Molecular imaging of T4 phage in mammalian tissues and cells

Kaźmierczak, Zuzanna; Piotrowicz, Agnieszka; Owczarek, Barbara; Hodyra, Katarzyna; Miernikiewicz, Paulina; Lecion, Dorota; Harhala, Marek; Górski, Andrzej; Dąbrowska, Krystyna

doi:10.4161/bact.28364

Cited by 38 publications

(35 citation statements)

References 20 publications

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“…presented one of the first images of free T2 phage particles within phagocytic cells, showing phages within endocytic vacuoles that fused with denser bodies of the cytoplasm as they migrated towards the cellular interior. Similar studies have shown evidence of filamentous phage and Myoviridae within mammalian cells, although these phages were either engineered to display exogenous protein markers or the mechanism of cellular entry was not clearly delineated . Several review articles have broached the topic of phage‐epithelial transcytosis, but few attempts have been made to experimentally validate whether phage transcytosis occurs naturally and via which route .…”

Section: Across the Cell Layermentioning

confidence: 99%

A bacteriophages journey through the human body

Barr

2017

Immunological Reviews

172

130

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The human body is colonized by a diverse collective of microorganisms, including bacteria, fungi, protozoa and viruses. The smallest entity of this microbial conglomerate are the bacterial viruses. Bacteriophages, or phages for short, exert significant selective pressure on their bacterial hosts, undoubtedly influencing the human microbiome and its impact on our health and well-being. Phages colonize all niches of the body, including the skin, oral cavity, lungs, gut, and urinary tract. As such our bodies are frequently and continuously exposed to diverse collections of phages. Despite the prevalence of phages throughout our bodies, the extent of their interactions with human cells, organs, and immune system is still largely unknown. Phages physically interact with our mucosal surfaces, are capable of bypassing epithelial cell layers, disseminate throughout the body and may manipulate our immune system. Here, I establish the novel concept of an "intra-body phageome," which encompasses the collection of phages residing within the classically "sterile" regions of the body. This review will take a phage-centric view of the microbiota, human body, and immune system with the ultimate goal of inspiring a greater appreciation for both the indirect and direct interactions between bacteriophages and their mammalian hosts.

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Section: Across the Cell Layermentioning

confidence: 99%

A bacteriophages journey through the human body

Barr

2017

Immunological Reviews

172

130

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“…2B). Previous studies showed that the phage particles transferred to the interior of macrophages and significantly reduced the damage caused by the cytotoxic effects of bacteria on phagocytes (4,16). Moreover, Barr et al (13) reported that interaction of phages and mucus-producing cells does occur in the elimination of bacteria, and this interaction is based on immunoglobulin-like domains.…”

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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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“…Phagocytosis of T4 phage by dendritic cells was observed with green‐fluorescence protein (GFP) inserted into the phage genome. Molecular imaging of the cells containing GFP phage particles confirmed that the phage particles were successfully ingested by the dendritic cells . In addition, dendritic cells were more sensitive and responsive upon uptake of active virion particles than that of synthetic particles .…”

Section: Phage Therapymentioning

confidence: 88%

“…Molecular imaging of the cells containing GFP phage particles confirmed that the phage particles were successfully ingested by the dendritic cells. 113 In addition, dendritic cells were more sensitive and responsive upon uptake of active virion particles than that of synthetic particles. 114,115 Following the ingestion of the phages, another study using macrophages confirmed phage degradation within splenocytes, which ended up in macrophage compartments.…”

Section: Phage Therapy and The Human Immune Systemmentioning

confidence: 99%

Interaction of phages, bacteria, and the human immune system: Evolutionary changes in phage therapy

Jariah

Hakim

2019

Reviews in Medical Virology

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Summary Phages and bacteria are known to undergo dynamic and co‐evolutionary arms race interactions in order to survive. Recent advances from in vitro and in vivo studies have improved our understanding of the complex interactions between phages, bacteria, and the human immune system. This insight is essential for the development of phage therapy to battle the growing problems of antibiotic resistance. It is also pivotal to prevent the development of phage‐resistance during the implementation of phage therapy in the clinic. In this review, we discuss recent progress of the interactions between phages, bacteria, and the human immune system and its clinical application for phage therapy. Proper phage therapy design will ideally produce large burst sizes, short latent periods, broad host ranges, and a low tendency to select resistance.

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Molecular imaging of T4 phage in mammalian tissues and cells

Cited by 38 publications

References 20 publications

A bacteriophages journey through the human body

A bacteriophages journey through the human body

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

Interaction of phages, bacteria, and the human immune system: Evolutionary changes in phage therapy

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