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Article: Measuring erythrocyte complement receptor 1 using flow cytometry

Kisserli, Aymric / Audonnet, Sandra / Duret, Valérie / Tabary, Thierry / Cohen, Jacques Henri Max / Mahmoudi, Rachid

Journal of visualized experiments. 2020 May 19, , no. 159

2020

Abstract: CR1 (CD35, Complement Receptor type 1 for C3b/C4b) is a high molecular weight membrane glycoprotein of about 200 kDa that controls complement activation, transports immune complexes, and participates in humoral and cellular immune responses. CR1 is ... ...

Abstract	CR1 (CD35, Complement Receptor type 1 for C3b/C4b) is a high molecular weight membrane glycoprotein of about 200 kDa that controls complement activation, transports immune complexes, and participates in humoral and cellular immune responses. CR1 is present on the surface of many cell types, including erythrocytes, and exhibits polymorphisms in length, structure (Knops, or KN, blood group), and density. The average density of CR1 per erythrocyte (CR1/E) is 500 molecules per erythrocyte. This density varies from one individual to another (100–1,200 CR1/E) and from one erythrocyte to another in the same individual. We present here a robust flow cytometry method to measure the density of CR1/E, including in subjects expressing a low density, with the help of an amplifying immunostaining system. This method has enabled us to show the lowering of CR1 erythrocyte expression in diseases such as Alzheimer's disease (AD), systemic lupus erythematosus (SLE), AIDS, or malaria.
Keywords	Alzheimer disease ; antigen-antibody complex ; blood groups ; cell-mediated immunity ; complement ; erythrocytes ; flow cytometry ; lupus erythematosus ; malaria ; membrane glycoproteins ; molecular weight
Language	English
Dates of publication	2020-0519
Size	p. e60810.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/60810
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Measuring Erythrocyte Complement Receptor 1 Using Flow Cytometry.

Kisserli, Aymric / Audonnet, Sandra / Duret, Valérie / Tabary, Thierry / Cohen, Jacques Henri Max / Mahmoudi, Rachid

Journal of visualized experiments : JoVE

2020 , Issue 159

Abstract	CR1 (CD35, Complement Receptor type 1 for C3b/C4b) is a high molecular weight membrane glycoprotein of about 200 kDa that controls complement activation, transports immune complexes, and participates in humoral and cellular immune responses. CR1 is present on the surface of many cell types, including erythrocytes, and exhibits polymorphisms in length, structure (Knops, or KN, blood group), and density. The average density of CR1 per erythrocyte (CR1/E) is 500 molecules per erythrocyte. This density varies from one individual to another (100-1,200 CR1/E) and from one erythrocyte to another in the same individual. We present here a robust flow cytometry method to measure the density of CR1/E, including in subjects expressing a low density, with the help of an amplifying immunostaining system. This method has enabled us to show the lowering of CR1 erythrocyte expression in diseases such as Alzheimer's disease (AD), systemic lupus erythematosus (SLE), AIDS, or malaria.
MeSH term(s)	Calibration ; Cell Count ; Erythrocytes/metabolism ; Flow Cytometry/methods ; Humans ; Receptors, Complement/blood ; Regression Analysis
Chemical Substances	Receptors, Complement
Language	English
Publishing date	2020-05-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/60810
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Article ; Online: Acquired decrease of the C3b/C4b receptor (CR1, CD35) and increased C4d deposits on erythrocytes from ICU COVID-19 patients.

Kisserli, Aymric / Schneider, Nathalie / Audonnet, Sandra / Tabary, Thierry / Goury, Antoine / Cousson, Joel / Mahmoudi, Rachid / Bani-Sadr, Firouze / Kanagaratnam, Lukshe / Jolly, Damien / Cohen, Jacques Hm

Immunobiology

2021 Volume 226, Issue 3, Page(s) 152093

Abstract: In order to study the mechanisms of COVID-19 damage following the complement activation phase occurring during the innate immune response to SARS-CoV-2, CR1 (the regulating complement activation factor, CD35, the C3b/C4b receptor), C4d deposits on ... ...

Abstract	In order to study the mechanisms of COVID-19 damage following the complement activation phase occurring during the innate immune response to SARS-CoV-2, CR1 (the regulating complement activation factor, CD35, the C3b/C4b receptor), C4d deposits on Erythrocytes (E), and the products of complement activation C3b/C3bi, were assessed in 52 COVID-19 patients undergoing O
MeSH term(s)	Antigen-Antibody Complex/metabolism ; COVID-19/immunology ; COVID-19/therapy ; Complement Activation ; Complement C4b/metabolism ; Erythrocytes/metabolism ; Erythrocytes/pathology ; France ; Gene Expression Regulation ; Humans ; Intensive Care Units ; Peptide Fragments/metabolism ; Receptors, Complement 3b/genetics ; Receptors, Complement 3b/metabolism ; Receptors, Complement 3b/therapeutic use ; SARS-CoV-2/physiology
Chemical Substances	Antigen-Antibody Complex ; CR1 protein, human ; Peptide Fragments ; Receptors, Complement 3b ; Complement C4b (80295-50-7) ; complement C4d (80295-52-9)
Language	English
Publishing date	2021-05-09
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	563292-4
ISSN	1878-3279 ; 0171-2985
ISSN (online)	1878-3279
ISSN	0171-2985
DOI	10.1016/j.imbio.2021.152093
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Article: High-resolution melting pcr for complement receptor 1 length polymorphism genotyping: an innovative tool for alzheimer's disease gene susceptibility assessment

Kisserli, Aymric / Tabary, Thierry / Cohen, Jacques Henri Max / Duret, Valérie / Mahmoudi, Rachid

Journal of visualized experiments. 2017 July 18, , no. 125

2017

Abstract: Complement receptor 1 (CR1), a transmembrane glycoprotein that plays a key role in the innate immune system, is expressed on many cell types, but especially on red blood cells (RBCs). As a receptor for the complement components C3b and C4b, CR1 regulates ...

Abstract	Complement receptor 1 (CR1), a transmembrane glycoprotein that plays a key role in the innate immune system, is expressed on many cell types, but especially on red blood cells (RBCs). As a receptor for the complement components C3b and C4b, CR1 regulates the activation of the complement cascade and promotes the phagocytosis of immune complexes and cellular debris, as well as the amyloid-beta (Aβ) peptide in Alzheimer's disease (AD). Several studies have confirmed AD-associated single nucleotide polymorphisms (SNPs), as well as a copy-number variation (CNV) in the CR1 gene. Here, we describe an innovative method for determining the length polymorphism of the CR1 receptor. The receptor includes three domains, called long homologous repeats (LHR)-LHR-A, LHR-C, and LHR-D-and an n domain, LHR-B, where n is an integer between 0 and 3. Using a single pair of specific primers, the genetic material is used to amplify a first fragment of the LHR-B domain (the variant amplicon B) and a second fragment of the LHR-C domain (the invariant amplicon). The variant amplicon B and the invariant amplicon display differences at five nucleotides outside of the hybridization areas of said primers. The numbers of variant amplicons B and of invariant amplicons is deduced using a quantitative tool (high-resolution melting (HRM) curves), and the ratio of the variant amplicon B to the invariant amplicon differs according to the CR1 length polymorphism. This method provides several advantages over the canonical phenotype method, as it does not require fresh material and is cheaper, faster, and therefore applicable to larger populations. Thus, the use of this method should be helpful to better understand the role of CR1 isoforms in the pathogenesis of diseases such as AD.
Keywords	Alzheimer disease ; antigen-antibody complex ; complement ; copy number variation ; erythrocytes ; genes ; genotyping ; glycoproteins ; melting ; nucleic acid hybridization ; nucleotides ; pathogenesis ; peptides ; phagocytosis ; phenotype ; single nucleotide polymorphism
Language	English
Dates of publication	2017-0718
Size	p. e56012.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/56012
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: High-resolution Melting PCR for Complement Receptor 1 Length Polymorphism Genotyping: An Innovative Tool for Alzheimer's Disease Gene Susceptibility Assessment.

Kisserli, Aymric / Tabary, Thierry / Cohen, Jacques Henri Max / Duret, Valérie / Mahmoudi, Rachid

Journal of visualized experiments : JoVE

2017 , Issue 125

Abstract	Complement receptor 1 (CR1), a transmembrane glycoprotein that plays a key role in the innate immune system, is expressed on many cell types, but especially on red blood cells (RBCs). As a receptor for the complement components C3b and C4b, CR1 regulates the activation of the complement cascade and promotes the phagocytosis of immune complexes and cellular debris, as well as the amyloid-beta (Aβ) peptide in Alzheimer's disease (AD). Several studies have confirmed AD-associated single nucleotide polymorphisms (SNPs), as well as a copy-number variation (CNV) in the CR1 gene. Here, we describe an innovative method for determining the length polymorphism of the CR1 receptor. The receptor includes three domains, called long homologous repeats (LHR)-LHR-A, LHR-C, and LHR-D-and an n domain, LHR-B, where n is an integer between 0 and 3. Using a single pair of specific primers, the genetic material is used to amplify a first fragment of the LHR-B domain (the variant amplicon B) and a second fragment of the LHR-C domain (the invariant amplicon). The variant amplicon B and the invariant amplicon display differences at five nucleotides outside of the hybridization areas of said primers. The numbers of variant amplicons B and of invariant amplicons is deduced using a quantitative tool (high-resolution melting (HRM) curves), and the ratio of the variant amplicon B to the invariant amplicon differs according to the CR1 length polymorphism. This method provides several advantages over the canonical phenotype method, as it does not require fresh material and is cheaper, faster, and therefore applicable to larger populations. Thus, the use of this method should be helpful to better understand the role of CR1 isoforms in the pathogenesis of diseases such as AD.
Language	English
Publishing date	2017-07-18
Publishing country	United States
Document type	Journal Article
ISSN	1940-087X
ISSN (online)	1940-087X
DOI	10.3791/56012
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Key role of the number of complement receptor 1 on erythrocytes for binding of Escherichia coli to erythrocytes and for leukocyte phagocytosis and oxidative burst in human whole blood.

Brekke, Ole-Lars / Christiansen, Dorte / Kisserli, Aymric / Fure, Hilde / Dahl, Jim Andre / Donvito, Béatrice / Reveil, Brigitte / Ludviksen, Judith Krey / Tabary, Thierry / Mollnes, Tom Eirik / Cohen, Jacques H M

Molecular immunology

2019 Volume 114, Page(s) 139–148

Abstract: Aim: To study the role of complement receptor 1 (CR1) for binding of Escherichia coli (E. coli) to erythrocytes, for leukocyte phagocytosis, oxidative burst and complement activation in human whole blood from a CR1 deficient (CR1D) patient and healthy ... ...

Abstract	Aim: To study the role of complement receptor 1 (CR1) for binding of Escherichia coli (E. coli) to erythrocytes, for leukocyte phagocytosis, oxidative burst and complement activation in human whole blood from a CR1 deficient (CR1D) patient and healthy controls with low, medium and high CR1 numbers. Methods: Alexa-labelled bacteria were used to quantify erythrocyte-bound bacteria, free bacteria in plasma and phagocytosis using flow cytometry. Complement activation in plasma was measured by enzyme-linked immunosorbent assay. The CR1 numbers as well as C3bc and C4bc deposition on erythrocytes were measured by flow cytometry. Cytokines were measured using multiplex technology, and bacterial growth was measured by colony forming units. CR1 was blocked using the anti-CR1 blocking mAb 3D9. Results: Approximately 85% of E. coli bound to erythrocytes after 15 min incubation in donor blood with high and medium CR1 numbers, 50% in the person with low CR1 numbers and virtually no detectable binding in the CR1D (r Conclusions: CR1D and low CR1 numbers prevented E. coli binding to erythrocytes, increased free bacteria in plasma, phagocytosis and oxidative burst, but did not affect plasma or surface complement activation and bacterial growth.
MeSH term(s)	Antigen-Antibody Complex/immunology ; Complement Activation/immunology ; Erythrocytes/immunology ; Erythrocytes/microbiology ; Escherichia coli/immunology ; Humans ; Leukocytes/immunology ; Leukocytes/microbiology ; Phagocytosis/immunology ; Receptors, Complement 3b/immunology ; Respiratory Burst/immunology
Chemical Substances	Antigen-Antibody Complex ; CR1 protein, human ; Receptors, Complement 3b
Language	English
Publishing date	2019-07-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2019.07.014
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Article ; Online: Inherited and Acquired Decrease in Complement Receptor 1 (CR1) Density on Red Blood Cells Associated with High Levels of Soluble CR1 in Alzheimer's Disease.

Mahmoudi, Rachid / Feldman, Sarah / Kisserli, Aymric / Duret, Valérie / Tabary, Thierry / Bertholon, Laurie-Anne / Badr, Sarah / Nonnonhou, Vignon / Cesar, Aude / Neuraz, Antoine / Novella, Jean Luc / Cohen, Jacques Henri Max

International journal of molecular sciences

2018 Volume 19, Issue 8

Abstract: The complement receptor 1 ( ...

Abstract	The complement receptor 1 (
MeSH term(s)	Aged ; Aged, 80 and over ; Alleles ; Alzheimer Disease/genetics ; Binding Sites/genetics ; Cohort Studies ; Erythrocytes/chemistry ; Erythrocytes/pathology ; Female ; Genotype ; Humans ; Male ; Methylation ; Multivariate Analysis ; Plaque, Amyloid/pathology ; Polymorphism, Restriction Fragment Length ; Protein Isoforms/blood ; Protein Isoforms/genetics ; Receptors, Complement 3b/blood ; Receptors, Complement 3b/genetics ; Risk Factors
Chemical Substances	CR1 protein, human ; Protein Isoforms ; Receptors, Complement 3b
Language	English
Publishing date	2018-07-25
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/ijms19082175
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Article ; Online: Discrimination between native and Tn6010-associated oqxAB in Klebsiella spp., Raoultella spp., and other Enterobacteriaceae by using a two-step strategy.

Guillard, Thomas / Lebreil, Anne-Laure / Hansen, Lars Hestbjerg / Kisserli, Aymric / Berger, Sibel / Lozniewski, Alain / Alauzet, Corentine / de Champs, Christophe

Antimicrobial agents and chemotherapy

2015 Volume 59, Issue 9, Page(s) 5838–5840

Abstract: We developed a two-step PCR-based strategy to detect genes encoding OqxAB, allowing a specific assignment of Tn6010-associated oqxAB in Enterobacteriaceae. Chromosomal location in this setup was confirmed by hybridization with I-CeuI-restricted genomes. ... ...

Abstract	We developed a two-step PCR-based strategy to detect genes encoding OqxAB, allowing a specific assignment of Tn6010-associated oqxAB in Enterobacteriaceae. Chromosomal location in this setup was confirmed by hybridization with I-CeuI-restricted genomes. This approach led us to find that Klebsiella sp. and Raoultella sp. reference strains chromosomally carried oqxAB.
MeSH term(s)	Anti-Bacterial Agents/pharmacology ; Enterobacteriaceae/drug effects ; Klebsiella/drug effects ; Microbial Sensitivity Tests ; Polymerase Chain Reaction
Chemical Substances	Anti-Bacterial Agents
Language	English
Publishing date	2015-06-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	217602-6
ISSN	1098-6596 ; 0066-4804
ISSN (online)	1098-6596
ISSN	0066-4804
DOI	10.1128/AAC.00669-15
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Article ; Online: Acquired decrease of the C3b/C4b Receptor (CR1, CD35) and C4d deposits on Erythrocytes from ICU COVID-19 Patients

KISSERLI, Aymric / SCHNEIDER, Nathalie / AUDONNET, Sandra / TABARY, Thierry / GOURY, Antoine / COUSSON, Joel / MAHMOUDI, Rachid / BANI-SADR, Firouze / KANAGARATNAM, Lukshe / JOLLY, Damien / COHEN, Jacques H M

medRxiv

Abstract: We determined CR1, CD35 the C3b, C4b receptor density, C3b/C3bi and C4d deposits densities on Erythrocytes (E) in 51 COVID-19 patients undergoing O2 therapy or assisted ventilation in ICU units in Rheims France. A clear acquired decrease of CR1 density ... ...

Abstract	We determined CR1, CD35 the C3b, C4b receptor density, C3b/C3bi and C4d deposits densities on Erythrocytes (E) in 51 COVID-19 patients undergoing O2 therapy or assisted ventilation in ICU units in Rheims France. A clear acquired decrease of CR1 density of E from COVID-19 patients was observed, particularly among fatal cases, and paralleling several severity parameters. Deposits of C4d largely above values observed in normal individuals, mostly without C3 deposits, have been observed in more than 80% of the patients, reminiscent of the sub endothelial pericapil-lary deposits in organ transplant rejection, already observed on E in parallel, as well as also observed on E in clinical SLE flares. Conversely, significant C3 deposits were only observed among 1/4 of the patients. The decrease of CR1/E density, and the detection of virus spike, C3 or C4 fragment on E, among COVID-19 pa-tients, are likely to be two aspects of the same phenomenon of immune complexes or complement fragment coated cell debris handling and clearance. Measurement of C4d deposit on E might represent a way for assessing inflammation and comple-ment activation occurring in organ capillaries. CR1/E decrease might represent a cumulative index of complement activation in COVID-19 patients. Taken together, these original findings stress on the participation of the complement regulatory pro-teins in that disease and evidence that E matter in immune mechanisms in COVID-19 patients. The use of CR1, or CR1-like molecules with the aim of down regulating complement activation and inflammation for therapy should also be considered.
Keywords	covid19
Language	English
Publishing date	2020-08-14
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2020.08.10.20162412
Database	COVID19

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Article ; Online: Inherited and Acquired Decrease in Complement Receptor 1 (CR1) Density on Red Blood Cells Associated with High Levels of Soluble CR1 in Alzheimer’s Disease

Rachid Mahmoudi / Sarah Feldman / Aymric Kisserli / Valérie Duret / Thierry Tabary / Laurie-Anne Bertholon / Sarah Badr / Vignon Nonnonhou / Aude Cesar / Antoine Neuraz / Jean Luc Novella / Jacques Henri Max Cohen

International Journal of Molecular Sciences, Vol 19, Iss 8, p

2018 Volume 2175

Abstract: The complement receptor 1 (CR1) gene was shown to be involved in Alzheimer’s disease (AD). We previously showed that AD is associated with low density of the long CR1 isoform, CR1*2 (S). Here, we correlated phenotype data (CR1 density per erythrocyte ( ... ...

Abstract	The complement receptor 1 (CR1) gene was shown to be involved in Alzheimer’s disease (AD). We previously showed that AD is associated with low density of the long CR1 isoform, CR1*2 (S). Here, we correlated phenotype data (CR1 density per erythrocyte (CR1/E), blood soluble CR1 (sCR1)) with genetic data (density/length polymorphisms) in AD patients and healthy controls. CR1/E was enumerated using flow cytometry, while sCR1 was quantified by ELISA. CR1 polymorphisms were assessed using restriction fragment length polymorphism (RFLP), pyrosequencing, and high-resolution melting PCR. In AD patients carrying the H allele (HindIII polymorphism) or the Q allele (Q981H polymorphism), CR1/E was significantly lower when compared with controls carrying the same alleles (p < 0.01), contrary to sCR1, which was significantly higher (p < 0.001). Using multivariate analysis, a reduction of 6.68 units in density was associated with an increase of 1% in methylation of CR1 (estimate −6.68; 95% confidence intervals (CIs) −12.37, −0.99; p = 0.02). Our data show that, in addition to inherited genetic factors, low density of CR1/E is also acquired. The involvement of CR1 in the pathogenesis of AD might be linked to insufficient clearance of amyloid deposits. These findings may open perspectives for new therapeutic strategies in AD.
Keywords	Alzheimer’s disease ; complement receptor 1 ; CR1 length polymorphism ; CR1 density ; complement C3b/C4b receptor ; complement ; dementia ; molecular biology ; neurosciences ; genetic risk ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	610
Language	English
Publishing date	2018-07-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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