Innate immune cells and bacterial infection in zebrafish

Astin, Jonathan W.; Keerthisinghe, Pramuk; Du, Lucia Y.; Sanderson, Leslie E.; Crosier, Kathryn K.E.; Crosier, Philip; Hall, Cathy W.

doi:10.1016/bs.mcb.2016.08.002

Cited by 26 publications

(29 citation statements)

References 79 publications

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“…This analysis revealed that the majority of mortality occurred within the first two days post-infection in all groups (Fig. 3c ), consistent with previous studies using the larval zebrafish hindbrain infection model 41 . Larvae kept under DL and DD conditions displayed a considerably lower survival rate following infection compared with the LD and LL groups (Fig.…”

Section: Resultssupporting

confidence: 91%

“…3a ). This is a well-established site for infection studies in larval zebrafish 23 , 24 , 41 , 42 . The infection dose of 1000 CFU was considered optimal as it resulted in ~50% survival under LD conditions (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Measurement of survival was performed as previously described 41 . Approximately 30 infected larvae were placed into 100 mm petri dishes to prevent overcrowding and monitored.…”

Section: Methodsmentioning

confidence: 99%

“…Individual CFU counts were performed as previously described 41 . In brief, infected larvae were washed in PBS three times to remove adherent bacteria.…”

Section: Methodsmentioning

confidence: 99%

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The innate immune cell response to bacterial infection in larval zebrafish is light-regulated

Darroch

Keerthisinghe

et al. 2017

Sci Rep

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The circadian clock, which evolved to help organisms harmonize physiological responses to external conditions (such as the light/dark cycle, LD), is emerging as an important regulator of the immune response to infection. Gaining a complete understanding of how the circadian clock influences the immune cell response requires animal models that permit direct observation of these processes within an intact host. Here, we investigated the use of larval zebrafish, a powerful live imaging system, as a new model to study the impact of a fundamental zeitgeber, light, on the innate immune cell response to infection. Larvae infected during the light phase of the LD cycle and in constant light condition (LL) demonstrated enhanced survival and bacterial clearance when compared with larvae infected during the dark phase of the LD cycle and in constant dark condition (DD). This increased survival was associated with elevated expression of the zebrafish orthologues of the mammalian pro-inflammatory cytokine genes, Tumour necrosis factor-α, Interleukin-8 and Interferon-γ, and increased neutrophil and macrophage recruitment. This study demonstrates for the first time that the larval zebrafish innate immune response to infection is enhanced during light exposure, suggesting that, similar to mammalian systems, the larval zebrafish response to infection is light-regulated.

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

“…Measurement of survival was performed as previously described 41 . Approximately 30 infected larvae were placed into 100 mm petri dishes to prevent overcrowding and monitored.…”

Section: Methodsmentioning

confidence: 99%

“…Individual CFU counts were performed as previously described 41 . In brief, infected larvae were washed in PBS three times to remove adherent bacteria.…”

Section: Methodsmentioning

confidence: 99%

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The innate immune cell response to bacterial infection in larval zebrafish is light-regulated

Darroch

Keerthisinghe

et al. 2017

Sci Rep

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“…For genetic depletion of macrophages, Tg(mpeg1:nfsB-mCherry) or Tg(mpeg1:nfsb-Cherry;lyz:EGFP) double-transgenic larvae (for neutrophil recruitment experiments) were treated with either 1 or 5 mM metronidazole at 30 hpf for 18 hours. For liposome clodronate-mediated ablation, liposomes (as supplied by ClodronateLiposomes.org) were injected as previously described (66). Of note, given the requirement of blood circulation for normal, definitive hematopoiesis (67), only liposome-injected larvae with unaffected blood flow were used for subsequent experiments.…”

Section: Macrophage Ablation Experimentsmentioning

confidence: 99%

Blocking fatty acid–fueled mROS production within macrophages alleviates acute gouty inflammation

Hall¹,

Sanderson²,

Lawrence³

et al. 2018

Journal of Clinical Investigation

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Gout is the most common inflammatory arthritis affecting men. Acute gouty inflammation is triggered by monosodium urate (MSU) crystal deposition in and around joints that activates macrophages into a proinflammatory state, resulting in neutrophil recruitment. A complete understanding of how MSU crystals activate macrophages in vivo has been difficult because of limitations of live imaging this process in traditional animal models. By live imaging the macrophage and neutrophil response to MSU crystals within an intact host (larval zebrafish), we reveal that macrophage activation requires mitochondrial ROS (mROS) generated through fatty acid oxidation. This mitochondrial source of ROS contributes to NF-κB-driven production of IL-1β and TNF-α, which promote neutrophil recruitment. We demonstrate the therapeutic utility of this discovery by showing that this mechanism is conserved in human macrophages and, via pharmacologic blockade, that it contributes to neutrophil recruitment in a mouse model of acute gouty inflammation. To our knowledge, this study is the first to uncover an immunometabolic mechanism of macrophage activation that operates during acute gouty inflammation. Targeting this pathway holds promise in the management of gout and, potentially, other macrophage-driven diseases.

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Isolation of Neutrophils from Larval Zebrafish and Their Transplantation into Recipient Larvae for Functional Studies

Darroch

Astin

Hall

2019

Methods in Molecular Biology

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Innate immune cells and bacterial infection in zebrafish

Cited by 26 publications

References 79 publications

The innate immune cell response to bacterial infection in larval zebrafish is light-regulated

The innate immune cell response to bacterial infection in larval zebrafish is light-regulated

Blocking fatty acid–fueled mROS production within macrophages alleviates acute gouty inflammation

Isolation of Neutrophils from Larval Zebrafish and Their Transplantation into Recipient Larvae for Functional Studies

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