ELISpot analysis of LPS-stimulated leukocytes: Human granulocytes selectively secrete IL-8, MIP-1β and TNF-α

Smedman, Christian; Gårdlund, Bengt; Nihlmark, Kopek; Gille-Johnson, Patrik; Andersson, Jan; Paulie, Staffan

doi:10.1016/j.jim.2009.04.001

Cited by 31 publications

(26 citation statements)

References 147 publications

(180 reference statements)

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“…Freshly-isolated neonatal and adult neutrophils stimulated with LPS had marked secretion of MIP-1 ␤ /CCL4, a chemokine involved in the recruitment of monocytes and neutrophils to inflammatory sites [43,46] , and is an observation consistent with existing literature [47,48] . We also now report that surviving neonatal neutrophils could be induced to secrete MIP-1 ␤ .…”

Section: Discussionsupporting

confidence: 88%

Neonatal Neutrophils with Prolonged Survival Secrete Mediators Associated with Chronic Inflammation

Nguyen

Schnulle²,

Chegini³

et al. 2010

Neonatology

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Background: The resolution of inflammation involves the efficient removal of apoptotic neutrophils (PMN). However, a subpopulation of PMN that are resistant to apoptosis may contribute to PMN persistence in tissues, an early hallmark of chronic inflammation. We previously made observations that neonatal PMN with prolonged survival had augmented expression of CD18/CD11b, an adhesion molecule critical to inflammation. Objectives: The objectives of this study were to test the hypothesis that surviving neonatal PMN retain the capacity to secrete key mediators associated with chronic inflammation. Methods: We profiled cytokine and chemokine secretion patterns of lipopolysaccharide (LPS)-stimulated neonatal and adult PMN using multicytokine array and ELISA. Results: We observed that surviving 24-hour neonatal PMN stimulated with LPS had enhanced secretion of interleukin (IL)-8, a chemokine involved in PMN activation and recruitment. In addition, 24-hour neonatal PMN secreted levels of monocyte inhibitory protein (MIP)-1β that were higher than those secreted by 0-hour PMN, but amounts of IL-1 receptor antagonist (IL-1Ra) were lower. Conclusions: The results of the present study extend previous observations of augmented function in surviving neonatal neutrophils, and further suggest their potential contribution to the pathogenesis of inflammatory disorders in neonates.

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Section: Discussionsupporting

confidence: 88%

Neonatal Neutrophils with Prolonged Survival Secrete Mediators Associated with Chronic Inflammation

Nguyen

Schnulle²,

Chegini³

et al. 2010

Neonatology

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“…[24][25][26] A great deal of controversy exists concerning the ability of human neutrophils to produce IL-10. [7][8][9][10][11][12]14,[27][28][29] This discordance may result from different factors, such as purification procedures, culture protocols, and presence of contaminating monocytes/eosinophils. In our laboratory, we obtained reproducible data on IL-10 secretion by neutrophils, both at the mRNA and protein levels, using real-time RT-PCR, ELISA, and western blotting analysis.…”

Section: Discussionmentioning

confidence: 99%

“…In the majority of experiments with negative results, neutrophils were treated with proinflammatory stimuli, such as fMLP, interferon-g, tumor necrosis factor, granulocyte-macrophages colony-stimulating factor, granulocyte colony-stimulating factor, LPS, Pam3CSK4, poly(I:C), or R-848. 10,27,28 In macrophages, LPS not only triggers nuclear factor-kB activation and subsequent production of proinflammatory cytokines but also initiates the negative feedback loop associated with IL-10 synthesis via a Janus-activated kinase/signal transducer and activator of transcription factor 3-dependent pathway. 39 The different ability of human monocytes and neutrophils to secrete IL-10 may be accounted for the differences in the chromatin status of the IL-10 locus.…”

Section: Discussionmentioning

confidence: 99%

Induction of human IL-10-producing neutrophils by LPS-stimulated Treg cells and IL-10

et al. 2016

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Recent evidence has revealed an unsuspected suppressive role played by neutrophils during microbial infections. An especially intriguing aspect of this role is the ability of neutrophils to produce interleukin (IL)-10 following interaction with lipopolysaccharide (LPS)-stimulated regulatory T (Treg) cells. The present study demonstrates that generation of IL-10 in neutrophils induced by LPS-stimulated Treg cells required direct cell-cell contact. This effect was dependent on the binding of CD11b and intercellular adhesion molecule 1. Neither stimulation of neutrophils with LPS nor their culture with unstimulated Treg cells, CD3/CD28 monoclonal antibodies-stimulated Treg cells, or T conventional cells affected intracellular IL-10 expression. IL-10-positive neutrophils were also induced by exogenous IL-10, providing an example of a positive feedback loop. Both LPS-stimulated Treg cells and exogenous IL-10 exclusively promoted posttranslational modifications of histones, H3K4me3 and H3Ac Lys4, that activate IL-10 genomic locus in neutrophils, while the promoter of IL-10 gene was inactive in resting, LPS-stimulated neutrophils, following blocking of direct interaction with LPS-stimulated Treg cells or in LPS-preactivated neutrophils incubated with LPS-stimulated Treg cells. We additionally confirmed the presence of IL-10-producing neutrophils in vivo in patients with periodontal abscess induced by Gram-negative bacteria, as opposed to neutrophils isolated from the site of aseptic inflammation in patients with neuromyelitis optica.

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“…In contrast, cytokine secretion does not reach detectable levels until hours after stimulation, indicating the involvement of different signaling pathway and perhaps secondary factors, which can be subject to regulation by the mobilization procedure. Various cytokines can be produced by neutrophils, 10,28,36 among which is the chemokine IL-8 (CXCL8). 37,38 Analysis of neutrophil gene expression profiles confirmed the presence of mRNA coding for various other chemokines, eg, CCL3, CCL4, and CCL5.…”

Section: Discussionmentioning

confidence: 99%

“…10,26 Although neutrophils have been previously described to also secrete CCL2 (monocyte chemoattractant protein 1), 27 CCL3 (macrophage inflammatory protein 1-␣), CCL4 (macrophage inflammatory protein 1-␤), CCL5 (regulated on activation normal T cell expressed and secreted), and TNF-␣, 26,28 depending on the (combination of) stimuli used, the most prominent cytokine generated by neutrophils is IL-8 (supplemental Table 2). …”

Section: Chemokine Expression In Mobilized Granulocytesmentioning

confidence: 99%

Toll-like receptor–induced reactivity and strongly potentiated IL-8 production in granulocytes mobilized for transfusion purposes

Drewniak

Tool

Geissler

et al. 2010

Blood

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ELISpot analysis of LPS-stimulated leukocytes: Human granulocytes selectively secrete IL-8, MIP-1β and TNF-α

Cited by 31 publications

References 147 publications

Neonatal Neutrophils with Prolonged Survival Secrete Mediators Associated with Chronic Inflammation

Neonatal Neutrophils with Prolonged Survival Secrete Mediators Associated with Chronic Inflammation

Induction of human IL-10-producing neutrophils by LPS-stimulated Treg cells and IL-10

Toll-like receptor–induced reactivity and strongly potentiated IL-8 production in granulocytes mobilized for transfusion purposes

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