The role of full-length apoE in clearance of Gram-negative bacteria and their endotoxins

Petruk, Ganna; Elvén, Malin; Hartman, Erik; Davoudi, Mina; Schmidtchen, Artur; Puthia, Manoj; Petrlova, Jitka

doi:10.1016/j.jlr.2021.100086

Cited by 19 publications

(23 citation statements)

References 70 publications

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“…Next, 50 μL of bacterial suspension was incubated with 2 μM of APOE, 2 μM of TCP-25 (used as a positive control), or buffer control (10 mM Tris buffer at pH 7.4) for 2 h at 37 °C. After 2 h, serial dilutions of the samples were plated on TH agar plates, incubated overnight at 37 °C, and followed by colony counting the next day [ 10 , 11 ].…”

Section: Methodsmentioning

confidence: 99%

“…Five μL of stained bacterial suspension were trapped between a slide and an 18 mm square coverslip. Ten view fields (1 × 1 mm) were examined from three independent sample preparations using a Zeiss AxioScope A.1 fluorescence microscope (objectives: Zeiss EC Plan-Neofluar 40×; camera: Zeiss AxioCam MRm; acquisition software: Zeiss Zen 2.6 [blue edition]) [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

“…Samples were adsorbed onto carbon-coated grids (Copper mesh, 400) for 60 s and stained with 7 µL of 2% uranyl acetate for 30 s. The grids were rendered hydrophilic via glow discharge at low air pressure. The size of aggregates was analyzed as the mean of gray value/μm ± standard deviation (SD) by ImageJ 1.52k, after all the images were converted to 8-bit and the threshold was manually adjusted [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

“…The signal was acquired by a Chemi-Doc (Bio-Rad) system. All the experiments were performed at least three times [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

“…It has been shown that APOE = driven peptides from the receptor- and heparin-binding region of APOE exhibit antibacterial, antiviral and immunomodulatory effects [ 6 , 7 , 8 , 9 ]. Moreover, we have previously reported that full-length APOE has antibacterial activity against Gram-negative bacteria, both in vitro and in vivo [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

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Antibacterial and Anti-Inflammatory Effects of Apolipoprotein E

et al. 2022

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Apolipoprotein E (APOE) is a lipid-transport protein that functions as a key mediator of lipid transport and cholesterol metabolism. Recent studies have shown that peptides derived from human APOE display anti-inflammatory and antimicrobial effects. Here, we applied in vitro assays and fluorescent microscopy to investigate the anti-bacterial effects of full-length APOE. The interaction of APOE with endotoxins from Escherichia coli was explored using surface plasmon resonance, binding assays, transmission electron microscopy and all-atom molecular dynamics (MD) simulations. We also studied the immunomodulatory activity of APOE using in vitro cell assays and an in vivo mouse model in combination with advanced imaging techniques. We observed that APOE exhibits anti-bacterial activity against several Gram-negative bacterial strains of Pseudomonas aeruginosa and Escherichia coli. In addition, we showed that APOE exhibits a significant binding affinity for lipopolysaccharide (LPS) and lipid A as well as heparin. MD simulations identified the low-density lipoprotein receptor (LDLR) binding region in helix 4 of APOE as a primary binding site for these molecules via electrostatic interactions. Together, our data suggest that APOE may have an important role in controlling inflammation during Gram-negative bacterial infection.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The signal was acquired by a Chemi-Doc (Bio-Rad) system. All the experiments were performed at least three times [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Antibacterial and Anti-Inflammatory Effects of Apolipoprotein E

et al. 2022

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SARS‐CoV‐2 spike protein aggregation is triggered by bacterial lipopolysaccharide

et al. 2022

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SARS-CoV-2 spike (S) protein is crucial for virus invasion in COVID-19. Here, we showed that lipopolysaccharide (LPS) can trigger S protein aggregation at high doses of LPS and S protein. We demonstrated the formation of S protein aggregates by microscopy analyses, aggregation and gel shift assays. LPS at high levels boosts the formation of S protein aggregates as detected by amytracker and thioflavin T dyes that specifically bind to aggregating proteins. We validated the role of LPS by blocking the formation of aggregates by the endotoxin-scavenging thrombin-derived peptide TCP-25. Aggregation-prone sequences in S protein are predicted to be nearby LPS binding sites, while molecular simulations showed stable formation of S protein-LPS higher-order oligomers. Collectively, our results provide evidence of LPS-induced S protein aggregation.

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Single‐cell transcriptomics reveals distinct cell response between acute and chronic pulmonary infection of Pseudomonas aeruginosa

et al. 2022

MedComm

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Knowledge of the changes in the immune microenvironment during pulmonary bacterial acute and chronic infections is limited. The dissection of immune system may provide a basis for effective therapeutic strategies against bacterial infection. Here, we describe a single immune cell atlas of mouse lungs after acute and chronic Pseudomonas aeruginosa infection using single‐cell transcriptomics, multiplex immunohistochemistry, and flow cytometry. Our single‐cell transcriptomic analysis revealed large‐scale comprehensive changes in immune cell composition and high variation in cell–cell interactions after acute and chronic P. aeruginosa infection. Bacterial infection reprograms the genetic architecture of immune cell populations. We identified specific immune cell types, including Cxcl2+ B cells and interstitial macrophages, in response to acute and chronic infection, such as their proportions, distribution, and functional status. Importantly, the patterns of immune cell response are drastically different between acute and chronic infection models. The distinct molecular signatures highlight the importance of the highly dynamic cell–cell interaction process in different pathological conditions, which has not been completely revealed previously. These findings provide a comprehensive and unbiased immune cellular landscape for respiratory pathogenesis in mice, enabling further understanding of immunologic mechanisms in infection and inflammatory diseases.

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The role of full-length apoE in clearance of Gram-negative bacteria and their endotoxins

Cited by 19 publications

References 70 publications

Antibacterial and Anti-Inflammatory Effects of Apolipoprotein E

Antibacterial and Anti-Inflammatory Effects of Apolipoprotein E

SARS‐CoV‐2 spike protein aggregation is triggered by bacterial lipopolysaccharide

Single‐cell transcriptomics reveals distinct cell response between acute and chronic pulmonary infection of Pseudomonas aeruginosa

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