IgE-dependent sensitization increases responsiveness to LPS but does not modify development of endotoxin tolerance in mast cells

Medina-Tamayo, Jaciel; Ibarra-Sánchez, Alfredo; Padilla-Trejo, Alejandro; González-Espinosa, Claudia

doi:10.1007/s00011-010-0230-4

Cited by 18 publications

(20 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Morphine effects did not involve TLR4 receptor internalization, which has been suggested as a desensitization event able to prevent LPS actions (34). Although some studies have shown that TNF is synthesized de novo after different stimuli in BMMCs (39), we have reported that IgE-dependent sensitization of BMMCs promotes the generation of a pool of TNF that is rapidly mobilized after TLR4 receptor stimulation (31). Accordingly, early LPS-induced TNF secretion in sensitized BMMCs was not sensitive to actinomycin D or brefeldin A pretreatments, but it was sensitive to morphine addition.…”

Section: Ige-sensitized Bmmcs As a Cellular Model To Study Opioid/ Tlmentioning

confidence: 73%

“…Age-matched cultures (5-8 wk old) were used in each experiment. Routinely, BMMCs were sensitized with 100 ng/ml monoclonal anti-DNP IgE (clone SPE-7 from Sigma-Aldrich) for 18 h at 37˚C, as this treatment leads to the production of a prestored, LPS-sensitive pool of TNF (31). Cells were then washed with fresh culture medium or Tyrode's BSA buffer (135 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 1.8 mM CaCl 2 , 5.6 mM glucose, 0.5 g/l BSA, and 20 mM HEPES [pH 7.4]) and were used for experiments.…”

Section: Bmmc Culturementioning

confidence: 99%

See 1 more Smart Citation

Morphine Prevents Lipopolysaccharide-Induced TNF Secretion in Mast Cells Blocking IκB Kinase Activation and SNAP-23 Phosphorylation: Correlation with the Formation of a β-Arrestin/TRAF6 Complex

Madera-Salcedo

Cruz

González-Espinosa

2013

The Journal of Immunology

View full text Add to dashboard Cite

We have previously shown that morphine pretreatment inhibits mast cell–dependent TNF production after LPS injection in the murine peritoneal cavity. In this study, we used bone marrow–derived mast cells (BMMCs) to investigate the molecular mechanisms of that inhibition. We found that morphine prevented LPS-induced TNF secretion in these cells. The observed inhibition was not due to morphine-induced TLR4 internalization and it was related to the blockage of preformed TNF secretion. LPS-induced TNF exocytosis in BMMCs was dependent on tetanus toxin–insensitive vesicle-associated membrane proteins and calcium mobilization, as well as PI3K, MAPK, and IκB kinase (IKK) activation. TNF secretion was also associated to the phosphorylation of synaptosomal-associated protein 23 (SNAP-23), which was found forming a complex with IKK in LPS-activated BMMCs. Morphine pretreatment prevented TLR4-dependent ERK and IKK phosphorylation. Analyzing the signaling events upstream of IKK activation, we found diminished TGF-β–activated kinase 1 (TAK1) phosphorylation and TNFR-associated factor (TRAF) 6 ubiquitination in BMMCs pretreated with morphine and stimulated with LPS. Morphine pretreatment provoked a marked increase in the formation of a molecular complex composed of TRAF6 and β-arrestin-2. Naloxone and a combination of μ and δ opioid receptor antagonists prevented morphine inhibitory actions. In conclusion, our results show that activation of μ and δ opioid receptors with morphine suppresses TLR4-induced TNF release in mast cells, preventing the IKK-dependent phosphorylation of SNAP-23, which is necessary for TNF exocytosis, and this inhibition correlates with the formation of a β-arrestin-2/TRAF6 complex. To our knowledge, these findings constitute the first evidence of molecular crosstalk between opioid receptors and the TLR4 signal transduction system in mast cells.

show abstract

Section: Ige-sensitized Bmmcs As a Cellular Model To Study Opioid/ Tlmentioning

confidence: 73%

Section: Bmmc Culturementioning

confidence: 99%

Morphine Prevents Lipopolysaccharide-Induced TNF Secretion in Mast Cells Blocking IκB Kinase Activation and SNAP-23 Phosphorylation: Correlation with the Formation of a β-Arrestin/TRAF6 Complex

Madera-Salcedo

Cruz

González-Espinosa

2013

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…Only cultures 98% positive for FcεRI receptor expression and showing a dose response of b-hexosaminidase release after IgE/Ag addition were used in the study. For each experiment, BMMCs were sensitized with 100 ng/ml a monoclonal anti-DNP IgE (clone SPE-7; SigmaAldrich) for 24 h at 37˚C, because this condition was shown to increase BMMCs' responsiveness to LPS (31,33). Routinely, after sensitization, cells were collected and resuspended in fresh culture medium or Tyrode's BSA buffer (135 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 1.8 mM CaCl 2 , 5.6 mM glucose, 0.5 g/l BSA, and 20 mM HEPES [pH 7.4]) for Western blot or immunoprecipitation experiments.…”

Section: Cellsmentioning

confidence: 99%

“…Two million IgE-sensitized WT and Fyn 2/2 BMMCs were collected and used for total RNA extraction and RT-PCR, as described (33). Briefly, total RNA was purified using TRI Reagent (Sigma-Aldrich) and suspended in 15 ml RNA secure solution (Ambion, Foster City, CA).…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

“…TNF primers were 59-TTCTG-TCTACTGAACTTCGGGGTGATCGGTCC-39 (sense) and 59-GTATGAGA-TAGCAAATCGGCTGACGGTGTGGG-39 (antisense), and GAPDH primers were (sense) 59-TGAAGGTCGGTGTGAACGGATTTGGC-39 and (antisense) 59-CATGTAGGCCATGAGGTCCACCAC-39 (32). Conditions used for amplification were reported previously (32,33), and the products were separated on 2% agarose gels with ethidium bromide dye.…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

See 1 more Smart Citation

Protein Tyrosine Kinase Fyn Regulates TLR4-Elicited Responses on Mast Cells Controlling the Function of a PP2A-PKCα/β Signaling Node Leading to TNF Secretion

Martín-Ávila

Medina-Tamayo

Ibarra-Sánchez

et al. 2016

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

Mast cells produce proinflammatory cytokines in response to TLR4 ligands, but the signaling pathways involved are not fully described. In this study, the participation of the Src family kinase Fyn in the production of TNF after stimulation with LPS was evaluated using bone marrow–derived mast cells from wild-type and Fyn-deficient mice. Fyn−/− cells showed higher LPS-induced secretion of preformed and de novo–synthesized TNF. In both cell types, TNF colocalized with vesicle-associated membrane protein (VAMP)3-positive compartments. Addition of LPS provoked coalescence of VAMP3 and its interaction with synaptosomal-associated protein 23; those events were increased in the absence of Fyn. Higher TNF mRNA levels were also observed in Fyn-deficient cells as a result of increased transcription and greater mRNA stability after LPS treatment. Fyn−/− cells also showed higher LPS-induced activation of TAK-1 and ERK1/2, whereas IκB kinase and IκB were phosphorylated, even in basal conditions. Increased responsiveness in Fyn−/− cells was associated with a lower activity of protein phosphatase 2A (PP2A) and augmented activity of protein kinase C (PKC)α/β, which was dissociated from PP2A and increased its association with the adapter protein neuroblast differentiation–associated protein (AHNAK, desmoyokin). LPS-induced PKCα/β activity was associated with VAMP3 coalescence in WT and Fyn-deficient cells. Reconstitution of MC-deficient Wsh mice with Fyn−/− MCs produced greater LPS-dependent production of TNF in the peritoneal cavity. Our data show that Fyn kinase is activated after TLR4 triggering and exerts an important negative control on LPS-dependent TNF production in MCs controlling the inactivation of PP2Ac and activation of PKCα/β necessary for the secretion of TNF by VAMP3+ carriers.

show abstract

Signaling through Toll‐like receptor 4 and mast cell‐dependent innate immunity responses

Avila

González-Espinosa

2011

IUBMB Life

View full text Add to dashboard Cite

SummarySignal transduction through Toll-like receptors (TLRs) has been one of the main topics in immunology research in recent years. Because of their signaling particularities based on the homotypic recognition of protein domains in multiple adaptors and selective activation of protein kinases, TLRs have become a paradigm to study ligand recognition coupled to dynamic and highly specific transcriptional and secretory responses in immune cells. Particularly, deleterious effects of Gram-negative bacteria-associated immune reactions has promoted intense research in the field, leading to the description of a number of canonical molecules connecting lipopolysaccharide-induced TLR4 activation with NFjB-dependent transcription. However, the diversity of immune cell phenotypes and the activity of distinct immune receptors in the same cell, strongly suggest that a number of elements in TLR4 signaling cascade, such as novel coreceptors, tyrosine kinases, and molecules regulating the secretion of preformed mediators remain to be described. Recent investigations have placed the mast cells, widely known by their role on allergic responses, as important effectors of innate immunity reactions against Gram-negative bacteria. Their remarkable capacity of cytokine storage, synthesis and release, and the large number of inflammatory reactions controlled by their activation, suggest the existence of new modulators of TLR4 signaling in this particular cell type.

show abstract

IgE-dependent sensitization increases responsiveness to LPS but does not modify development of endotoxin tolerance in mast cells

Abstract: IgE-induced sensitization primes mast cells for higher response to LPS through pre-activation of NFκB transcription factor. IgE-dependent sensitization does not modify events leading to endotoxin tolerance.

Cited by 18 publications

References 37 publications

Morphine Prevents Lipopolysaccharide-Induced TNF Secretion in Mast Cells Blocking IκB Kinase Activation and SNAP-23 Phosphorylation: Correlation with the Formation of a β-Arrestin/TRAF6 Complex

Morphine Prevents Lipopolysaccharide-Induced TNF Secretion in Mast Cells Blocking IκB Kinase Activation and SNAP-23 Phosphorylation: Correlation with the Formation of a β-Arrestin/TRAF6 Complex

Protein Tyrosine Kinase Fyn Regulates TLR4-Elicited Responses on Mast Cells Controlling the Function of a PP2A-PKCα/β Signaling Node Leading to TNF Secretion

Signaling through Toll‐like receptor 4 and mast cell‐dependent innate immunity responses

Contact Info

Product

Resources

About