Lectinophagocytosis of encapsulated Klebsiella pneumoniae mediated by surface lectins of guinea pig alveolar macrophages and human monocyte-derived macrophages

Athamna, Abed; Ofek, Itzhak; Keisari, Yona; Sd, Markowitz; Dutton, Guy G.S.; Sharon, Nathan

doi:10.1128/iai.59.5.1673-1682.1991

Cited by 94 publications

(39 citation statements)

References 41 publications

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“…Previous phagocytosis assays using adherent cells demand timely and cumbersome microscopic counting to determine the actual number of phagocytes with internalized targets (2,10,17,20). Our method, utilizing a simple labeling procedure and the power of flow cytometric analysis, quantifies the percent of phagocytic cells while easily distinguishing between phagocytosed cells and surface-bound cells.…”

Section: Discussionmentioning

confidence: 99%

A novel flow cytometric method for quantifying phagocytosis of apoptotic cells

et al. 1997

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Many eukaryotic cell types are capable of specific recognition and phagocytosis of apoptotic cells, and there is increasing interest in the mechanisms involved in this process. To facilitate analysis of these mechanisms, we designed a novel fluorescence-based method to quantify phagocytosis in vitro using endothelial cell engulfment of apoptotic cells as a model. The B-cell line WEHI-231 was labeled with the fluorophore 5-(&-6)-carboxytetramethyl-rhodamine-succinimidyl-ester (TAMRA) and then induced to undergo apoptosis by crosslinking cell surface immunoglobulin. An endothelial cell line was subsequently allowed to ingest these TAMRA-labeled apoptotic lymphocytes. After 24 h, nonbound lymphocytes were removed and the mono-layers were dissociated. Any nonphagocytosed lymphocytes that remained tightly bound to the endothelial cells were then indirectly immunofluorescein labeled for the pan leukocyte-specific marker CD45. Flow cytometric analysis of the cells distinguished three endothellal cell populations: 1) endothelial cells with surface bound lymphocytes (TAMRA+ CD45+); 2) endothelial cells containing phagocytosed apoptotic lymphocytes (TAMRA+ CD45-); and 3) endothelial cells that were not associated with lymphocytes. The identification of these populations was verified by confocal microscopy of sorted cells. The method described herein will facilitate detailed studies on phagocytic recognition of apoptotic cells and should have broad applications to other phagocytic cell systems.

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Section: Discussionmentioning

confidence: 99%

A novel flow cytometric method for quantifying phagocytosis of apoptotic cells

et al. 1997

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“…Macrophage lectin receptors recognize repeated mannose or rhamnose sugars to induce phagocytosis. However, K1, K2, K5 and K57 capsule types lack these repeated sugars, therefore facilitating the evasion of lectinophagocytosis and macrophage killing [85][86][87][88][89][90][91]. K1 isolates were found to be more resistant than cKp isolates to extracellular and intracellular killing by neutrophils [92], which could aid in systemic dissemination [93] as well as limiting exposure to extracellular antibiotics [94].…”

Section: Defining Hvkp Genotype and Capsule Typementioning

confidence: 99%

Hypervirulent Klebsiella pneumoniae – clinical and molecular perspectives

2019

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Hypervirulent Klebsiella pneumoniae (hvKp) has emerged as a concerning global pathogen. hvKp is more virulent than classical K. pneumoniae (cKp) and capable of causing community‐acquired infections, often in healthy individuals. hvKp is carried in the gastrointestinal tract, which contributes to its spread in the community and healthcare settings. First recognized in Asia, hvKp arose as a leading cause of pyogenic liver abscesses. In the decades since, hvKp has spread globally and causes a variety of infections. In addition to liver abscesses, hvKp is distinct from cKp in its ability to metastasize to distant sites, including most commonly the eye, lung and central nervous system (CNS). hvKp has also been implicated in primary extrahepatic infections including bacteremia, pneumonia and soft tissue infections. The genetic determinants of hypervirulence are often found on large virulence plasmids as well as chromosomal mobile genetic elements which can be used as biomarkers to distinguish hvKp from cKp clinical isolates. These distinct virulence determinants of hvKp include up to four siderophore systems for iron acquisition, increased capsule production, K1 and K2 capsule types, and the colibactin toxin. Additionally, hvKp strains demonstrate hypermucoviscosity, a phenotypic description of hvKp in laboratory conditions that has become a distinguishing feature of many hypervirulent isolates. Alarmingly, multidrug‐resistant hypervirulent strains have emerged, creating a new challenge in combating this already dangerous pathogen.

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“…The monolayers of macrophages were incubated with H. pylori bacteria ( 107/well) in PRMI-1640 medium without serum for 30 min at 4"C, washed three times with medium to remove non-bound bacteria, and fixed with methanol. The numbers of macrophage-attached bacteria were determined by ELISA using anti-H. pylori antibodies (3,4). Standard curves showing the relationship between the absorbance value (OD 492) and the number of H. p.v/ori strain 17874 or 25 bacteria (from 2.5X lo4 to I.OXIOx) were used.…”

Section: Attuchtiient To Und Ingestion Qf H Pylori Hactcriti Hj Mumentioning

confidence: 99%

The role of heparan sulphate‐binding activity of Helicobacter pylori bacteria in their adhesion to murine macrophages

Chmiela

Paziak-Domańska

Rudnicka

et al. 1995

APMIS

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M., Paziak-Domanska, B., Rudnicka, W. & Wadstrom, T. The role of heparan sulphatebinding activity of Helicobacter pylori bacteria in their adhesion to murine macrophages. APMIS 103: 469474, 1995.The aim of this study was to determine the role of heparan sulphate (HS)-binding activity of Helicobucter pylori microbes in their adhesion to and ingestion by inflammatory peritoneal macrophages. Two H . pylori strains expressing sialic acid-specific haemagglutinins but differing in the expression of heparan sulphate-binding capacity were chosen for investigation. The attachment to an ingestion by macrophages of the H. pylori bacteria were estimated by ELISA using anti-H. pylori antibodies. The adhesion of both H. pylori strains could be inhibited by pretreatment of the bacteria with heparin (H), HS or fetuin, as well as by preincubation of the macrophages with heparinase or neuraminidase. However, detailed analysis of the data on the inhibition of bacterial adhesion to macrophages led to the conclusion that the attachment of H. pylori 25 bacteria, which expressed a high heparan sulphate binding, was mainly determined by HS-binding structures. In contrast, the adhesion to macrophages of H. pylori bacteria 17874 microbes, which expressed a weak heparan sulphate binding, was more dependent on the exhibition of sialic acid-dependent haemagglutinins. The described variation in H. pylori bacterial surface structures mediating their adhesion to macrophages could suggest a similar variation in bacterial adhesion to stomach mucosa and maybe in the pathogenicity of H. pylori strains.

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Lectinophagocytosis of encapsulated Klebsiella pneumoniae mediated by surface lectins of guinea pig alveolar macrophages and human monocyte-derived macrophages

Cited by 94 publications

References 41 publications

A novel flow cytometric method for quantifying phagocytosis of apoptotic cells

A novel flow cytometric method for quantifying phagocytosis of apoptotic cells

Hypervirulent Klebsiella pneumoniae – clinical and molecular perspectives

The role of heparan sulphate‐binding activity of Helicobacter pylori bacteria in their adhesion to murine macrophages

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