A first step toward liposome-mediated intracellular bacteriophage therapy

Nieth, Anita; Verseux, Cyprien; Barnert, Sabine; Süss, Regine; Römer, Winfried

doi:10.1517/17425247.2015.1043125

Cited by 72 publications

(57 citation statements)

References 50 publications

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“…Recent studies have investigated the effect of experimental phage therapy on intracellular killing of bacteria in patients' peripheral blood monocytes, polymorphonuclear neutrophils, and murine macrophages (3-6). We set out to study the intracellular killing potential of such lytic phages in nonprofessional phagocytic cells (7,8). We investigated this question using phage vB_SauM_JS25, a broad-spectrum virulent phage belonging to the family Myoviridae (9).…”

Section: Abstract Mac-t Cells Staphylococcus Aureus Bacteriophagesmentioning

confidence: 99%

Intracellular Staphylococcus aureus Control by Virulent Bacteriophages within MAC-T Bovine Mammary Epithelial Cells

Zhang¹,

Sun²,

Wei³

et al. 2017

Antimicrob Agents Chemother

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Bacteriophages (phages) are known to effectively kill extracellular multiplying bacteria. The present study demonstrated that phages penetrated bovine mammary epithelial cells and cleared intracellular Staphylococcus aureus in a timedependent manner. In particular, phage vB_SauM_JS25 reached the nucleus within 3 h postincubation. The phages had an endocytotic efficiency of 12%. This ability to kill intracellular host bacteria suggests the utility of phage-based therapies and may protect patients from recurrent infection and treatment failure. KEYWORDS MAC-T cells, Staphylococcus aureus, bacteriophagesA lthough traditionally considered an extracellular pathogen, Staphylococcus aureus is able to internalize into host cells, including professional (i.e., macrophages and neutrophils) and nonprofessional (i.e., bovine mammary epithelial cells) phagocytic cells, which is associated with chronic and recurrent infections (1, 2). Phage therapy appears to be a potent and safe alternative tool for treating such bacterial infections. Recent studies have investigated the effect of experimental phage therapy on intracellular killing of bacteria in patients' peripheral blood monocytes, polymorphonuclear neutrophils, and murine macrophages (3-6). We set out to study the intracellular killing potential of such lytic phages in nonprofessional phagocytic cells (7,8). We investigated this question using phage vB_SauM_JS25, a broad-spectrum virulent phage belonging to the family Myoviridae (9).Mammary epithelial cells play an essential role in the surveillance of mammary tissue during infection by assisting immune cell recruitment and bacterial recognition (10). Can phages penetrate within nonprofessional phagocytic cells and eliminate intracellular S. aureus? To answer this, bovine mammary epithelial (MAC-T) cells were cultured in 24-well plates (10 5 cells/well), incubated overnight (37°C, 5% CO 2 ) in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (Sigma, Milan, Italy) and then infected with S. aureus JYG2 (10 6 bacteria/well). One hour after infection at 37°C in 5% CO 2 , cells were washed three times with phosphatebuffered saline (PBS), and extracellular bacteria were killed with lysostaphin (20 g/ml). Two hours later, cells were washed and treated with phage vB_SauM_JS25 (10 8 PFU/well) and incubated for another 3 or 12 h. Then cells were washed three times with PBS and treated with citric acid buffer (40 mM citric acid, 10 mM KCl, 135 mM NaCl, pH 3.0) for 2 min to inactivate any phage particles that remained on the surface (11). The cells were washed two times with medium to remove the acid buffer, followed by digestion with 0.25% trypsogen and lysis with Triton X-100 (final concentration, 0.1%)

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Section: Abstract Mac-t Cells Staphylococcus Aureus Bacteriophagesmentioning

confidence: 99%

Intracellular Staphylococcus aureus Control by Virulent Bacteriophages within MAC-T Bovine Mammary Epithelial Cells

Zhang¹,

Sun²,

Wei³

et al. 2017

Antimicrob Agents Chemother

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“…2,3 Alternatively, hydrogel-assisted swelling facilitated construction of vesicles under physiological conditions with various lipid compositions. 8,15 The technique has since expanded the possibility to study complex lipid-protein interactions, 16,17 vesicles for applications in drug delivery 18 and use of vesicles as micro-reactors. 19,20 However no studies have reported multicompartment formation in vesicles using this methodology.…”

Section: Introductionmentioning

confidence: 99%

Engineering bio-mimicking functional vesicles with multiple compartments for quantifying molecular transport

et al. 2020

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“…P‐glycoprotein carries a role in crossing the blood‐brain barriers and in the uptake into cancer cells. GUVs formed using the gel‐assisted hydration could be used for medical purposes such as delivering bacteriophages to cells to fight again antibiotic‐resistant bacteria . Bacteriophages cannot penetrate cells but by encapsulating bacteriophages in liposomes GUVs, they could be delivered to eukaryotic cells.…”

Section: Gel‐assisted Hydrationmentioning

confidence: 99%

Self‐Assembly of Giant Unilamellar Vesicles by Film Hydration Methodologies

2019

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Self‐assembly of lipids or polymeric amphiphiles into vesicular structures has been achieved by various methods since the first generation of liposomes in the 1960s. Vesicles can be obtained with diameters from the nanometer to the micrometer regime. From the perspective of cell mimicking, vesicles with diameters of several micrometers are most relevant. These vesicles are called giant unilamellar vesicles (GUVs). Commonly used methods to form GUVs are solvent‐displacement techniques, especially since the development of microfluidics. These methodologies however, trap undesirable organic solvents in their membrane as well as other potentially undesired additives (surfactants, polyelectrolytes, polymers, etc.). In contrast to those strategies, summarized herein are solvent‐free approaches as suitable clean alternatives. The vesicles are formed from a dry thin layer of the lipid or amphiphilic polymers and are hydrated in aqueous media using the entropically favored self‐assembly of amphiphiles into GUVs. The rearrangement of the amphiphilic films into vesicular structures is usually aided by shear forces such as an alternative current (electroformation) or the swelling of water‐soluble polymeric supports (gel‐assisted hydration).

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A first step toward liposome-mediated intracellular bacteriophage therapy

Abstract: These are important milestones in the development of an intracellular bacteriophage therapy that might be useful in the fight against multi-drug-resistant intracellular pathogens like Mycobacterium tuberculosis.

Cited by 72 publications

References 50 publications

Intracellular Staphylococcus aureus Control by Virulent Bacteriophages within MAC-T Bovine Mammary Epithelial Cells

Intracellular Staphylococcus aureus Control by Virulent Bacteriophages within MAC-T Bovine Mammary Epithelial Cells

Engineering bio-mimicking functional vesicles with multiple compartments for quantifying molecular transport

Self‐Assembly of Giant Unilamellar Vesicles by Film Hydration Methodologies

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