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Article ; Online: Monocyte-crosstalk drives interferon-mediated signaling following SARS-CoV-2 exposure.

Theobald, Sebastian J / Rybniker, Jan

2022 Volume 18, Issue 9, Page(s) e11256

Abstract: Cells of the innate immune system represent the first line of defense against SARS-CoV-2 and play an essential role in activating adaptive immunity, which mediates long-term protection. In addition, the same cells are key drivers of tissue damage by ... ...

Abstract	Cells of the innate immune system represent the first line of defense against SARS-CoV-2 and play an essential role in activating adaptive immunity, which mediates long-term protection. In addition, the same cells are key drivers of tissue damage by causing the hyperinflammatory state and cytokine storm that makes COVID-19 a deadly disease. Thus, careful dissection of the host-pathogen interaction on a cellular level is essential to understanding SARS-CoV-2 pathogenesis and developing new treatment modalities against COVID-19. In their recent work, Goffinet and colleagues (Kazmierski et al, 2022) investigate the cell-intrinsic responses of human primary peripheral blood mononuclear cells (PBMCs) exposed to SARS coronaviruses.
MeSH term(s)	COVID-19/drug therapy ; Humans ; Immunity, Innate ; Interferons ; Leukocytes, Mononuclear ; Monocytes ; SARS-CoV-2
Chemical Substances	Interferons (9008-11-1)
Language	English
Publishing date	2022-09-12
Publishing country	England
Document type	Journal Article ; Comment
ZDB-ID	2193510-5
ISSN	1744-4292 ; 1744-4292
ISSN (online)	1744-4292
ISSN	1744-4292
DOI	10.15252/msb.202211256
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Article: Modeling Human Cytomegalovirus in Humanized Mice for Vaccine Testing.

Koenig, Johannes / Theobald, Sebastian J / Stripecke, Renata

Vaccines

2020 Volume 8, Issue 1

Abstract: Human cytomegalovirus (HCMV or HHV-5) is a globally spread pathogen with strictly human tropism that establishes a lifelong persistence. After primary infection, high levels of long-term T and B cell responses are elicited, but the virus is not cleared. ... ...

Abstract	Human cytomegalovirus (HCMV or HHV-5) is a globally spread pathogen with strictly human tropism that establishes a lifelong persistence. After primary infection, high levels of long-term T and B cell responses are elicited, but the virus is not cleared. HCMV persists mainly in hematopoietic reservoirs, whereby occasional viral reactivation and spread are well controlled in immunocompetent hosts. However, when the immune system cannot control viral infections or reactivations, such as with newborns, patients with immune deficiencies, or immune-compromised patients after transplantations, the lytic outbursts can be severely debilitating or lethal. The development of vaccines for immunization of immune-compromised hosts has been challenging. Several vaccine candidates did not reach the potency expected in clinical trials and were not approved. Before anti-HCMV vaccines can be tested pre-clinically in immune-compromised hosts, reliable in vivo models recapitulating HCMV infection might accelerate their clinical translation. Therefore, immune-deficient mouse strains implanted with human cells and tissues and developing a human immune system (HIS) are being explored to test anti-HCMV vaccines. HIS-mice resemble immune-compromised hosts as they are equipped with antiviral human T and B cells, but the immune reactivity is overall low. Several groups have independently shown that HCMV infections and reactivations can be mirrored in HIS mice. However, these models and the analyses employed varied widely. The path forward is to improve human immune reconstitution and standardize the analyses of adaptive responses so that HIS models can be forthrightly used for testing novel generations of anti-HCMV vaccines in the preclinical pipeline.
Language	English
Publishing date	2020-02-17
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines8010089
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Article ; Online: Discovery of dual-active ethionamide boosters inhibiting the Mycobacterium tuberculosis ESX-1 secretion system.

Gries, Raphael / Chhen, Jason / van Gumpel, Edeltraud / Theobald, Sebastian J / Sonnenkalb, Lindsay / Utpatel, Christian / Metzen, Fabian / Koch, Manuel / Dallenga, Tobias / Djaout, Kamel / Baulard, Alain / Dal Molin, Michael / Rybniker, Jan

Cell chemical biology

2024 Volume 31, Issue 4, Page(s) 699–711.e6

Abstract: Drug-resistant Mycobacterium tuberculosis (Mtb) remains a major public health concern requiring complementary approaches to standard anti-tuberculous regimens. Anti-virulence molecules or compounds that enhance the activity of antimicrobial prodrugs are ... ...

Abstract	Drug-resistant Mycobacterium tuberculosis (Mtb) remains a major public health concern requiring complementary approaches to standard anti-tuberculous regimens. Anti-virulence molecules or compounds that enhance the activity of antimicrobial prodrugs are promising alternatives to conventional antibiotics. Exploiting host cell-based drug discovery, we identified an oxadiazole compound (S3) that blocks the ESX-1 secretion system, a major virulence factor of Mtb. S3-treated mycobacteria showed impaired intracellular growth and a reduced ability to lyse macrophages. RNA sequencing experiments of drug-exposed bacteria revealed strong upregulation of a distinct set of genes including ethA, encoding a monooxygenase activating the anti-tuberculous prodrug ethionamide. Accordingly, we found a strong ethionamide boosting effect in S3-treated Mtb. Extensive structure-activity relationship experiments revealed that anti-virulence and ethionamide-boosting activity can be uncoupled by chemical modification of the primary hit molecule. To conclude, this series of dual-active oxadiazole compounds targets Mtb via two distinct mechanisms of action.
MeSH term(s)	Humans ; Mycobacterium tuberculosis ; Ethionamide/pharmacology ; Type VII Secretion Systems ; Tuberculosis ; Oxadiazoles/pharmacology ; Bacterial Proteins/genetics
Chemical Substances	Ethionamide (OAY8ORS3CQ) ; Type VII Secretion Systems ; Oxadiazoles ; Bacterial Proteins
Language	English
Publishing date	2024-01-04
Publishing country	United States
Document type	Journal Article
ISSN	2451-9448
ISSN (online)	2451-9448
DOI	10.1016/j.chembiol.2023.12.007
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Article ; Online: RHBDL4-triggered downregulation of COPII adaptor protein TMED7 suppresses TLR4-mediated inflammatory signaling.

Knopf, Julia D / Steigleder, Susanne S / Korn, Friederike / Kühnle, Nathalie / Badenes, Marina / Tauber, Marina / Theobald, Sebastian J / Rybniker, Jan / Adrain, Colin / Lemberg, Marius K

Nature communications

2024 Volume 15, Issue 1, Page(s) 1528

Abstract: The toll-like receptor 4 (TLR4) is a central regulator of innate immunity that primarily recognizes bacterial lipopolysaccharide cell wall constituents to trigger cytokine secretion. We identify the intramembrane protease RHBDL4 as a negative regulator ... ...

Abstract	The toll-like receptor 4 (TLR4) is a central regulator of innate immunity that primarily recognizes bacterial lipopolysaccharide cell wall constituents to trigger cytokine secretion. We identify the intramembrane protease RHBDL4 as a negative regulator of TLR4 signaling. We show that RHBDL4 triggers degradation of TLR4's trafficking factor TMED7. This counteracts TLR4 transport to the cell surface. Notably, TLR4 activation mediates transcriptional upregulation of RHBDL4 thereby inducing a negative feedback loop to reduce TLR4 trafficking to the plasma membrane. This secretory cargo tuning mechanism prevents the over-activation of TLR4-dependent signaling in an in vitro Mycobacterium tuberculosis macrophage infection model and consequently alleviates septic shock in a mouse model. A hypomorphic RHBDL4 mutation linked to Kawasaki syndrome, an ill-defined inflammatory disorder in children, further supports the pathophysiological relevance of our findings. In this work, we identify an RHBDL4-mediated axis that acts as a rheostat to prevent over-activation of the TLR4 pathway.
MeSH term(s)	Animals ; Child ; Humans ; Mice ; Adaptor Proteins, Signal Transducing/metabolism ; Cell Membrane/metabolism ; Down-Regulation ; Lipopolysaccharides/metabolism ; Signal Transduction ; Toll-Like Receptor 4/metabolism
Chemical Substances	Adaptor Proteins, Signal Transducing ; Lipopolysaccharides ; TLR4 protein, human ; Toll-Like Receptor 4 ; Rhbdl3 protein, mouse (EC 3.4.21.105)
Language	English
Publishing date	2024-03-07
Publishing country	England
Document type	Journal Article
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-024-45615-2
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Article ; Online: Modeling Human Cytomegalovirus in Humanized Mice for Vaccine Testing

Johannes Koenig / Sebastian J. Theobald / Renata Stripecke

Vaccines, Vol 8, Iss 1, p

2020 Volume 89

Abstract	Human cytomegalovirus (HCMV or HHV-5) is a globally spread pathogen with strictly human tropism that establishes a lifelong persistence. After primary infection, high levels of long-term T and B cell responses are elicited, but the virus is not cleared. HCMV persists mainly in hematopoietic reservoirs, whereby occasional viral reactivation and spread are well controlled in immunocompetent hosts. However, when the immune system cannot control viral infections or reactivations, such as with newborns, patients with immune deficiencies, or immune-compromised patients after transplantations, the lytic outbursts can be severely debilitating or lethal. The development of vaccines for immunization of immune-compromised hosts has been challenging. Several vaccine candidates did not reach the potency expected in clinical trials and were not approved. Before anti-HCMV vaccines can be tested pre-clinically in immune-compromised hosts, reliable in vivo models recapitulating HCMV infection might accelerate their clinical translation. Therefore, immune-deficient mouse strains implanted with human cells and tissues and developing a human immune system (HIS) are being explored to test anti-HCMV vaccines. HIS-mice resemble immune-compromised hosts as they are equipped with antiviral human T and B cells, but the immune reactivity is overall low. Several groups have independently shown that HCMV infections and reactivations can be mirrored in HIS mice. However, these models and the analyses employed varied widely. The path forward is to improve human immune reconstitution and standardize the analyses of adaptive responses so that HIS models can be forthrightly used for testing novel generations of anti-HCMV vaccines in the preclinical pipeline.
Keywords	cmv ; hhv-5 ; humanized mice ; his ; transplantation ; t cells ; antibodies ; dendritic cells ; vaccines ; Medicine ; R
Subject code	570
Language	English
Publishing date	2020-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: A comparative analysis of remdesivir and other repurposed antivirals against SARS‐CoV‐2

Alexander Simonis / Sebastian J Theobald / Gerd Fätkenheuer / Jan Rybniker / Jakob J Malin

EMBO Molecular Medicine, Vol 13, Iss 1, Pp n/a-n/a (2021)

2021

Abstract: Abstract The ongoing SARS‐CoV‐2 pandemic stresses the need for effective antiviral drugs that can quickly be applied in order to reduce morbidity, mortality, and ideally viral transmission. By repurposing of broadly active antiviral drugs and compounds ... ...

Abstract	Abstract The ongoing SARS‐CoV‐2 pandemic stresses the need for effective antiviral drugs that can quickly be applied in order to reduce morbidity, mortality, and ideally viral transmission. By repurposing of broadly active antiviral drugs and compounds that are known to inhibit viral replication of related viruses, several advances could be made in the development of treatment strategies against COVID‐19. The nucleoside analog remdesivir, which is known for its potent in vitro activity against Ebolavirus and other RNA viruses, was recently shown to reduce the time to recovery in patients with severe COVID‐19. It is to date the only approved antiviral for treating COVID‐19. Here, we provide a mechanism and evidence‐based comparative review of remdesivir and other repurposed drugs with proven in vitro activity against SARS‐CoV‐2.
Keywords	antivirals ; COVID‐19 ; remdesivir ; SARS‐CoV‐2 ; Medicine (General) ; R5-920 ; Genetics ; QH426-470
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
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Article ; Online: Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice.

Theobald, Sebastian J / Fiestas, Elena / Schneider, Andreas / Ostermann, Benjamin / Danisch, Simon / von Kaisenberg, Constantin / Rybniker, Jan / Hammerschmidt, Wolfgang / Zeidler, Reinhard / Stripecke, Renata

Cells

2023 Volume 13, Issue 1

Abstract: Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B ... ...

Abstract	Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern and an additional hurdle for antibody discovery in humans is the source of human tissues with an immunological microenvironment. Here, we show an efficient method to pharm human antibodies using immortalized B cells recovered from Nod.Rag.Gamma (NRG) mice reconstituting the human immune system (HIS). Humanized HIS mice were immunized either with autologous engineered dendritic cells expressing the human cytomegalovirus gB envelope protein (HCMV-gB) or with Epstein-Barr virus-like particles (EB-VLP). Human B cells recovered from spleen of HIS mice were efficiently immortalized with EBV in vitro. We show that these immortalized B cells secreted human IgGs with neutralization capacities against prototypic HCMV-gB and EBV-gp350. Taken together, we show that HIS mice can be successfully used for the generation and pharming fully human IgGs. This technology can be further explored to generate antibodies against emerging infections for diagnostic or therapeutic purposes.
MeSH term(s)	Humans ; Animals ; Mice ; Spleen ; Herpesvirus 4, Human ; Epstein-Barr Virus Infections ; Antibodies, Viral ; Immunoglobulin G ; Cancer Vaccines ; Cytomegalovirus
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Cancer Vaccines
Language	English
Publishing date	2023-12-21
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells13010020
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Article ; Online: A comparative analysis of remdesivir and other repurposed antivirals against SARS-CoV-2.

Simonis, Alexander / Theobald, Sebastian J / Fätkenheuer, Gerd / Rybniker, Jan / Malin, Jakob J

EMBO molecular medicine

2020 Volume 13, Issue 1, Page(s) e13105

Abstract: The ongoing SARS-CoV-2 pandemic stresses the need for effective antiviral drugs that can quickly be applied in order to reduce morbidity, mortality, and ideally viral transmission. By repurposing of broadly active antiviral drugs and compounds that are ... ...

Abstract	The ongoing SARS-CoV-2 pandemic stresses the need for effective antiviral drugs that can quickly be applied in order to reduce morbidity, mortality, and ideally viral transmission. By repurposing of broadly active antiviral drugs and compounds that are known to inhibit viral replication of related viruses, several advances could be made in the development of treatment strategies against COVID-19. The nucleoside analog remdesivir, which is known for its potent in vitro activity against Ebolavirus and other RNA viruses, was recently shown to reduce the time to recovery in patients with severe COVID-19. It is to date the only approved antiviral for treating COVID-19. Here, we provide a mechanism and evidence-based comparative review of remdesivir and other repurposed drugs with proven in vitro activity against SARS-CoV-2.
MeSH term(s)	Adenosine Monophosphate/analogs & derivatives ; Adenosine Monophosphate/pharmacology ; Adenosine Monophosphate/therapeutic use ; Alanine/analogs & derivatives ; Alanine/pharmacology ; Alanine/therapeutic use ; Amides/pharmacology ; Amides/therapeutic use ; Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Benzamidines ; Drug Repositioning/methods ; Esters/pharmacology ; Esters/therapeutic use ; Guanidines/pharmacology ; Guanidines/therapeutic use ; Guanine/pharmacology ; Guanine/therapeutic use ; Humans ; Indoles/pharmacology ; Indoles/therapeutic use ; Lopinavir/pharmacology ; Lopinavir/therapeutic use ; Protease Inhibitors/pharmacology ; Protease Inhibitors/therapeutic use ; Pyrazines/pharmacology ; Pyrazines/therapeutic use ; Ribavirin/pharmacology ; Ribavirin/therapeutic use ; Ritonavir/pharmacology ; Ritonavir/therapeutic use ; SARS-CoV-2/drug effects ; SARS-CoV-2/physiology ; Virus Internalization/drug effects ; Virus Replication/drug effects ; COVID-19 Drug Treatment
Chemical Substances	Amides ; Antiviral Agents ; Benzamidines ; Esters ; Guanidines ; Indoles ; Protease Inhibitors ; Pyrazines ; camostat (0FD207WKDU) ; Lopinavir (2494G1JF75) ; penciclovir (359HUE8FJC) ; remdesivir (3QKI37EEHE) ; Adenosine Monophosphate (415SHH325A) ; Ribavirin (49717AWG6K) ; Guanine (5Z93L87A1R) ; umifenovir (93M09WW4RU) ; favipiravir (EW5GL2X7E0) ; Ritonavir (O3J8G9O825) ; Alanine (OF5P57N2ZX) ; nafamostat (Y25LQ0H97D)
Keywords	covid19
Language	English
Publishing date	2020-11-03
Publishing country	England
Document type	Comparative Study ; Journal Article ; Review
ZDB-ID	2467145-9
ISSN	1757-4684 ; 1757-4676
ISSN (online)	1757-4684
ISSN	1757-4676
DOI	10.15252/emmm.202013105
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Article ; Online: Viral Glycoproteins Induce NLRP3 Inflammasome Activation and Pyroptosis in Macrophages.

Eisfeld, Hannah S / Simonis, Alexander / Winter, Sandra / Chhen, Jason / Ströh, Luisa J / Krey, Thomas / Koch, Manuel / Theobald, Sebastian J / Rybniker, Jan

Viruses

2021 Volume 13, Issue 10

Abstract: Infections with viral pathogens are widespread and can cause a variety of different diseases. In-depth knowledge about viral triggers initiating an immune response is necessary to decipher viral pathogenesis. Inflammasomes, as part of the innate immune ... ...

Abstract	Infections with viral pathogens are widespread and can cause a variety of different diseases. In-depth knowledge about viral triggers initiating an immune response is necessary to decipher viral pathogenesis. Inflammasomes, as part of the innate immune system, can be activated by viral pathogens. However, viral structural components responsible for inflammasome activation remain largely unknown. Here we analyzed glycoproteins derived from SARS-CoV-1/2, HCMV and HCV, required for viral entry and fusion, as potential triggers of NLRP3 inflammasome activation and pyroptosis in THP-1 macrophages. All tested glycoproteins were able to potently induce NLRP3 inflammasome activation, indicated by ASC-SPECK formation and secretion of cleaved IL-1β. Lytic cell death via gasdermin D (GSDMD), pore formation, and pyroptosis are required for IL-1β release. As a hallmark of pyroptosis, we were able to detect cleavage of GSDMD and, correspondingly, cell death in THP-1 macrophages. CRISPR-Cas9 knockout of NLRP3 and GSDMD in THP-1 macrophages confirmed and strongly support the evidence that viral glycoproteins can act as innate immunity triggers. With our study, we decipher key mechanisms of viral pathogenesis by showing that viral glycoproteins potently induce innate immune responses. These insights could be beneficial in vaccine development and provide new impulses for the investigation of vaccine-induced innate immunity.
MeSH term(s)	Cell Line, Tumor ; Cytomegalovirus/immunology ; Hepacivirus/immunology ; Humans ; Immunity, Innate/immunology ; Inflammasomes/immunology ; Interleukin-1beta/biosynthesis ; Interleukin-1beta/immunology ; Macrophages/immunology ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology ; Pyroptosis/immunology ; SARS Virus/immunology ; SARS-CoV-2/immunology ; THP-1 Cells ; Viral Envelope Proteins/immunology ; Viral Fusion Proteins/immunology
Chemical Substances	IL1B protein, human ; Inflammasomes ; Interleukin-1beta ; NLR Family, Pyrin Domain-Containing 3 Protein ; NLRP3 protein, human ; Viral Envelope Proteins ; Viral Fusion Proteins
Language	English
Publishing date	2021-10-15
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13102076
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Article ; Online: Identification of FasL as a crucial host factor driving COVID-19 pathology and lethality.

Albert, Marie-Christine / Uranga-Murillo, Iratxe / Arias, Maykel / De Miguel, Diego / Peña, Natacha / Montinaro, Antonella / Varanda, Ana Beatriz / Theobald, Sebastian J / Areso, Itziar / Saggau, Julia / Koch, Manuel / Liccardi, Gianmaria / Peltzer, Nieves / Rybniker, Jan / Hurtado-Guerrero, Ramón / Merino, Pedro / Monzón, Marta / Badiola, Juan J / Reindl-Schwaighofer, Roman /

Sanz-Pamplona, Rebeca / Cebollada-Solanas, Alberto / Megyesfalvi, Zsolt / Dome, Balazs / Secrier, Maria / Hartmann, Boris / Bergmann, Michael / Pardo, Julián / Walczak, Henning

Cell death and differentiation

2024

Abstract: The dysregulated immune response and inflammation resulting in severe COVID-19 are still incompletely understood. Having recently determined that aberrant death-ligand-induced cell death can cause lethal inflammation, we hypothesized that this process ... ...

Abstract	The dysregulated immune response and inflammation resulting in severe COVID-19 are still incompletely understood. Having recently determined that aberrant death-ligand-induced cell death can cause lethal inflammation, we hypothesized that this process might also cause or contribute to inflammatory disease and lung failure following SARS-CoV-2 infection. To test this hypothesis, we developed a novel mouse-adapted SARS-CoV-2 model (MA20) that recapitulates key pathological features of COVID-19. Concomitantly with occurrence of cell death and inflammation, FasL expression was significantly increased on inflammatory monocytic macrophages and NK cells in the lungs of MA20-infected mice. Importantly, therapeutic FasL inhibition markedly increased survival of both, young and old MA20-infected mice coincident with substantially reduced cell death and inflammation in their lungs. Intriguingly, FasL was also increased in the bronchoalveolar lavage fluid of critically-ill COVID-19 patients. Together, these results identify FasL as a crucial host factor driving the immuno-pathology that underlies COVID-19 severity and lethality, and imply that patients with severe COVID-19 may significantly benefit from therapeutic inhibition of FasL.
Language	English
Publishing date	2024-03-21
Publishing country	England
Document type	Journal Article
ZDB-ID	1225672-9
ISSN	1476-5403 ; 1350-9047
ISSN (online)	1476-5403
ISSN	1350-9047
DOI	10.1038/s41418-024-01278-6
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