Expression of NanoLuc Luciferase in Listeria innocua for Development of Biofilm Assay

Berlec, Aleš; Janež, Nika; Sterniša, Meta; Klančnik, Anja; Sabotič, Jerica

doi:10.3389/fmicb.2021.636421

Cited by 9 publications

(5 citation statements)

References 41 publications

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“…Bacterial culture. Lm EGD 44 , PNF8-GFP 45 and Lm Mu strains 14 were stored as frozen glycerol stocks (∼1x10 9 /ml) at -80°C. Bacteria were cultured in BHI medium and harvested during log phase growth for inoculation of tissue and mice 46 .…”

Section: Methodsmentioning

confidence: 99%

Listeria motility increases the efficiency of goblet cell invasion during intestinal infection

Wortel

Kim

Liu

et al. 2022

Preprint

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Listeria monocytogenes (Lm) is a food-borne pathogen that causes severe bacterial gastroenteritis, with high rates of hospitalization and mortality. Lm is ubiquitous in soil, water and livestock, and can survive and proliferate at low temperatures. Following oral ingestion of contaminated food, Lm crosses the epithelial through intestinal goblet cells in a mechanism depending on Lm InlA and host E-cadherin. Importantly, human infections typically occur with Lm growing at or below room temperature, which are flagellated and motile. Even though many important human bacterial pathogens are flagellated, little is known regarding the effect of bacterial motility on invasion and immune evasion. Here, we used complementary imaging and computer modeling approaches to test the hypothesis that bacterial motility helps Lm locate and engage target cells permissive for invasion. Imaging explanted mouse and human intestine, we confirmed that Lm grown at room temperature uses motility to scan the epithelial surface and preferentially attach to target cells. Furthermore, we integrated quantitative parameters from our imaging experiments to construct a versatile "layered" cellular Potts model (L-CPM) that simulates host-pathogen dynamics. Simulated data are consistent with the hypothesis that bacterial motility enhances invasion by allowing bacteria to search the epithelial surface for their preferred invasion targets. Indeed, our model consistently predicts that motile bacteria have invaded ~2-fold more at the 1-hour mark. This invasion advantage persists even in the presence of host phagocytes, with the balance between invasion and phagocytosis governed almost entirely by bacterial motility. In conclusion, our simulations provide insight into host pathogen interactions and challenge fundamental assumptions regarding how phagocytes might limit bacterial invasion early during infection.

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Section: Methodsmentioning

confidence: 99%

Listeria motility increases the efficiency of goblet cell invasion during intestinal infection

Wortel

Kim

Liu

et al. 2022

Preprint

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show abstract

“…27 L. innocua is a non-pathogenic microorganism often investigated as a surrogate for Listeria monocytogenes, an important foodborne pathogen. 28 P. fluorescens is a relevant spoilage microorganism in dairy products. 29 To obtain the inoculum, the bacteria were activated twice in brain heart infusion broth (BHI, Kasvi, Italy), incubated for 24 h at 23 ºC for P. fluorescens and 37 ºC for the others, streaked over plate count agar (PCA, Oxoid, England) and incubated again at the same conditions.…”

Section: Biological Assays In Vitromentioning

confidence: 99%

Green Synthesis of Cellulose-Based Biofilms Doped with AgNPs with Antibacterial Activity

Pereira¹,

Silva²,

Marques³

et al. 2023

J. Braz. Chem. Soc.

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In this work, cellulose-based composite biofilms doped with silver nanoparticles (AgNPs) have been produced by eco-friendly process. The AgNPs were synthesized from Ilex paraguariensis with an average size of 45 nm, a characteristic plasmon band at 450 nm and zeta potential of -23 mV. The biofilm has been synthesized by casting method, using different volumes of AgNPs suspension with glycerol plasticizing agent (T6-T10) or not (T1-T5). The AgNPs were successfully incorporated into the biofilms that was confirmed by scanning electron microscopy. The mechanical and optical structural and the antimicrobial properties of the biofilms have been determined. In the presence of AgNPs, the mechanical properties, thermal stability and water vapor barrier of the biofilms have been not altered. However, the addition of glycerol decreased the tensile strength and the modulus of elasticity, in addition to increasing the elongation of the biofilms and the water vapor barrier. The biofilm T10 showed a transmittance of ca. 10%, with high potential to be used as a UV light blocker. Finally, the biofilms doped with AgNPs showed activity against the bacterium Pseudomonas fluorescens. Given the above, this work presented a new biofilm with activity against the bacterium Pseudomonus fluorescens obtained by eco-friendly process.

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“…Lm EGD [33], PNF8-GFP [34] and Lm Mu strains [14] were stored as frozen glycerol stocks (*1x10 9 /ml) at -80℃. Bacteria were cultured in BHI medium and harvested during log phase growth for inoculation of tissue and mice [35].…”

Section: Bacterial Culturementioning

confidence: 99%

Listeria motility increases the efficiency of epithelial invasion during intestinal infection

et al. 2022

View full text Add to dashboard Cite

Listeria monocytogenes (Lm) is a food-borne pathogen that causes severe bacterial gastroenteritis, with high rates of hospitalization and mortality. Lm is ubiquitous in soil, water and livestock, and can survive and proliferate at low temperatures. Following oral ingestion of contaminated food, Lm crosses the epithelium through intestinal goblet cells in a mechanism mediated by Lm InlA binding host E-cadherin. Importantly, human infections typically occur with Lm growing at or below room temperature, which is flagellated and motile. Even though many important human bacterial pathogens are flagellated, little is known regarding the effect of Lm motility on invasion and immune evasion. Here, we used complementary imaging and computer modeling approaches to test the hypothesis that bacterial motility helps Lm locate and engage target cells permissive for invasion. Imaging explanted mouse and human intestine, we showed that Lm grown at room temperature uses motility to scan the epithelial surface and preferentially attach to target cells. Furthermore, we integrated quantitative parameters from our imaging experiments to construct a versatile “layered” cellular Potts model (L-CPM) that simulates host-pathogen dynamics. Simulated data are consistent with the hypothesis that bacterial motility enhances invasion by allowing bacteria to search the epithelial surface for their preferred invasion targets. Indeed, our model consistently predicts that motile bacteria invade twice as efficiently over the first hour of infection. We also examined how bacterial motility affected interactions with host cellular immunity. In a mouse model of persistent infection, we found that neutrophils migrated to the apical surface of the epithelium 5 hours post infection and interacted with Lm. Yet in contrast to the view that neutrophils “hunt” for bacteria, we found that these interactions were driven by motility of Lm—which moved at least ~50x faster than neutrophils. Furthermore, our L-CPM predicts that motile bacteria maintain their invasion advantage even in the presence of host phagocytes, with the balance between invasion and phagocytosis governed almost entirely by bacterial motility. In conclusion, our simulations provide insight into host pathogen interaction dynamics at the intestinal epithelial barrier early during infection.

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Expression of NanoLuc Luciferase in Listeria innocua for Development of Biofilm Assay

Cited by 9 publications

References 41 publications

Listeria motility increases the efficiency of goblet cell invasion during intestinal infection

Listeria motility increases the efficiency of goblet cell invasion during intestinal infection

Green Synthesis of Cellulose-Based Biofilms Doped with AgNPs with Antibacterial Activity

Listeria motility increases the efficiency of epithelial invasion during intestinal infection

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