18 results on '"Kuishu Ren"'
Search Results
2. Resilience of S309 and AZD7442 monoclonal antibody treatments against infection by SARS-CoV-2 Omicron lineage strains
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James Brett Case, Samantha Mackin, John M. Errico, Zhenlu Chong, Emily A. Madden, Bradley Whitener, Barbara Guarino, Michael A. Schmid, Kim Rosenthal, Kuishu Ren, Ha V. Dang, Gyorgy Snell, Ana Jung, Lindsay Droit, Scott A. Handley, Peter J. Halfmann, Yoshihiro Kawaoka, James E. Crowe, Daved H. Fremont, Herbert W. Virgin, Yueh-Ming Loo, Mark T. Esser, Lisa A. Purcell, Davide Corti, and Michael S. Diamond
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Science - Abstract
SARS-CoV-2 variants of concern are less susceptible to therapeutic neutralizing antibodies, given mutations in the surface glycoprotein S. Here, Case et al. show that therapeutic antibodies S309 and AZD7442 reduce lung infection with SARSCoV-2 Omicron lineages in humanized mouse model despite the loss of neutralizing potency in vitro.
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- 2022
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3. Qualification of a Biolayer Interferometry Assay to Support AZD7442 Resistance Monitoring
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Tyler Brady, Tianhui Zhang, Kevin M. Tuffy, Nantaporn Haskins, Qun Du, Jia Lin, Gilad Kaplan, Steven Novick, Tiffany L. Roe, Kuishu Ren, Kim Rosenthal, Patrick M. McTamney, Michael E. Abram, Katie Streicher, and Elizabeth J. Kelly
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severe acute respiratory syndrome coronavirus 2 ,coronavirus disease 2019 ,monoclonal antibody ,spike protein ,receptor binding domain ,Microbiology ,QR1-502 - Abstract
ABSTRACT AZD7442, a combination of two long-acting monoclonal antibodies (tixagevimab [AZD8895] and cilgavimab [AZD1061]), has been authorized for the prevention and treatment of coronavirus disease 2019 (COVID-19). The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants requires methods capable of quickly characterizing resistance to AZD7442. To support AZD7442 resistance monitoring, a biolayer interferometry (BLI) assay was developed to screen the binding of tixagevimab and cilgavimab to SARS-CoV-2 spike proteins to reduce the number of viral variants for neutralization susceptibility verification. Six spike variants were chosen to assess the assay’s performance: four with decreased affinity for tixagevimab (F486S:D614G and F486W:D614G proteins) or cilgavimab (S494L:D614G and K444R:D614G proteins) and two reference proteins (wild-type HexaPro and D614G protein). Equilibrium dissociation constant (KD) values from each spike protein were used to determine shifts in binding affinity. The assay’s precision, range, linearity, and limits of quantitation were established. Qualification acceptance criteria determined whether the assay was fit for purpose. By bypassing protein purification, the BLI assay provided increased screening throughput. Although limited correlation between pseudotype neutralization and BLI data (50% inhibitory concentration versus KD) was observed for full immunoglobulins (IgGs), the correlations for antibody fragments (Fabs) were stronger and reflected a better comparison of antibody binding kinetics with neutralization potency. Therefore, despite strong assay performance characteristics, the use of full IgGs limited the screening utility of the assay; however, the Fab approach warrants further exploration as a rapid, high-throughput variant-screening method for future resistance-monitoring programs. IMPORTANCE SARS-CoV-2 variants harbor multiple substitutions in their spike trimers, potentially leading to breakthrough infections and clinical resistance to immune therapies. For this reason, a BLI assay was developed and qualified to evaluate the reliability of screening SARS-CoV-2 spike trimer variants against anti-SARS-CoV-2 monoclonal antibodies (MAbs) tixagevimab and cilgavimab, the components of AZD7442, prior to in vitro pseudovirus neutralization susceptibility verification testing. The assay bypasses protein purification with rapid assessment of the binding affinity of each MAb for each recombinant protein, potentially providing an efficient preliminary selection step, thus allowing a reduced testing burden in the more technically complex viral neutralization assays. Despite precise and specific measures, an avidity effect associated with MAb binding to the trimer confounded correlation with neutralization potency, negating the assay’s utility as a surrogate for neutralizing antibody potency. Improved correlation with Fabs suggests that assay optimization could overcome any avidity limitation, warranting further exploration to support future resistance-monitoring programs.
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- 2022
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4. The ChAdOx1 vectored vaccine, AZD2816, induces strong immunogenicity against SARS-CoV-2 beta (B.1.351) and other variants of concern in preclinical studies
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Alexandra J Spencer, Susan Morris, Marta Ulaszewska, Claire Powers, Reshma Kailath, Cameron Bissett, Adam Truby, Nazia Thakur, Joseph Newman, Elizabeth R Allen, Indra Rudiansyah, Chang Liu, Wanwisa Dejnirattisai, Juthathip Mongkolsapaya, Hannah Davies, Francesca R Donnellan, David Pulido, Thomas P. Peacock, Wendy S. Barclay, Helen Bright, Kuishu Ren, Gavin Screaton, Patrick McTamney, Dalan Bailey, Sarah C Gilbert, and Teresa Lambe
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Vaccines ,SARS CoV2 ,variants of concern ,Antibodies ,T cells ,Medicine ,Medicine (General) ,R5-920 - Abstract
Summary: Background: There is an ongoing global effort to design, manufacture, and clinically assess vaccines against SARS-CoV-2. Over the course of the ongoing pandemic a number of new SARS-CoV-2 virus isolates or variants of concern (VoC) have been identified containing mutations in key proteins. Methods: In this study we describe the generation and preclinical assessment of a ChAdOx1-vectored vaccine (AZD2816) which expresses the spike protein of the Beta VoC (B.1.351). Findings: We demonstrate that AZD2816 is immunogenic after a single dose. When AZD2816 is used as a booster dose in animals primed with a vaccine encoding the original spike protein (ChAdOx1 nCoV-19/ [AZD1222]), an increase in binding and neutralising antibodies against Beta (B.1.351), Gamma (P.1) and Delta (B.1.617.2) is observed following each additional dose. In addition, a strong and polyfunctional T cell response was measured all booster regimens. Interpretation: Real world data is demonstrating that one or more doses of licensed SARS-CoV-2 vaccines confer reduced protection against hospitalisation and deaths caused by divergent VoC, including Omicron. Our data support the ongoing clinical development and testing of booster vaccines to increase immunity against highly mutated VoC. Funding: This research was funded by AstraZeneca with supporting funds from MRC and BBSRC.
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- 2022
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5. Inferior immunogenicity and efficacy of respiratory syncytial virus fusion protein-based subunit vaccine candidates in aged versus young mice.
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Corinne Cayatte, Angie Snell Bennett, Gaurav Manohar Rajani, Leigh Hostetler, Sean K Maynard, Michelle Lazzaro, Patrick McTamney, Kuishu Ren, Terrence O'Day, Michael P McCarthy, and Kirsten Schneider-Ohrum
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Medicine ,Science - Abstract
Respiratory syncytial virus (RSV) is recognized as an important cause of lower and upper respiratory tract infections in older adults, and a successful vaccine would substantially lower morbidity and mortality in this age group. Recently, two vaccine candidates based on soluble purified glycoprotein F (RSV F), either alone or adjuvanted with glucopyranosyl lipid A formulated in a stable emulsion (GLA-SE), failed to reach their primary endpoints in clinical efficacy studies, despite demonstrating the desired immunogenicity profile and efficacy in young rodent models. Here, one of the RSV F vaccine candidates (post-fusion conformation, RSV post-F), and a stabilized pre-fusion form of RSV F (RSV pre-F, DS-Cav1) were evaluated in aged BALB/c mice. Humoral and cellular immunogenicity elicited after immunization of naïve, aged mice was generally lower compared to young animals. In aged mice, RSV post-F vaccination without adjuvant poorly protected the respiratory tract from virus replication, and addition of GLA-SE only improved protection in the lungs, but not in nasal turbinates. RSV pre-F induced higher neutralizing antibody titers compared to RSV post-F (as previously reported) but interestingly, RSV F-specific CD8 T cell responses were lower compared to RSV post-F responses regardless of age. The vaccines were also tested in RSV seropositive aged mice, in which both antigen forms similarly boosted neutralizing antibody titers, although GLA-SE addition boosted neutralizing activity only in RSV pre-F immunized animals. Cell-mediated immune responses in the aged mice were only slightly boosted and well below levels induced in seronegative young mice. Taken together, the findings suggest that the vaccine candidates were not able to induce a strong anti-RSV immune response in recipient mice with an aged immune system, in agreement with recent human clinical trial results. Therefore, the aged mouse model could be a useful tool to evaluate improved vaccine candidates, targeted to prevent RSV disease in older adults.
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- 2017
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6. A Multivalent Polyomavirus Vaccine Elicits Durable Neutralizing Antibody Responses in Macaques
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Alberto Peretti, Diana G. Scorpio, Wing-Pui Kong, Yuk-Ying S. Pang, Michael McCarthy, Kuishu Ren, Moriah Jackson, Barney S. Graham, Christopher B. Buck, Patrick M. McTamney, and Diana V. Pastrana
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Infectious Diseases ,General Veterinary ,General Immunology and Microbiology ,Public Health, Environmental and Occupational Health ,Molecular Medicine - Abstract
In 2019, there were about 100,000 kidney transplants globally, with more than a quarter of them performed in the United States. Unfortunately, some engrafted organs are lost to polyomavirus-associated nephropathy (PyVAN) caused by BK and JC viruses (BKPyV and JCPyV). Transplant patients are routinely monitored for BKPyV viremia, which is an accepted hallmark of nascent nephropathy. If viremia is detected, a reduction in immunosuppressive therapy is standard care, but the intervention comes with increased risk of immune rejection of the engrafted organ. Recent reports have suggested that transplant recipients with high levels of polyomavirus-neutralizing antibodies are protected against PyVAN. Virus-like particle (VLP) vaccines, similar to approved human papillomaviruses vaccines, have an excellent safety record and are known to induce high levels of neutralizing antibodies associated and long-lasting protection from infection. In this study, we demonstrate that VLPs representing BKPyV genotypes I, II, and IV, as well as JCPyV genotype 2 produced in insect cells elicit robust antibody titers. In rhesus macaques, all monkeys developed neutralizing antibody titers above a previously proposed protective threshold of 10,000. A second inoculation, administered 19 weeks after priming, boosted titers to a plateau of ≥25,000 that was maintained for almost two years. No vaccine-related adverse events were observed in any macaques. A multivalent BK/JC VLP immunogen did not show inferiority compared to the single-genotype VLP immunogens. Considering these encouraging results, we believe a clinical trial administering the multivalent VLP vaccine in patients waiting to receive a kidney transplant is warranted to evaluate its ability to reduce or eliminate PyVAN.HIGHLIGHTSRecombinant virus-like particle vaccine was safely administered to rhesus macaquesVaccination generated high-titer neutralizing antibody responsesMultivalent BK/JC polyomavirus vaccine was as effective as monovalent vaccinesHigh neutralizing titers were sustained for 92 weeks without appreciable decline
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- 2022
7. A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
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Valeria Riguero, Jared Delmar, Andrew Dippel, Patrick McTamney, Ethan Luo, Antonio Martinez, Kuishu Ren, Nydia van Dyk, and Ellen O'Connor
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Biotechnology - Published
- 2023
8. Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein
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Kuishu Ren, Shao Huan Samuel Weng, Sonja Hess, Raffaello Cimbro, Mahboobe Ghaedi, William F Dall'Acqua, Yasuhiro Ikeda, Nicole Kallewaard-Lelay, Lori Clarke, Chelsea C. Boo, Lisa H. Cazares, Sarav Rajan, Patrick M. McTamney, Raghothama Chaerkady, and Hsiang En Hsu
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Resolution (mass spectrometry) ,Viral protein ,viruses ,Pneumonia, Viral ,010402 general chemistry ,Orbitrap ,medicine.disease_cause ,01 natural sciences ,Mass Spectrometry ,Article ,Analytical Chemistry ,law.invention ,Betacoronavirus ,Limit of Detection ,law ,medicine ,Coronavirus Nucleocapsid Proteins ,Humans ,Nanotechnology ,Amino Acid Sequence ,Pandemics ,Chromatography, High Pressure Liquid ,Detection limit ,Chromatography ,SARS-CoV-2 ,Chemistry ,010401 analytical chemistry ,COVID-19 ,Nucleocapsid Proteins ,Phosphoproteins ,0104 chemical sciences ,Nucleoprotein ,Reverse transcription polymerase chain reaction ,Targeted mass spectrometry ,Spike Glycoprotein, Coronavirus ,RNA extraction ,Coronavirus Infections - Abstract
There is an urgent need for robust and high-throughput methods for SARS-CoV-2 detection in suspected patient samples to facilitate disease management, surveillance, and control. Although nucleic acid detection methods such as reverse transcription polymerase chain reaction (RT-PCR) are the gold standard, during the current pandemic, the deployment of RT-PCR tests has been extremely slow, and key reagents such as PCR primers and RNA extraction kits are at critical shortages. Rapid point-of-care viral antigen detection methods have been previously employed for the diagnosis of respiratory viruses such as influenza and respiratory syncytial viruses. Therefore, the direct detection of SARS-CoV-2 viral antigens in patient samples could also be used for diagnosis of active infection, and alternative methodologies for specific and sensitive viral protein detection should be explored. Targeted mass spectrometry techniques have enabled the identification and quantitation of a defined subset of proteins/peptides at single amino acid resolution with attomole level sensitivity and high reproducibility. Herein, we report a targeted mass spectrometry assay for the detection of SARS-CoV-2 spike protein and nucleoprotein in a relevant biological matrix. Recombinant full-length spike protein and nucleoprotein were digested and proteotypic peptides were selected for parallel reaction monitoring (PRM) quantitation using a high-resolution Orbitrap instrument. A spectral library, which contained seven proteotypic peptides (four from spike protein and three from nucleoprotein) and the top three to four transitions, was generated and evaluated. From the original spectral library, we selected two best performing peptides for the final PRM assay. The assay was evaluated using mock test samples containing inactivated SARS-CoV-2 virions, added to in vitro derived mucus. The PRM assay provided a limit of detection of ∼200 attomoles and a limit of quantitation of ∼ 390 attomoles. Extrapolating from the test samples, the projected titer of virus particles necessary for the detection of SARS-CoV-2 spike and nucleoprotein detection was approximately 2 × 105 viral particles/mL, making it an attractive alternative to RT-PCR assays. Potentially, mass spectrometry-based methods for viral antigen detection may deliver higher throughput and could serve as a complementary diagnostic tool to RT-PCR. Furthermore, this assay could be used to evaluate the presence of SARS-CoV-2 in archived or recently collected biological fluids, in vitro-derived research materials, and wastewater samples.
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- 2020
9. The ChAdOx1 vectored vaccine, AZD2816, induces strong immunogenicity against SARS-CoV-2 beta (B.1.351) and other variants of concern in preclinical studies
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Alexandra J Spencer, Susan Morris, Marta Ulaszewska, Claire Powers, Reshma Kailath, Cameron Bissett, Adam Truby, Nazia Thakur, Joseph Newman, Elizabeth R Allen, Indra Rudiansyah, Chang Liu, Wanwisa Dejnirattisai, Juthathip Mongkolsapaya, Hannah Davies, Francesca R Donnellan, David Pulido, Thomas P. Peacock, Wendy S. Barclay, Helen Bright, Kuishu Ren, Gavin Screaton, Patrick McTamney, Dalan Bailey, Sarah C Gilbert, and Teresa Lambe
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COVID-19 Vaccines ,SARS-CoV-2 ,ChAdOx1 nCoV-19 ,COVID-19 ,Humans ,General Medicine ,General Biochemistry, Genetics and Molecular Biology - Abstract
Background There is an ongoing global effort to design, manufacture, and clinically assess vaccines against SARS-CoV-2. Over the course of the ongoing pandemic a number of new SARS-CoV-2 virus isolates or variants of concern (VoC) have been identified containing mutations in key proteins. Methods In this study we describe the generation and preclinical assessment of a ChAdOx1-vectored vaccine (AZD2816) which expresses the spike protein of the Beta VoC (B.1.351). Findings We demonstrate that AZD2816 is immunogenic after a single dose. When AZD2816 is used as a booster dose in animals primed with a vaccine encoding the original spike protein (ChAdOx1 nCoV-19/ [AZD1222]), an increase in binding and neutralising antibodies against Beta (B.1.351), Gamma (P.1) and Delta (B.1.617.2) is observed following each additional dose. In addition, a strong and polyfunctional T cell response was measured all booster regimens. Interpretation Real world data is demonstrating that one or more doses of licensed SARS-CoV-2 vaccines confer reduced protection against hospitalisation and deaths caused by divergent VoC, including Omicron. Our data support the ongoing clinical development and testing of booster vaccines to increase immunity against highly mutated VoC. Funding This research was funded by AstraZeneca with supporting funds from MRC and BBSRC.
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- 2021
10. AZD7442 demonstrates prophylactic and therapeutic efficacy in non-human primates and extended half-life in humans
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Ana I. Kuehne, Katie Streicher, Richard Roque, Yueh-Ming Loo, Jerome Bouquet, Daniel J Flores, Robert A. Gasser, Michael E Abram, Nicholas J. Bond, Andrew S. Herbert, Mark T. Esser, John M. Dye, Elizabeth J. Kelly, Rosalinda H Arends, Kim Rosenthal, Nicole L. Kallewaard, Owen Cornwell, Kevin M Tuffy, Hanne Andersen, Yue Huang, James E. Crowe, Helen Bright, Anastasia Aksyuk, Menelas N. Pangalos, Robert H. Carnahan, Patrick M. McTamney, James Dunyak, Seme Diallo, Kuishu Ren, Anton I. Rosenbaum, and Lily Cheng
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Lung ,business.industry ,medicine.drug_class ,Half-life ,Mucous membrane of nose ,Monoclonal antibody ,Epitope ,Virus ,Titer ,medicine.anatomical_structure ,Immunology ,Medicine ,business ,Receptor - Abstract
Despite the success of SARS-CoV-2 vaccines, there remains a need for more prevention and treatment options for individuals remaining at risk of COVID-19. Monoclonal antibodies (mAbs) against the viral spike protein have potential to both prevent and treat COVID-19, and reduce the risk of severe disease and death. Here, we describe AZD7442, a combination of two mAbs, AZD8895 (tixagevimab) and AZD1061 (cilgavimab), that simultaneously bind to distinct non-overlapping epitopes on the spike protein receptor binding domain to potently neutralize SARS-CoV-2. Initially isolated from individuals with prior SARS-CoV-2 infection, the two mAbs were designed to extend their half-lives and abrogate effector functions. The AZD7442 mAbs individually prevent the spike protein from binding to angiotensin-converting enzyme 2 receptor, blocking virus cell entry. Together, these two mAbs create a higher barrier to viral escape and a wider breadth of coverage, neutralizing all known SARS-CoV-2 variants of concern. In a non-human primate model of SARS-CoV-2 infection, prophylactic AZD7442 administration prevented infection, while therapeutic administration accelerated virus clearance from lung. In an ongoing Phase I study in healthy participants (NCT04507256), 300 mg intramuscular AZD7442 provided SARS-CoV-2 serum geometric mean neutralizing titers >10-fold above those of convalescent sera for ≥3 months, which remained 3-fold above those of convalescent sera 9 months post-AZD7442 administration. Approximately 1–2% of serum AZD7442 levels were detected in nasal mucosa, a site of SARS-CoV-2 infection. Extrapolation of the time course of serum AZD7442 concentrations suggests AZD7442 may provide up to 12 months of protection and benefit individuals at high-risk of COVID-19.
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- 2021
11. The ChAdOx1 vectored vaccine, AZD2816, induces strong immunogenicity against SARS-CoV-2 Beta (B.1.351) and other variants of concern in preclinical studies
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Francesca R. Donnellan, Gavin R. Screaton, Wanwisa Dejnirattisai, Juthathip Mongkolsapaya, Nazia Thakur, A Truby, Susan J. Morris, Wendy S. Barclay, Sarah C. Gilbert, Reshma Kailath, David Pulido, Cameron Bissett, Thomas P. Peacock, Joseph Newman, Alexandra J. Spencer, Teresa Lambe, Chang Liu, Marta Ulaszewska, Elizabeth R. Allen, Claire Powers, Dalan Bailey, P McTammy, Hannah Davies, Helen Bright, and Kuishu Ren
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biology ,business.industry ,Immunogenicity ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Booster dose ,Virology ,Virus ,Neutralization ,Pandemic ,biology.protein ,Medicine ,Antibody ,business ,Original antigenic sin - Abstract
There is an ongoing global effort to design, manufacture, and clinically assess vaccines against SARS-CoV-2. Over the course of the ongoing pandemic a number of new SARS-CoV-2 virus isolates or variants of concern (VoC) have been identified containing mutations in key proteins. In this study we describe the generation and preclinical assessment of a ChAdOx1-vectored vaccine (AZD2816) which expresses the spike protein of the Beta VoC (B.1.351). We demonstrate that AZD2816 is immunogenic after a single dose. When AZD2816 is used as a booster dose in animals primed with a vaccine encoding the original spike protein (ChAdOx1 nCoV-19/ [AZD1222]), high titre binding and neutralising antibodies against Beta (B.1.351), Gamma (P.1) and Delta (B.1.617.2) are induced. In addition, a strong and polyfunctional T cell response was measured in these booster regimens. These data support the ongoing clinical development and testing of this new variant vaccine.
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- 2021
12. Potently neutralizing human antibodies that block SARS-CoV-2 receptor binding and protect animals
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David R. Martinez, Ralph S. Baric, Elaine C. Chen, Luke Myers, Natasha M. Kafai, Yueh-Ming Loo, Ahmed O. Hassan, Samuel Day, Nicole L. Kallewaard, Larissa B. Thackray, Rita E. Chen, James Brett Case, James E. Crowe, Seth J. Zost, Rachel S. Nargi, Pavlo Gilchuk, Lisa E. Gralinski, Kuishu Ren, Elad Binshtein, Lauren E. Williamson, Taylor Jones, Rachel E. Sutton, Naveenchandra Suryadevara, James J. Steinhardt, Alexandra Schäfer, Emma S. Winkler, Julie M. Fox, Michael S. Diamond, Andrew Trivette, Robert H. Carnahan, and Joseph X. Reidy
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chemistry.chemical_classification ,biology ,medicine.drug_class ,SARS-CoV-2 ,COVID-19 ,Antibodies, Monoclonal ,SARS-CoV ,Adaptive Immunity ,Monoclonal antibody ,Virology ,Neutralization ,Virus ,Epitope ,Article ,Coronavirus ,chemistry ,Antigen ,Humoral immunity ,biology.protein ,medicine ,Antibody ,Glycoprotein ,Human - Abstract
The COVID-19 pandemic is a major threat to global health for which there are only limited medical countermeasures, and we lack a thorough understanding of mechanisms of humoral immunity1,2. From a panel of monoclonal antibodies (mAbs) targeting the spike (S) glycoprotein isolated from the B cells of infected subjects, we identified several mAbs that exhibited potent neutralizing activity with IC50values as low as 0.9 or 15 ng/mL in pseudovirus or wild-type (wt) SARS-CoV-2 neutralization tests, respectively. The most potent mAbs fully block the receptor-binding domain of S (SRBD) from interacting with human ACE2. Competition-binding, structural, and functional studies allowed clustering of the mAbs into defined classes recognizing distinct epitopes within major antigenic sites on the SRBD. Electron microscopy studies revealed that these mAbs recognize distinct conformational states of trimeric S protein. Potent neutralizing mAbs recognizing unique sites, COV2-2196 and COV2-2130, bound simultaneously to S and synergistically neutralized authentic SARS-CoV-2 virus. In two murine models of SARS-CoV-2 infection, passive transfer of either COV2-2916 or COV2-2130 alone or a combination of both mAbs protected mice from severe weight loss and reduced viral burden and inflammation in the lung. These results identify protective epitopes on the SRBDand provide a structure-based framework for rational vaccine design and the selection of robust immunotherapeutic cocktails.
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- 2020
13. Prevalence and Significance of Substitutions in the Fusion Protein of Respiratory Syncytial Virus Resulting in Neutralization Escape From Antibody MEDI8897
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Nicole L. Kallewaard, Susan R. Palaszynski, Steve Wong, Fiona Fernandes, Weijia Wang, Seme Diallo, Nancy Ulbrandt, Mark T. Esser, Bin Lu, Catherine Svabek, Patrick M. Mctamney, Kuishu Ren, Qing Zhu, Hong Jing, JoAnn Suzich, and Brian Moldt
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0301 basic medicine ,Protein Conformation ,medicine.drug_class ,viruses ,030106 microbiology ,Population ,Virus Attachment ,Respiratory Syncytial Virus Infections ,Antibodies, Viral ,Crystallography, X-Ray ,Virus Replication ,medicine.disease_cause ,Monoclonal antibody ,Neutralization ,Virus ,03 medical and health sciences ,Gene Frequency ,Neutralization Tests ,Drug Resistance, Viral ,Prevalence ,medicine ,Humans ,Immunologic Factors ,Immunology and Allergy ,education ,Immune Evasion ,Biological Products ,Mutation ,education.field_of_study ,Binding Sites ,biology ,Antibodies, Neutralizing ,Virology ,Fusion protein ,United States ,030104 developmental biology ,Infectious Diseases ,Amino Acid Substitution ,Viral replication ,Respiratory Syncytial Virus, Human ,biology.protein ,Mutant Proteins ,Antibody ,Viral Fusion Proteins - Abstract
Background Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infection among infants and young children. To date, no vaccine is approved for the broad population of healthy infants. MEDI8897, a potent anti-RSV fusion antibody with extended serum half-life, is currently under clinical investigation as a potential passive RSV vaccine for all infants. As a ribonucleic acid virus, RSV is prone to mutation, and the possibility of viral escape from MEDI8897 neutralization is a potential concern. Methods We generated RSV monoclonal antibody (mAb)-resistant mutants (MARMs) in vitro and studied the effect of the amino acid substitutions identified on binding and viral neutralization susceptibility to MEDI8897. The impact of resistance-associated mutations on in vitro growth kinetics and the prevalence of these mutations in currently circulating strains of RSV in the United States was assessed. Results Critical residues identified in MARMs for MEDI8897 neutralization were located in the MEDI8897 binding site defined by crystallographic analysis. Substitutions in these residues affected the binding of mAb to virus, without significant impact on viral replication in vitro. The frequency of natural resistance-associated polymorphisms was low. Conclusions Results from this study provide insights into the mechanism of MEDI8897 escape and the complexity of monitoring for emergence of resistance.
- Published
- 2018
14. Potently neutralizing and protective human antibodies against SARS-CoV-2
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Taylor Jones, Pavlo Gilchuk, Benjamin K. Mueller, Rachel S. Nargi, Rita E. Chen, Yueh-Ming Loo, Natasha M. Kafai, Broc T. McCune, Larissa B. Thackray, Joseph P. Nkolola, Alexandra Schäfer, Rachel E. Sutton, James Brett Case, Michael S. Diamond, Luke Myers, Andrew Trivette, Elad Binshtein, Emma S. Winkler, Nicole L. Kallewaard, Samuel Day, Swathi Shrihari, Jens Meiler, Shamus P. Keeler, Kuishu Ren, Noe B. Mercado, Lauren E. Williamson, Seth J. Zost, Abishek Chandrashekar, Dan H. Barouch, Robert H. Carnahan, David R. Martinez, Ralph S. Baric, James E. Crowe, Elaine C. Chen, Joseph X. Reidy, Naveenchandra Suryadevara, James J. Steinhardt, Ahmed O. Hassan, Julie M. Fox, Michael J. Holtzman, and Lisa E. Gralinski
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0301 basic medicine ,Male ,medicine.drug_class ,Pneumonia, Viral ,Inflammation ,Biology ,Cross Reactions ,Peptidyl-Dipeptidase A ,Monoclonal antibody ,Antibodies, Viral ,Severe Acute Respiratory Syndrome ,Binding, Competitive ,Virus ,Epitope ,Article ,Cell Line ,03 medical and health sciences ,Betacoronavirus ,Mice ,0302 clinical medicine ,Neutralization Tests ,medicine ,Animals ,Humans ,Pandemics ,Multidisciplinary ,SARS-CoV-2 ,Antibodies, Monoclonal ,COVID-19 ,Middle Aged ,Virology ,Antibodies, Neutralizing ,Macaca mulatta ,Disease Models, Animal ,030104 developmental biology ,Severe acute respiratory syndrome-related coronavirus ,Cell culture ,Humoral immunity ,Spike Glycoprotein, Coronavirus ,biology.protein ,Epitopes, B-Lymphocyte ,Female ,Pre-Exposure Prophylaxis ,Angiotensin-Converting Enzyme 2 ,Antibody ,medicine.symptom ,Coronavirus Infections ,Viral load ,030217 neurology & neurosurgery - Abstract
The ongoing pandemic of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a major threat to global health1 and the medical countermeasures available so far are limited2,3. Moreover, we currently lack a thorough understanding of the mechanisms of humoral immunity to SARS-CoV-24. Here we analyse a large panel of human monoclonal antibodies that target the spike (S) glycoprotein5, and identify several that exhibit potent neutralizing activity and fully block the receptor-binding domain of the S protein (SRBD) from interacting with human angiotensin-converting enzyme 2 (ACE2). Using competition-binding, structural and functional studies, we show that the monoclonal antibodies can be clustered into classes that recognize distinct epitopes on the SRBD, as well as distinct conformational states of the S trimer. Two potently neutralizing monoclonal antibodies, COV2-2196 and COV2-2130, which recognize non-overlapping sites, bound simultaneously to the S protein and neutralized wild-type SARS-CoV-2 virus in a synergistic manner. In two mouse models of SARS-CoV-2 infection, passive transfer of COV2-2196, COV2-2130 or a combination of both of these antibodies protected mice from weight loss and reduced the viral burden and levels of inflammation in the lungs. In addition, passive transfer of either of two of the most potent ACE2-blocking monoclonal antibodies (COV2-2196 or COV2-2381) as monotherapy protected rhesus macaques from SARS-CoV-2 infection. These results identify protective epitopes on the SRBD and provide a structure-based framework for rational vaccine design and the selection of robust immunotherapeutic agents. An analysis identifies human monoclonal antibodies that potently neutralize wild-type SARS-CoV-2 and protect animals from disease, including two that synergize in a cocktail, suggesting that these could be candidates for use as therapeutic agents for the treatment of COVID-19 in humans.
- Published
- 2019
15. Immunization with Low Doses of Recombinant Postfusion or Prefusion Respiratory Syncytial Virus F Primes for Vaccine-Enhanced Disease in the Cotton Rat Model Independently of the Presence of a Th1-Biasing (GLA-SE) or Th2-Biasing (Alum) Adjuvant
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Lily Cheng, Krystal Nacel, Kirsten Schneider-Ohrum, Corinne Cayatte, Michael P. McCarthy, Terrence O'Day, Patrick M. Mctamney, Kuishu Ren, Gaurav Manohar Rajani, Angie Snell Bennett, Leigh Hostetler, and Gregory A. Prince
- Subjects
0301 basic medicine ,viruses ,medicine.medical_treatment ,Immunology ,Respiratory Syncytial Virus Infections ,Antibodies, Viral ,Microbiology ,Virus ,03 medical and health sciences ,Th2 Cells ,0302 clinical medicine ,Adjuvants, Immunologic ,Antigen ,Virology ,Vaccines and Antiviral Agents ,Respiratory Syncytial Virus Vaccines ,medicine ,Animals ,Antibody-dependent enhancement ,Sigmodontinae ,030212 general & internal medicine ,Cotton rat ,Alum adjuvant ,Vaccines, Synthetic ,biology ,Th1 Cells ,respiratory system ,biology.organism_classification ,Antibodies, Neutralizing ,Antibody-Dependent Enhancement ,Recombinant Proteins ,Disease Models, Animal ,Lipid A ,030104 developmental biology ,Insect Science ,biology.protein ,Alum Compounds ,Antibody ,Viral Fusion Proteins ,Adjuvant - Abstract
Respiratory syncytial virus (RSV) infection of children previously immunized with a nonlive, formalin-inactivated (FI)-RSV vaccine has been associated with serious enhanced respiratory disease (ERD). Consequently, detailed studies of potential ERD are a critical step in the development of nonlive RSV vaccines targeting RSV-naive children and infants. The fusion glycoprotein (F) of RSV in either its postfusion (post-F) or prefusion (pre-F) conformation is a target for neutralizing antibodies and therefore an attractive antigen candidate for a pediatric RSV subunit vaccine. Here, we report the evaluation of RSV post-F and pre-F in combination with glucopyranosyl lipid A (GLA) integrated into stable emulsion (SE) (GLA-SE) and alum adjuvants in the cotton rat model. Immunization with optimal doses of RSV F antigens in the presence of GLA-SE induced high titers of virus-neutralizing antibodies and conferred complete lung protection from virus challenge, with no ERD signs in the form of alveolitis. To mimic a waning immune response, and to assess priming for ERD under suboptimal conditions, an antigen dose de-escalation study was performed in the presence of either GLA-SE or alum. At low RSV F doses, alveolitis-associated histopathology was unexpectedly observed with either adjuvant at levels comparable to FI-RSV-immunized controls. This occurred despite neutralizing-antibody titers above the minimum levels required for protection and with no/low virus replication in the lungs. These results emphasize the need to investigate a pediatric RSV vaccine candidate carefully for priming of ERD over a wide dose range, even in the presence of strong neutralizing activity, Th1 bias-inducing adjuvant, and protection from virus replication in the lower respiratory tract. IMPORTANCE RSV disease is of great importance worldwide, with the highest burden of serious disease occurring upon primary infection in infants and children. FI-RSV-induced enhanced disease, observed in the 1960s, presented a major and ongoing obstacle for the development of nonlive RSV vaccine candidates. The findings presented here underscore the need to evaluate a nonlive RSV vaccine candidate during preclinical development over a wide dose range in the cotton rat RSV enhanced-disease model, as suboptimal dosing of several RSV F subunit vaccine candidates led to the priming for ERD. These observations are relevant to the validity of the cotton rat model itself and to safe development of nonlive RSV vaccines for seronegative infants and children.
- Published
- 2017
16. Evidence for sex steroid inhibition of lipoprotein lipase in men: comparison of abdominal and femoral adipose tissue
- Author
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Martha P. McMurry, Karon J. Felten, Per-Henrik Iverius, Kuishu Ren, A. Wayne Meikle, Maria E. Ramirez, and Gail A. Wiebke
- Subjects
Adult ,Male ,medicine.medical_specialty ,Time Factors ,Endocrinology, Diabetes and Metabolism ,Adipose tissue ,Biology ,Endocrinology ,Internal medicine ,Abdomen ,medicine ,Humans ,Testosterone ,Femur ,Gonadal Steroid Hormones ,Lipoprotein lipase ,Anthropometry ,Estradiol ,Heparin ,Weight change ,Body Weight ,Middle Aged ,medicine.disease ,Obesity ,Body Height ,Lipoprotein Lipase ,Adipose Tissue ,Sex steroid ,Dihydrotestosterone ,lipids (amino acids, peptides, and proteins) ,medicine.drug - Abstract
Plasma estradiol has been suggested to suppress adipose tissue lipoprotein lipase (LPL) activity in women. The present study explores the regulation of LPL by sex steroids in sedentary obese men (N = 24) at their usual weight. Femoral adipose tissue LPL activity, eluted with serum and heparin or extracted with detergent, showed significant inverse correlations with plasma levels of testosterone, bioavailable testosterone, dihydrotestosterone, and estradiol. Both measures of femoral LPL activity were also correlated with the weight change occurring despite efforts to maintain a constant weight. Abdominal LPL activity showed significant but weaker inverse correlations with bioavailable testosterone only. Multivariate analysis of potential predictors for eluted femoral LPL activity showed that plasma testosterone, dihydrotestosterone, and estradiol were interdependent, whereas the rate of weight change was an independent variable. In the regression equation, only bioavailable testosterone and weight change were retained, explaining 63% of the variability (R = .79, P = .0002). These results suggest that sex steroids suppress adipose tissue LPL activity in men, and more so in the thigh than in the abdomen, thereby possibly contributing to a central fat accumulation. The data are compatible with a model from male animals suggesting that testosterone effects on adipose tissue LPL are mediated by estradiol formed locally.
- Published
- 1997
17. Increased lipase inhibition in uremia: Identification of pre-β-HDL as a major inhibitor in normal and uremic plasma
- Author
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Alfred K. Cheung, Kuishu Ren, Per Henrik Iverius, and Charles J. Parker
- Subjects
medicine.medical_specialty ,Population ,030204 cardiovascular system & hematology ,High-Density Lipoproteins, Pre-beta ,Lipase inhibitor activity ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Humans ,Lipase ,Enzyme Inhibitors ,education ,Triglycerides ,030304 developmental biology ,Uremia ,Hypertriglyceridemia ,0303 health sciences ,education.field_of_study ,Lipoprotein lipase ,biology ,Chemistry ,medicine.disease ,Endocrinology ,Nephrology ,Lipase inhibitors ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Hepatic lipase ,Lipoproteins, HDL - Abstract
Increased lipase inhibition in uremia: Identification of pre-β-HDL as a major inhibitor in normal and uremic plasma. The hypertriglyceridemia commonly observed in uremia has been attributed to an abnormally high inhibitor activity in plasma for lipoprotein lipase (LPL) and hepatic lipase (HL), both of which have a key role in lipoprotein metabolism. The purpose of this investigation was to establish a relationship between plasma lipase inhibitor activity and hypertriglyceridemia, identify the main plasma lipase inhibitor, and determine the basis for the greater inhibitor activity in uremia. In a mixed population of normal (N = 8) and uremic subjects (N = 12), log-transformed plasma triglycerides correlated with both inhibitor activity and uremic status. However, inhibitor activity was the only retained predictor variable for triglycerides in a multiple linear regression model (r = 0.91; P < 0.0001). An inhibitor isolated from normal plasma was identified as a particle containing apolipoprotein A-I (apo A-I) and 3% phospholipid. This particle, which has pre-β electrophoretic mobility and a Stokes' radius of 54 Å, therefore corresponds to a form of the previously described pre-β-HDL (free apo A-I) in the non-lipoprotein fraction of plasma. Comparison of normal and uremic plasma indicated that the greater lipase inhibitor activity in the latter could be attributed to an increased concentration of apo A-I in the non-lipoprotein fraction of plasma (pre-β-HDL), as well as to increased inhibition by the uremic lipoproteins. The increased plasma lipase inhibitor activity may be important in the pathogenesis of hypertriglyceridemia in chronic renal failure.
- Full Text
- View/download PDF
18. Immunization with Low Doses of Recombinant Postfusion or Prefusion Respiratory Syncytial Virus F Primes for Vaccine-Enhanced Disease in the Cotton Rat Model Independently of the Presence of a Th1-Biasing (GLA-SE) or Th2-Biasing (Alum) Adjuvant.
- Author
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Schneider-Ohrum, Kirsten, Cayatte, Corinne, Bennett, Angie Snell, Rajani, Gaurav Manohar, McTamney, Patrick, Nacel, Krystal, Hostetler, Leigh, Lily Cheng, Kuishu Ren, O'Day, Terrence, Prince, Gregory A., and McCarthy, Michael P.
- Subjects
- *
RESPIRATORY syncytial virus infections , *RECOMBINANT viruses , *VIRAL vaccines , *LABORATORY rats , *FORMALDEHYDE , *JUVENILE diseases , *THERAPEUTICS - Abstract
Respiratory syncytial virus (RSV) infection of children previously immunized with a nonlive, formalin-inactivated (FI)-RSV vaccine has been associated with serious enhanced respiratory disease (ERD). Consequently, detailed studies of potential ERD are a critical step in the development of nonlive RSV vaccines targeting RSV-naive children and infants. The fusion glycoprotein (F) of RSV in either its postfusion (post-F) or prefusion (pre-F) conformation is a target for neutralizing antibodies and therefore an attractive antigen candidate for a pediatric RSV subunit vaccine. Here, we report the evaluation of RSV post-F and pre-F in combination with glucopyranosyl lipid A (GLA) integrated into stable emulsion (SE) (GLA-SE) and alum adjuvants in the cotton rat model. Immunization with optimal doses of RSV F antigens in the presence of GLA-SE induced high titers of virus-neutralizing antibodies and conferred complete lung protection from virus challenge, with no ERD signs in the form of alveolitis. To mimic a waning immune response, and to assess priming for ERD under suboptimal conditions, an antigen dose de-escalation study was performed in the presence of either GLA-SE or alum. At low RSV F doses, alveolitis-associated histopathology was unexpectedly observed with either adjuvant at levels comparable to FI-RSV-immunized controls. This occurred despite neutralizing-antibody titers above the minimum levels required for protection and with no/low virus replication in the lungs. These results emphasize the need to investigate a pediatric RSV vaccine candidate carefully for priming of ERD over a wide dose range, even in the presence of strong neutralizing activity, Th1 bias-inducing adjuvant, and protection from virus replication in the lower respiratory tract. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
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