteins ,GENETIC mutation ,MONOCLONAL antibodies - Abstract
The atomic-level mechanisms by which G protein-coupled receptors (GPCRs) transmit extracellular ligand binding events through their transmembrane helices to activate intracellular G proteins remain unclear. Using a comprehensive library of mutations covering all 352 residues of the GPCR CXC chemokine receptor 4 (CXCR4), we identified 41 amino acids that are required for signaling induced by the chemokine ligand CXCL12 (stromal cell-derived factor 1). CXCR4 variants with each of these mutations do not signal properly but remain folded, based on receptor surface trafficking, reactivity to conformationally sensitive monoclonal antibodies, and ligand binding. When visualized on the structure of CXCR4, the majority of these residues form a continuous intramolecular signaling chain through the transmembrane helices; this chain connects chemokine binding residues on the extracellular side of CXCR4 to G protein-coupling residues on its intracellular side. Integrated into a cohesive model of signal transmission, these CXCR4 residues cluster into five functional groups that mediate (i) chemokine engagement, (ii) signal initiation, (iii) signal propagation, (iv) microswitch activation, and (v) G protein coupling. Propagation of the signal passes through a "hydrophobic bridge" on helix VI that coordinates with nearly every known GPCR signaling motif. Our results agree with known conserved mechanisms of GPCR activation and significantly expand on understanding the structural principles of CXCR4 signaling. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
24. Human Antibodies that Recognize Novel Immunodominant Quaternary Epitopes on the HIV-1 Env Protein.
- Author
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Hicar, Mark D., Chen, Xuemin, Sulli, Chidananda, Barnes, Trevor, Goodman, Jason, Sojar, Hakimuddin, Briney, Bryan, Willis, Jordan, Chukwuma, Valentine U., Kalams, Spyros A., Doranz, Benjamin J., Spearman, Paul, and Jr.Crowe, James E.
- Subjects
VIRAL envelope proteins ,IMMUNOGLOBULINS ,HIV ,EPITOPES ,OLIGOMERIZATION ,GLYCOSYLATION - Abstract
Numerous broadly neutralizing antibodies (Abs) target epitopes that are formed or enhanced during mature HIV envelope formation (i.e. quaternary epitopes). Generally, it is thought that Env epitopes that induce broadly neutralizing Abs are difficult to access and poorly immunogenic because of the characteristic oligomerization, conformational flexibility, sequence diversity and extensive glycosylation of Env protein. To enhance for isolation of quaternary epitope-targeting Abs (QtAbs), we previously used HIV virus-like particles (VLPs) to bind B cells from long-term non-progressor subjects to identify a panel of monoclonal Abs. When expressed as recombinant full-length Abs, a subset of these novel Abs exhibited the binding profiles of QtAbs, as they either failed to bind to monomeric Env protein or showed much higher affinity for Env trimers and VLPs. These QtAbs represented a significant proportion of the B-cell response identified with VLPs. The Ab genes of these clones were highly mutated, but they did not neutralize common HIV strains. We sought to further define the epitopes targeted by these QtAbs. Competition-binding and mapping studies revealed these Abs targeted four separate epitopes; they also failed to compete for binding by Abs to known major neutralizing epitopes. Detailed epitope mapping studies revealed that two of the four epitopes were located in the gp41 subunit of Env. These QtAbs bound pre-fusion forms of antigen and showed differential binding kinetics depending on whether oligomers were produced as recombinant gp140 trimers or as full-length Env incorporated into VLPs. Antigenic regions within gp41 present unexpectedly diverse structural epitopes, including these QtAb epitopes, which may be targeted by the naturally occurring Ab response to HIV infection. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
25. Lower IgG somatic hypermutation rates during acute dengue virus infection is compatible with a germinal center-independent B cell response.
- Author
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Godoy-Lozano, Elizabeth Ernestina, Téllez-Sosa, Juan, Sánchez-González, Gilberto, Sámano-Sánchez, Hugo, Aguilar-Salgado, Andrés, Salinas-Rodríguez, Aarón, Cortina-Ceballos, Bernardo, Vivanco-Cid, Héctor, Hernández-Flores, Karina, Pfaff, Jennifer M., Kahle, Kristen M., Doranz, Benjamin J., Gómez-Barreto, Rosa Elena, Valdovinos-Torres, Humberto, López-Martínez, Irma, Rodriguez, Mario H., and Martínez-Barnetche, Jesús
- Subjects
B cells ,DENGUE viruses ,ANTISENSE DNA ,NUCLEOTIDE sequencing ,ANTISENSE nucleic acids - Abstract
Background: The study of human B cell response to dengue virus (DENV) infection is critical to understand serotype-specific protection and the cross-reactive sub-neutralizing response. Whereas the first is beneficial and thus represents the ultimate goal of vaccination, the latter has been implicated in the development of severe disease, which occurs in a small, albeit significant, fraction of secondary DENV infections. Both primary and secondary infections are associated with the production of poly-reactive and cross-reactive IgG antibodies. Methods: To gain insight into the effect of DENV infection on the B cell repertoire, we used VH region high-throughput cDNA sequencing of the peripheral blood IgG B cell compartment of 19 individuals during the acute phase of infection. For 11 individuals, a second sample obtained 6 months later was analyzed for comparison. Probabilities of sequencing antibody secreting cells or memory B cells were estimated using second-order Monte Carlo simulation. Results: We found that in acute disease there is an increase in IgG B cell diversity and changes in the relative use of segments IGHV1-2, IGHV1-18, and IGHV1-69. Somewhat unexpectedly, an overall low proportion of somatic hypermutated antibody genes was observed during the acute phase plasmablasts, particularly in secondary infections and those cases with more severe disease. Conclusions: Our data are consistent with an innate-like antiviral recognition system mediated by B cells using defined germ-line coded B cell receptors, which could provide a rapid germinal center-independent antibody response during the early phase of infection. A model describing concurrent T-dependent and T-independent B cell responses in the context of DENV infection is proposed, which incorporates the selection of B cells using hypomutated IGHV segments and their potential role in poly/cross-reactivity. Its formal demonstration could lead to a definition of its potential implication in antibody-dependent enhancement, and may contribute to rational vaccine development efforts. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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- View/download PDF
26. Rapid isolation of dengue-neutralizing antibodies from single cell-sorted human antigen-specific memory B-cell cultures.
- Author
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Cox, Kara S., Tang, Aimin, Chen, Zhifeng, Horton, Melanie S., Yan, Hao, Wang, Xin-Min, Dubey, Sheri A., DiStefano, Daniel J., Ettenger, Andrew, Fong, Rachel H., Doranz, Benjamin J., Casimiro, Danilo R., and Vora, Kalpit A.
- Published
- 2016
- Full Text
- View/download PDF
27. Neutralizing Monoclonal Antibodies Block Chikungunya Virus Entry and Release by Targeting an Epitope Critical to Viral Pathogenesis.
- Author
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Jin, Jing, Liss, Nathan M., Chen, Dong-Hua, Liao, Maofu, Fox, Julie M., Shimak, Raeann M., Fong, Rachel H., Chafets, Daniel, Bakkour, Sonia, Keating, Sheila, Fomin, Marina E., Muench, Marcus O., Sherman, Michael B., Doranz, Benjamin J., Diamond, Michael S., and Simmons, Graham
- Abstract
Summary We evaluated the mechanism by which neutralizing human monoclonal antibodies inhibit chikungunya virus (CHIKV) infection. Potently neutralizing antibodies (NAbs) blocked infection at multiple steps of the virus life cycle, including entry and release. Cryo-electron microscopy structures of Fab fragments of two human NAbs and chikungunya virus-like particles showed a binding footprint that spanned independent domains on neighboring E2 subunits within one viral spike, suggesting a mechanism for inhibiting low-pH-dependent membrane fusion. Detailed epitope mapping identified amino acid E2-W64 as a critical interaction residue. An escape mutation (E2-W64G) at this residue rendered CHIKV attenuated in mice. Consistent with these data, CHIKV-E2-W64G failed to emerge in vivo under the selection pressure of one of the NAbs, IM-CKV063. As our study suggests that antibodies engaging the residue E2-W64 can potently inhibit CHIKV at multiple stages of infection, antibody-based therapies or immunogens that target this region might have protective value. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
28. Cryo-EM structures elucidate neutralizing mechanisms of anti-chikungunya human monoclonal antibodies with therapeutic activity.
- Author
-
Feng Long, Fong, Rachel H., Austin, Stephen K., Zhenguo Chen, Klose, Thomas, Fokine, Andrei, Yue Liu, Porta, Jason, Sapparapu, Gopal, Akahata, Wataru, Doranz, Benjamin J., Crowe Jr., James E., Diamond, Michael S., and Rossmann, Michael G.
- Subjects
CHIKUNGUNYA virus ,IMMUNOGLOBULINS ,MOLECULAR structure of glycoproteins ,MICROSCOPY ,NEUTRALIZATION (Chemistry) - Abstract
Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes severe acute and chronic disease in humans. Although highly inhibitory murine and human monoclonal antibodies (mAbs) have been generated, the structural basis of their neutralizing activity remains poorly characterized. Here, we determined the cryo-EM structures of chikungunya virus-like particles complexed with antibody fragments (Fab) of two highly protective human mAbs, 4J21 and 5M16, that block virus fusion with host membranes. Both mAbs bind primarily to sites within the A and B domains, as well as to the B domain's β-ribbon connector of the viral glycoprotein E2. The footprints of these antibodies on the viral surface were consistent with results from loss-of-binding studies using an alanine scanning mutagenesis- based epitope mapping approach. The Fab fragments stabilized the position of the B domain relative to the virus, particularly for the complex with 5M16. This finding is consistent with a mechanism of neutralization in which anti-CHIKV mAbs that bridge the A and B domains impede movement of the B domain away from the underlying fusion loop on the E1 glycoprotein and therefore block the requisite pH-dependent fusion of viral and host membranes. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
29. Measuring Membrane Protein Interactions Using Optical Biosensors.
- Author
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Rucker, Joseph, Davidoff, Candice, and Doranz, Benjamin J.
- Published
- 2010
- Full Text
- View/download PDF
30. Sites of vulnerability in HCV E1E2 identified by comprehensive functional screening.
- Author
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Pfaff-Kilgore, Jennifer M., Davidson, Edgar, Kadash-Edmondson, Kathryn, Hernandez, Mayda, Rosenberg, Erin, Chambers, Ross, Castelli, Matteo, Clementi, Nicola, Mancini, Nicasio, Bailey, Justin R., Crowe, James E., Law, Mansun, and Doranz, Benjamin J.
- Abstract
The E1 and E2 envelope proteins of hepatitis C virus (HCV) form a heterodimer that drives virus-host membrane fusion. Here, we analyze the role of each amino acid in E1E2 function, expressing 545 individual alanine mutants of E1E2 in human cells, incorporating them into infectious viral pseudoparticles, and testing them against 37 different monoclonal antibodies (MAbs) to ascertain full-length translation, folding, heterodimer assembly, CD81 binding, viral pseudoparticle incorporation, and infectivity. We propose a model describing the role of each critical residue in E1E2 functionality and use it to examine how MAbs neutralize infection by exploiting functionally critical sites of vulnerability on E1E2. Our results suggest that E1E2 is a surprisingly fragile protein complex where even a single alanine mutation at 92% of positions disrupts its function. The amino-acid-level targets identified are highly conserved and functionally critical and can be exploited for improved therapies and vaccines. [Display omitted] • Test 545 hepatis C virus (HCV) E1E2 envelope mutants for infectivity, antibody binding • Identify residues important for HCV E1 and E2 folding, E1E2 interaction, infectivity • HCV E1E2 is a fragile protein complex where most mutations compromise function • Functional residues of E1E2 are highly conserved across genotypes Pfaff-Kilgore et al. describe the role of individual amino acids in hepatis C virus E1E2 protein function by generating a comprehensive E1E2 mutation library and testing 545 clones for E1E2 folding, assembly, and infectivity. Their model describes the role of residues in E1E2 functionality and examines how antibodies neutralize infection. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
31. A high-throughput shotgun mutagenesis approach to mapping B-cell antibody epitopes.
- Author
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Davidson, Edgar and Doranz, Benjamin J.
- Subjects
MUTAGENESIS ,B cells ,MONOCLONAL antibodies ,DRUG development ,PROTEIN folding ,BINDING sites ,EPITOPES ,G protein coupled receptors - Abstract
Characterizing the binding sites of monoclonal antibodies (m Abs) on protein targets, their 'epitopes', can aid in the discovery and development of new therapeutics, diagnostics and vaccines. However, the speed of epitope mapping techniques has not kept pace with the increasingly large numbers of m Abs being isolated. Obtaining detailed epitope maps for functionally relevant antibodies can be challenging, particularly for conformational epitopes on structurally complex proteins. To enable rapid epitope mapping, we developed a high-throughput strategy, shotgun mutagenesis, that enables the identification of both linear and conformational epitopes in a fraction of the time required by conventional approaches. Shotgun mutagenesis epitope mapping is based on large-scale mutagenesis and rapid cellular testing of natively folded proteins. Hundreds of mutant plasmids are individually cloned, arrayed in 384-well microplates, expressed within human cells, and tested for m Ab reactivity. Residues are identified as a component of a m Ab epitope if their mutation (e.g. to alanine) does not support candidate m Ab binding but does support that of other conformational m Abs or allows full protein function. Shotgun mutagenesis is particularly suited for studying structurally complex proteins because targets are expressed in their native form directly within human cells. Shotgun mutagenesis has been used to delineate hundreds of epitopes on a variety of proteins, including G protein-coupled receptor and viral envelope proteins. The epitopes mapped on dengue virus pr M/ E represent one of the largest collections of epitope information for any viral protein, and results are being used to design better vaccines and drugs. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
32. Dengue virus envelope protein domain I/II hinge determines long-lived serotype-specific dengue immunity.
- Author
-
Messer, William B., de Alwis, Ruklanthi, Yount, Boyd L., Royal, Scott R., Huynh, Jeremy P., Smith, Scott A., Crowe Jr., James E., Doranz, Benjamin J., Kahle, Kristen M., Pfaff, Jennifer M., White, Laura J., Sariol, Carlos A., de Silva, Aravinda M., and Baric, Ralph S.
- Subjects
DENGUE viruses ,IMMUNOGLOBULINS ,GLYCOPROTEINS ,DENGUE hemorrhagic fever ,VACCINES - Abstract
The four dengue virus (DENV) serotypes, DENV-1, -2, -3, and -4, are endemic throughout tropical and subtropical regions of the world, with an estimated 390 million acute infections annually. Infection confers long-term protective immunity against the infecting serotype, but secondary infection with a different serotype carries a greater risk of potentially fatal severe dengue disease, including dengue hemorrhagic fever and dengue shock syndrome. The single most effective measure to control this threat to global health is a tetravalent DENV vaccine. To date, attempts to develop a protective vaccine have progressed slowly, partly because the targets of type-specific human neutralizing antibodies (NAbs), which are critical for long-term protection, remain poorly defined, impeding our understanding of natural immunity and hindering effective vaccine development. Here, we show that the envelope glycoprotein domain I/II hinge of DENV-3 and DENV-4 is the primary target of the long-term type-specific NAb response in humans. Transplantation of a DENV-4 hinge into a recombinant DENV-3 virus showed that the hinge determines the serotype-specific neutralizing potency of primary human and nonhuman primate DENV immune sera and that the hinge region both induces NAbs and is targeted by protective NAbs in rhesus macaques. These results suggest that the success of live dengue vaccines may depend on their ability to stimulate NAbs that target the envelope glycoprotein domain I/II hinge region. More broadly, this study shows that complex conformational antibody epitopes can be transplanted between live viruses, opening up similar possibilities for improving the breadth and specificity of vaccines for influenza, HIV, hepatitis C virus, and other clinically important viral pathogens. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
33. Atomic-level functional model of dengue virus Envelope protein infectivity.
- Author
-
Christian, Elizabeth A., Kahle, Kristen M., Mattia, Kimberly, Puffer, Bridget A., Pfaff, Jennifer M., Miller, Adam, Paes, Cheryl, Davidson, Edgar, and Doranz, Benjamin J.
- Subjects
DENGUE viruses ,FLAVIVIRUSES ,ATOMIC interactions ,VACCINATION ,CELL membranes - Abstract
A number of structures have been solved for the Envelope (E) protein from dengue virus and closely related flaviviruses, providing detailed pictures of the conformational states of the protein at different stages of infectivity. However, the key functional residues responsible for mediating the dynamic changes between these structures remain largely unknown. Using a comprehensive library of functional point mutations covering all 390 residues of the dengue virus E protein ectodomain, we identified residues that are critical for virus infectivity, but that do not affect E protein expression, folding, virion assembly, or budding. The locations and atomic interactions of these critical residues within different structures representing distinct fusogenic conformations help to explain how E protein (i) regulates fusion-loop exposure by shielding, tethering, and triggering its release; (ii) enables hinge movements between E domain interfaces during triggered structural transformations; and (iii) drives membrane fusion through late-stage zipper contacts with stem. These results provide structural targets for drug and vaccine development and integrate the findings from structural studies and isolated mutagenesis efforts into a cohesive model that explains how specific residues in this class II viral fusion protein enable virus infectivity. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
34. Dengue Reporter Virus Particles for Measuring Neutralizing Antibodies against Each of the Four Dengue Serotypes.
- Author
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Mattia, Kimberly, Puffer, Bridget A., Williams, Katherine L., Gonzalez, Ritela, Murray, Meredith, Sluzas, Emily, Pagano, Dan, Ajith, Sandya, Bower, Megan, Berdougo, Eli, Harris, Eva, and Doranz, Benjamin J.
- Subjects
DENGUE ,IMMUNOGLOBULINS ,SEROTYPES ,DISEASE progression ,VACCINES - Abstract
The lack of reliable, high-throughput tools for characterizing anti-dengue virus (DENV) antibodies in large numbers of serum samples has been an obstacle in understanding the impact of neutralizing antibodies on disease progression and vaccine efficacy. A reporter system using pseudoinfectious DENV reporter virus particles (RVPs) was previously developed by others to facilitate the genetic manipulation and biological characterization of DENV virions. In the current study, we demonstrate the diagnostic utility of DENV RVPs for measuring neutralizing antibodies in human serum samples against all four DENV serotypes, with attention to the suitability of DENV RVPs for large-scale, long-term studies. DENV RVPs used against human sera yielded serotype-specific responses and reproducible neutralization titers that were in statistical agreement with Plaque Reduction Neutralization Test (PRNT) results. DENV RVPs were also used to measure neutralization titers against the four DENV serotypes in a panel of human sera from a clinical study of dengue patients. The high-throughput capability, stability, rapidity, and reproducibility of assays using DENV RVPs offer advantages for detecting immune responses that can be applied to large-scale clinical studies of DENV infection and vaccination. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
35. Probenecid Inhibits the Human Bitter Taste Receptor TAS2R16 and Suppresses Bitter Perception of Salicin.
- Author
-
Greene, Tiffani A., Alarcon, Suzanne, Thomas, Anu, Berdougo, Eli, Doranz, Benjamin J., Breslin, Paul A. S., and Rucker, Joseph B.
- Subjects
G protein coupled receptors ,PROBENECID (Drug) ,BITTERNESS (Taste) ,GLUCOSIDES ,THIOUREA - Abstract
Bitter taste stimuli are detected by a diverse family of G protein-coupled receptors (GPCRs) expressed in gustatory cells. Each bitter taste receptor (TAS2R) responds to an array of compounds, many of which are toxic and can be found in nature. For example, human TAS2R16 (hTAS2R16) responds to b-glucosides such as salicin, and hTAS2R38 responds to thioureacontaining molecules such as glucosinolates and phenylthiocarbamide (PTC). While many substances are known to activate TAS2Rs, only one inhibitor that specifically blocks bitter receptor activation has been described. Here, we describe a new inhibitor of bitter taste receptors, p-(dipropylsulfamoyl)benzoic acid (probenecid), that acts on a subset of TAS2Rs and inhibits through a novel, allosteric mechanism of action. Probenecid is an FDA-approved inhibitor of the Multidrug Resistance Protein 1 (MRP1) transporter and is clinically used to treat gout in humans. Probenecid is also commonly used to enhance cellular signals in GPCR calcium mobilization assays. We show that probenecid specifically inhibits the cellular response mediated by the bitter taste receptor hTAS2R16 and provide molecular and pharmacological evidence for direct interaction with this GPCR using a non-competitive (allosteric) mechanism. Through a comprehensive analysis of hTAS2R16 point mutants, we define amino acid residues involved in the probenecid interaction that result in decreased sensitivity to probenecid while maintaining normal responses to salicin. Probenecid inhibits hTAS2R16, hTAS2R38, and hTAS2R43, but does not inhibit the bitter receptor hTAS2R31 or non-TAS2R GPCRs. Additionally, structurally unrelated MRP1 inhibitors, such as indomethacin, fail to inhibit hTAS2R16 function. Finally, we demonstrate that the inhibitory activity of probenecid in cellular experiments translates to inhibition of bitter taste perception of salicin in humans. This work identifies probenecid as a pharmacological tool for understanding the cell biology of bitter taste and as a lead for the development of broad specificity bitter blockers to improve nutrition and medical compliance. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
36. Potent neutralization of SARS-CoV-2 variants of concern by an antibody with an uncommon genetic signature and structural mode of spike recognition.
- Author
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Kramer, Kevin J., Johnson, Nicole V., Shiakolas, Andrea R., Suryadevara, Naveenchandra, Periasamy, Sivakumar, Raju, Nagarajan, Williams, Jazmean K., Wrapp, Daniel, Zost, Seth J., Walker, Lauren M., Wall, Steven C., Holt, Clinton M., Hsieh, Ching-Lin, Sutton, Rachel E., Paulo, Ariana, Nargi, Rachel S., Davidson, Edgar, Doranz, Benjamin J., Crowe, James E., and Bukreyev, Alexander
- Abstract
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineages that are more transmissible and resistant to currently approved antibody therapies poses a considerable challenge to the clinical treatment of coronavirus disease (COVID-19). Therefore, the need for ongoing discovery efforts to identify broadly reactive monoclonal antibodies to SARS-CoV-2 is of utmost importance. Here, we report a panel of SARS-CoV-2 antibodies isolated using the linking B cell receptor to antigen specificity through sequencing (LIBRA-seq) technology from an individual who recovered from COVID-19. Of these antibodies, 54042-4 shows potent neutralization against authentic SARS-CoV-2 viruses, including variants of concern (VOCs). A cryoelectron microscopy (cryo-EM) structure of 54042-4 in complex with the SARS-CoV-2 spike reveals an epitope composed of residues that are highly conserved in currently circulating SARS-CoV-2 lineages. Further, 54042-4 possesses uncommon genetic and structural characteristics that distinguish it from other potently neutralizing SARS-CoV-2 antibodies. Together, these findings provide motivation for the development of 54042-4 as a lead candidate to counteract current and future SARS-CoV-2 VOCs. [Display omitted] • LIBRA-seq identifies 54042-4, a potently neutralizing SARS-CoV-2 antibody • 54042-4 maintains potent neutralization against Alpha, Beta, Gamma, and Delta VOCs • The epitope of 54042-4 is highly conserved among current SARS-CoV-2 isolates Kramer et al. demonstrate that antibody 54042-4 recognizes residues highly conserved across global SARS-CoV-2 isolates. Antibody 54042-4 potently neutralizes all known circulating variants of concern (VOCs) and could be developed as a clinical candidate to treat COVID-19 infection. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
37. Cell-Free Assay of G-Protein-Coupled Receptors Using Fluorescence Polarization.
- Author
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Jones, Jessi Wildeson, Greene, Tiffani A., Grygon, Christine A., Doranz, Benjamin J., and Brown, Martha P.
- Subjects
NANOTECHNOLOGY ,G proteins ,CELL receptors ,PROSPECTING ,FIRE assay ,MEMBRANE proteins - Abstract
A recently developed nanotechnology, the Integral Molecular lipoparticle, provides an essentially soluble cell-free system in which G-protein-coupled receptors (GPCRs) in their native conformations are concentrated within virus-like particles. As a result, the lipoparticle provides a means to overcome 2 common obstacles to the development of homogeneous, nonradioactive GPCR ligand-binding assays: membrane protein solubilization and low receptor density. The work reported here describes the first application of this nanotechnology to a fluorescence polarization (FP) molecular binding assay format. The GPCR chosen for these studies was the well-studied chemokine receptor CXCR4 for which a peptide ligand (T-22) has been previously characterized. The EC
50 determined for the CXCR4-T-22 peptide interaction via FP with CXCR4 lipoparticles (15 nM) is consistent with the IC50 determined for the unlabeled T-22 peptide via competitive binding (59 nM). [ABSTRACT FROM AUTHOR]- Published
- 2008
- Full Text
- View/download PDF
38. Convergent antibody responses to the SARS-CoV-2 spike protein in convalescent and vaccinated individuals.
- Author
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Chen, Elaine C., Gilchuk, Pavlo, Zost, Seth J., Suryadevara, Naveenchandra, Winkler, Emma S., Cabel, Carly R., Binshtein, Elad, Chen, Rita E., Sutton, Rachel E., Rodriguez, Jessica, Day, Samuel, Myers, Luke, Trivette, Andrew, Williams, Jazmean K., Davidson, Edgar, Li, Shuaizhi, Doranz, Benjamin J., Campos, Samuel K., Carnahan, Robert H., and Thorne, Curtis A.
- Abstract
Unrelated individuals can produce genetically similar clones of antibodies, known as public clonotypes, which have been seen in responses to different infectious diseases, as well as healthy individuals. Here we identify 37 public clonotypes in memory B cells from convalescent survivors of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection or in plasmablasts from an individual after vaccination with mRNA-encoded spike protein. We identify 29 public clonotypes, including clones recognizing the receptor-binding domain (RBD) in the spike protein S1 subunit (including a neutralizing, angiotensin-converting enzyme 2 [ACE2]-blocking clone that protects in vivo) and others recognizing non-RBD epitopes that bind the S2 domain. Germline-revertant forms of some public clonotypes bind efficiently to spike protein, suggesting these common germline-encoded antibodies are preconfigured for avid recognition. Identification of large numbers of public clonotypes provides insight into the molecular basis of efficacy of SARS-CoV-2 vaccines and sheds light on the immune pressures driving the selection of common viral escape mutants. [Display omitted] • Public clonotypes are shared among SARS-CoV-2-vaccinated and -infected individuals • Group 3 antibodies share the same binding site as CR3022 but neutralize variants • Germline-revertant forms of public clonotypes bind efficiently to spike protein • Public clonotype responses inform research on new viral variants of concern Chen et al. describe neutralizing and non-neutralizing public antibodies elicited to SARS-CoV-2 spike in vaccinated and naturally infected individuals. Germline-revertant forms of some public clonotypes bind efficiently to spike protein, suggesting common germline-encoded properties for recognition. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
39. Author Correction: Molecular determinants and mechanism for antibody cocktail preventing SARS-CoV-2 escape.
- Author
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Ku, Zhiqiang, Xie, Xuping, Davidson, Edgar, Ye, Xiaohua, Su, Hang, Menachery, Vineet D., Li, Yize, Yuan, Zihao, Zhang, Xianwen, Muruato, Antonio E., i Escuer, Ariadna Grinyo, Tyrell, Breanna, Doolan, Kyle, Doranz, Benjamin J., Wrapp, Daniel, Bates, Paul F., McLellan, Jason S., Weiss, Susan R., Zhang, Ningyan, and Shi, Pei-Yong
- Subjects
SARS-CoV-2 ,IMMUNOGLOBULINS - Abstract
The original version of this Article contained an error in the Acknowledgements, which incorrectly listed the NIH grant "AI142759". This has been corrected in both the PDF and HTML versions of the Article. [Extracted from the article]
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- 2021
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40. Convergence of a common solution for broad ebolavirus neutralization by glycan cap-directed human antibodies.
- Author
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Murin, Charles D., Gilchuk, Pavlo, Ilinykh, Philipp A., Huang, Kai, Kuzmina, Natalia, Shen, Xiaoli, Bruhn, Jessica F., Bryan, Aubrey L., Davidson, Edgar, Doranz, Benjamin J., Williamson, Lauren E., Copps, Jeffrey, Alkutkar, Tanwee, Flyak, Andrew I., Bukreyev, Alexander, Crowe, James E., and Ward, Andrew B.
- Abstract
Antibodies that target the glycan cap epitope on the ebolavirus glycoprotein (GP) are common in the adaptive response of survivors. A subset is known to be broadly neutralizing, but the details of their epitopes and basis for neutralization are not well understood. Here, we present cryoelectron microscopy (cryo-EM) structures of diverse glycan cap antibodies that variably synergize with GP base-binding antibodies. These structures describe a conserved site of vulnerability that anchors the mucin-like domains (MLDs) to the glycan cap, which we call the MLD anchor and cradle. Antibodies that bind to the MLD cradle share common features, including use of IGHV1-69 and IGHJ6 germline genes, which exploit hydrophobic residues and form β-hairpin structures to mimic the MLD anchor, disrupt MLD attachment, destabilize GP quaternary structure, and block cleavage events required for receptor binding. Our results provide a molecular basis for ebolavirus neutralization by broadly reactive glycan cap antibodies. [Display omitted] • Glycan cap antibody-mediated GP destabilization correlates with synergy • Cryo-EM structures reveal antibodies target a highly conserved epitope • Antibodies use long CDRH3 loops to displace and mimic portions of the glycan cap • Glycan cap antibodies block cleavage events required for viral entry A rare subset of ebolavirus antibodies targeting the glycan cap are broadly neutralizing. Murin et al. report cryo-EM structures and custom in vitro assays identifying a conserved site of vulnerability in the glycan cap and detail mechanisms of action, including structural mimicry, trimer instability, and blocking cleavage. [ABSTRACT FROM AUTHOR]
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- 2021
- Full Text
- View/download PDF
41. CD4-independent, CCR5-dependent infection of brain capillary endothelial cells by a neurovirulent...
- Author
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Edinger, Aimee L., Mankowski, Joseph L., Doranz, Benjamin J., Margulies, Barry J., Lee, Benhur, Rucker, Joseph, Sharron, Matthew, Hoffman, Trevor L., Berson, Joanne F., Zink, M. Christine, Hirsch, Vanessa M., Clements, Janice E., and Doms, Robert W.
- Subjects
CAPILLARIES ,SIMIAN viruses - Abstract
Provides information on findings that rhesus macaque brain capillary endothelial cells (BCECs) express chemokine receptors involved in HIV and simian immunodeficiency virus (SIV) entry including CCR5, CCR3, CXCR4, and STRL33, but not CCR2b, GPR1, or GPR15. Affect of CD4-independent infection by HIV-1 and SIV strains on BCECs; What was observed about the envelope (env) proteins; Indications of study.
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- 1997
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42. Antibody Strategies for Membrane Protein Targets.
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Banik, Soma S. R. and Doranz, Benjamin J.
- Subjects
MEMBRANE proteins ,LIPOPROTEIN antibodies ,IMMUNOGLOBULINS ,CLINICAL medicine ,THERAPEUTICS - Abstract
The article discusses the antibody strategies for membrane protein. It provides an overview on membrane-embedded proteins which is most important class of therapeutic targets as well as the innovative approach to generate antibodies against the membrane proteins. It also provides information on lipoparticles which is used to produce antibodies against several classes of membrane proteins.
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- 2009
43. 1177-P: Discovery of New Therapeutic Antibodies to Two Metabolically Relevant Targets—GPCR CB1 and Transporter GLUT4—Using Virus-Like Particles.
- Author
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TUCKER, DAVID, CHARPENTIER, TOM, MOLINO, NICHOLAS, ROSENBERG, ERIN, SCRENCI, BRAD, STAFFORD, LEWIS J., SULLI, CHIDANANDA, CHAMBERS, ROSS, DORANZ, BENJAMIN J., and RUCKER, JOSEPH B.
- Abstract
Integral membrane proteins are important drug targets, and monoclonal antibodies (mAbs) against them are highly sought for therapeutic and diagnostic purposes. However, the complex structure of multipass membrane proteins makes discovery of these mAbs particularly challenging. By employing strategies such as sequential immunization with DNA (prime) and virus-like particles (boost), and using divergent species (chickens) as immunization hosts, we routinely isolate diverse panels of mAbs against structurally complex and highly conserved membrane protein targets. Here, we report the isolation and characterization of mAbs recognizing two distinct metabolically relevant targets: the 12-transmembrane insulin-responsive glucose transporter GLUT4 and the 7-transmembrane GPCR cannabinoid receptor-1 (CB1). In both cases, we isolated diverse panels of mAbs against native conformational epitopes. Affinities ranged from 2.6 nM (anti-CB1 IM-102) to 1 pM (anti-GLUT4 LM048). Two anti-GLUT4 mAbs discriminated between inward-open and outward-open conformational states, including one mAb (LM048) that inhibited transporter function. Similarly, we isolated several anti-CB1 mAbs that antagonized CB1-mediated calcium flux. Lead mAbs were confirmed to be specific for their target proteins, based on reactivity profiling in the Membrane Proteome Array of >5,600 human membrane proteins. Identified mAbs represent novel and valuable molecular tools for understanding and potentially treating common metabolic diseases, such as type 2 diabetes mellitus and nonalcoholic steatohepatitis/fatty liver disease (NASH/NAFLD). Furthermore, the technologies underpinning their discovery can be rapidly and routinely employed to facilitate discovery of mAbs against other therapeutically relevant membrane protein targets. Disclosure: D. Tucker: None. T. Charpentier: None. N. Molino: None. E. Rosenberg: None. B. Screnci: None. L.J. Stafford: None. C. Sulli: None. R. Chambers: None. B.J. Doranz: None. J.B. Rucker: None. Funding: National Institutes of Health [ABSTRACT FROM AUTHOR]
- Published
- 2019
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44. In Vivo Delivery of Synthetic Human DNA-Encoded Monoclonal Antibodies Protect against Ebolavirus Infection in a Mouse Model.
- Author
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Patel, Ami, Park, Daniel H., Davis, Carl W., Smith, Trevor R.F., Leung, Anders, Tierney, Kevin, Bryan, Aubrey, Davidson, Edgar, Yu, Xiaoying, Racine, Trina, Reed, Charles, Gorman, Marguerite E., Wise, Megan C., Elliott, Sarah T.C., Esquivel, Rianne, Yan, Jian, Chen, Jing, Muthumani, Kar, Doranz, Benjamin J., and Saphire, Erica Ollmann
- Abstract
Summary Synthetically engineered DNA-encoded monoclonal antibodies (DMAbs) are an in vivo platform for evaluation and delivery of human mAb to control against infectious disease. Here, we engineer DMAbs encoding potent anti- Zaire ebolavirus (EBOV) glycoprotein (GP) mAbs isolated from Ebola virus disease survivors. We demonstrate the development of a human IgG1 DMAb platform for in vivo EBOV-GP mAb delivery and evaluation in a mouse model. Using this approach, we show that DMAb-11 and DMAb-34 exhibit functional and molecular profiles comparable to recombinant mAb, have a wide window of expression, and provide rapid protection against lethal mouse-adapted EBOV challenge. The DMAb platform represents a simple, rapid, and reproducible approach for evaluating the activity of mAb during clinical development. DMAbs have the potential to be a mAb delivery system, which may be advantageous for protection against highly pathogenic infectious diseases, like EBOV, in resource-limited and other challenging settings. Graphical Abstract Highlights • DMAbs are an in vivo approach for mAb development/delivery against Zaire ebolavirus • DMAbs can be functionally equivalent to recombinant mAb • Reproducible and cost-effective mouse model for in vivo mAb evaluation • Enables evaluation of fully human mAbs rapidly in vivo Monoclonal antibodies are an important approach for emerging infectious disease prevention. Patel et al. demonstrate engineering and in vivo delivery of DNA-encoded monoclonal antibodies (DMAbs) targeting the Zaire ebolavirus (EBOV) glycoprotein. DMAbs protect against lethal mouse-adapted EBOV and are useful for rapid evaluation of fully human mAbs in live animal models. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
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