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Article ; Online: A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging.

Mamede, João I / Griffin, Joseph / Gambut, Stéphanie / Hope, Thomas J

2021 Volume 13, Issue 3

Abstract: During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to ...

Abstract	During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes to follow individual virus particles while they progress throughout infection. The inclusion of such fusion motifs or surrogates frequently disrupts viral infectivity or results in a change of the wild-type phenotype. Here, we detail the construction and functional characterization of two new constructs where we fused fluorescent proteins to the N-terminus of HIV-1 Integrase. In the first, IN is recruited into assembling particles via a codon optimized Gag to complement other viral constructs, while the second is fused to a Gag-Pol expression vector fully capable of integration. Our data shows that N-terminal tagged IN is functional for integration by both recovery of integration of catalytically inactive IN and by the successful infectivity of viruses carrying only labeled IN. These tools will be important to study the individual behavior of viral particles and associate such behavior to infectivity.
MeSH term(s)	Cell Line ; Cell Line, Tumor ; Fluorescence ; HEK293 Cells ; HIV Infections/virology ; HIV-1/genetics ; HeLa Cells ; Humans ; Integrases/genetics ; Optical Imaging/methods ; Viral Proteins/genetics ; Virion/genetics ; Virus Replication/genetics
Chemical Substances	Viral Proteins ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2021-02-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13030386
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging

Mamede, João I / Griffin, Joseph / Gambut, Stéphanie / Hope, Thomas J

Viruses. 2021 Feb. 28, v. 13, no. 3

2021

Abstract	During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes to follow individual virus particles while they progress throughout infection. The inclusion of such fusion motifs or surrogates frequently disrupts viral infectivity or results in a change of the wild-type phenotype. Here, we detail the construction and functional characterization of two new constructs where we fused fluorescent proteins to the N-terminus of HIV-1 Integrase. In the first, IN is recruited into assembling particles via a codon optimized Gag to complement other viral constructs, while the second is fused to a Gag-Pol expression vector fully capable of integration. Our data shows that N-terminal tagged IN is functional for integration by both recovery of integration of catalytically inactive IN and by the successful infectivity of viruses carrying only labeled IN. These tools will be important to study the individual behavior of viral particles and associate such behavior to infectivity.
Keywords	fluorescence ; fluorescence microscopy ; fluorescent dyes ; genetic vectors ; integrases ; pathogenicity ; phenotype ; viruses
Language	English
Dates of publication	2021-0228
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
Note	NAL-light
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13030386
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Secretion of the epithelial sodium channel chaperone PCSK9 from the cortical collecting duct links sodium retention with hypercholesterolemia in nephrotic syndrome.

Molina-Jijon, Eduardo / Gambut, Stéphanie / Macé, Camille / Avila-Casado, Carmen / Clement, Lionel C

Kidney international

2020 Volume 98, Issue 6, Page(s) 1449–1460

Abstract: The proprotein PCSK9 functions as a chaperone for the epithelial sodium channel in the cortical collecting duct (CCD), is highly expressed in the liver, and plays a significant role in the pathogenesis of hypercholesterolemia. Lower levels of PCSK9 ... ...

Abstract	The proprotein PCSK9 functions as a chaperone for the epithelial sodium channel in the cortical collecting duct (CCD), is highly expressed in the liver, and plays a significant role in the pathogenesis of hypercholesterolemia. Lower levels of PCSK9 expression also occur in the normal kidney and intestine. Here, we found increased PCSK9 expression in the CCD of biopsies of patients with primary glomerular disease and explored a possible relationship with hypercholesterolemia of nephrotic syndrome. Significantly elevated serum PCSK9 and cholesterol levels were noted in two models of focal and segmental glomerulosclerosis, the Rrm2b-/- mouse and the Buffalo/Mna rat. Increased expression of PCSK9 in the kidney occurred when liver expression was reduced in both models. The impact of reduced or increased PCSK9 in the CCD on hypercholesterolemia in nephrotic syndrome was next studied. Mice with selective deficiency of PCSK9 expression in the collecting duct failed to develop hypercholesterolemia after injection of nephrotoxic serum. Blocking epithelial sodium channel activity with Amiloride in Rrm2b-/- mice resulted in increased expression of its chaperone PCSK9 in the CCD, followed by elevated plasma levels and worsening hypercholesterolemia. Thus, our data suggest that PCSK9 in the kidney plays a role in the initiation of hypercholesterolemia in nephrotic syndrome and make a case for depletion of PCSK9 early in patients with nephrotic syndrome to prevent the development of hypercholesterolemia.
MeSH term(s)	Animals ; Aquaporin 2 ; Epithelial Sodium Channels/genetics ; Humans ; Hypercholesterolemia/genetics ; Male ; Mice ; Mice, Inbred C57BL ; Nephrotic Syndrome ; Proprotein Convertase 9/genetics ; Rats ; Sodium/metabolism
Chemical Substances	Aquaporin 2 ; Epithelial Sodium Channels ; Sodium (9NEZ333N27) ; PCSK9 protein, human (EC 3.4.21.-) ; PCSK9 protein, rat (EC 3.4.21.-) ; Pcsk9 protein, mouse (EC 3.4.21.-) ; Proprotein Convertase 9 (EC 3.4.21.-)
Language	English
Publishing date	2020-08-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	120573-0
ISSN	1523-1755 ; 0085-2538
ISSN (online)	1523-1755
ISSN	0085-2538
DOI	10.1016/j.kint.2020.06.045
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Cytokine storm-based mechanisms for extrapulmonary manifestations of SARS-CoV-2 infection.

Del Nogal Avila, Maria / Das, Ranjan / Kharlyngdoh, Joubert / Molina-Jijon, Eduardo / Donoro Blazquez, Hector / Gambut, Stéphanie / Crowley, Michael / Crossman, David K / Gbadegesin, Rasheed A / Chugh, Sunveer S / Chugh, Sunjeet S / Avila-Casado, Carmen / Macé, Camille / Clement, Lionel C / Chugh, Sumant S

JCI insight

2023 Volume 8, Issue 10

Abstract: Viral illnesses like SARS-CoV-2 have pathologic effects on nonrespiratory organs in the absence of direct viral infection. We injected mice with cocktails of rodent equivalents of human cytokine storms resulting from SARS-CoV-2/COVID-19 or rhinovirus ... ...

Abstract	Viral illnesses like SARS-CoV-2 have pathologic effects on nonrespiratory organs in the absence of direct viral infection. We injected mice with cocktails of rodent equivalents of human cytokine storms resulting from SARS-CoV-2/COVID-19 or rhinovirus common cold infection. At low doses, COVID-19 cocktails induced glomerular injury and albuminuria in zinc fingers and homeoboxes 2 (Zhx2) hypomorph and Zhx2+/+ mice to mimic COVID-19-related proteinuria. Common Cold cocktail induced albuminuria selectively in Zhx2 hypomorph mice to model relapse of minimal change disease, which improved after depletion of TNF-α, soluble IL-4Rα, or IL-6. The Zhx2 hypomorph state increased cell membrane to nuclear migration of podocyte ZHX proteins in vivo (both cocktails) and lowered phosphorylated STAT6 activation (COVID-19 cocktail) in vitro. At higher doses, COVID-19 cocktails induced acute heart injury, myocarditis, pericarditis, acute liver injury, acute kidney injury, and high mortality in Zhx2+/+ mice, whereas Zhx2 hypomorph mice were relatively protected, due in part to early, asynchronous activation of STAT5 and STAT6 pathways in these organs. Dual depletion of cytokine combinations of TNF-α with IL-2, IL-13, or IL-4 in Zhx2+/+ mice reduced multiorgan injury and eliminated mortality. Using genome sequencing and CRISPR/Cas9, an insertion upstream of ZHX2 was identified as a cause of the human ZHX2 hypomorph state.
MeSH term(s)	Humans ; Mice ; Animals ; Homeodomain Proteins/genetics ; Albuminuria ; Tumor Necrosis Factor-alpha ; Common Cold ; Cytokine Release Syndrome ; COVID-19 ; SARS-CoV-2/metabolism ; Transcription Factors/genetics
Chemical Substances	Homeodomain Proteins ; Tumor Necrosis Factor-alpha ; ZHX2 protein, human ; Transcription Factors
Language	English
Publishing date	2023-05-22
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.166012
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: De novo emergence of SARS-CoV-2 spike mutations in immunosuppressed patients.

Simons, Lacy M / Ozer, Egon A / Gambut, Stephanie / Dean, Taylor J / Zhang, Li / Bhimalli, Pavan / Schneider, Jeffrey R / Mamede, João I / Ison, Michael G / Karmali, Reem / Gordon, Leo I / Lorenzo-Redondo, Ramon / Hultquist, Judd F

Transplant infectious disease : an official journal of the Transplantation Society

2022 Volume 24, Issue 6, Page(s) e13914

Abstract: Background: The continuing evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with decreased susceptibility to neutralizing antibodies is of clinical importance. Several spike mutations associated with immune escape have ... ...

Abstract	Background: The continuing evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with decreased susceptibility to neutralizing antibodies is of clinical importance. Several spike mutations associated with immune escape have evolved independently in association with different variants of concern (VOCs). How and when these mutations arise is still unclear. We hypothesized that such mutations might arise in the context of persistent viral replication in immunosuppressed hosts. Methods: Nasopharyngeal specimens were collected longitudinally from two immunosuppressed patients with persistent SARS-CoV-2 infection. Plasma was collected from these same patients late in disease course. SARS-CoV-2 whole genome sequencing was performed to assess the emergence and frequency of mutations over time. Select Spike mutations were assessed for their impact on viral entry and antibody neutralization in vitro. Results: Our sequencing results revealed the intrahost emergence of spike mutations that are associated with circulating VOCs in both immunosuppressed patients (del241-243 and E484Q in one patient, and E484K in the other). These mutations decreased antibody-mediated neutralization of pseudotyped virus particles in cell culture, but also decreased efficiency of spike-mediated cell entry. Conclusions: These observations demonstrate the de novo emergence of SARS-CoV-2 spike mutations with enhanced immune evasion in immunosuppressed patients with persistent infection. These data suggest one potential mechanism for the evolution of VOCs and emphasize the importance of continued efforts to develop antiviral drugs for suppression of viral replication in hospitalized settings.
MeSH term(s)	Humans ; SARS-CoV-2/genetics ; COVID-19 ; Mutation ; Antiviral Agents ; Immunocompromised Host ; Antibodies, Neutralizing ; Antibodies, Viral
Chemical Substances	Antiviral Agents ; Antibodies, Neutralizing ; Antibodies, Viral
Language	English
Publishing date	2022-08-01
Publishing country	Denmark
Document type	Journal Article
ZDB-ID	1476094-0
ISSN	1399-3062 ; 1398-2273
ISSN (online)	1399-3062
ISSN	1398-2273
DOI	10.1111/tid.13914
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 5100: Show issues

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bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (2.OG)
ab Jg. 2022: Lesesaal (EG)

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Article: Recognition of HIV-1 capsid by PQBP1 licenses an innate immune sensing of nascent HIV-1 DNA

Molecular cell. 2022 Aug. 04, v. 82, no. 15

2022

Abstract: We have previously described polyglutamine-binding protein 1 (PQBP1) as an adapter required for the cyclic GMP-AMP synthase (cGAS)-mediated innate response to the human immunodeficiency virus 1 (HIV-1) and other lentiviruses. Cytoplasmic HIV-1 DNA is a ... ...

Abstract	We have previously described polyglutamine-binding protein 1 (PQBP1) as an adapter required for the cyclic GMP-AMP synthase (cGAS)-mediated innate response to the human immunodeficiency virus 1 (HIV-1) and other lentiviruses. Cytoplasmic HIV-1 DNA is a transient and low-abundance pathogen-associated molecular pattern (PAMP), and the mechanism for its detection and verification is not fully understood. Here, we show a two-factor authentication strategy by the innate surveillance machinery to selectively respond to the low concentration of HIV-1 DNA, while distinguishing these species from extranuclear DNA molecules. We find that, upon HIV-1 infection, PQBP1 decorates the intact viral capsid, and this serves as a primary verification step for the viral nucleic acid cargo. As reverse transcription and capsid disassembly initiate, cGAS is recruited to the capsid in a PQBP1-dependent manner. This positions cGAS at the site of PAMP generation and sanctions its response to a low-abundance DNA PAMP.
Keywords	DNA ; HIV infections ; Human immunodeficiency virus 1 ; capsid ; monitoring ; pathogen-associated molecular patterns ; reverse transcription
Language	English
Dates of publication	2022-0804
Size	p. 2871-2884.e6.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1415236-8
ISSN	1097-4164 ; 1097-2765
ISSN (online)	1097-4164
ISSN	1097-2765
DOI	10.1016/j.molcel.2022.06.010
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Bulk IgG glycosylation predicts COVID-19 severity and vaccine antibody response.

Cell reports

2022 Volume 41, Issue 11, Page(s) 111799

Abstract: Although vaccination efforts have expanded, there are still gaps in our understanding surrounding the immune response to SARS-CoV-2. Measuring IgG Fc glycosylation provides insight into an infected individual's inflammatory state, among other functions. ... ...

Abstract	Although vaccination efforts have expanded, there are still gaps in our understanding surrounding the immune response to SARS-CoV-2. Measuring IgG Fc glycosylation provides insight into an infected individual's inflammatory state, among other functions. We set out to interrogate bulk IgG glycosylation changes from SARS-CoV-2 infection and vaccination, using plasma from mild or hospitalized COVID-19 patients, and from vaccinated individuals. Inflammatory glycans are elevated in hospitalized COVID-19 patients and increase over time, while mild patients have anti-inflammatory glycans that increase over time, including increased sialic acid correlating with RBD antibody levels. Vaccinated individuals with low RBD antibody levels and low neutralization have the same IgG glycan traits as hospitalized COVID-19 patients. In addition, a small vaccinated cohort reveals a decrease in inflammatory glycans associated with peak IgG concentrations and neutralization. This report characterizes the bulk IgG glycome associated with COVID-19 severity and vaccine responsiveness and can help guide future studies into SARS-CoV-2 protective immunity.
MeSH term(s)	Humans ; Antibody Formation ; Glycosylation ; COVID-19 ; SARS-CoV-2 ; Vaccines ; Immunoglobulin G ; Antibodies, Viral
Chemical Substances	Vaccines ; Immunoglobulin G ; Antibodies, Viral
Language	English
Publishing date	2022-11-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2022.111799
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Recognition of HIV-1 capsid by PQBP1 licenses an innate immune sensing of nascent HIV-1 DNA.

Molecular cell

2022 Volume 82, Issue 15, Page(s) 2871–2884.e6

Abstract	We have previously described polyglutamine-binding protein 1 (PQBP1) as an adapter required for the cyclic GMP-AMP synthase (cGAS)-mediated innate response to the human immunodeficiency virus 1 (HIV-1) and other lentiviruses. Cytoplasmic HIV-1 DNA is a transient and low-abundance pathogen-associated molecular pattern (PAMP), and the mechanism for its detection and verification is not fully understood. Here, we show a two-factor authentication strategy by the innate surveillance machinery to selectively respond to the low concentration of HIV-1 DNA, while distinguishing these species from extranuclear DNA molecules. We find that, upon HIV-1 infection, PQBP1 decorates the intact viral capsid, and this serves as a primary verification step for the viral nucleic acid cargo. As reverse transcription and capsid disassembly initiate, cGAS is recruited to the capsid in a PQBP1-dependent manner. This positions cGAS at the site of PAMP generation and sanctions its response to a low-abundance DNA PAMP.
MeSH term(s)	Capsid/metabolism ; DNA/metabolism ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; HIV-1/genetics ; Humans ; Immunity, Innate ; Nucleotidyltransferases/metabolism ; Pathogen-Associated Molecular Pattern Molecules/metabolism
Chemical Substances	DNA-Binding Proteins ; PQBP1 protein, human ; Pathogen-Associated Molecular Pattern Molecules ; DNA (9007-49-2) ; Nucleotidyltransferases (EC 2.7.7.-)
Language	English
Publishing date	2022-07-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1415236-8
ISSN	1097-4164 ; 1097-2765
ISSN (online)	1097-4164
ISSN	1097-2765
DOI	10.1016/j.molcel.2022.06.010
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 5055: Show issues

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Jg. 1995 - 2021: Lesesall (2.OG)
ab Jg. 2022: Lesesaal (EG)

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Article ; Online: A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging.

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Article: A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging

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Article ; Online: Secretion of the epithelial sodium channel chaperone PCSK9 from the cortical collecting duct links sodium retention with hypercholesterolemia in nephrotic syndrome.

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Article ; Online: Cytokine storm-based mechanisms for extrapulmonary manifestations of SARS-CoV-2 infection.

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Article ; Online: De novo emergence of SARS-CoV-2 spike mutations in immunosuppressed patients.

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Article: Recognition of HIV-1 capsid by PQBP1 licenses an innate immune sensing of nascent HIV-1 DNA

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In stock of ZB MED Bonn / Germany

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Article ; Online: Bulk IgG glycosylation predicts COVID-19 severity and vaccine antibody response.

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Article ; Online: Recognition of HIV-1 capsid by PQBP1 licenses an innate immune sensing of nascent HIV-1 DNA.

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