Modified Method for Isolation of Peripheral Blood Neutrophils from Bovines and Humans

Akhtar, Raheela; Chaudhary, Z. I.; He, Yongqun

doi:10.5455/ijavms.20101202023635

Cited by 4 publications

(4 citation statements)

References 12 publications

(6 reference statements)

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“…Neutrophils were isolated as previously described ( 7 , 97 ). Briefly, 10 ml of blood was drawn from healthy donors into heparinized Vacutainer tubes and mixed 1:1 with sterile 1× phosphate-buffered saline (PBS) (approval UTK IRB-18-04604-XP) ( 97 ). After mixing, 10 ml of lymphocyte separation medium was underlaid.…”

Section: Methodsmentioning

confidence: 99%

A secreted sirtuin from Campylobacter jejuni contributes to neutrophil activation and intestinal inflammation during infection

Callahan,

Hancock,

Doster

et al. 2023

Sci. Adv.

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Histone modifications control numerous processes in eukaryotes, including inflammation. Some bacterial pathogens alter the activity or expression of host-derived factors, including sirtuins, to modify histones and induce responses that promote infection. In this study, we identified a deacetylase encoded by Campylobacter jejuni which has sirtuin activities and contributes to activation of human neutrophils by the pathogen. This sirtuin is secreted from the bacterium into neutrophils, where it associates with and deacetylates host histones to promote neutrophil activation and extracellular trap production. Using the murine model of campylobacteriosis, we found that a mutant of this bacterial sirtuin efficiently colonized the gastrointestinal tract but was unable to induce cytokine production, gastrointestinal inflammation, and tissue pathology. In conclusion, these results suggest that secreted bacterial sirtuins represent a previously unreported class of bacterial effector and that bacterial-mediated modification of host histones is responsible for the inflammation and pathology that occurs during campylobacteriosis.

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Section: Methodsmentioning

confidence: 99%

A secreted sirtuin from Campylobacter jejuni contributes to neutrophil activation and intestinal inflammation during infection

Callahan,

Hancock,

Doster

et al. 2023

Sci. Adv.

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show abstract

“…Neutrophils were isolated as previously described (Callahan et al, 2020, 2021b). Briefly, 10 ml of blood was drawn from healthy donors into heparinized Vacutainer tubes and mixed 1:1 with sterile 1× PBS (approval UTK IRB-18-04604-XP) (Akhtar et al, 2010). After mixing, 10 ml of lymphocyte separation medium was underlaid.…”

Section: Methodsmentioning

confidence: 99%

“…:3>21D Isolation of primary human neutrophils Neutrophils were isolated as previously described (Callahan et al, 2020(Callahan et al, , 2021b. Briefly, 10 ml of blood was drawn from healthy donors into heparinized Vacutainer tubes and mixed 1:1 with sterile 1× PBS (approval UTK IRB-18-04604-XP) (Akhtar et al, 2010).…”

Section: Bacterial Cultures and Culture Conditionsmentioning

confidence: 99%

Battle for the histones: a secreted bacterial sirtuin from Campylobacter jejuni activates neutrophils and induces inflammation during infection

Callahan

Hancock

Doster

et al. 2022

Preprint

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Histone modifications alter numerous cornerstone processes in eukaryotes, including metabolism, physiology, and immunity. Numerous bacterial pathogens can alter expression of host-derived sirtuins to deacetylate histones in order to promote infection, yet, a bacterial-derived sirtuin has yet to be investigated to deacetylate host histones. Using Campylobacter jejuni, the leading cause of bacterial-derived gastroenteritis, we found a secreted sirtuin, SliP, which binds to and deacetylates neutrophil histones. We found neutrophil activation and extrusion of neutrophil extracellular traps was SliP dependent, whereby sliP mutants are unable to activate neutrophils or promote NETosis. Leveraging the mouse model of campylobacteriosis, we further demonstrate the sliP mutant can efficiently infect IL-10-/- mice, but induction of proinflammatory cytokine production and gastrointestinal pathology is SliP-dependent. In conclusion, we investigate a unique bacterial effector which targets host histones and is responsible for the inflammatory response and tissue pathology observed during campylobacteriosis.HighlightsC. jejuni encodes a secreted effector, SliP, which functions as a canonical sirtuinSliP binds to and deacetylates neutrophil histone H3 during bacterial infectionC. jejuni-induced neutrophil activation and NETosis are SliP-dependentInflammation and tissue pathology during C. jejuni infection is SliP-dependent

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“…Total cellular RNA was extracted from isolated granulocytes (protocol by Raheela Akhtar et al) [15] by TRIzol Reagent (Life Technologies, Inc., Gaithersburg, MD) and was further purified with the RNeasy kit (Qiagen). The yielded RNA was converted to cDNA using a cDNA synthesis kit (Qiagen) with both oligo (dT) and random hexamer primers.…”

Section: Rna Extraction Reverse Transcription and Quantitative Real-mentioning

confidence: 99%

Role of S100A12/RAGE axis in eccentric exercise-induced delayed-onset muscle soreness in male non-athletes

Khorramdelazad

Rohani²,

Jafarzadeh

et al. 2015

Sport Sci Health

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Modified Method for Isolation of Peripheral Blood Neutrophils from Bovines and Humans

Cited by 4 publications

References 12 publications

A secreted sirtuin from Campylobacter jejuni contributes to neutrophil activation and intestinal inflammation during infection

A secreted sirtuin from Campylobacter jejuni contributes to neutrophil activation and intestinal inflammation during infection

Battle for the histones: a secreted bacterial sirtuin from Campylobacter jejuni activates neutrophils and induces inflammation during infection

Role of S100A12/RAGE axis in eccentric exercise-induced delayed-onset muscle soreness in male non-athletes

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