232 results on '"Jun-Gyu Park"'
Search Results
2. A Noble Extract of Pseudomonas sp. M20A4R8 Efficiently Controlling the Influenza Virus-Induced Cell Death
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Su-Bin Jung, Grace Choi, Hyo-Jin Kim, Kyeong-Seo Moon, Gun Lee, Kyeong-Hak Na, Yong Min Kwon, Jimin Moon, Mi Yeong Shin, Jae-Yeong Yu, Yeong-Bin Baek, Jun-Gyu Park, and Sang-Ik Park
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Pseudomonas ,influenza virus ,broad-spectrum therapeutics ,marine bacterium ,cell death ,Biology (General) ,QH301-705.5 - Abstract
Epidemic diseases that arise from infectious RNA viruses, particularly influenza viruses, pose a constant threat to the global economy and public health. Viral evolution has undermined the efficacy of acquired immunity from vaccines and the antiviral effects of FDA-approved drugs. As such, there is an urgent need to develop new antiviral lead agents. Natural compounds, owing to their historical validation of application and safety, have become a promising solution. In this light, a novel marine bacterium, Pseudomonas sp. M20A4R8, has been found to exhibit significant antiviral activity [half maximal inhibitory concentration (IC50) = 1.3 µg/mL, selectivity index (SI) = 919.4] against influenza virus A/Puerto Rico/8/34, surpassing the activity of chloroquine. The antiviral response via M20A4R8 extract was induced during post-entry stages of the influenza virus, indicating suitability for post-application after the establishment of viral infection. Furthermore, post-treatment with M20A4R8 extract protected the host from virus-induced apoptosis, suggesting its potential use in acute respiratory disease complexes resulting from immune effectors’ overstimulation and autophagy-mediated self-apoptosis. The extract demonstrated an outstanding therapeutic index against influenza virus A/Wisconsin/15/2009 (IC50 = 8.1 µg/mL, SI = 146.2) and B/Florida/78/2015 Victoria lineage (IC50 = 3.5 µg/mL, SI = 343.8), indicating a broad anti-influenza virus activity with guaranteed safety and effectiveness. This study provides a new perspective on mechanisms for preventing a broad spectrum of viral infections through antiviral agents from novel and natural origins. Future studies on a single or combined compound from the extract hold promise, encouraging its use in preclinical challenge tests with various influenza virus strains.
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- 2024
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3. Modeling SARS-CoV-2 and influenza infections and antiviral treatments in human lung epithelial tissue equivalents
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Hoda Zarkoob, Anna Allué-Guardia, Yu-Chi Chen, Andreu Garcia-Vilanova, Olive Jung, Steven Coon, Min Jae Song, Jun-Gyu Park, Fatai Oladunni, Jesse Miller, Yen-Ting Tung, Ivan Kosik, David Schultz, James Iben, Tianwei Li, Jiaqi Fu, Forbes D. Porter, Jonathan Yewdell, Luis Martinez-Sobrido, Sara Cherry, Jordi B. Torrelles, Marc Ferrer, and Emily M. Lee
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Biology (General) ,QH301-705.5 - Abstract
Human alveolar and tracheobronchial epithelial air liquid interface (ALI) tissues are used as models to examine cellular responses to SARS-CoV-2 and influenza A virus infections and as antiviral drug screening assay platforms.
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- 2022
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4. Immunization with Recombinant Accessory Protein-Deficient SARS-CoV-2 Protects against Lethal Challenge and Viral Transmission
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Chengjin Ye, Jun-Gyu Park, Kevin Chiem, Piyush Dravid, Anna Allué-Guardia, Andreu Garcia-Vilanova, Paula Pino Tamayo, Vinay Shivanna, Amit Kapoor, Mark R. Walter, James J. Kobie, Richard K. Plemper, Jordi B. Torrelles, and Luis Martinez-Sobrido
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SARS-CoV-2 ,live-attenuated vaccine ,immune protection ,viral shedding ,viral transmission ,coronavirus ,Microbiology ,QR1-502 - Abstract
ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to a worldwide coronavirus disease 2019 (COVID-19) pandemic. Despite the high efficacy of the authorized vaccines, there may be uncertain and unknown side effects or disadvantages associated with current vaccination approaches. Live-attenuated vaccines (LAVs) have been shown to elicit robust and long-term protection by the induction of host innate and adaptive immune responses. In this study, we sought to verify an attenuation strategy by generating 3 double open reading frame (ORF)-deficient recombinant SARS-CoV-2s (rSARS-CoV-2s) simultaneously lacking two accessory ORF proteins (ORF3a/ORF6, ORF3a/ORF7a, and ORF3a/ORF7b). We report that these double ORF-deficient rSARS-CoV-2s have slower replication kinetics and reduced fitness in cultured cells compared with their parental wild-type (WT) counterpart. Importantly, these double ORF-deficient rSARS-CoV-2s showed attenuation in both K18 hACE2 transgenic mice and golden Syrian hamsters. A single intranasal dose vaccination induced high levels of neutralizing antibodies against SARS-CoV-2 and some variants of concern and activated viral component-specific T cell responses. Notably, double ORF-deficient rSARS-CoV-2s were able to protect, as determined by the inhibition of viral replication, shedding, and transmission, against challenge with SARS-CoV-2 in both K18 hACE2 mice and golden Syrian hamsters. Collectively, our results demonstrate the feasibility of implementing the double ORF-deficient strategy to develop safe, immunogenic, and protective LAVs to prevent SARS-CoV-2 infection and associated COVID-19. IMPORTANCE Live-attenuated vaccines (LAVs) are able to induce robust immune responses, including both humoral and cellular immunity, representing a very promising option to provide broad and long-term immunity. To develop LAVs for SARS-CoV-2, we engineered attenuated recombinant SARS-CoV-2 (rSARS-CoV-2) that simultaneously lacks the viral open reading frame 3a (ORF3a) in combination with either ORF6, ORF7a, or ORF7b (Δ3a/Δ6, Δ3a/Δ7a, and Δ3a/Δ7b, respectively) proteins. Among them, the rSARS-CoV-2 Δ3a/Δ7b was completely attenuated and able to provide 100% protection against an otherwise lethal challenge in K18 hACE2 transgenic mice. Moreover, the rSARS-CoV-2 Δ3a/Δ7b conferred protection against viral transmission between golden Syrian hamsters.
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- 2023
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5. Identification of a conserved S2 epitope present on spike proteins from all highly pathogenic coronaviruses
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Rui P Silva, Yimin Huang, Annalee W Nguyen, Ching-Lin Hsieh, Oladimeji S Olaluwoye, Tamer S Kaoud, Rebecca E Wilen, Ahlam N Qerqez, Jun-Gyu Park, Ahmed M Khalil, Laura R Azouz, Kevin C Le, Amanda L Bohanon, Andrea M DiVenere, Yutong Liu, Alison G Lee, Dzifa A Amengor, Sophie R Shoemaker, Shawn M Costello, Eduardo A Padlan, Susan Marqusee, Luis Martinez-Sobrido, Kevin N Dalby, Sheena D'Arcy, Jason S McLellan, and Jennifer A Maynard
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coronavirus ,MERS-CoV ,spike ,SARS-CoV-2 ,S2 core ,S2 hinge ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
To address the ongoing SARS-CoV-2 pandemic and prepare for future coronavirus outbreaks, understanding the protective potential of epitopes conserved across SARS-CoV-2 variants and coronavirus lineages is essential. We describe a highly conserved, conformational S2 domain epitope present only in the prefusion core of β-coronaviruses: SARS-CoV-2 S2 apex residues 980–1006 in the flexible hinge. Antibody RAY53 binds the native hinge in MERS-CoV and SARS-CoV-2 spikes on the surface of mammalian cells and mediates antibody-dependent cellular phagocytosis and cytotoxicity against SARS-CoV-2 spike in vitro. Hinge epitope mutations that ablate antibody binding compromise pseudovirus infectivity, but changes elsewhere that affect spike opening dynamics, including those found in Omicron BA.1, occlude the epitope and may evade pre-existing serum antibodies targeting the S2 core. This work defines a third class of S2 antibody while providing insights into the potency and limitations of S2 core epitope targeting.
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- 2023
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6. Monitoring SARS-CoV-2 Infection Using a Double Reporter-Expressing Virus
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Kevin Chiem, Jun-Gyu Park, Desarey Morales Vasquez, Richard K. Plemper, Jordi B. Torrelles, James J. Kobie, Mark R. Walter, Chengjin Ye, and Luis Martinez-Sobrido
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coronavirus ,fluorescent ,infection ,luciferase ,recombinant ,reporter ,Microbiology ,QR1-502 - Abstract
ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the highly contagious agent responsible for the coronavirus disease 2019 (COVID-19) pandemic. An essential requirement for understanding SARS-CoV-2 biology and the impact of antiviral therapeutics is a robust method to detect the presence of the virus in infected cells or animal models. Despite the development and successful generation of recombinant (r)SARS-CoV-2-expressing fluorescent or luciferase reporter genes, knowledge acquired from their use in in vitro assays and/or in live animals is limited to the properties of the fluorescent or luciferase reporter genes. Herein, for the first time, we engineered a replication-competent rSARS-CoV-2 that expresses both fluorescent (mCherry) and luciferase (Nluc) reporter genes (rSARS-CoV-2/mCherry-Nluc) to overcome limitations associated with the use of a single reporter gene. In cultured cells, rSARS-CoV-2/mCherry-Nluc displayed similar viral fitness as rSARS-CoV-2 expressing single reporter fluorescent and luciferase genes (rSARS-CoV-2/mCherry and rSARS-CoV-2/Nluc, respectively) or wild-type (WT) rSARS-CoV-2, while maintaining comparable expression levels of both reporter genes. In vivo, rSARS-CoV-2/mCherry-Nluc has similar pathogenicity in K18 human angiotensin-converting enzyme 2 (hACE2) transgenic mice than rSARS-CoV-2 expressing individual reporter genes or WT rSARS-CoV-2. Importantly, rSARS-CoV-2/mCherry-Nluc facilitates the assessment of viral infection and transmission in golden Syrian hamsters using in vivo imaging systems (IVIS). Altogether, this study demonstrates the feasibility of using this novel bioreporter-expressing rSARS-CoV-2 for the study of SARS-CoV-2 in vitro and in vivo. IMPORTANCE Despite the availability of vaccines and antivirals, the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to ravage health care institutions worldwide. Previously, we generated replication-competent recombinant (r)SARS-CoV-2 expressing fluorescent or luciferase reporter proteins to track viral infection in vitro and/or in vivo. However, these rSARS-CoV-2 are restricted to express only a single fluorescent or a luciferase reporter gene, limiting or preventing their use in specific in vitro assays and/or in vivo studies. To overcome this limitation, we have engineered a rSARS-CoV-2 expressing both fluorescent (mCherry) and luciferase (Nluc) genes and demonstrated its feasibility to study the biology of SARS-CoV-2 in vitro and/or in vivo, including the identification and characterization of neutralizing antibodies and/or antivirals. Using rodent models, we visualized SARS-CoV-2 infection and transmission through in vivo imaging systems (IVIS).
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- 2022
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7. SARS-CoV-2 Nsp14 protein associates with IMPDH2 and activates NF-κB signaling
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Tai-Wei Li, Adam D. Kenney, Jun-Gyu Park, Guillaume N. Fiches, Helu Liu, Dawei Zhou, Ayan Biswas, Weiqiang Zhao, Jianwen Que, Netty Santoso, Luis Martinez-Sobrido, Jacob S. Yount, and Jian Zhu
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SARS-CoV-2 ,NF-κB ,Nsp14 ,IL-8 ,IMPDH2 ,ribavirin ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection leads to NF-κB activation and induction of pro-inflammatory cytokines, though the underlying mechanism for this activation is not fully understood. Our results reveal that the SARS-CoV-2 Nsp14 protein contributes to the viral activation of NF-κB signaling. Nsp14 caused the nuclear translocation of NF-κB p65. Nsp14 induced the upregulation of IL-6 and IL-8, which also occurred in SARS-CoV-2 infected cells. IL-8 upregulation was further confirmed in lung tissue samples from COVID-19 patients. A previous proteomic screen identified the putative interaction of Nsp14 with host Inosine-5’-monophosphate dehydrogenase 2 (IMPDH2), which is known to regulate NF-κB signaling. We confirmed the Nsp14-IMPDH2 protein interaction and identified that IMPDH2 knockdown or chemical inhibition using ribavirin (RIB) and mycophenolic acid (MPA) abolishes Nsp14- mediated NF-κB activation and cytokine induction. Furthermore, IMPDH2 inhibitors (RIB, MPA) or NF-κB inhibitors (bortezomib, BAY 11-7082) restricted SARS-CoV-2 infection, indicating that IMPDH2-mediated activation of NF-κB signaling is beneficial to viral replication. Overall, our results identify a novel role of SARS-CoV-2 Nsp14 in inducing NF-κB activation through IMPDH2 to promote viral infection.
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- 2022
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8. Potent universal beta-coronavirus therapeutic activity mediated by direct respiratory administration of a Spike S2 domain-specific human neutralizing monoclonal antibody.
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Michael S Piepenbrink, Jun-Gyu Park, Ashlesha Deshpande, Andreas Loos, Chengjin Ye, Madhubanti Basu, Sanghita Sarkar, Ahmed Magdy Khalil, David Chauvin, Jennifer Woo, Philip Lovalenti, Nathaniel B Erdmann, Paul A Goepfert, Vu L Truong, Richard A Bowen, Mark R Walter, Luis Martinez-Sobrido, and James J Kobie
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) marks the third novel β-coronavirus to cause significant human mortality in the last two decades. Although vaccines are available, too few have been administered worldwide to keep the virus in check and to prevent mutations leading to immune escape. To determine if antibodies could be identified with universal coronavirus activity, plasma from convalescent subjects was screened for IgG against a stabilized pre-fusion SARS-CoV-2 spike S2 domain, which is highly conserved between human β-coronavirus. From these subjects, several S2-specific human monoclonal antibodies (hmAbs) were developed that neutralized SARS-CoV-2 with recognition of all variants of concern (VoC) tested (Beta, Gamma, Delta, Epsilon, and Omicron). The hmAb 1249A8 emerged as the most potent and broad hmAb, able to recognize all human β-coronavirus and neutralize SARS-CoV and MERS-CoV. 1249A8 demonstrated significant prophylactic activity in K18 hACE2 mice infected with SARS-CoV-2 lineage A and lineage B Beta, and Omicron VoC. 1249A8 delivered as a single 4 mg/kg intranasal (i.n.) dose to hamsters 12 hours following infection with SARS-CoV-2 Delta protected them from weight loss, with therapeutic activity further enhanced when combined with 1213H7, an S1-specific neutralizing hmAb. As little as 2 mg/kg of 1249A8 i.n. dose 12 hours following infection with SARS-CoV Urbani strain, protected hamsters from weight loss and significantly reduced upper and lower respiratory viral burden. These results indicate in vivo cooperativity between S1 and S2 specific neutralizing hmAbs and that potent universal coronavirus neutralizing mAbs with therapeutic potential can be induced in humans and can guide universal coronavirus vaccine development.
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- 2022
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9. Vaccinia Virus Strain MVA Expressing a Prefusion-Stabilized SARS-CoV-2 Spike Glycoprotein Induces Robust Protection and Prevents Brain Infection in Mouse and Hamster Models
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María M. Lorenzo, Alejandro Marín-López, Kevin Chiem, Luis Jimenez-Cabello, Irfan Ullah, Sergio Utrilla-Trigo, Eva Calvo-Pinilla, Gema Lorenzo, Sandra Moreno, Chengjin Ye, Jun-Gyu Park, Alejandro Matía, Alejandro Brun, Juana M. Sánchez-Puig, Aitor Nogales, Walther Mothes, Pradeep D. Uchil, Priti Kumar, Javier Ortego, Erol Fikrig, Luis Martinez-Sobrido, and Rafael Blasco
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poxvirus ,modified vaccinia virus Ankara ,vaccine ,SARS-CoV-2 ,COVID-19 ,recombinant viral vectors ,Medicine - Abstract
The COVID-19 pandemic has underscored the importance of swift responses and the necessity of dependable technologies for vaccine development. Our team previously developed a fast cloning system for the modified vaccinia virus Ankara (MVA) vaccine platform. In this study, we reported on the construction and preclinical testing of a recombinant MVA vaccine obtained using this system. We obtained recombinant MVA expressing the unmodified full-length SARS-CoV-2 spike (S) protein containing the D614G amino-acid substitution (MVA-Sdg) and a version expressing a modified S protein containing amino-acid substitutions designed to stabilize the protein a in a pre-fusion conformation (MVA-Spf). S protein expressed by MVA-Sdg was found to be expressed and was correctly processed and transported to the cell surface, where it efficiently produced cell–cell fusion. Version Spf, however, was not proteolytically processed, and despite being transported to the plasma membrane, it failed to induce cell–cell fusion. We assessed both vaccine candidates in prime-boost regimens in the susceptible transgenic K18-human angiotensin-converting enzyme 2 (K18-hACE2) in mice and in golden Syrian hamsters. Robust immunity and protection from disease was induced with either vaccine in both animal models. Remarkably, the MVA-Spf vaccine candidate produced higher levels of antibodies, a stronger T cell response, and a higher degree of protection from challenge. In addition, the level of SARS-CoV-2 in the brain of MVA-Spf inoculated mice was decreased to undetectable levels. Those results add to our current experience and range of vaccine vectors and technologies for developing a safe and effective COVID-19 vaccine.
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- 2023
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10. Lethality of SARS-CoV-2 infection in K18 human angiotensin-converting enzyme 2 transgenic mice
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Fatai S. Oladunni, Jun-Gyu Park, Paula A. Pino, Olga Gonzalez, Anwari Akhter, Anna Allué-Guardia, Angélica Olmo-Fontánez, Shalini Gautam, Andreu Garcia-Vilanova, Chengjin Ye, Kevin Chiem, Colwyn Headley, Varun Dwivedi, Laura M. Parodi, Kendra J. Alfson, Hilary M. Staples, Alyssa Schami, Juan I. Garcia, Alison Whigham, Roy Neal Platt, Michal Gazi, Jesse Martinez, Colin Chuba, Stephanie Earley, Oscar H. Rodriguez, Stephanie Davis Mdaki, Katrina N. Kavelish, Renee Escalona, Cory R. A. Hallam, Corbett Christie, Jean L. Patterson, Tim J. C. Anderson, Ricardo Carrion, Edward J. Dick, Shannan Hall-Ursone, Larry S. Schlesinger, Xavier Alvarez, Deepak Kaushal, Luis D. Giavedoni, Joanne Turner, Luis Martinez-Sobrido, and Jordi B. Torrelles
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Science - Abstract
Here, the authors characterize tissue-level SARS-CoV-2 infection and pathogenesis in transgenic mice expressing human angiotensin converting enzyme 2 (hACE2) by the human cytokeratin 18 promoter (K18-hACE2) and show that infection induces lethality, making the K18-hACE2 model suitable for vaccine and therapeutic evaluation.
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- 2020
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11. Structural basis of RNA cap modification by SARS-CoV-2
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Thiruselvam Viswanathan, Shailee Arya, Siu-Hong Chan, Shan Qi, Nan Dai, Anurag Misra, Jun-Gyu Park, Fatai Oladunni, Dmytro Kovalskyy, Robert A. Hromas, Luis Martinez-Sobrido, and Yogesh K. Gupta
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Science - Abstract
Specific non-structural proteins (nsp) of SARS coronaviruses are involved in methylation of virally encoded mRNAs to mimic cellular mRNAs for protection against host innate immune restriction. Here, the authors present a high resolution structure of SARS-CoV-2 nsp16/nsp10 ternary complex in the presence of cognate RNA substrate analogue and methyl donor, S-adenosyl methionine, revealing unique ligand-binding sites that may represent alternative targets for antiviral development.
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- 2020
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12. Bi-Reporter Vaccinia Virus for Tracking Viral Infections In Vitro and In Vivo
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Kevin Chiem, Maria M. Lorenzo, Javier Rangel-Moreno, Maria De La Luz Garcia-Hernandez, Jun-Gyu Park, Aitor Nogales, Rafael Blasco, and Luis Martínez-Sobrido
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vaccinia virus ,reporter genes ,fluorescence ,Scarlet ,GFP ,bioluminescence ,Microbiology ,QR1-502 - Abstract
ABSTRACT Recombinant viruses expressing reporter genes allow visualization and quantification of viral infections and can be used as valid surrogates to identify the presence of the virus in infected cells and animal models. However, one of the limitations of recombinant viruses expressing reporter genes is the use of either fluorescent or luciferase proteins that are used alternatively for different purposes. Vaccinia virus (VV) is widely used as a viral vector, including recombinant (r)VV singly expressing either fluorescent or luciferase reporter genes that are useful for specific purposes. In this report, we engineered two novel rVV stably expressing both fluorescent (Scarlet or GFP) and luciferase (Nluc) reporter genes from different loci in the viral genome. In vitro, these bi-reporter-expressing rVV have similar growth kinetics and plaque phenotype than those of the parental WR VV isolate. In vivo, rVV Nluc/Scarlet and rVV Nluc/GFP effectively infected mice and were easily detected using in vivo imaging systems (IVIS) and ex vivo in the lungs from infected mice. Importantly, we used these bi-reporter-expressing rVV to assess viral pathogenesis, infiltration of immune cells in the lungs, and to directly identify the different subsets of cells infected by VV in the absence of antibody staining. Collectively, these rVV expressing two reporter genes open the feasibility to study the biology of viral infections in vitro and in vivo, including host-pathogen interactions and dynamics or tropism of viral infections. IMPORTANCE Despite the eradication of variola virus (VARV), the causative agent of smallpox, poxviruses still represent an important threat to human health due to their possible use as bioterrorism agents and the emergence of zoonotic poxvirus diseases. Recombinant vaccinia viruses (rVV) expressing easily traceable fluorescent or luciferase reporter genes have significantly contributed to the progress of poxvirus research. However, rVV expressing one marker gene have several constraints for in vitro and in vivo studies, since both fluorescent and luciferase proteins impose certain limitations for specific applications. To overcome these limitations, we generated optimized rVV stably expressing both fluorescent (Scarlet or GFP) and luciferase (Nluc) reporter genes to easily track viral infection in vitro and in vivo. This new generation of double reporter-expressing rVV represent an excellent option to study viral infection dynamics in cultured cells and validated animal models of infection.
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- 2021
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13. Immunogenicity and protective efficacy of an intranasal live-attenuated vaccine against SARS-CoV-2
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Jun-Gyu Park, Fatai S. Oladunni, Mohammed A. Rohaim, Jayde Whittingham-Dowd, James Tollitt, Matthew D.J. Hodges, Nadin Fathallah, Muhsref Bakri Assas, Wafaa Alhazmi, Abdullah Almilaibary, Munir Iqbal, Pengxiang Chang, Renee Escalona, Vinay Shivanna, Jordi B. Torrelles, John J. Worthington, Lucy H. Jackson-Jones, Luis Martinez-Sobrido, and Muhammad Munir
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immunology ,virology ,Science - Abstract
Summary: Global deployment of an effective and safe vaccine is necessary to curtail the coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we evaluated a Newcastle disease virus (NDV)-based vectored-vaccine in mice and hamsters for its immunogenicity, safety, and protective efficacy against SARS-CoV-2. Intranasal administration of recombinant (r)NDV-S vaccine expressing spike (S) protein of SARS-CoV-2 to mice induced high levels of SARS-CoV-2-specific neutralizing immunoglobulin A (IgA) and IgG2a antibodies and T-cell-mediated immunity. Hamsters immunized with two doses of vaccine showed complete protection from lung infection, inflammation, and pathological lesions following SARS-CoV-2 challenge. Importantly, administration of two doses of intranasal rNDV-S vaccine significantly reduced the SARS-CoV-2 shedding in nasal turbinate and lungs in hamsters. Collectively, intranasal vaccination has the potential to control infection at the site of inoculation, which should prevent both clinical disease and virus transmission to halt the spread of the COVID-19 pandemic.
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- 2021
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14. Development of a live attenuated trivalent porcine rotavirus A vaccine against disease caused by recent strains most prevalent in South Korea
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Jun-Gyu Park, Mia Madel Alfajaro, Eun-Hyo Cho, Ji-Yun Kim, Mahmoud Soliman, Yeong-Bin Baek, Chul-Ho Park, Ju-Hwan Lee, Kyu-Yeol Son, Kyoung-Oh Cho, and Mun-Il Kang
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Veterinary medicine ,SF600-1100 - Abstract
Abstract Porcine rotaviruses cause severe economic losses in the Korean swine industry due to G- and P-genotype mismatches between the predominant field and vaccine strains. Here, we developed a live attenuated trivalent porcine group A rotavirus vaccine using 80 cell culture passages of the representative Korean predominant strains G8P[7] 174-1, G9P[23] PRG942, and G5P[7] K71. Vaccination with the trivalent vaccine or its individual components induced no diarrhea during the first 2 weeks post-vaccination, i.e., the vaccines were attenuated. Challenge of trivalent-vaccinated or component-vaccinated piglets with homologous virulent strain(s) did not induce diarrhea for 2 weeks post-challenge. Immunization with the trivalent vaccine or its individual components also alleviated the histopathological lesions in the small intestines caused by challenge with the corresponding original virulent strain(s). Fecal secretory IgAs specific for each of vaccine strains were detected starting at 14 days post-vaccination (dpv), and IgA levels gradually increased up to 28 dpv. Oral immunization with the trivalent vaccine or its individual components induced high levels of serum virus-neutralizing antibody by 7 dpv. No diarrhea was observed in any experimental piglets during five consecutive passages of each vaccine strain. Our data indicated that the live attenuated trivalent vaccine was safe and effective at protecting piglets from diarrhea induced by challenge exposure of homologous virulent strains. This trivalent vaccine will potentially contribute toward controlling porcine rotavirus disease in South Korea and other countries where rotavirus infections with similar G and P genotypes are problematic.
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- 2019
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15. GMP Manufacturing and IND-Enabling Studies of a Recombinant Hyperimmune Globulin Targeting SARS-CoV-2
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Rena A. Mizrahi, Wendy Y. Lin, Ashley Gras, Ariel R. Niedecken, Ellen K. Wagner, Sheila M. Keating, Nikita Ikon, Vishal A. Manickam, Michael A. Asensio, Jackson Leong, Angelica V. Medina-Cucurella, Emily Benzie, Kyle P. Carter, Yao Chiang, Robert C. Edgar, Renee Leong, Yoong Wearn Lim, Jan Fredrik Simons, Matthew J. Spindler, Kacy Stadtmiller, Nicholas Wayham, Dirk Büscher, Jose Vicente Terencio, Clara Di Germanio, Steven M. Chamow, Charles Olson, Paula A. Pino, Jun-Gyu Park, Amberlee Hicks, Chengjin Ye, Andreu Garcia-Vilanova, Luis Martinez-Sobrido, Jordi B. Torrelles, David S. Johnson, and Adam S. Adler
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recombinant hyperimmune ,GMP manufacturing ,SARS-CoV-2 ,Medicine - Abstract
Conventionally, hyperimmune globulin drugs manufactured from pooled immunoglobulins from vaccinated or convalescent donors have been used in treating infections where no treatment is available. This is especially important where multi-epitope neutralization is required to prevent the development of immune-evading viral mutants that can emerge upon treatment with monoclonal antibodies. Using microfluidics, flow sorting, and a targeted integration cell line, a first-in-class recombinant hyperimmune globulin therapeutic against SARS-CoV-2 (GIGA-2050) was generated. Using processes similar to conventional monoclonal antibody manufacturing, GIGA-2050, comprising 12,500 antibodies, was scaled-up for clinical manufacturing and multiple development/tox lots were assessed for consistency. Antibody sequence diversity, cell growth, productivity, and product quality were assessed across different manufacturing sites and production scales. GIGA-2050 was purified and tested for good laboratory procedures (GLP) toxicology, pharmacokinetics, and in vivo efficacy against natural SARS-CoV-2 infection in mice. The GIGA-2050 master cell bank was highly stable, producing material at consistent yield and product quality up to >70 generations. Good manufacturing practices (GMP) and development batches of GIGA-2050 showed consistent product quality, impurity clearance, potency, and protection in an in vivo efficacy model. Nonhuman primate toxicology and pharmacokinetics studies suggest that GIGA-2050 is safe and has a half-life similar to other recombinant human IgG1 antibodies. These results supported a successful investigational new drug application for GIGA-2050. This study demonstrates that a new class of drugs, recombinant hyperimmune globulins, can be manufactured consistently at the clinical scale and presents a new approach to treating infectious diseases that targets multiple epitopes of a virus.
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- 2022
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16. Rescue of SARS-CoV-2 from a Single Bacterial Artificial Chromosome
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Chengjin Ye, Kevin Chiem, Jun-Gyu Park, Fatai Oladunni, Roy Nelson Platt, Tim Anderson, Fernando Almazan, Juan Carlos de la Torre, and Luis Martinez-Sobrido
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BAC ,COVID-19 ,SARS-CoV-2 ,coronavirus ,hamsters ,recombinant virus ,Microbiology ,QR1-502 - Abstract
ABSTRACT Infectious coronavirus (CoV) disease 2019 (COVID-19) emerged in the city of Wuhan (China) in December 2019, causing a pandemic that has dramatically impacted public health and socioeconomic activities worldwide. A previously unknown coronavirus, severe acute respiratory syndrome CoV-2 (SARS-CoV-2), has been identified as the causative agent of COVID-19. To date, there are no U.S. Food and Drug Administration (FDA)-approved vaccines or therapeutics available for the prevention or treatment of SARS-CoV-2 infection and/or associated COVID-19 disease, which has triggered a large influx of scientific efforts to develop countermeasures to control SARS-CoV-2 spread. To contribute to these efforts, we have developed an infectious cDNA clone of the SARS-CoV-2 USA-WA1/2020 strain based on the use of a bacterial artificial chromosome (BAC). Recombinant SARS-CoV-2 (rSARS-CoV-2) was readily rescued by transfection of the BAC into Vero E6 cells. Importantly, BAC-derived rSARS-CoV-2 exhibited growth properties and plaque sizes in cultured cells comparable to those of the natural SARS-CoV-2 isolate. Likewise, rSARS-CoV-2 showed levels of replication similar to those of the natural isolate in nasal turbinates and lungs of infected golden Syrian hamsters. This is, to our knowledge, the first BAC-based reverse genetics system for the generation of infectious rSARS-CoV-2 that displays features in vivo similar to those of a natural viral isolate. This SARS-CoV-2 BAC-based reverse genetics will facilitate studies addressing several important questions in the biology of SARS-CoV-2, as well as the identification of antivirals and development of vaccines for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. IMPORTANCE The pandemic coronavirus (CoV) disease 2019 (COVID-19) caused by severe acute respiratory syndrome CoV-2 (SARS-CoV-2) is a major threat to global human health. To date, there are no approved prophylactics or therapeutics available for COVID-19. Reverse genetics is a powerful approach to understand factors involved in viral pathogenesis, antiviral screening, and vaccine development. In this study, we describe the feasibility of generating recombinant SARS-CoV-2 (rSARS-CoV-2) by transfection of a single bacterial artificial chromosome (BAC). Importantly, rSARS-CoV-2 possesses the same phenotype as the natural isolate in vitro and in vivo. This is the first description of a BAC-based reverse genetics system for SARS-CoV-2 and the first time that an rSARS-CoV-2 isolate has been shown to be phenotypically identical to a natural isolate in a validated animal model of SARS-CoV-2 infection. The BAC-based reverse genetics approach will facilitate the study of SARS-CoV-2 and the development of prophylactics and therapeutics for the treatment of COVID-19.
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- 2020
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17. Oncolytic Vaccinia Virus Augments T Cell Factor 1-Positive Stem-like CD8+ T Cells, Which Underlies the Efficacy of Anti-PD-1 Combination Immunotherapy
- Author
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Yun-Hui Jeon, Namhee Lee, Jiyoon Yoo, Solchan Won, Suk-kyung Shin, Kyu-Hwan Kim, Jun-Gyu Park, Min-Gang Kim, Hang-Rae Kim, Keunhee Oh, and Dong-Sup Lee
- Subjects
oncolytic vaccinia virus ,cancer antigen-specific T cells ,stem-like CD8+ T cells ,lymphoid organs ,immune checkpoint blockade ,Biology (General) ,QH301-705.5 - Abstract
Oncolytic virotherapy has garnered attention as an antigen-agnostic therapeutic cancer vaccine that induces cancer-specific T cell responses without additional antigen loading. As anticancer immune responses are compromised by a lack of antigenicity and chronic immunosuppressive microenvironments, an effective immuno-oncology modality that converts cold tumors into hot tumors is crucial. To evaluate the immune-activating characteristics of oncolytic vaccinia virus (VACV; JX-594, pexastimogene devacirepvec), diverse murine syngeneic cancer models with different tissue types and immune microenvironments were used. Intratumorally administered mJX-594, a murine variant of JX-594, potently increased CD8+ T cells, including antigen-specific cancer CD8+ T cells, and decreased immunosuppressive cells irrespective of tissue type or therapeutic efficacy. Remodeling of tumors into inflamed ones by mJX-594 led to a response to combined anti-PD-1 treatment, but not to mJX-594 or anti-PD-1 monotherapy. mJX-594 treatment increased T cell factor 1-positive stem-like T cells among cancer-specific CD8+ T cells, and anti-PD-1 combination treatment further increased proliferation of these cells, which was important for therapeutic efficacy. The presence of functional cancer-specific CD8+ T cells in the spleen and bone marrow for an extended period, which proliferated upon encountering cancer antigen-loaded splenic dendritic cells, further indicated that long-term durable anticancer immunity was elicited by oncolytic VACV.
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- 2022
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18. Porcine sapovirus Cowden strain enters LLC-PK cells via clathrin- and cholesterol-dependent endocytosis with the requirement of dynamin II
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Mahmoud Soliman, Deok-Song Kim, Chonsaeng Kim, Ja-Young Seo, Ji-Yun Kim, Jun-Gyu Park, Mia Madel Alfajaro, Yeong-Bin Baek, Eun-Hyo Cho, Sang-Ik Park, Mun-Il Kang, Kyeong-Ok Chang, Ian Goodfellow, and Kyoung-Oh Cho
- Subjects
Veterinary medicine ,SF600-1100 - Abstract
Abstract Caliciviruses in the genus Sapovirus are a significant cause of viral gastroenteritis in humans and animals. However, the mechanism of their entry into cells is not well characterized. Here, we determined the entry mechanism of porcine sapovirus (PSaV) strain Cowden into permissive LLC-PK cells. The inhibition of clathrin-mediated endocytosis using chlorpromazine, siRNAs, and a dominant negative (DN) mutant blocked entry and infection of PSaV Cowden strain, confirming a role for clathrin-mediated internalization. Entry and infection were also inhibited by the cholesterol-sequestering drug methyl-β-cyclodextrin and was restored by the addition of soluble cholesterol, indicating that cholesterol also contributes to entry and infection of this strain. Furthermore, the inhibition of dynamin GTPase activity by dynasore, siRNA depletion of dynamin II, or overexpression of a DN mutant of dynamin II reduced the entry and infection, suggesting that dynamin mediates the fission and detachment of clathrin- and cholesterol-pits for entry of this strain. In contrast, the inhibition of caveolae-mediated endocytosis using nystatin, siRNAs, or a DN mutant had no inhibitory effect on entry and infection of this strain. It was further determined that cell entry of PSaV Cowden strain required actin rearrangements for vesicle internalization, endosomal trafficking from early to late endosomes through microtubules, and late endosomal acidification for uncoating. We conclude that PSaV strain Cowden is internalized into LLC-PK cells by clathrin- and cholesterol-mediated endocytosis that requires dynamin II and actin rearrangement, and that the uncoating occurs in the acidified late endosomes after trafficking from the early endosomes through microtubules.
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- 2018
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19. Characteristics of Biogas Production from Organic Wastes Mixed at Optimal Ratios in an Anaerobic Co-Digestion Reactor
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Young-Ju Song, Kyung-Su Oh, Beom Lee, Dae-Won Pak, Ji-Hwan Cha, and Jun-Gyu Park
- Subjects
sewage sludge ,food waste ,livestock manure ,anaerobic co-digestion ,mixing ratio ,Technology - Abstract
This study determined the optimal mixing ratio of food waste and livestock manure for efficient co-digestion of sewage sludge by applying the biochemical methane potential (BMP) test, Design Expert software, and continuous reactor operation. The BMP test of sewage sludge revealed a maximum methane yield of 334 mL CH4/g volatile solids (VS) at an organic loading rate (OLR) of 4 kg VS/(m3·d). For food waste, the maximum methane yield was 573 mL CH4/g VS at an OLR of 6 kg VS/(m3·d). Livestock manure showed the lowest methane yield. The BMP tests with various mixing ratios confirmed that a higher mixing ratio of food waste resulted in a higher methane yield, which showed improved biodegradability and an improved VS removal rate. The optimal mixing ratio of 2:1:1 for sewage sludge, food waste, and livestock manure was determined using Design Expert 10. Using continuous co-digestion reactor operation under an optimal mixing ratio, greater organic matter removal and methane yield was possible. The process stability of co-digestion of optimally mixed substrate was improved compared with that of operations with each substrate alone. Therefore, co-digestion could properly maintain the balance of each stage of anaerobic digestion reactions by complementing the characteristics of each substrate under a higher OLR.
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- 2021
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20. Author Correction: Immune cell composition in normal human kidneys
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Jun-Gyu Park, Myeongsu Na, Min-Gang Kim, Su Hwan Park, Hack June Lee, Dong Ki Kim, Cheol Kwak, Yon Su Kim, Sunghoe Chang, Kyung Chul Moon, Dong-Sup Lee, and Seung Seok Han
- Subjects
Medicine ,Science - Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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- 2021
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21. Potent Inhibition of Zika Virus Replication by Aurintricarboxylic Acid
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Jun-Gyu Park, Ginés Ávila-Pérez, Ferralita Madere, Thomas A. Hilimire, Aitor Nogales, Fernando Almazán, and Luis Martínez-Sobrido
- Subjects
Flavivirus ,Zika virus ,aurintricarboxylic acid ,antivirals ,prophylactic ,therapeutic ,Microbiology ,QR1-502 - Abstract
Zika virus (ZIKV) is one of the recently emerging vector-borne viruses in humans and is responsible for severe congenital abnormalities such as microcephaly in the Western Hemisphere. Currently, only a few vaccine candidates and therapeutic drugs are being developed for the treatment of ZIKV infections, and as of yet none are commercially available. The polyanionic aromatic compound aurintricarboxylic acid (ATA) has been shown to have a broad-spectrum antimicrobial and antiviral activity. In this study, we evaluated ATA as a potential antiviral drug against ZIKV replication. The antiviral activity of ATA against ZIKV replication in vitro showed median inhibitory concentrations (IC50) of 13.87 ± 1.09 μM and 33.33 ± 1.13 μM in Vero and A549 cells, respectively; without showing any cytotoxic effect in both cell lines (median cytotoxic concentration (CC50) > 1,000 μM). Moreover, ATA protected both cell types from ZIKV-induced cytopathic effect (CPE) and apoptosis in a time- and concentration-dependent manner. In addition, pre-treatment of Vero cells with ATA for up to 72 h also resulted in effective suppression of ZIKV replication with similar IC50. Importantly, the inhibitory effect of ATA on ZIKV infection was effective against strains of the African and Asian/American lineages, indicating that this inhibitory effect was not strain dependent. Overall, these results demonstrate that ATA has potent inhibitory activity against ZIKV replication and may be considered as a potential anti-ZIKV therapy for future clinical evaluation.
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- 2019
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22. Identification of Inhibitors of ZIKV Replication
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Desarey Morales Vasquez, Jun-Gyu Park, Ginés Ávila-Pérez, Aitor Nogales, Juan Carlos de la Torre, Fernando Almazan, and Luis Martinez-Sobrido
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flavivirus ,Zika virus ,antivirals ,therapeutic ,drug treatment ,ReFRAME library ,Microbiology ,QR1-502 - Abstract
Zika virus (ZIKV) was identified in 1947 in the Zika forest of Uganda and it has emerged recently as a global health threat, with recurring outbreaks and its associations with congenital microcephaly through maternal fetal transmission and Guillain-Barré syndrome. Currently, there are no United States (US) Food and Drug Administration (FDA)-approved vaccines or antivirals to treat ZIKV infections, which underscores an urgent medical need for the development of disease intervention strategies to treat ZIKV infection and associated disease. Drug repurposing offers various advantages over developing an entirely new drug by significantly reducing the timeline and resources required to advance a candidate antiviral into the clinic. Screening the ReFRAME library, we identified ten compounds with antiviral activity against the prototypic mammarenavirus lymphocytic choriomeningitis virus (LCMV). Moreover, we showed the ability of these ten compounds to inhibit influenza A and B virus infections, supporting their broad-spectrum antiviral activity. In this study, we further evaluated the broad-spectrum antiviral activity of the ten identified compounds by testing their activity against ZIKV. Among the ten compounds, Azaribine (SI-MTT = 146.29), AVN-944 (SI-MTT = 278.16), and Brequinar (SI-MTT = 157.42) showed potent anti-ZIKV activity in post-treatment therapeutic conditions. We also observed potent anti-ZIKV activity for Mycophenolate mofetil (SI-MTT = 20.51), Mycophenolic acid (SI-MTT = 36.33), and AVN-944 (SI-MTT = 24.51) in pre-treatment prophylactic conditions and potent co-treatment inhibitory activity for Obatoclax (SI-MTT = 60.58), Azaribine (SI-MTT = 91.51), and Mycophenolate mofetil (SI-MTT = 73.26) in co-treatment conditions. Importantly, the inhibitory effect of these compounds was strain independent, as they similarly inhibited ZIKV strains from both African and Asian/American lineages. Our results support the broad-spectrum antiviral activity of these ten compounds and suggest their use for the development of antiviral treatment options of ZIKV infection.
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- 2020
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23. Ghost-MTD: Moving Target Defense via Protocol Mutation for Mission-Critical Cloud Systems
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Jun-Gyu Park, Yangjae Lee, Ki-Wan Kang, Sang-Hoon Lee, and Ki-Woong Park
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moving target defense ,deception ,protocol mutation ,mission-critical cloud systems ,Technology - Abstract
Research on various security technologies has been actively underway to protect systems from attackers. However, attackers can secure enough time to reconnoiter and attack the target system owing to its static nature. This develops asymmetric warfare in which attackers outwit defenders. Moving target defense (MTD) technologies, which obfuscate the attack surface by modifying the main properties of the potential target system, have been gaining attention as an active cyber security technology. Particularly, network-based MTD (NMTD) technologies, which dynamically mutate the network configuration information, such as IP and ports of the potential target system, can dramatically increase the time required for an attacker to analyze the system. Therefore, this system defense technology has been actively researched. However, increasing the analysis complexity of the target system is limited in conventional NMTD because the variation of system properties (e.g., IP, port) that can be mutated is restricted by the system configuration environment. Therefore, there is a need for an MTD technique that effectively delays an attacker during the system analysis by increasing the variation of system properties. Additionally, in terms of practicality, minimizing the computational overhead arising by the MTD technology and solving the compatibility problem with existing communication protocols are critical issues that cannot be overlooked. In this study, we propose a technology called Ghost-MTD (gMTD). gMTD allows only the user who is aware of protocol mutation patterns to correctly communicate with the service modules of the server system through protocol mutation using the pre-shared one-time bit sequence. Otherwise, gMTD deceives the attackers who attempt to infiltrate the system by redirecting their messages to a decoy-hole module. The experimental results show that the proposed technology enables protocol mutation and validation with a very low performance overhead of only 3.28% to 4.97% using an m-bit (m ≥ 4) length one-time bit sequence and can be applied to real systems regardless of the specific communication protocols.
- Published
- 2020
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24. A Broad and Potent H1-Specific Human Monoclonal Antibody Produced in Plants Prevents Influenza Virus Infection and Transmission in Guinea Pigs
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Jun-Gyu Park, Chengjin Ye, Michael S. Piepenbrink, Aitor Nogales, Haifeng Wang, Michael Shuen, Ashley J. Meyers, Luis Martinez-Sobrido, and James J. Kobie
- Subjects
orthomyxovirus ,influenza virus ,antivirals ,plantibody ,prophylactic ,therapeutic ,monoclonal antibody ,antibody treatment ,neutralizing antibody ,virus infection ,virus transmission ,Microbiology ,QR1-502 - Abstract
Although seasonal influenza vaccines block most predominant influenza types and subtypes, humans still remain vulnerable to waves of seasonal and new potential pandemic influenza viruses for which no immunity may exist because of viral antigenic drift and/or shift. Previously, we described a human monoclonal antibody (hMAb), KPF1, which was produced in human embryonic kidney 293T cells (KPF1-HEK) with broad and potent neutralizing activity against H1N1 influenza A viruses (IAV) in vitro, and prophylactic and therapeutic activities in vivo. In this study, we produced hMAb KPF1 in tobacco plants (KPF1-Antx) and demonstrated how the plant-produced KPF1-Antx hMAb possesses similar biological activity compared with the mammalian-produced KPF1-HEK hMAb. KPF1-Antx hMAb showed broad binding to recombinant HA proteins and H1N1 IAV, including A/California/04/2009 (pH1N1) in vitro, which was comparable to that observed with KPF1-HEK hMAb. Importantly, prophylactic administration of KPF1-Antx hMAb to guinea pigs prevented pH1N1 infection and transmission in both prophylactic and therapeutic experiments, substantiating its clinical potential to prevent and treat H1N1 infections. Collectively, this study demonstrated, for the first time, a plant-produced influenza hMAb with in vitro and in vivo activity against influenza virus. Because of the many advantages of plant-produced hMAbs, such as rapid batch production, low cost, and the absence of mammalian cell products, they represent an alternative strategy for the production of immunotherapeutics for the treatment of influenza viral infections, including emerging seasonal and/or pandemic strains.
- Published
- 2020
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25. Activation of PI3K, Akt, and ERK during early rotavirus infection leads to V-ATPase-dependent endosomal acidification required for uncoating.
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Mahmoud Soliman, Ja-Young Seo, Deok-Song Kim, Ji-Yun Kim, Jun-Gyu Park, Mia Madel Alfajaro, Yeong-Bin Baek, Eun-Hyo Cho, Joseph Kwon, Jong-Soon Choi, Mun-Il Kang, Sang-Ik Park, and Kyoung-Oh Cho
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The cellular PI3K/Akt and/or MEK/ERK signaling pathways mediate the entry process or endosomal acidification during infection of many viruses. However, their roles in the early infection events of group A rotaviruses (RVAs) have remained elusive. Here, we show that late-penetration (L-P) human DS-1 and bovine NCDV RVA strains stimulate these signaling pathways very early in the infection. Inhibition of both signaling pathways significantly reduced production of viral progeny due to blockage of virus particles in the late endosome, indicating that neither of the two signaling pathways is involved in virus trafficking. However, immunoprecipitation assays using antibodies specific for pPI3K, pAkt, pERK and the subunit E of the V-ATPase co-immunoprecipitated the V-ATPase in complex with pPI3K, pAkt, and pERK. Moreover, Duolink proximity ligation assay revealed direct association of the subunit E of the V-ATPase with the molecules pPI3K, pAkt, and pERK, indicating that both signaling pathways are involved in V-ATPase-dependent endosomal acidification. Acidic replenishment of the medium restored uncoating of the RVA strains in cells pretreated with inhibitors specific for both signaling pathways, confirming the above results. Isolated components of the outer capsid proteins, expressed as VP4-VP8* and VP4-VP5* domains, and VP7, activated the PI3K/Akt and MEK/ERK pathways. Furthermore, psoralen-UV-inactivated RVA and CsCl-purified RVA triple-layered particles triggered activation of the PI3K/Akt and MEK/ERK pathways, confirming the above results. Our data demonstrate that multistep binding of outer capsid proteins of L-P RVA strains with cell surface receptors phosphorylates PI3K, Akt, and ERK, which in turn directly interact with the subunit E of the V-ATPase to acidify the late endosome for uncoating of RVAs. This study provides a better understanding of the RVA-host interaction during viral uncoating, which is of importance for the development of strategies aiming at controlling or preventing RVA infections.
- Published
- 2018
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26. Feline calicivirus- and murine norovirus-induced COX-2/PGE2 signaling pathway has proviral effects.
- Author
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Mia Madel Alfajaro, Eun-Hyo Cho, Jun-Gyu Park, Ji-Yun Kim, Mahmoud Soliman, Yeong-Bin Baek, Mun-Il Kang, Sang-Ik Park, and Kyoung-Oh Cho
- Subjects
Medicine ,Science - Abstract
Cyclooxygenases (COXs)/prostaglandin E2 (PGE2) signaling pathways are known to modulate a variety of homeostatic processes and are involved in various pathophysiological conditions. COXs/PGE2 signaling pathways have also been demonstrated to have proviral or antiviral effects, which appeared different even in the same virus family. A porcine sapovirus Cowden strain, a member of genus Sapovirus within the Caliciviridae family, induces strong COX-2/PGE2 but transient COX-1/PGE2 signaling to enhance virus replication. However, whether infections of other viruses in the different genera activate COXs/PGE2 signaling, and thus affect the replication of viruses, remains unknown. In the present study, infections of cells with the feline calicivirus (FCV) F9 strain in the genus Vesivirus and murine norovirus (MNV) CW-1 strain in the genus Norovirus only activated the COX-2/PGE2 signaling in a time-dependent manner. Treatment with pharmacological inhibitors or transfection of small interfering RNAs (siRNAs) against COX-2 enzyme significantly reduced the production of PGE2 as well as FCV and MNV replications. The inhibitory effects of these pharmacological inhibitors against COX-2 enzyme on the replication of both viruses were restored by the addition of PGE2. Silencing of COX-1 via siRNAs and inhibition of COX-1 via an inhibitor also decrease the production of PGE2 and replication of both viruses, which can be attributed to the inhibition COX-1/PGE2 signaling pathway. These data indicate that the COX-2/PGE2 signaling pathway has proviral effects for the replication of FCV and MNV, and pharmacological inhibitors against these enzymes serve as potential therapeutic candidates for treating FCV and MNV infections.
- Published
- 2018
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27. Improvement of Waste Dehydrated Sludge for Anaerobic Digestion through High-Temperature and High-Pressure Solubilization
- Author
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Eui-Hwan Hong, Jun-Gyu Park, Beom Lee, Wei-Qi Shi, and Hang-Bae Jun
- Subjects
anaerobic digestion ,waste dehydrated sludge ,high temperature ,high pressure ,solubilization ,Technology - Abstract
Biochemical methane potential tests and lab-scale continuous experiments were conducted to improve the yield and energy efficiency of anaerobic digestion through thermal hydrolysis pre-treatment. Methane generation, yield, and solubilization efficiency were evaluated through lab-scale tests. The pre-treated samples presented 50% biodegradability at 140 °C and 61.5% biodegradability at 165 °C. The increase in biodegradability was insignificant at 165 °C or higher temperature, and it was confirmed that the optimum conditions were achieved at 165 °C and 20 min of solubilization. The lab-scale continuous experiments confirmed that polymers were decomposed into low-molecular-weight compounds due to thermal hydrolysis, and pH decreased. NH4HCO3 produced by thermal hydrolysis acted as an alkali to enable a more stable operation compared to that before thermal hydrolysis. Total chemical oxygen demand as chromium (TCODCr) and soluble chemical oxygen demand as chromium (SCODCr) indicated 35.4% and 23.1% removal efficiency in terms of organic matter removal, respectively. Methane yield was approximately 0.35 kg m−3 at 2.0−4.0 kg (m3 d)−1 and 0.26 kg m−3 at 5.0 kg (m3 d)−1. The solubilization rate of 40.9% by thermal hydrolysis was confirmed through the lab-scale tests to determine its full-scale applicability.
- Published
- 2019
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28. Changes of Bacterial Communities in an Anaerobic Digestion and a Bio-Electrochemical Anaerobic Digestion Reactors According to Organic Load
- Author
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Jun-Gyu Park, Won-Beom Shin, Wei-Qi Shi, and Hang-Bae Jun
- Subjects
bio-electrochemical anaerobic digestion (BEAD) ,bacterial communities ,bulk solution ,organic loading rate ,food waste ,Technology - Abstract
Bacterial communities change in bulk solution of anaerobic digestion (AD) and bio-electrochemical anaerobic digestion reactors (BEAD) were monitored at each organic loading rate (OLR) to investigate the effect of voltage supply on bacterial species change in bulk solution. Chemical oxygen demand (COD) degradation and methane production from AD and BEAD reactors were also analyzed by gradually increasing food waste OLR. The BEAD reactor maintained stable COD removal and methane production at 6.0 kg/m3·d. The maximum OLR of AD reactor for optimal operation was 4.0 kg/m3·d. pH and alkalinity decline and volatile fatty acid (VFA) accumulation, which are the problem in high load anaerobic digestion of readily decomposable food wastes, were again the major factors destroying the optimal operation condition of the AD reactor at 6.0 kg/m3·d. Contrarily, the electrochemically activated dense communities of exoelectrogenic bacteria and VFA-oxidizing bacteria prevented VFAs from accumulating inside the BEAD reactor. This maintained stable pH and alkalinity conditions, ultimately contributing to stable methane production.
- Published
- 2019
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29. Both α2,3- and α2,6-linked sialic acids on O-linked glycoproteins act as functional receptors for porcine Sapovirus.
- Author
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Deok-Song Kim, Myra Hosmillo, Mia Madel Alfajaro, Ji-Yun Kim, Jun-Gyu Park, Kyu-Yeol Son, Eun-Hye Ryu, Frederic Sorgeloos, Hyung-Jun Kwon, Su-Jin Park, Woo Song Lee, Duck Cho, Joseph Kwon, Jong-Soon Choi, Mun-Il Kang, Ian Goodfellow, and Kyoung-Oh Cho
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Sapovirus, a member of the Caliciviridae family, is an important cause of acute gastroenteritis in humans and pigs. Currently, the porcine sapovirus (PSaV) Cowden strain remains the only cultivable member of the Sapovirus genus. While some caliciviruses are known to utilize carbohydrate receptors for entry and infection, a functional receptor for sapovirus is unknown. To characterize the functional receptor of the Cowden strain of PSaV, we undertook a comprehensive series of protein-ligand biochemical assays in mock and PSaV-infected cell culture and/or piglet intestinal tissue sections. PSaV revealed neither hemagglutination activity with red blood cells from any species nor binding activity to synthetic histo-blood group antigens, indicating that PSaV does not use histo-blood group antigens as receptors. Attachment and infection of PSaV were markedly blocked by sialic acid and Vibrio cholerae neuraminidase (NA), suggesting a role for α2,3-linked, α2,6-linked or α2,8-linked sialic acid in virus attachment. However, viral attachment and infection were only partially inhibited by treatment of cells with sialidase S (SS) or Maackia amurensis lectin (MAL), both specific for α2,3-linked sialic acid, or Sambucus nigra lectin (SNL), specific for α2,6-linked sialic acid. These results indicated that PSaV recognizes both α2,3- and α2,6-linked sialic acids for viral attachment and infection. Treatment of cells with proteases or with benzyl 4-O-β-D-galactopyranosyl-β-D-glucopyranoside (benzylGalNAc), which inhibits O-linked glycosylation, also reduced virus binding and infection, whereas inhibition of glycolipd synthesis or N-linked glycosylation had no such effect on virus binding or infection. These data suggest PSaV binds to cellular receptors that consist of α2,3- and α2,6-linked sialic acids on glycoproteins attached via O-linked glycosylation.
- Published
- 2014
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30. Lectin histochemistry in the small intestines of piglets naturally infected with porcine epidemic diarrhea virus.
- Author
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Bohye Kim, Sungwoong Jang, Hyewon Jang, Joong-Sun Kim, Tae-Il Jeon, Jun-Gyu Park, In-Sik Shin, Kyoung-Oh Cho, and Changjong Moon
- Subjects
PORCINE epidemic diarrhea virus ,CELL receptors ,SMALL intestine ,CHORIONIC villi ,WHEAT germ - Abstract
Importance: Porcine epidemic diarrhea virus (PEDV) binds to particular cell surface receptors to penetrate cells. The virus specifically identifies certain carbohydrate structures present on the surface of the cell to facilitate the binding process. Nevertheless, the influence of viral infections on specific alterations of glycoconjugates in the small intestines remains unexplored. Objective: This work aimed to examine the alterations in glycoconjugates in the small intestines of piglets naturally infected with PEDV using lectin histochemistry. Methods: Six piglets including three PEDV-infected and three non-infected piglets were evaluated. Small intestinal samples were histopathologically examined, and lectin histochemistry was performed. Results: Piglets infected with PEDV had significant histological abnormalities in their small intestines, such as pronounced villous atrophy, varying degrees of villous fusion, and diverse mucosal alterations. Specific regions of the duodenum, jejunum, and ileum showed discernible variations in glycoconjugate distribution, as determined by lectin histochemistry. Compared with the controls, the PEDV-infected piglets showed significant changes in N-acetylglucosamine- and galactose-binding lectins (particularly wheat germ agglutinin and Arachis hypogaea (peanut) agglutinin) in multiple intestinal regions. Conclusions and Relevance: These findings can enhance understanding of how viruses such as PEDV impact the glycoconjugate composition of the small intestines and emphasize the potential connection between the pathogenesis of PEDV and glycoconjugate. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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31. FACT subunit SUPT16H associates with BRD4 and contributes to silencing of interferon signaling
- Author
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Dawei Zhou, Zhenyu Wu, Jun-Gyu Park, Guillaume N Fiches, Tai-Wei Li, Qin Ma, Huachao Huang, Ayan Biswas, Luis Martinez-Sobrido, Netty G Santoso, and Jian Zhu
- Subjects
Cell Cycle Proteins ,Article ,gene silencing ,Genetics ,Humans ,NK cell ,acetylation ,SARS-CoV-2 ,Zika Virus Infection ,SUPT16H ,High Mobility Group Proteins ,COVID-19 ,Nuclear Proteins ,Epithelial Cells ,Zika Virus ,interferon ,antiviral immunity ,DNA-Binding Proteins ,Killer Cells, Natural ,FACT complex ,BRD4 ,Interferons ,Transcriptional Elongation Factors ,TIP60 ,Signal Transduction ,Transcription Factors - Abstract
FACT (FAcilitates Chromatin Transcription) is a heterodimeric protein complex composed of SUPT16H and SSRP1, and a histone chaperone participating in chromatin remodeling during gene transcription. FACT complex is profoundly regulated, and contributes to both gene activation and suppression. Here we reported that SUPT16H, a subunit of FACT, is acetylated in both epithelial and natural killer (NK) cells. The histone acetyltransferase TIP60 contributes to the acetylation of SUPT16H middle domain (MD) at lysine 674 (K674). Such acetylation of SUPT16H is recognized by bromodomain protein BRD4, which promotes protein stability of SUPT16H in both epithelial and NK cells. We further demonstrated that SUPT16H-BRD4 associates with histone modification enzymes (HDAC1, EZH2), and further regulates their activation status and/or promoter association as well as affects the relevant histone marks (H3ac, H3K9me3 and H3K27me3). BRD4 is known to profoundly regulate interferon (IFN) signaling, while such function of SUPT16H has never been explored. Surprisingly, our results revealed that SUPT16H genetic knockdown via RNAi or pharmacological inhibition by using its inhibitor, curaxin 137 (CBL0137), results in the induction of IFNs and interferon-stimulated genes (ISGs). Through this mechanism, depletion or inhibition of SUPT16H is shown to efficiently inhibit infection of multiple viruses, including Zika, influenza, and SARS-CoV-2. Furthermore, we demonstrated that depletion or inhibition of SUPT16H also causes the remarkable activation of IFN signaling in NK cells, which promotes the NK-mediated killing of virus-infected cells in a co-culture system using human primary NK cells. Overall, our studies unraveled the previously un-appreciated role of FACT complex in coordinating with BRD4 and regulating IFN signaling in both epithelial and NK cells, and also proposed the novel application of the FACT inhibitor CBL0137 to treat viral infections.
- Published
- 2022
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32. Multimodal Wrinkle Micro‐Nanoarchitectonics by Patterned Surface Material Properties for Transformative Structural Coloration
- Author
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Jun Gyu Park, Yeong Hoon Jeong, Sungjoon Ji, Juyeol Bae, Qitao Zhou, and Taesung Kim
- Subjects
Atomic and Molecular Physics, and Optics ,Electronic, Optical and Magnetic Materials - Published
- 2023
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33. Back Cover: Targeted Protein Upregulation of STING for Boosting the Efficacy of Immunotherapy (Angew. Chem. Int. Ed. 18/2023)
- Author
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Wansang Cho, Solchan Won, Yoona Choi, Sihyeong Yi, Jong Beom Park, Jun‐Gyu Park, Caroline E. Kim, Chintam Narayana, Ju Hee Kim, Junhyeong Yim, Young Il Choi, Dong‐Sup Lee, and Seung Bum Park
- Subjects
General Chemistry ,Catalysis - Published
- 2023
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34. Rücktitelbild: Targeted Protein Upregulation of STING for Boosting the Efficacy of Immunotherapy (Angew. Chem. 18/2023)
- Author
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Wansang Cho, Solchan Won, Yoona Choi, Sihyeong Yi, Jong Beom Park, Jun‐Gyu Park, Caroline E. Kim, Chintam Narayana, Ju Hee Kim, Junhyeong Yim, Young Il Choi, Dong‐Sup Lee, and Seung Bum Park
- Subjects
General Medicine - Published
- 2023
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35. Identification of a conserved S2 epitope present on spike proteins from all highly pathogenic coronaviruses
- Author
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Annalee W Nguyen, Yimin Huang, Rui P Silva, Ching-Lin Hsieh, Oladimeji S Olaluwoye, Tamer S Kaoud, Rebecca E Wilen, Ahlam N Qerqez, Jun-Gyu Park, Ahmed M Khalil, Laura R Azouz, Kevin C Le, Amanda L Bohanon, Andrea M DiVenere, Yutong Liu, Alison G Lee, Dzifa A Amengor, Sophie R Shoemaker, Shawn M Costello, Eduardo A Padlan, Susan Marqusee, Luis Martinez-Sobrido, Kevin N Dalby, Sheena D'Arcy, Jason S McLellan, and Jennifer A Maynard
- Subjects
General Immunology and Microbiology ,General Neuroscience ,General Medicine ,General Biochemistry, Genetics and Molecular Biology - Abstract
To address the ongoing SARS-CoV-2 pandemic and prepare for future coronavirus outbreaks, understanding the protective potential of epitopes conserved across SARS-CoV-2 variants and coronavirus lineages is essential. We describe a highly conserved, conformational S2 domain epitope present only in the prefusion core of β-coronaviruses: SARS-CoV-2 S2 apex residues 980–1006 in the flexible hinge. Antibody RAY53 binds the native hinge in MERS-CoV and SARS-CoV-2 spikes on the surface of mammalian cells and mediates antibody-dependent cellular phagocytosis and cytotoxicity against SARS-CoV-2 spike in vitro. Hinge epitope mutations that ablate antibody binding compromise pseudovirus infectivity, but changes elsewhere that affect spike opening dynamics, including those found in Omicron BA.1, occlude the epitope and may evade pre-existing serum antibodies targeting the S2 core. This work defines a third class of S2 antibody while providing insights into the potency and limitations of S2 core epitope targeting.
- Published
- 2023
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- View/download PDF
36. SARS-CoV-2 prefusion spike protein stabilized by six rather than two prolines is more potent for inducing antibodies that neutralize viral variants of concern
- Author
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Mijia Lu, Michelle Chamblee, Yuexiu Zhang, Chengjin Ye, Piyush Dravid, Jun-Gyu Park, KC Mahesh, Sheetal Trivedi, Satyapramod Murthy, Himanshu Sharma, Cole Cassady, Supranee Chaiwatpongsakorn, Xueya Liang, Jacob S. Yount, Prosper N. Boyaka, Mark E. Peeples, Luis Martinez-Sobrido, Amit Kapoor, and Jianrong Li
- Subjects
Multidisciplinary ,COVID-19 Vaccines ,Proline ,SARS-CoV-2 ,COVID-19 ,Viral Vaccines ,Vesiculovirus ,Antibodies, Viral ,Antibodies, Neutralizing ,Mice ,Viral Proteins ,Cricetinae ,Spike Glycoprotein, Coronavirus ,Vaccine Development ,Animals ,Humans ,Vesicular Stomatitis - Abstract
The spike (S) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the main target for neutralizing antibodies (NAbs). The S protein trimer is anchored in the virion membrane in its prefusion (preS) but metastable form. The preS protein has been stabilized by introducing two or six proline substitutions, to generate stabilized, soluble 2P or HexaPro (6P) preS proteins. Currently, it is not known which form is the most immunogenic. Here, we generated recombinant vesicular stomatitis virus (rVSV) expressing preS-2P, preS-HexaPro, and native full-length S, and compared their immunogenicity in mice and hamsters. The rVSV-preS-HexaPro produced and secreted significantly more preS protein compared to rVSV-preS-2P. Importantly, rVSV-preS-HexaPro triggered significantly more preS-specific serum IgG antibody than rVSV-preS-2P in both mice and hamsters. Antibodies induced by preS-HexaPro neutralized the B.1.1.7, B.1.351, P.1, B.1.427, and B.1.617.2 variants approximately two to four times better than those induced by preS-2P. Furthermore, preS-HexaPro induced a more robust Th1-biased cellular immune response than preS-2P. A single dose (10 4 pfu) immunization with rVSV-preS-HexaPro and rVSV-preS-2P provided complete protection against challenge with mouse-adapted SARS-CoV-2 and B.1.617.2 variant, whereas rVSV-S only conferred partial protection. When the immunization dose was lowered to 10 3 pfu, rVSV-preS-HexaPro induced two- to sixfold higher antibody responses than rVSV-preS-2P in hamsters. In addition, rVSV-preS-HexaPro conferred 70% protection against lung infection whereas only 30% protection was observed in the rVSV-preS-2P. Collectively, our data demonstrate that both preS-2P and preS-HexaPro are highly efficacious but preS-HexaPro is more immunogenic and protective, highlighting the advantages of using preS-HexaPro in the next generation of SARS-CoV-2 vaccines.
- Published
- 2023
37. Author response: Identification of a conserved S2 epitope present on spike proteins from all highly pathogenic coronaviruses
- Author
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Annalee W Nguyen, Yimin Huang, Rui P Silva, Ching-Lin Hsieh, Oladimeji S Olaluwoye, Tamer S Kaoud, Rebecca E Wilen, Ahlam N Qerqez, Jun-Gyu Park, Ahmed M Khalil, Laura R Azouz, Kevin C Le, Amanda L Bohanon, Andrea M DiVenere, Yutong Liu, Alison G Lee, Dzifa A Amengor, Sophie R Shoemaker, Shawn M Costello, Eduardo A Padlan, Susan Marqusee, Luis Martinez-Sobrido, Kevin N Dalby, Sheena D'Arcy, Jason S McLellan, and Jennifer A Maynard
- Published
- 2023
- Full Text
- View/download PDF
38. Systematic comparison between BNT162b2 and CoronaVac in the seroprotection against SARS-CoV-2 Alpha, Beta, Gamma, and Delta variants
- Author
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Wing Ying Au, Chengjin Ye, Sydney Leigh Briner, Gianmarco Domenico Suarez, Jeewon Han, Xinzhou Xu, Jun-Gyu Park, Melinda Ann Brindley, Luis Martinez-Sobrido, and Peter Pak-Hang Cheung
- Subjects
Microbiology (medical) ,COVID-19 Vaccines ,Infectious Diseases ,SARS-CoV-2 ,COVID-19 ,Humans ,Antibodies, Viral ,BNT162 Vaccine - Published
- 2022
- Full Text
- View/download PDF
39. A highly efficacious live attenuated mumps virus–based SARS-CoV-2 vaccine candidate expressing a six-proline stabilized prefusion spike
- Author
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Yuexiu Zhang, Mijia Lu, K C Mahesh, Eunsoo Kim, Mohamed M. Shamseldin, Chengjin Ye, Piyush Dravid, Michelle Chamblee, Jun-Gyu Park, Jesse M. Hall, Sheetal Trivedi, Supranee Chaiwatpongsakorn, Adam D. Kenny, Satyapramod Srinivasa Murthy, Himanshu Sharma, Xueya Liang, Jacob S. Yount, Amit Kapoor, Luis Martinez-Sobrido, Purnima Dubey, Prosper N. Boyaka, Mark E. Peeples, and Jianrong Li
- Subjects
COVID-19 Vaccines ,Multidisciplinary ,Mesocricetus ,Proline ,SARS-CoV-2 ,COVID-19 ,Vaccine Efficacy ,Antibodies, Viral ,Vaccines, Attenuated ,Antibodies, Neutralizing ,Mice ,Immunogenicity, Vaccine ,Mumps virus ,Spike Glycoprotein, Coronavirus ,Animals ,Measles-Mumps-Rubella Vaccine - Abstract
With the rapid increase in SARS-CoV-2 cases in children, a safe and effective vaccine for this population is urgently needed. The MMR (measles/mumps/rubella) vaccine has been one of the safest and most effective human vaccines used in infants and children since the 1960s. Here, we developed live attenuated recombinant mumps virus (rMuV)–based SARS-CoV-2 vaccine candidates using the MuV Jeryl Lynn (JL2) vaccine strain backbone. The soluble prefusion SARS-CoV-2 spike protein (preS) gene, stablized by two prolines (preS-2P) or six prolines (preS-6P), was inserted into the MuV genome at the P–M or F–SH gene junctions in the MuV genome. preS-6P was more efficiently expressed than preS-2P, and preS-6P expression from the P–M gene junction was more efficient than from the F–SH gene junction. In mice, the rMuV-preS-6P vaccine was more immunogenic than the rMuV-preS-2P vaccine, eliciting stronger neutralizing antibodies and mucosal immunity. Sera raised in response to the rMuV-preS-6P vaccine neutralized SARS-CoV-2 variants of concern, including the Delta variant equivalently. Intranasal and/or subcutaneous immunization of IFNAR1 −/− mice and golden Syrian hamsters with the rMuV-preS-6P vaccine induced high levels of neutralizing antibodies, mucosal immunoglobulin A antibody, and T cell immune responses, and were completely protected from challenge by both SARS-CoV-2 USA-WA1/2020 and Delta variants. Therefore, rMuV-preS-6P is a highly promising COVID-19 vaccine candidate, warranting further development as a tetravalent MMR vaccine, which may include protection against SARS-CoV-2.
- Published
- 2022
- Full Text
- View/download PDF
40. Jet-nozzle based improvement of dissolved H2 concentration for efficient in-situ biogas upgrading in an up-flow anaerobic sludge blanket (UASB) reactor
- Author
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Hye-Jeong Kwon, A-In Cheon, Hang-Bae Jun, and Jun-Gyu Park
- Subjects
Mass transfer coefficient ,Jet (fluid) ,060102 archaeology ,Renewable Energy, Sustainability and the Environment ,Methanogenesis ,020209 energy ,Energy conversion efficiency ,Nozzle ,06 humanities and the arts ,02 engineering and technology ,Blanket ,Pulp and paper industry ,Biogas ,Methanation ,0202 electrical engineering, electronic engineering, information engineering ,Environmental science ,0601 history and archaeology - Abstract
Improvement of dissolved concentration and mass transfer coefficient (KLa) of H2 is a main strategy for efficient biogas upgrading process. In this study, Jet nozzle reactor (JNR) was applied to improve dissolved H2 (DH2) concentration and KLa for successful in-situ biogas upgrading in an up-flow anaerobic sludge blanket (UASB) reactor. JNR could improve the DH2 concentration and KLa to 1.71 and 1.72 times higher than those of control reactor, respectively. Increased DH2 concentration and KLa contributed to increase CH4 content to 96.1% in UASB combined with JNR (JN-UASB). JN-UASB achieved maximum H2 conversion efficiency of 97.9% and minimum H2 content of 3.9% in biogas. Community of hydrogenotrophic methanogens and efficiency of H2 conversion in JN-UASB were 1.1 and 1.3 times higher than those in control UASB (C-UASB), respectively. This study confirmed that DH2 concentration and KLa increased by JNR affected the improvement of H2 methanation efficiency via rapid hydrogenotrophic methanogenesis. The findings of this study provide technical strategy and further research direction to upgrade biogas.
- Published
- 2021
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- View/download PDF
41. Safety analysis of ex vivo-expanded canine natural killer cells in a xenogeneic mouse model of graft-versus-host disease
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Sang-Ki Kim, Yu-Jin Lim, Kyoung-Oh Cho, Mee-Sun Yoon, Dong-Jun Shin, Soo-Hyeon Lee, Se-Cheol Park, Cheol-Jung Kim, Yeong-Bin Baek, Jun-Gyu Park, and Jeong Won Hong
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Necrosis ,CD3 Complex ,CD3 ,medicine.medical_treatment ,Transplantation, Heterologous ,Immunology ,Receptors, Antigen, T-Cell ,Graft vs Host Disease ,Mice, SCID ,Peripheral blood mononuclear cell ,Mice ,03 medical and health sciences ,Dogs ,0302 clinical medicine ,Cancer immunotherapy ,Mice, Inbred NOD ,medicine ,Animals ,Immunology and Allergy ,biology ,Cell Biology ,Immunotherapy ,medicine.disease ,Killer Cells, Natural ,030104 developmental biology ,Graft-versus-host disease ,030220 oncology & carcinogenesis ,Leukocytes, Mononuclear ,biology.protein ,CD5 ,medicine.symptom ,Ex vivo - Abstract
Canine natural killer (NK) cells are large, granular lymphocytes that are neither B lymphocytes nor T lymphocytes. However, it has been reported that canine NK cells share some of the phenotypic characteristics of T lymphocytes, such as CD3 and CD5. Studies are needed to assess the safety of canine NK cells for immunotherapy, especially because the safety of using allogeneic NK cells as an immunotherapy for dogs has yet to be shown. In this study, the safety of cultured canine NK cells was assessed using a xenogeneic mouse model of graft-versus-host disease (GVHD). Mice were injected with either canine peripheral blood mononuclear cells (PBMCs) or cultured NK cells for 2 or 3 weeks. Data were then collected on changes in mice body weights, disease severity scores, and survival rates. Histopathological and immunohistochemical evaluations were also performed. All mice injected with canine PBMCs died within 45 days after injection. Severe clinical signs were caused by GVHD. The histopathological and immunohistochemical evaluations showed that mice injected with canine PBMCs had multiple lesions, including necrosis in their lungs, livers, kidneys, and stomachs, and the injected cells were present around the lesions. By contrast, no mice injected with cultured NK cells without removing the CD3+TCR– cells exhibited any clinical abnormalities. Moreover, they all survived the 90-day experimental period without exhibiting any histopathological changes. Accordingly, the results of this study suggest that canine NK cells do not cause significant side effects such as GVHD and allogeneic NK cells can safely be used for cancer immunotherapy in dogs.
- Published
- 2021
- Full Text
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42. Direct Single-Step Printing of Conductive Grids on Curved Surfaces Using Template-Guided Foaming
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Taesung Kim, Jun Gyu Park, Youngchul Chae, Juyeol Bae, Janghyun Ju, and Ronghui Wu
- Subjects
Materials science ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Grid ,01 natural sciences ,Evaporation (deposition) ,Silver nanoparticle ,0104 chemical sciences ,Nanomaterials ,Template ,General Materials Science ,0210 nano-technology ,Lithography ,Electrical conductor ,Transparent conducting film - Abstract
Advanced transparent conductors have been studied intensively in the aspects of materials, structures, and printing methods. The material and structural advancements have been successfully accomplished with various conductive nanomaterials and spring-like structures for better electrical conductivity and high mechanical flexibility of the transparent conductors. However, the capability to print submicrometer conductive patterns directly and conformally on curved surfaces with low processing cost and high throughput remains a technological challenge to achieve, primarily because of the original two-dimensional (2D) nature of conventional lithography processes. In our study, we exploit a liquid-mediated patterning approach in the development of flexible templates, enabling printing of curvilinear silver grids in a single-step and strain-free manner at a submicrometer resolution within several minutes with minimum loss of noble metals. The template can guide arrays of receding liquid-air interfaces on curved substrates during liquid evaporation, thereby generating ordered 2D foam structures that can confine and assemble silver nanoparticles in grid patterns. The printed silver grids exhibit suitable optical, electrical, and Joule-heating performances, enabling their application in transparent heaters. Our technique has the potential to extend the existing 2D micro/nanofluidic liquid-mediated patterning approach to three-dimensional (3D) control of liquid-air interfaces for low-cost all-liquid-processed functional 3D optoelectronics in the future.
- Published
- 2021
- Full Text
- View/download PDF
43. Animal Models of COVID-19: Transgenic Mouse Model
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Jun-Gyu Park, Paula A. Pino, Anwari Akhter, Xavier Alvarez, Jordi B. Torrelles, and Luis Martinez-Sobrido
- Subjects
Mice, Inbred C57BL ,Disease Models, Animal ,Mice ,SARS-CoV-2 ,Animals ,COVID-19 ,Cytokines ,Mice, Transgenic ,Chemokines ,Antibodies, Neutralizing ,Lung ,Article - Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), emerged in December 2019 in Wuhan, China, and rapidly spread throughout the world, threatening global public health. An animal model is a valuable and a crucial tool that allows understanding of nature in the pathogenesis of SARS-CoV-2 and its associated COVID-19 disease. Here we introduce detailed protocols of SARS-CoV-2 infection and COVID-19 disease using C57BL/6 (B6) transgenic mice expressing the human angiotensin-converting enzyme 2 (hACE2) from the human cytokeratin 18 promoter (K18 hACE2). To mimic natural SARS-CoV-2 infection, K18 hACE2 transgenic mice are infected intranasally under anesthesia. Upon infection, viral pathogenesis is determined by monitoring changes in body weight (morbidity) and monitoring survival (mortality), cytokine/chemokine responses, gross-lung pathology, histopathology, and viral replication in tissues. The presence of the virus and viral replication is evaluated by immunohistochemistry (IHC) and viral titrations, respectively, from the upper (nasal turbinate) and the lower (lungs) respiratory tracts, and nervous system (brain). Also, the immune response to SARS-CoV-2 infection is measured by cytokine/chemokine enzyme-linked immunosorbent assay (ELISA) from lung, spleen and brain homogenates to characterize the cytokine storm that hallmarks as one of the major causes of death caused by SARS-CoV-2 infection. This small rodent animal model based on the use of K18 hACE2 transgenic mice represents an excellent option to understand the pathogenicity of natural SARS-CoV-2 strains and its recently described Variants of Concern (VoC), and will be applicable to the identification and characterization of prophylactic (vaccine) and therapeutic (antiviral and/or neutralizing monoclonal antibodies) strategies for the prevention or treatment of SARS-CoV-2 infection or its associated COVID-19 disease.
- Published
- 2022
44. Kidney residency of VISTA-positive macrophages accelerates repair from ischemic injury
- Author
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Eun Young Choi, Jun Gyu Park, Dong Ki Kim, Min Gang Kim, Hyun Mu Shin, Seung Seok Han, Gwanghun Kim, Seung Hee Yang, Yun Hui Jeon, Cho Rong Lee, Murim Choi, Kwon Wook Joo, Cheol Kwak, Yon Su Kim, Hang Rae Kim, and Dong Sup Lee
- Subjects
0301 basic medicine ,Phagocytosis ,CD14 ,T cell ,030232 urology & nephrology ,Inflammation ,Kidney ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Ischemia ,medicine ,Animals ,Chemistry ,Macrophages ,Internship and Residency ,Immune checkpoint ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Nephrology ,Apoptosis ,Reperfusion Injury ,medicine.symptom - Abstract
Tissue-resident macrophages have unique tissue-specific functions in maintaining homeostasis and resolving inflammation. However, the repair role and relevant molecules of kidney-resident macrophages after ischemic injury remain unresolved. To this end, mice without kidney-resident R1 macrophages but containing infiltrating monocyte-derived R2 macrophages were generated using differential cellular kinetics following clodronate liposome treatment. When ischemia-reperfusion injury was induced in these mice, late phase repair was reduced. Transcriptomic and flow cytometric analyses identified that V-domain Ig suppressor of T cell activation (VISTA), an inhibitory immune checkpoint molecule, was constitutively expressed in kidney-resident R1 macrophages, but not in other tissue-resident macrophages. Here, VISTA functioned as a scavenger of apoptotic cells and served as a checkpoint to control kidney-infiltrating T cells upon T cell receptor-mediated stimulation. Together these functions improved the repair process after ischemia-reperfusion injury. CD14+ CD33+ mononuclear phagocytes of human kidney also expressed VISTA, which has similar functions to the mouse counterpart. Thus, VISTA is upregulated in kidney macrophages in a tissue-dependent manner and plays a repair role during ischemic injury.
- Published
- 2020
- Full Text
- View/download PDF
45. Effects of voltage supply on the methane production rates and pathways in an anaerobic digestion reactor using different electron donors
- Author
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Hang-Bae Jun, Wei-Qi Shi, Tae-Young Heo, Jun-Gyu Park, and Hye-Jeong Kwon
- Subjects
Anaerobic sludge ,Renewable Energy, Sustainability and the Environment ,Chemistry ,Methanogenesis ,Energy Engineering and Power Technology ,02 engineering and technology ,Electron ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,01 natural sciences ,Methane ,0104 chemical sciences ,Anaerobic digestion ,Electron transfer ,chemistry.chemical_compound ,Fuel Technology ,Environmental chemistry ,Methane production ,0210 nano-technology ,Voltage - Abstract
The effects of voltage on the methane production rate (k) of various types of anaerobic sludge biomasses are quantitatively demonstrated. Voltage is supplied to the anaerobic digestion (AD) sludges independently and quickly improves the methane production rates and specific methanogenic activities (SMAs), regardless of bio-electrochemical activation of the sludge. This supports previous findings that bio-electrochemical AD (BEAD) shows higher stabilization and methane production rates than those of AD. However, in reactors fed with H2/CO2, SMA and k are not significantly increased by the voltage supply, indicating that direct electron transfer for hydrogenotrophic methanogenesis is not a major methane formation pathway. The voltage supply contributed to indirect electron transfer for H2 production. Therefore, indirect electron transfer via H2 is a significant factor in hydrogenotrophic methanogenesis. These findings indicate changes in the general pathways of AD achieved using a voltage supply and provide a better understanding of previous BEAD studies.
- Published
- 2020
- Full Text
- View/download PDF
46. Heterogeneous semiconductor nanowire array for sensitive broadband photodetector by crack photolithography-based micro-/nanofluidic platforms
- Author
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Taesung Kim, Qitao Zhou, and Jun Gyu Park
- Subjects
Microelectromechanical systems ,Nanoelectromechanical systems ,Materials science ,Fabrication ,business.industry ,General Chemical Engineering ,Nanowire ,Photodetector ,02 engineering and technology ,General Chemistry ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,law.invention ,Semiconductor ,law ,Optoelectronics ,Photolithography ,0210 nano-technology ,business ,Microfabrication - Abstract
The in situ growth of nanowires (NWs) into nano-/microelectromechanical systems (NEMS/MEMS) by solution processing is attractive for its relative simplicity and economic value. We present innovative, versatile microfabrication that produces multiple, heterogeneous semiconductor NWs. A crack photolithography-based micro-/nanofluidic platform has been developed. This platform offers the in situ solution growth of NWs while enabling the control of quantity, dimensions, orientation, alignment, position, and material. The generation of grain boundary (GB)-rich CH3NH3PbI3 NWs using the micro-/nanofluidic device is exemplary. High-quality single-crystal CH3NH3PbI3 NWs were derived by injection of a CH3NH3PbI3 solution. We produced a parallel NW array of CH3NH3PbI3 NW for visible light detection and ZnO NW for ultraviolet detection, thereby demonstrating an unprecedented broadband composite photodetector. The micro-/nanofluidic fabrication platform enables the production of multiple, heterogeneous semiconductor NW arrays on one substrate, offering the potential to elicit synergistic performance and functional enhancements from various NWs.
- Published
- 2020
- Full Text
- View/download PDF
47. Numb-associated kinases are required for SARS-CoV-2 infection and are cellular targets for therapy
- Author
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Marwah Karim, Sirle Saul, Luca Ghita, Malaya Kumar Sahoo, Chengjin Ye, Nishank Bhalla, Jing Jin, Jun-Gyu Park, Belén Martinez-Gualda, Michael Patrick East, Gary L. Johnson, Benjamin A. Pinsky, Luis Martinez-Sobrido, Christopher R. M. Asquith, Aarthi Narayanan, Steven De Jonghe, and Shirit Einav
- Subjects
viruses ,fungi ,virus diseases ,respiratory tract diseases - Abstract
The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to pose serious threats to global health. We previously reported that AAK1, BIKE and GAK, members of the Numb-associated kinase family, control intracellular trafficking of multiple RNA viruses during viral entry and assembly/egress. Here, using both genetic and pharmacological approaches, we probe the functional relevance of NAKs for SARS-CoV-2 infection. siRNA-mediated depletion of AAK1, BIKE, GAK, and STK16, the fourth member of the NAK family, suppressed SARS-CoV-2 infection in human lung epithelial cells. Both known and novel small molecules with potent AAK1/BIKE, GAK or STK16 activity suppressed SARS-CoV-2 infection. Moreover, combination treatment with the approved anti-cancer drugs, sunitinib and erlotinib, with potent anti-AAK1/BIKE and GAK activity, respectively, demonstrated synergistic effect against SARS-CoV-2 infection in vitro. Time-of-addition experiments revealed that pharmacological inhibition of AAK1 and BIKE suppressed viral entry as well as late stages of the SARS-CoV-2 life cycle. Lastly, suppression of NAKs expression by siRNAs inhibited entry of both wild type and SARS-CoV-2 pseudovirus. These findings provide insight into the roles of NAKs in SARS-CoV-2 infection and establish a proof-of-principle that pharmacological inhibition of NAKs can be potentially used as a host-targeted approach to treat SARS-CoV-2 with potential implications to other coronaviruses.
- Published
- 2022
- Full Text
- View/download PDF
48. Potent universal-coronavirus therapeutic activity mediated by direct respiratory administration of a Spike S2 domain-specific human neutralizing monoclonal antibody
- Author
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Michael S. Piepenbrink, Jun-Gyu Park, Ashlesha Desphande, Andreas Loos, Chengjin Ye, Madhubanti Basu, Sanghita Sarkar, David Chauvin, Jennifer Woo, Philip Lovalenti, Nathaniel B. Erdmann, Paul A. Goepfert, Vu L. Truong, Richard A. Bowen, Mark R. Walter, Luis Martinez-Sobrido, and James J. Kobie
- Subjects
viruses ,virus diseases - Abstract
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) marks the third novel β-coronavirus to cause significant human mortality in the last two decades. Although vaccines are available, too few have been administered worldwide to keep the virus in check and to prevent mutations leading to immune escape. To determine if antibodies could be identified with universal coronavirus activity, plasma from convalescent subjects was screened for IgG against a stabilized pre-fusion SARS-CoV-2 spike S2 domain, which is highly conserved between human β-coronavirus. From these subjects, several S2-specific human monoclonal antibodies (hmAbs) were developed that neutralized SARS-CoV-2 with recognition of all variants of concern (VoC) tested (Beta, Gamma, Delta, Epsilon, and Omicron). The hmAb 1249A8 emerged as the most potent and broad hmAb, able to recognize all human β-coronavirus and neutralize SARS-CoV and MERS-CoV. 1249A8 demonstrated significant prophylactic activity in K18 hACE2 mice infected with SARS-CoV-2 lineage A and lineage B Beta, and Omicron VoC. 1249A8 delivered as a single 4 mg/kg intranasal (i.n.) dose to hamsters 12 hours following infection with SARS-CoV-2 Delta protected them from weight loss, with therapeutic activity further enhanced when combined with 1213H7, an S1-specific neutralizing hmAb. As little as 2 mg/kg of 1249A8 i.n. dose 12 hours following infection with SARS-CoV Urbani strain, protected hamsters from weight loss and significantly reduced upper and lower respiratory viral burden. These results indicate in vivo cooperativity between S1 and S2 specific neutralizing hmAbs and that potent universal coronavirus neutralizing mAbs with therapeutic potential can be induced in humans and can guide universal coronavirus vaccine development.
- Published
- 2022
49. Numb-associated kinases are required for SARS-CoV-2 infection and are cellular targets for antiviral strategies
- Author
-
Marwah Karim, Sirle Saul, Luca Ghita, Malaya Kumar Sahoo, Chengjin Ye, Nishank Bhalla, Chieh-Wen Lo, Jing Jin, Jun-Gyu Park, Belén Martinez-Gualda, Michael Patrick East, Gary L. Johnson, Benjamin A. Pinsky, Luis Martinez-Sobrido, Christopher R.M. Asquith, Aarthi Narayanan, Steven De Jonghe, and Shirit Einav
- Subjects
Pharmacology ,SARS-CoV-2 ,Virology ,Humans ,Membrane Proteins ,Nerve Tissue Proteins ,Protein Serine-Threonine Kinases ,Virus Internalization ,Antiviral Agents ,Pandemics ,Transcription Factors ,COVID-19 Drug Treatment - Abstract
The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to pose serious threats to global health. We previously reported that AAK1, BIKE and GAK, members of the Numb-associated kinase family, control intracellular trafficking of multiple RNA viruses during viral entry and assembly/egress. Here, using both genetic and pharmacological approaches, we probe the functional relevance of NAKs for SARS-CoV-2 infection. siRNA-mediated depletion of AAK1, BIKE, GAK, and STK16, the fourth member of the NAK family, suppressed SARS-CoV-2 infection in human lung epithelial cells. Both known and novel small molecules with potent AAK1/BIKE, GAK or STK16 activity suppressed SARS-CoV-2 infection. Moreover, combination treatment with the approved anti-cancer drugs, sunitinib and erlotinib, with potent anti-AAK1/BIKE and GAK activity, respectively, demonstrated synergistic effect against SARS-CoV-2 infection in vitro. Time-of-addition experiments revealed that pharmacological inhibition of AAK1 and BIKE suppressed viral entry as well as late stages of the SARS-CoV-2 life cycle. Lastly, suppression of NAKs expression by siRNAs inhibited entry of both wild type and SARS-CoV-2 pseudovirus. These findings provide insight into the roles of NAKs in SARS-CoV-2 infection and establish a proof-of-principle that pharmacological inhibition of NAKs can be potentially used as a host-targeted approach to treat SARS-CoV-2 with potential implications to other coronaviruses.
- Published
- 2022
50. Immunization with recombinant accessory protein-deficient SARS-CoV-2 protects against lethal challenge and viral transmission
- Author
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Chengjin Ye, Jun-Gyu Park, Kevin Chiem, Piyush Dravid, Anna Allué-Guardia, Andreu Garcia-Vilanova, Amit Kapoor, Mark R. Walter, James J. Kobie, Richard K. Plemper, Jordi B. Torrelles, and Luis Martinez-Sobrido
- Subjects
viruses ,virus diseases ,Article - Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has led to a worldwide Coronavirus Disease 2019 (COVID-19) pandemic. Despite high efficacy of the authorized vaccines, protection against the surging variants of concern (VoC) was less robust. Live-attenuated vaccines (LAV) have been shown to elicit robust and long-term protection by induction of host innate and adaptive immune responses. We sought to develop a COVID-19 LAV by generating 3 double open reading frame (ORF)-deficient recombinant (r)SARS-CoV-2 simultaneously lacking two accessory open reading frame (ORF) proteins (ORF3a/ORF6, ORF3a/ORF7a, and ORF3a/ORF7b). Here, we report that these double ORF-deficient rSARS-CoV-2 have slower replication kinetics and reduced fitness in cultured cells as compared to their parental wild-type (WT) counterpart. Importantly, these double ORF-deficient rSARS-CoV-2 showed attenuation in both K18 hACE2 transgenic mice and golden Syrian hamsters. A single intranasal dose vaccination induced high levels of neutralizing antibodies against different SARS-CoV-2 VoC, and also activated viral component-specific T-cell responses. Notably, the double ORF-deficient rSARS-CoV-2 were able to protect, as determined by inhibition of viral replication, shedding, and transmission, against challenge with SARS-CoV-2. Collectively, our results demonstrate the feasibility to implement these double ORF-deficient rSARS-CoV-2 as safe, stable, immunogenic and protective LAV for the prevention of SARS-CoV-2 infection and associated COVID-19 disease.
- Published
- 2022
- Full Text
- View/download PDF
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