JAGN1 is required for fungal killing in neutrophil extracellular traps: Implications for severe congenital neutropenia

Khandagale, Avinash; Lazzaretto, Beatrice; Carlsson, Göran; Sundin, Mikael; Shafeeq, Sulman; Römling, Ute; Fadeel, Bengt

doi:10.1002/jlb.4a0118-030rr

Cited by 28 publications

(29 citation statements)

References 59 publications

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“…The latter study suggests that it is highly relevant to study responses of immune cells to NETs in the absence and presence of LPS. Furthermore, we believe it is critically important to study primary human immune cells to complement findings obtained using animal models (69).…”

Section: Discussionmentioning

confidence: 99%

Intra- and Extracellular Degradation of Neutrophil Extracellular Traps by Macrophages and Dendritic Cells

Lazzaretto

Fadeel

2019

The Journal of Immunology

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Neutrophil extracellular traps (NETs) composed of nuclear DNA associated with histones and granule proteins are involved in the extracellular killing of pathogens. Excessive NET formation has been implicated in several noninfectious pathological conditions. The disposal of NETs is, therefore, important to prevent inadvertent effects resulting from the continued presence of NETs in the extracellular environment. In this study, we investigated the interaction of NETs released by freshly isolated, PMA-stimulated primary human neutrophils with primary human monocyte–derived macrophages or dendritic cells (DCs). NETs were internalized by macrophages, and removal of the protein component prevented engulfment of NETs, whereas complexation with LL-37 restored the uptake of “naked” (protein-free) NETs. NETs were also found to dampen the bacterial LPS-induced maturation of DCs. Cytokine profiling was conducted by using a multiplex array following the interaction of NETs with macrophages or DCs, and NETs alone were found to be noninflammatory, whereas immunomodulatory effects were noted in the presence of LPS with significant upregulation of IL-1β secretion, and a marked suppression of other LPS-induced factors including vascular endothelial growth factor (VEGF) in both cell types. Moreover, macrophage digestion of NETs was dependent on TREX1 (also known as DNaseIII), but not DNaseII, whereas extracellular DNase1L3-mediated degradation of NETs was observed for DCs. Collectively, these findings shed light on the interactions between NETs and phagocytic cells and provide new insights regarding the clearance of NETs, double-edged swords of innate immunity.

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Section: Discussionmentioning

confidence: 99%

Intra- and Extracellular Degradation of Neutrophil Extracellular Traps by Macrophages and Dendritic Cells

Lazzaretto

Fadeel

2019

The Journal of Immunology

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“…12 More recent studies have shown that JAGN1 and its murine homologue are required for the anti-fungal actions of neutrophils. 13,14 Nevertheless, the precise role of JAGN1 in the regulation of cell death remains poorly understood.…”

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confidence: 99%

Severe congenital neutropenia‐associated JAGN1 mutations unleash a calpain‐dependent cell death programme in myeloid cells

et al. 2020

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Severe congenital neutropenia (SCN) of autosomal recessive inheritance, also known as Kostmann disease, is characterised by a lack of neutrophils and a propensity for life-threatening infections. Using whole-exome sequencing, we identified homozygous JAGN1 mutations (p.Gly14Ser and p.Glu21Asp) in three patients with Kostmann-like SCN, thus confirming the recent attribution of JAGN1 mutations to SCN. Using the human promyelocytic cell line HL-60 as a model, we found that overexpression of patient-derived JAGN1 mutants, but not silencing of JAGN1, augmented cell death in response to the pro-apoptotic stimuli, etoposide, staurosporine, and thapsigargin. Furthermore, cells expressing mutant JAGN1 were remarkably susceptible to agonists that normally trigger degranulation and succumbed to a calcium-dependent cell death programme. This mode of cell death was completely prevented by pharmacological inhibition of calpain but unaffected by caspase inhibition. In conclusion, our results confirmed the association between JAGN1 mutations and SCN and showed that SCN-associated JAGN1 mutations unleash a calcium-and calpain-dependent cell death in myeloid cells.

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“…As neutrophils have a short lifespan after isolation, HL-60 cells that can differentiate into neutrophil-like cells in response to 1.25% DMSO were used as a model [21] to investigate the impact of the lipoxin A4 receptor on NET expression. We induced the differentiation of HL-60 cells into neutrophil-like cells and the silencing of the lipoxin A4 receptor in neutrophil-like HL-60 cells ( t (4) =15.378, P < 0.001) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The human acute promyelocytic leukemia cell line HL-60 (provided by the Cell Bank of the Chinese Academy of Sciences) was maintained in RIPM 1640 medium supplemented with 10% heat-inactivated FBS (v/v) in 5% CO 2 at 37 °C. Cells were seeded at a density of 5 × 10 5 cells/ml and cultured for 5 days in the above-mentioned cell medium supplemented with 1.25% DMSO to induce their differentiation into neutrophil-like cells [21]. Fresh medium was added on the third day of culture to prevent cell overgrowth and depletion of nutrients.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the efficiency of lipoxin A4 receptor gene down-regulation, the reverse-transcribed cDNA was used as a template in qPCR reaction mixtures containing SYBR Green Real-time PCR Master Mix (TaKaRa) and 0.4 μM forward and reverse primers. The following primers were utilized: GAPDH forward [21] (5′-CCC CTT CAT TGA CCT CAA CTA C-3′), GAPDH reverse (5′-GAG TCC TTC CAC GAT ACC AAA G-3′); lipoxin A4 receptor forward [43] (5′-GTG ATC TGG GTG GCT GGA TT-3′); and lipoxin A4 receptor reverse (5′-AGG GCC AGG TTC AGG TAA CA-3′). Data are shown as relative mRNA levels normalized to GAPDH.…”

Section: Methodsmentioning

confidence: 99%

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Role of the lipoxin A4 receptor in the development of neutrophil extracellular traps in Leishmania infantum infection

et al. 2019

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Background Neutrophils play an immunomodulatory role through the release of neutrophil extracellular traps (NETs). NETs are released in response to Leishmania infection, but the mechanism of NET extrusion has not been elucidated. The lipoxin A4 receptor on neutrophils is crucial for the inflammatory response and immune regulation of many diseases, including Leishmania infection. Therefore, in the present study, we tried to explore whether Leishmania infantum promastigotes stimulate neutrophil activation and NET release via activating the lipoxin A4 receptor. Results Leishmania infantum promastigotes stimulated neutrophil activity, but blocking of the lipoxin A4 receptor with its antagonist Boc prior to L. infantum stimulation abrogated these effects. Neutrophils showed citrullinated histone H3 expression and simultaneous NET extrusion on L. infantum stimulation, but a decline in both was observed on blocking of the lipoxin A4 receptor. Moreover, differentiated HL-60 cells with lipoxin A4 receptor silencing showed a decrease in citrullinated histone H3 expression as compared to the unsilenced HL-60 samples on stimulation with promastigotes. Conclusions Leishmania infantum promastigotes altered the characteristics of neutrophils and induced NET extrusion by activating the lipoxin A4 receptor. The lipoxin A4 receptor may have potential as a therapeutic target in relation to NET extrusion in the treatment of leishmaniasis, but its mechanisms of action need to be explored in more depth.

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JAGN1 is required for fungal killing in neutrophil extracellular traps: Implications for severe congenital neutropenia

Cited by 28 publications

References 59 publications

Intra- and Extracellular Degradation of Neutrophil Extracellular Traps by Macrophages and Dendritic Cells

Intra- and Extracellular Degradation of Neutrophil Extracellular Traps by Macrophages and Dendritic Cells

Severe congenital neutropenia‐associated JAGN1 mutations unleash a calpain‐dependent cell death programme in myeloid cells

Role of the lipoxin A4 receptor in the development of neutrophil extracellular traps in Leishmania infantum infection

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