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Article ; Online: Kaposi's sarcoma-associated herpesvirus replication and transcription activator protein activates CD274/PD-L1 gene promoter.

Miyazawa, Masanori / Yamamoto, Yuichiro / Katayama, Kazuhiro / Sugimoto, Yoshikazu / Noguchi, Kohji

2022 Volume 114, Issue 4, Page(s) 1718–1728

Abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) is responsible for the pathogenesis of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman disease. The expression of immunosuppressive genes, such as IL-10 and CD274/PD-L1 is observed ... ...

Abstract	Kaposi's sarcoma-associated herpesvirus (KSHV) is responsible for the pathogenesis of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman disease. The expression of immunosuppressive genes, such as IL-10 and CD274/PD-L1 is observed during KSHV-associated pathogenesis, and the modulation of the host immune system by KSHV contributes to establishing viral persistence in the host. Understanding the mechanism that allows the virus to evade host cell immunity would be helpful in order to develop therapeutic strategies for KSHV malignancy. In this study, we show that KSHV replication and transcriptional activator (K-RTA), an essential activator of the viral lytic cycle, transactivates the CD274/PD-L1 gene promoter. Mechanistically, we demonstrate that the binding of K-RTA to the cellular specificity protein 1 (SP1) is critical for K-RTA-mediated CD274/PD-L1 promoter activation. These findings suggest that K-RTA cooperates with intracellular SP1 to activate the expression of CD274/PD-L1, which helps the virus regulate immune checkpoints to escape and survive.
MeSH term(s)	Humans ; B7-H1 Antigen/genetics ; B7-H1 Antigen/metabolism ; Herpesvirus 8, Human/genetics ; Herpesvirus 8, Human/metabolism ; Immediate-Early Proteins/genetics ; Trans-Activators/genetics ; Trans-Activators/metabolism ; Transcription Factors/metabolism ; Virus Replication/genetics ; Promoter Regions, Genetic
Chemical Substances	B7-H1 Antigen ; CD274 protein, human ; Immediate-Early Proteins ; Trans-Activators ; Transcription Factors
Language	English
Publishing date	2022-12-16
Publishing country	England
Document type	Journal Article
ZDB-ID	2115647-5
ISSN	1349-7006 ; 1349-7006
ISSN (online)	1349-7006
ISSN	1349-7006
DOI	10.1111/cas.15673
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Overcoming antibody-resistant SARS-CoV-2 variants with bispecific antibodies constructed using non-neutralizing antibodies.

Inoue, Tetsuya / Yamamoto, Yuichiro / Sato, Kaoru / Okemoto-Nakamura, Yuko / Shimizu, Yoshimi / Ogawa, Motohiko / Onodera, Taishi / Takahashi, Yoshimasa / Wakita, Takaji / Kaneko, Mika K / Fukasawa, Masayoshi / Kato, Yukinari / Noguchi, Kohji

iScience

2024 Volume 27, Issue 4, Page(s) 109363

Abstract: A current challenge is the emergence of SARS-CoV-2 variants, such as BQ.1.1 and XBB.1.5, that can evade immune defenses, thereby limiting antibody drug effectiveness. Emergency-use antibody drugs, including the widely effective bebtelovimab, are losing ... ...

Abstract	A current challenge is the emergence of SARS-CoV-2 variants, such as BQ.1.1 and XBB.1.5, that can evade immune defenses, thereby limiting antibody drug effectiveness. Emergency-use antibody drugs, including the widely effective bebtelovimab, are losing their benefits. One potential approach to address this issue are bispecific antibodies which combine the targeting abilities of two antibodies with distinct epitopes. We engineered neutralizing bispecific antibodies in the IgG-scFv format from two initially non-neutralizing antibodies, CvMab-6 (which binds to the receptor-binding domain [RBD]) and CvMab-62 (targeting a spike protein S2 subunit epitope adjacent to the known anti-S2 antibody epitope). Furthermore, we created a bispecific antibody by incorporating the scFv of bebtelovimab with our anti-S2 antibody, demonstrating significant restoration of effectiveness against bebtelovimab-resistant BQ.1.1 variants. This study highlights the potential of neutralizing bispecific antibodies, which combine existing less effective anti-RBD antibodies with anti-S2 antibodies, to revive the effectiveness of antibody therapeutics compromised by immune-evading variants.
Language	English
Publishing date	2024-02-29
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2024.109363
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Article ; Online: SARS-CoV-2 Spike Protein Mutation at Cysteine-488 Impairs Its Golgi Localization and Intracellular S1/S2 Processing.

Yamamoto, Yuichiro / Inoue, Tetsuya / Inoue, Miyu / Murae, Mana / Fukasawa, Masayoshi / Kaneko, Mika K / Kato, Yukinari / Noguchi, Kohji

International journal of molecular sciences

2022 Volume 23, Issue 24

Abstract: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds to the cellular receptor-angiotensin-converting enzyme-2 (ACE2) as the first step in viral cell entry. SARS-CoV-2 spike protein expression in the ACE2-expressing cell ... ...

Abstract	The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds to the cellular receptor-angiotensin-converting enzyme-2 (ACE2) as the first step in viral cell entry. SARS-CoV-2 spike protein expression in the ACE2-expressing cell surface induces cell-cell membrane fusion, thus forming syncytia. To exert its fusogenic activity, the spike protein is typically processed at a specific site (the S1/S2 site) by cellular proteases such as furin. The C488 residue, located at the spike-ACE2 interacting surface, is critical for the fusogenic and infectious roles of the SARS-CoV-2 spike protein. We have demonstrated that the C488 residue of the spike protein is involved in subcellular targeting and S1/S2 processing. C488 mutant spike localization to the Golgi apparatus and cell surface were impaired. Consequently, the S1/S2 processing of the spike protein, probed by anti-Ser-686-cleaved spike antibody, markedly decreased in C488 mutant spike proteins. Moreover, brefeldin-A-mediated endoplasmic-reticulum-to-Golgi traffic suppression also suppressed spike protein S1/S2 processing. As brefeldin A treatment and C488 mutation inhibited S1/S2 processing and syncytia formation, the C488 residue of spike protein is required for functional spike protein processing.
Language	English
Publishing date	2022-12-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms232415834
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Article ; Online: Anti-HBV drug entecavir ameliorates DSS-induced colitis through PD-L1 induction.

Yamamoto, Yuichiro / Carreras, Joaquim / Shimizu, Takanobu / Kakizaki, Masatoshi / Kikuti, Yara Yukie / Roncador, Giovanna / Nakamura, Naoya / Kotani, Ai

Pharmacological research

2022 Volume 179, Page(s) 105918

Abstract: PD-L1-mediated signaling is one of the major processes that regulate local inflammatory responses in the gut. To date, protective effects against colitis through direct Fc-fused PD-L1 administration or indirect PD-L1 induction by probiotics have been ... ...

Abstract	PD-L1-mediated signaling is one of the major processes that regulate local inflammatory responses in the gut. To date, protective effects against colitis through direct Fc-fused PD-L1 administration or indirect PD-L1 induction by probiotics have been reported. We have previously shown that the anti-HBV drug entecavir (ETV) induces PD-L1 expression in human hepatocytes. In the present study, we investigated whether ETV induces PD-L1 expression in intestinal cells and provides a protective effect against DSS-induced colitis. ETV induced PD-L1 expression in epithelial cells, rather than T and B cells, improving the symptoms of colitis. In the mechanistic analysis, Th17 cell differentiation was inhibited and B cell infiltration into the lamina propria was reduced. In addition, PD-L1 expression was positively correlated with Foxp3 or CSF1-R. In conclusion, ETV upregulated PD-L1 expression in epithelial cells and ameliorated inflammation in DSS-induced colitis. These results suggest that ETV may be a potential therapeutic agent as a PD-L1 enhancer for the treatment of human IBD.
MeSH term(s)	Animals ; B7-H1 Antigen/metabolism ; Colitis/chemically induced ; Colitis/drug therapy ; Colitis/metabolism ; Dextran Sulfate/pharmacology ; Guanine/analogs & derivatives ; Humans ; Mice ; Mice, Inbred C57BL ; Pharmaceutical Preparations/metabolism ; T-Lymphocytes, Regulatory ; Th17 Cells
Chemical Substances	B7-H1 Antigen ; Pharmaceutical Preparations ; entecavir (5968Y6H45M) ; Guanine (5Z93L87A1R) ; Dextran Sulfate (9042-14-2)
Language	English
Publishing date	2022-01-11
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1003347-6
ISSN	1096-1186 ; 0031-6989 ; 1043-6618
ISSN (online)	1096-1186
ISSN	0031-6989 ; 1043-6618
DOI	10.1016/j.phrs.2021.105918
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Article: The function of SARS-CoV-2 spike protein is impaired by disulfide-bond disruption with mutation at cysteine-488 and by thiol-reactive N-acetyl-cysteine and glutathione

Murae, Mana / Shimizu, Yoshimi / Yamamoto, Yuichiro / Kobayashi, Asuka / Houri, Masumi / Inoue, Tetsuya / Irie, Takuya / Gemba, Ryutaro / Kondo, Yosuke / Nakano, Yoshio / Miyazaki, Satoru / Yamada, Daisuke / Saitoh, Akiyoshi / Ishii, Isao / Onodera, Taishi / Takahashi, Yoshimasa / Wakita, Takaji / Fukasawa, Masayoshi / Noguchi, Kohji

Biochemical and biophysical research communications. 2022 Mar. 15, v. 597

2022

Abstract: Viral spike proteins play important roles in the viral entry process, facilitating attachment to cellular receptors and fusion of the viral envelope with the cell membrane. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds ... ...

Abstract	Viral spike proteins play important roles in the viral entry process, facilitating attachment to cellular receptors and fusion of the viral envelope with the cell membrane. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds to the cellular receptor angiotensin converting enzyme-2 (ACE2) via its receptor-binding domain (RBD). The cysteine residue at position 488, consisting of a disulfide bridge with cysteine 480 is located in an important structural loop at ACE2-binding surface of RBD, and is highly conserved among SARS-related coronaviruses. We showed that the substitution of Cys-488 with alanine impaired pseudotyped SARS-CoV-2 infection, syncytium formation, and cell-cell fusion triggered by SARS-CoV-2 spike expression. Consistently, in vitro binding of RBD and ACE2, spike-mediated cell-cell fusion, and pseudotyped viral infection of VeroE6/TMPRSS2 cells were inhibited by the thiol-reactive compounds N-acetylcysteine (NAC) and a reduced form of glutathione (GSH). Furthermore, we demonstrated that the activity of variant spikes from the SARS-CoV-2 alpha and delta strains were also suppressed by NAC and GSH. Taken together, these data indicate that Cys-488 in spike RBD is required for SARS-CoV-2 spike functions and infectivity, and could be a target of anti-SARS-CoV-2 therapeutics.
Keywords	Severe acute respiratory syndrome coronavirus 2 ; acetylcysteine ; alanine ; cell membranes ; cysteine ; disulfide bonds ; giant cells ; glutathione ; mutation ; pathogenicity ; research ; therapeutics
Language	English
Dates of publication	2022-0315
Size	p. 30-36.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2022.01.106
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: An Automatic Image Collection System for Multicenter Clinical Studies.

Takeda, Toshihiro / Manabe, Shirou / Hattori, Atsushi / Yamaguchi, Junji / Konishi, Shozo / Yamamoto, Yuichiro / Takahashi, Daiyo / Matsumura, Yasushi

Studies in health technology and informatics

2020 Volume 270, Page(s) 23–27

Abstract: The acquisition of medical images from multiple medial institutions has become important for high-quality clinical studies. In recent years, electronic data submission has enabled the transmission of image data to independent institutions more quickly ... ...

Abstract	The acquisition of medical images from multiple medial institutions has become important for high-quality clinical studies. In recent years, electronic data submission has enabled the transmission of image data to independent institutions more quickly and easily than before. However, the selection, anonymization, and transmission of medical images still require human resources in the form of clinical research collaborators. In this study, we developed an image collection system that works with the electronic data capture (EDC) system. In this image collection system, medical images are selected based on EDC input information, patient ID is anonymized to a subject ID issued by the EDC, and the selected anonymized images are transferred to the research institute without human intervention. In the research institute, clinical information registered by the EDC and clinical images collected by the image collection system are managed by the same subject ID and can be used for clinical studies. In October 2019, our image collection system was introduced to 13 medical institutions and has now begun collecting medical images from the in-hospital picture archiving and communication system (PACS) of those institutions.
MeSH term(s)	Automation ; Humans ; Image Processing, Computer-Assisted ; Radiology Information Systems
Language	English
Publishing date	2020-06-13
Publishing country	Netherlands
Document type	Journal Article
ISSN	1879-8365
ISSN (online)	1879-8365
DOI	10.3233/SHTI200115
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Extracellular vesicles secreted by HBV-infected cells modulate HBV persistence in hydrodynamic HBV transfection mouse model.

Kakizaki, Masatoshi / Yamamoto, Yuichiro / Otsuka, Motoyuki / Kitamura, Kouichi / Ito, Masatoshi / Kawai, Hideki Derek / Muramatsu, Masamichi / Kagawa, Tatehiro / Kotani, Ai

The Journal of biological chemistry

2020 Volume 295, Issue 35, Page(s) 12449–12460

Abstract: Hepatitis B, a viral infection that affects the liver, is thought to affect over 257 million people worldwide, and long-term infection can lead to life-threatening issues such as cirrhosis or liver cancer. Chronic hepatitis B develops by the interaction ... ...

Abstract	Hepatitis B, a viral infection that affects the liver, is thought to affect over 257 million people worldwide, and long-term infection can lead to life-threatening issues such as cirrhosis or liver cancer. Chronic hepatitis B develops by the interaction between hepatitis B virus (HBV) and host immune response. However, questions of how HBV-infected cells thwart immune system defenses remain unanswered. Extracellular vesicles (EVs) are used for cellular communication, carrying cargoes such as RNAs, proteins, and lipids and delivering them intracellularly after being endocytosed by target cells. HBV-infected liver cells secrete several types of EVs into body fluids such as complete and incomplete virions, and exosomes. We previously demonstrated that monocytes that incorporated EVs moved to immunoregulatory phenotypes via up-regulation of PD-L1, an immunocheckpoint molecule, and down-regulation of CD69, a leukocyte activation molecule. In this study, we transfected mice with HBV using hydrodynamic injection and studied the effects of EVs secreted by HBV-infected liver cells. EVs secreted from cells with HBV replication strongly suppressed the immune response, inhibiting the eradication of HBV-replicating cells in the mice transfected with HBV. EVs were systemically incorporated in multiple organs, including liver, bone marrow (BM), and intestine. Intriguingly, the BM cells that incorporated EVs acquired intestinal tropism and the dendritic cell populations in the intestine increased. These findings suggest that the EVs secreted by HBV-infected liver cells exert immunosuppressive functions, and that an association between the liver, bone marrow, and intestinal tract exists through EVs secreted from HBV-infected cells.
MeSH term(s)	Animals ; Antigens, CD/genetics ; Antigens, CD/metabolism ; Antigens, Differentiation, T-Lymphocyte/genetics ; Antigens, Differentiation, T-Lymphocyte/metabolism ; B7-H1 Antigen/genetics ; B7-H1 Antigen/metabolism ; Disease Models, Animal ; Extracellular Vesicles/genetics ; Extracellular Vesicles/pathology ; Extracellular Vesicles/virology ; Hep G2 Cells ; Hepatitis B virus/genetics ; Hepatitis B virus/metabolism ; Hepatitis B, Chronic/genetics ; Hepatitis B, Chronic/metabolism ; Hepatitis B, Chronic/pathology ; Humans ; Hydrodynamics ; Lectins, C-Type/genetics ; Lectins, C-Type/metabolism ; Mice ; Transfection
Chemical Substances	Antigens, CD ; Antigens, Differentiation, T-Lymphocyte ; B7-H1 Antigen ; CD69 antigen ; Cd274 protein, mouse ; Lectins, C-Type
Language	English
Publishing date	2020-07-10
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.RA120.014317
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Article ; Online: The immunological function of extracellular vesicles in hepatitis B virus-infected hepatocytes.

Kakizaki, Masatoshi / Yamamoto, Yuichiro / Yabuta, Suemi / Kurosaki, Natsumi / Kagawa, Tatehiro / Kotani, Ai

PloS one

2018 Volume 13, Issue 12, Page(s) e0205886

Abstract: Hepatitis B virus (HBV) generates large amounts of complete and incomplete viral particles. Except for the virion, which acts as infectious particles, the function of those particles remains elusive. Extracellular vesicles (EVs) have been revealed to ... ...

Abstract	Hepatitis B virus (HBV) generates large amounts of complete and incomplete viral particles. Except for the virion, which acts as infectious particles, the function of those particles remains elusive. Extracellular vesicles (EVs) have been revealed to have biological functions. The EVs which size are less than 100 nm in diameter, were collected from HBV infected-patients. These vesicles contain, complete and incomplete virions, and exosomes, which have been recently shown to be critical as intercellular communicators. Here, the effects of the exosome, the complete, and the incomplete particles on the target cells were investigated. These particles are endocytosed by monocyte/macrophages and function primarily to upregulate PD-L1. The functions and composition of the EVs were affected by nucleotide reverse transcriptase inhibitors (NRTIs), suggesting that the EVs are involved in the pathogenesis of HBV hepatitis and clinical course of those patients treated by NRTIs.
MeSH term(s)	B7-H1 Antigen/biosynthesis ; Cell Line ; Extracellular Vesicles/metabolism ; Extracellular Vesicles/ultrastructure ; Extracellular Vesicles/virology ; Hemangioendothelioma ; Hepatitis B/metabolism ; Hepatitis B/pathology ; Hepatitis B virus/metabolism ; Up-Regulation
Chemical Substances	B7-H1 Antigen ; CD274 protein, human
Language	English
Publishing date	2018-12-31
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0205886
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Overcoming antibody-resistant SARS-CoV-2 variants with bispecific antibodies constructed using non-neutralizing antibodies

Inoue, Tetsuya / Yamamoto, Yuichiro / Sato, Kaoru / Nakamura, Yuko / Shimizu, Yoshimi / Ogawa, Motohiko / Onodera, Taishi / Takahashi, Yoshimasa / Wakita, Takaji / Kaneko, Mika K / Fukasawa, Masayoshi / Kato, Yukinari / Noguchi, Kohji

bioRxiv

Abstract	A current challenge is the emergence of SARS-CoV-2 variants, such as BQ.1.1 and XBB.1.5, that can evade immune defenses, thereby limiting antibody drug effectiveness. Emergency-use antibody drugs, including the widely effective bebtelovimab, are losing their benefits. One potential approach to address this issue are bispecific antibodies which combine the targeting abilities of two antibodies with distinct epitopes. We engineered neutralizing bispecific antibodies in the IgG-scFv format from two initially non-neutralizing antibodies, CvMab-6 (which binds to the receptor-binding domain [RBD]) and CvMab-62 (targeting a spike protein S2 subunit epitope adjacent to the known anti-S2 antibody epitope). Furthermore, we created a bispecific antibody by incorporating the scFv of bebtelovimab with our anti-S2 antibody, demonstrating significant restoration of effectiveness against bebtelovimab-resistant BQ.1.1 variants. This study highlights the potential of neutralizing bispecific antibodies, which combine existing less effective anti-RBD antibodies with anti-S2 antibodies, to revive the effectiveness of antibody therapeutics compromised by immune-evading variants.
Keywords	covid19
Language	English
Publishing date	2023-10-27
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2023.10.26.564289
Database	COVID19

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Article ; Online: The function of SARS-CoV-2 spike protein is impaired by disulfide-bond disruption with mutation at cysteine-488 and by thiol-reactive N-acetyl-cysteine and glutathione.

Biochemical and biophysical research communications

2022 Volume 597, Page(s) 30–36

Abstract	Viral spike proteins play important roles in the viral entry process, facilitating attachment to cellular receptors and fusion of the viral envelope with the cell membrane. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds to the cellular receptor angiotensin converting enzyme-2 (ACE2) via its receptor-binding domain (RBD). The cysteine residue at position 488, consisting of a disulfide bridge with cysteine 480 is located in an important structural loop at ACE2-binding surface of RBD, and is highly conserved among SARS-related coronaviruses. We showed that the substitution of Cys-488 with alanine impaired pseudotyped SARS-CoV-2 infection, syncytium formation, and cell-cell fusion triggered by SARS-CoV-2 spike expression. Consistently, in vitro binding of RBD and ACE2, spike-mediated cell-cell fusion, and pseudotyped viral infection of VeroE6/TMPRSS2 cells were inhibited by the thiol-reactive compounds N-acetylcysteine (NAC) and a reduced form of glutathione (GSH). Furthermore, we demonstrated that the activity of variant spikes from the SARS-CoV-2 alpha and delta strains were also suppressed by NAC and GSH. Taken together, these data indicate that Cys-488 in spike RBD is required for SARS-CoV-2 spike functions and infectivity, and could be a target of anti-SARS-CoV-2 therapeutics.
Language	English
Publishing date	2022-01-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2022.01.106
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