Role of Vesicle-Associated Membrane Protein-2, Through Q-Soluble<i>N</i>-Ethylmaleimide-Sensitive Factor Attachment Protein Receptor/R-Soluble<i>N</i>-Ethylmaleimide-Sensitive Factor Attachment Protein Receptor Interaction, in the Exocytosis of Specific and Tertiary Granules of Human Neutrophils

Mollinedo, Faustino; Martı́n-Martı́n, Belén; Calafat, Jero; Nabokina, Svetlana M.; Lazo, Pedro A.

doi:10.4049/jimmunol.170.2.1034

Cited by 73 publications

(88 citation statements)

References 57 publications

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“…S100A8 (red) was also present in the nuclei of CD11b ϩ cells (green), which were likely macrophages (E), and both cellular populations decreased during convalescence (F). sequestration and production of antibacterial peroxide species during the inflammatory response (36,37). Myeloperoxidase (MPO) and nitric oxide synthase 2 (NOS2) also limit bacterial growth by production of hypochlorous acid and NO, respectively (38)(39)(40).…”

Section: Discussionmentioning

confidence: 99%

Comparative Proteomic Analysis Reveals Activation of Mucosal Innate Immune Signaling Pathways during Cholera

et al. 2015

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i Vibrio cholerae O1 is a major cause of acute watery diarrhea in over 50 countries. Evidence suggests that V. cholerae O1 may activate inflammatory pathways, and a recent study of a Bangladeshi population showed that variants in innate immune genes play a role in mediating susceptibility to cholera. We analyzed human proteins present in the small intestine of patients infected with V. cholerae O1 to characterize the host response to this pathogen. We collected duodenal biopsy specimens from patients with acute cholera after stabilization and again 30 days after initial presentation. Peptides extracted from biopsy specimens were sequenced and quantified using label-free mass spectrometry and SEQUEST. Twenty-seven host proteins were differentially abundant between the acute and convalescent stages of infection; the majority of these have known roles in innate defense, cytokine production, and apoptosis. Immunostaining confirmed that two proteins, WARS and S100A8, were more abundant in lamina propria cells during the acute stage of cholera. Analysis of the differentially abundant proteins revealed the activation of key regulators of inflammation by the innate immune system, including Toll-like receptor 4, nuclear factor kappa-light-chain-enhancer of activated B cells, mitogen-activated protein kinases, and caspase-dependent inflammasomes. Interleukin-12␤ (IL-12␤) was a regulator of several proteins that were activated during cholera, and we confirmed that IL-12␤ was produced by lymphocytes recovered from duodenal biopsy specimens of cholera patients. Our study shows that a broad inflammatory response is generated in the gut early after onset of cholera, which may be critical in the development of long-term mucosal immunity against V. cholerae O1. Vibrio cholerae O1, which causes 3 to 5 million cases of cholera annually, is a noninvasive pathogen that does not penetrate the intestinal mucosa of human hosts (1, 2). Cholera is considered a prototypical, noninflammatory diarrheal illness, and there are no gross changes in the integrity of mucosal tissue during V. cholerae O1 infection (2, 3). However, there is mounting evidence that V. cholerae O1 triggers inflammatory responses in the gut (2, 4, 5). Studies of duodenal tissue extracted from cholera patients indicate that V. cholerae O1 infection causes upregulation of innate host defenses in the intestinal mucosa, including neutrophil-derived antibacterial proteins, proinflammatory cytokines, and BPIFB1 (also known as LPLUNC1), which is an immune-modifying defense protein that may attenuate the host response to bacterial lipopolysaccharide (LPS) (4-6). A candidate gene study showed that a variant in the promoter region of LPLUNC1 is associated with cholera susceptibility in a population in Bangladesh, an area where cholera is an ancient disease (7).Genome-wide association studies also underscore the importance of the innate immune system in influencing the clinical manifestations of V. cholerae O1 infection (8). A recent analysis of signals of natural selection in in...

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Section: Discussionmentioning

confidence: 99%

Comparative Proteomic Analysis Reveals Activation of Mucosal Innate Immune Signaling Pathways during Cholera

et al. 2015

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“…Jurkat cells (2 Â 10 7 ) were lysed with 200 ml of lysis buffer (30 mM Tris-HCl, pH 7.5, 1% Triton X-100, 10% glycerol, 150 mM NaCl, 2 mM Na 3 VO 4 , 2 mM PMSF) and lysates were precleared for 4 h at 4 1C with 300 ml of 20% protein A-Sepharose, and immunoprecipitated with anti-JNK antibody precoupled to protein A-Sepharose as previously described (Mollinedo et al, 2003). Samples were subjected to SDS-PAGE, and proteins were transferred to nitrocellulose and immunoblotted.…”

Section: Co-immunoprecipitationmentioning

confidence: 99%

Proapoptotic role of Hsp90 by its interaction with c-Jun N-terminal kinase in lipid rafts in edelfosine-mediated antileukemic therapy

et al. 2007

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Heat shock protein 90 (Hsp90) is a survival signaling chaperone and a cancer chemotherapeutic target. However, we have found that inhibitors of Hsp90 diminished the apoptotic response induced in leukemic cells by the antitumor alkyl-lysophospholipid analog edelfosine, which acts through lipid raft reorganization. Edelfosine treatment recruited Hsp90, c-Jun N-terminal kinase (JNK) and apoptotic molecules in lipid rafts, but not the JNK regulators apoptosis signal-regulating kinase 1 (ASK1) and Daxx, or the survival signaling molecules extracellular signal-regulated kinase (ERK) and Akt. Following edelfosine treatment, Hsp90 bound to JNK in lipid rafts and Hsp90-JNK clusters were identified at the plasma membrane by immunoelectron microscopy. Hsp90 inhibition reduced JNK protein level in lipid rafts and turned proapoptotic persistent JNK activation into a transient response in edelfosine-treated cells. Decrease in edelfosine-induced JNK activation and apoptosis by Hsp90 inhibition was prevented through proteasome inhibition, suggesting that Hsp90 inhibition diminishes apoptosis by promoting JNK protein degradation. Expression of ASK1 dominant negative mutant did not affect JNK activation and apoptosis following edelfosine treatment. These data indicate that lipid raft-recruited JNK is ASK1-independent and becomes a novel Hsp90 client protein. Our results reveal a new chaperoning role of Hsp90 on JNKmediated apoptosis following its recruitment in lipid rafts.

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“…Immunoelectron Microscopy-Resting human neutrophils incubated with L. major or L. donovani for 3 h were fixed and processed for ultrathin cryosectioning as previously described (14,23). For immunolabeling, ultrathin cryosections were incubated with anti-lactoferrin (Cappel Laboratories, West Chester, PA), anti-myeloperoxidase (DAKO, Glostrup, Denmark), or anti-gelatinase (24) (provided by N. Borregaard, The National University Hospital, Copenhagen, Denmark) rabbit antibodies, followed with 10-nm protein A-conjugated colloidal gold (EM Laboratories, Utrecht University, The Netherlands).…”

Section: Methodsmentioning

confidence: 99%

“…CD11b and CD66b are present in the membranes of both specific and tertiary granules (14,18,26), whereas CD63 is present in the membranes of azurophilic granules (27) in resting human neutrophils. All these antigens are incorporated into the cell surface facing the extracellular medium following exocytosis of the above neutrophil granules (13,14,18). Leishmania interaction with neutrophils did not induce statistically significant CD11b, CD66b, or CD63 cell surface up-regulation (Fig.…”

Section: Major Parasite Ingestion Does Not Mobilize Specific or Tementioning

confidence: 99%

Selective Fusion of Azurophilic Granules with Leishmania-containing Phagosomes in Human Neutrophils

Mollinedo

Janssen

Iglesia-Vicente

et al. 2010

Journal of Biological Chemistry

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Leishmania parasites use polymorphonuclear neutrophils as intermediate hosts before their ultimate delivery to macrophages following engulfment of parasite-infected neutrophils. This leads to a silent and unrecognized entry of Leishmania into the macrophage host cell. Neutrophil function depends on its cytoplasmic granules, but their mobilization and role in how Leishmania parasites evade intracellular killing in neutrophils remain undetermined. Here, we have found by ultrastructural approaches that neutrophils ingested Leishmania major promastigotes, and azurophilic granules fused in a preferential way with parasite-containing phagosomes, without promoting parasite killing. Azurophilic granules, identified by the granule marker myeloperoxidase, also fused with Leishmania donovaniengulfed vacuoles in human neutrophils. In addition, the azurophilic membrane marker CD63 was also detected in the vacuole surrounding the parasite, and in the fusion of azurophilic granules with the parasite-engulfed phagosome. Tertiary and specific granules, involved in vacuole acidification and superoxide anion generation, hardly fused with Leishmania-containing phagosomes. L. major interaction with neutrophils did not elicit production of reactive oxygen species or mobilization of tertiary and specific granules. By using immunogold electron microscopy approaches in the engulfment of L. major and L. donovani by human neutrophils, we did not find a significant contribution of endoplasmic reticulum to the formation of Leishmania-containing vacuoles. Live Leishmania parasites were required to be optimally internalized by neutrophils. Our data suggest that Leishmania promastigotes modulate their uptake by neutrophils, and regulate granule fusion processes in a rather selective way to favor parasite survival in human neutrophils.

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Role of Vesicle-Associated Membrane Protein-2, Through Q-SolubleN-Ethylmaleimide-Sensitive Factor Attachment Protein Receptor/R-SolubleN-Ethylmaleimide-Sensitive Factor Attachment Protein Receptor Interaction, in the Exocytosis of Specific and Tertiary Granules of Human Neutrophils

Cited by 73 publications

References 57 publications

Comparative Proteomic Analysis Reveals Activation of Mucosal Innate Immune Signaling Pathways during Cholera

Comparative Proteomic Analysis Reveals Activation of Mucosal Innate Immune Signaling Pathways during Cholera

Proapoptotic role of Hsp90 by its interaction with c-Jun N-terminal kinase in lipid rafts in edelfosine-mediated antileukemic therapy

Selective Fusion of Azurophilic Granules with Leishmania-containing Phagosomes in Human Neutrophils

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