The intracellular lifestyle of Francisella noatunensis in Atlantic cod (Gadus morhua L.) leucocytes

Furevik, Anette; Pettersen, Eirin Fausa; Colquhoun, Duncan J.; Wergeland, Heidrun I.

doi:10.1016/j.fsi.2010.11.019

Cited by 27 publications

(20 citation statements)

References 27 publications

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“…Our study showed that the bacterium infected and replicated well within these cells, and the results concerning the infection pattern were in full accordance with earlier findings where intracellular bacteria were dispersed in the cytosol 24 hpi and intracellular replication was shown (Furevik et al 2011, Vestvik et al 2013.…”

Section: Discussionsupporting

confidence: 93%

“…noatunensis strain used in this study (NCMB 14265 T ) was isolated from farmed Atlantic cod (Mikalsen et al 2007) and supplied by Dr. Colquhoun at the National Veterinary Institute, Oslo, Norway. Bacteria were grown as des cribed by Furevik et al (2011) using Bacto TM Eugon Broth (Becton-Dickson) containing 2 mM FeCl 3 * 6H 2 O (Sigma-Aldrich) at 20°C with gentle shaking (150 rpm). A growth curve was established by measuring optical density at 600 nm (OD 600nm ) using a Hitachi U-1100 spectrophotometer.…”

Section: Bacteriamentioning

confidence: 99%

“…After incubation, extracellular bacteria were removed by washing in L-15+ followed by addition of L-15+ with FBS and subsequent incubation. The bacterial dose used for infection was based upon previous studies (Furevik et al 2011). In non-infected controls, only L-15+ medium was added to the wells.…”

Section: In Vitro Infection Of Macrophagesmentioning

confidence: 99%

“…Infected leucocytes and non-infected controls were washed 3 times in PBS and fixed in 1 ml of 3.7% (w/v) paraformaldehyde (Sigma-Aldrich), and the cell membrane was permeabilized by 0.1% (v/v) Triton X-100 as described by Furevik et al (2011). One ml rabbit-anti-Francisella noatunensis serum (dilu ted 1:30 000 in PBS containing 0.5% w/v BSA) was added to each well and incubated for 1 h at 18°C.…”

Section: Immunostaining Of Bacteria Within Leucocytes For Microcopymentioning

confidence: 99%

“…noatunensis has the ability to reside and replicate within Atlantic cod adherent head kidney leucocytes and can inhibit the potent killing mechanism known as respiratory burst (Vestvik et al 2013). The existence within immune cells that normally eradicate bacteria shows that the bacterium has potent virulence mechanisms en abling its intracellular existence (Bakkemo et al 2011, Furevik et al 2011, Vestvik et al 2013. The intracellular location of the bacteria will most likely protect them from antibacterial agents with poor penetration into phagocytic cells, and basic knowledge of the concentrations of antibacterial agents within Atlantic cod leucocytes is therefore needed.…”

Section: Introductionmentioning

confidence: 99%

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Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication

Kaldestad¹,

Haugland²,

Rønneseth³

et al. 2014

Dis. Aquat. Org.

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The granuloma disease caused by Francisella noatunensis subsp. noatunensis in farmed Atlantic cod has not been successfully treated by use of antibacterials, even when antibacterial resistance testing indicates a sufficient effect. The reason for this treatment failure may be the intracellular existence of the bacteria within immune cells, mainly macrophages. To investigate the effect of antibacterials on intracellular Francisella replication, we established a protocol for the detection of drugs within Atlantic cod immune cells using high-performance liquid chromatography (HPLC). When the uptake and intracellular concentrations of oxolinic acid and flumequine were analysed in isolated adherent head kidney leucocytes (HKLs) by HPLC, we found that uptake was rapid and the intracellular concentrations reflected the extracellular exposure concentrations. To investigate the effect of the antibacterial compounds on intracellular bacterial replication, adherent HKLs experimentally infected with the bacteria were analysed using flow cytometry and intracellular labelling of bacteria by specific antibodies. We found that flumequine did not inhibit intracellular bacterial replication. Unexpectedly, the results indicated that the intracellularly effiacy of the drug was reduced. The HPLC method used proved to be highly applicable for accurate determination of intracellular drug concentrations. When combined with sensitive and specific flow cytometry analyses for identification and measurement of intracellular bacterial replication, we suggest that this approach can be very valuable for the design of antibacterial treatments of intracellular pathogens.

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Section: Discussionsupporting

confidence: 93%

Section: Bacteriamentioning

confidence: 99%

Section: In Vitro Infection Of Macrophagesmentioning

confidence: 99%

Section: Immunostaining Of Bacteria Within Leucocytes For Microcopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication

Kaldestad¹,

Haugland²,

Rønneseth³

et al. 2014

Dis. Aquat. Org.

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Francisella Inflammasomes: Integrated Responses to a Cytosolic Stealth Bacterium

Wallet¹,

Lagrange²,

Henry³

2016

Current Topics in Microbiology and Immunology

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Francisella tularensis is a facultative intracellular bacterium causing tularemia, a zoonotic disease. Francisella replicates in the macrophage cytosol and eventually triggers cytosolic immune responses. In murine macrophages, Francisella novicida and Francisella tularensis live vaccine strain lyse in the host cytosol and activate the cytosolic DNA receptor Aim2. Here, we review the mechanisms leading or contributing to Aim2 inflammasome activation, including the role of TLRs and of IFN signaling and the implication of the guanylate-binding proteins 2 and 5 in triggering cytosolic bacteriolysis. Furthermore, we present how this cytosolic Gram-negative bacterium escapes recognition by caspase-11 but can trigger a non-canonical caspase-8 inflammasome. In addition, we highlight the differences in inflammasome activation in murine and human cells with pyrin, NLRP3, and AIM2 involved in sensing Francisella in human phagocytes. From a bacterial prospective, we describe the hiding strategy of Francisella to escape recognition by innate sensors and to resist to bacteriolysis in the host cytosol. Finally, we discuss the inability of the inflammasome sensors to detect F. tularensis subspecies tularensis strains, making them highly pathogenic stealth microbes.

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Disentangling the immune response and host-pathogen interactions in Francisella noatunensis infected Atlantic cod

Solbakken

Jentoft

Reitan

et al. 2019

Comparative Biochemistry and Physiology Part D: Genomics and Pr

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The intracellular lifestyle of Francisella noatunensis in Atlantic cod (Gadus morhua L.) leucocytes

Cited by 27 publications

References 27 publications

Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication

Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication

Francisella Inflammasomes: Integrated Responses to a Cytosolic Stealth Bacterium

Disentangling the immune response and host-pathogen interactions in Francisella noatunensis infected Atlantic cod

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