Analysis of the antimicrobial mechanism of porcine beta defensin 2 against E. coli by electron microscopy and differentially expressed genes

Chen, Ruibo; Zhang, Kun; Zhang, Heng; Gao, Chunyu; Li, Chunli

doi:10.1038/s41598-018-32822-3

Cited by 21 publications

(16 citation statements)

References 57 publications

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“…Similarly, MIC of rPBD114 protein was carried out and rPBD114 protein exhibited antimicrobial activity against E. coli but not Gram-positive bacteria and fungus. However, a great number of studies have shown that defensins have a wide range of antimicrobial activities [34–37]. The antimicrobial activity of rPBD114 protein in our study suggested that Pichia pastoris X 33 would be more tolerant to expression of rPBD114.…”

Section: Discussionmentioning

confidence: 50%

Differential expression, molecular cloning, and characterization of porcine beta defensin 114

Xie

Chen

et al. 2019

J Animal Sci Biotechnol

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Background β-defensins have attracted considerable research interest because of their roles in protecting hosts from various pathogens. This study was conducted to investigate the expression profiles of the porcine β-defensin 114 ( PBD114 ) in different breeds and in response to infections. Moreover, the function of PBD114 protein was partially investigated. Methods Six Tibetan pigs (TP) and six DLY (Duroc×Landrace×Yorkshire) pigs were slaughtered to explore the expression profiles of PBD114 in different breeds and tissues. For infection models, sixteen DLY pigs were divided into two groups and challenged either with sterile saline or E. coli K88. The recombinant protein PBD114 (rPBD114) was obtained by using a heterologous expression system in E. coli . Results PBD114 gene was highly expressed in tissues such as the intestine, liver, spleen, and thymus. Interestingly, the expression level of PBD114 gene was higher in the TP pigs than in the DLY pigs ( P < 0.05), and was significantly elevated upon E. coli K88 challenge ( P < 0.05). The nucleotide sequences of PBD114 from Tibetan and DLY pigs was identical, and both showed a 210-bp open reading frame encoding a 69-amino acid mature peptide. To explaore the function of PBD114 protein, PBD114 gene was successfully expressed in E. coli Origami B (DE3) and the molecular weight of the rPBD114 was estimated by SDS-PAGE to be 25 kDa. The rPBD114 was purified and mass spectrometry verified the protein as PBD114. Importantly, rPBD114 showed antimicrobial activities against E. coli DH5α and E. coli K88, and the minimal inhibitory concentrations (MICs) were 64 and 128 μg/mL, respectively. Hemolytic and cytotoxicity assays showed that rPBD114 did not affect cell viability under physiological concentrations. Conclusions PBD114 is an infection response gene that is differentially-expressed between different porcine breeds and tissues. The antimicrobial activity of PBD114 protein, against pathogens such as the E. coli K88, suggested that it may serve as a candidate for the substitution of conventionally used antibiotics. Electronic supplementary material The online version of this article (10.1186/s40104-019-0367-0) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 50%

Differential expression, molecular cloning, and characterization of porcine beta defensin 114

Xie

Chen

et al. 2019

J Animal Sci Biotechnol

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“…Such changes were attributed by others to bacteria damage and death caused by the contact with antimicrobial materials. [29][30][31] In turn, S. aureus was less sensitive to N,N-dimethyl-N-n-octylammonium groups. In few places, damaged and dead bacteria were seen on FrGO I fibers but they were overgrown by other microorganisms without noticeable damage, as evidenced in Figure 5(G).…”

Section: Resultsmentioning

confidence: 99%

Antibacterial electroconductive rGO modified cotton fabric

Mizerska

Fortuniak

Makowski

et al. 2021

Polymers for Advanced Techs

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Cotton fabric was modified with organosilicon coating containing graphene oxide (GO), which was then thermally reduced to reduced graphene oxide. The thermal reduction imparted electrical conductivity and hydrophobicity to the fabric. In order to augment the reduction of GO in the coating, the organosilicon sol with active reducing ─SiH groups, was synthesized from alkoxysilanes. Chemical modification of the coating with tert-butylamine groups and N,N-dimethyl-N-n-octylammonium groups allowed to achieve antibacterial activity of the electroconductive fabric against Escherichia coli and Staphylococcus aureus. Although the modification worsened the electrical conductivity, the coated fabric, especially that modified with tertbutylamine groups, remained electroconductive.

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“…The ability of avian defensins to chemoattract immune cells also contribute to their antibacterial activities [ 51 ]. Although the antibacterial mechanism of PBD-2 against Staphylococcus aureus and E. coli has been characterized as being membrane disruption and regulation of DNA transcription and translation after PBD-2 is present in the cytoplasm [ 55 , 56 ], the antibacterial mechanism of PBD-2 against G. parasuis is unclear and requires further studies.…”

Section: Discussionmentioning

confidence: 99%

Pigs Overexpressing Porcine β-Defensin 2 Display Increased Resilience to Glaesserella parasuis Infection

et al. 2020

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As the causative agent of Glässer’s disease, Glaesserella (Haemophilus) parasuis has led to serious economic losses to the swine industry worldwide. Due to the low cross-protection of vaccines and increasing antimicrobial resistance of G. parasuis, it is important to develop alternative approaches to prevent G. parasuis infection. Defensins are host defense peptides that have been suggested to be promising substitutes for antibiotics in animal production, while porcine β-defensin 2 (PBD-2) is a potent antimicrobial peptide discovered in pigs. Our previous study generated transgenic (TG) pigs overexpressing PBD-2, which displayed enhanced resistance to Actinobacillus pleuropneumoniae. In this study, the antibacterial activities of PBD-2 against G. parasuis are determined in vitro and in the TG pig model. The concentration-dependent bactericidal activity of synthetic PBD-2 against G. parasuis was measured by bacterial counting. Moreover, after being infected with G. parasuis via a cohabitation challenge model, TG pigs overexpressing PBD-2 displayed significantly milder clinical signs and less severe gross pathological changes than their wild-type (WT) littermates. The TG pigs also exhibited alleviated lung and brain lesions, while bacterial loads in the lung and brain tissues of the TG pigs were significantly lower than those of the WT pigs. Additionally, lung and brain homogenates from TG pigs possessed enhanced antibacterial activity against G. parasuis when compared with those from the WT pigs. Altogether, these proved that overexpression of PBD-2 could also endow pigs with increased resilience to G. parasuis infection, which further confirmed the potential of using the PBD-2 coding gene to develop disease-resistant pigs and provided a novel strategy to combat G. parasuis as well.

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Analysis of the antimicrobial mechanism of porcine beta defensin 2 against E. coli by electron microscopy and differentially expressed genes

Cited by 21 publications

References 57 publications

Differential expression, molecular cloning, and characterization of porcine beta defensin 114

Differential expression, molecular cloning, and characterization of porcine beta defensin 114

Antibacterial electroconductive rGO modified cotton fabric

Pigs Overexpressing Porcine β-Defensin 2 Display Increased Resilience to Glaesserella parasuis Infection

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