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We are analyzing https://link.springer.com/article/10.1186/ar4483.

Title:
CARD8 is a negative regulator for NLRP3 inflammasome, but mutant NLRP3 in cryopyrin-associated periodic syndromes escapes the restriction | Arthritis Research & Therapy
Description:
Introduction NLRP3 plays a role in sensing various pathogen components or stresses in the innate immune system. Once activated, NLRP3 associates with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and procaspase-1 to form a large protein complex termed inflammasome. Although some investigators have proposed a model of NLRP3-inflammasome containing an adaptor protein caspase recruitment domain-containing protein 8 (CARD8), the role of this molecule remains obscure. This study aimed to clarify the interaction between CARD8 and wild-type NLRP3 as well as mutant forms of NLRP3 linked with cryopyrin-associated periodic syndromes (CAPS). Methods In here HEK293 expression system, cells were transfected with the cDNAs for inflammasome components. Also used were peripheral blood mononuclear cells (PBMCs) and human monocyte-derived macrophages (HMDMs) from healthy volunteers. The interaction of CARD8 and NLRP3 was studied by immunoprecipitation. The effect of CARD8 expression on IL-1β secretion was assessed by ELISA. CARD8 knockdown experiments were carried out by transfection of the specific siRNA into HMDMs. Results In HEK293 cells, CARD8 interacted with wild-type NLRP3, but not with CAPS-associated mutant NLRP3. CARD8 significantly reduced IL-1β secretion from cells transfected with wild-type NLRP3, but not if they were transfected with mutant NLRP3. In addition, association of endogenously expressed CARD8 with NLRP3 was confirmed in resting PBMCs, and CARD8 knockdown resulted in higher amount of IL-1β secretion from HMDMs. Conclusions Until specific stimuli activate NLRP3, CARD8 holds NLRP3, and is supposed to prevent activation by subtle stimuli. However, CAPS-associated mutant NLRP3 is unable to bind with CARD8, which might be relevant to the pathogenesis of CAPS.
Website Age:
28 years and 1 months (reg. 1997-05-29).

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🌠 Phenomenal Traffic: 5M - 10M visitors per month


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Keywords {🔍}

card, nlrp, cells, article, inflammasome, pubmed, asc, caspase, ilβ, figure, protein, google, scholar, domain, cas, expression, transfected, interaction, hek, secretion, transfection, recruitment, results, hours, human, hmdms, cell, mutant, procaspase, activation, specklike, study, western, atp, domaincontaining, capsassociated, blotting, supernatants, medium, lps, authors, full, proteins, proilβ, usa, blood, elisa, syndrome, endogenous, central,

Topics {✒️}

real-time quantitative rt-pcr fr/issaid/infevers/search atp-induced il-1β secretion cold-induced urticarial rash muckle-wells autoinflammatory disorder muckle-wells syndrome caused article download pdf atp-stimulated il-1β secretion nf-κb-activating ligands yukichi hara & tetsuo kubota national bio-resource project inhibit il-1β secretion reduce il-1β secretion human monocyte-derived macrophages il-1β secretion resulted anti-cleaved il-1β nucleotide binding-oligomerization domain ice-cold lysis buffer isotype-matched control antibody muckle-wells syndrome full size image nlrp3-gfp-speck positive cells caspase-1 processes proil-1β il-1β-processing inflammasome 50 ng/ml gm-csf speck-positive cells divided vsv-tagged card8 interacts flag-tagged truncated nlrp3 modulates asc-induced apoptosis c-terminal card domain calcium-sensing receptor regulates monogenic autoinflammatory diseases il-1β secretion activated-caspase-1 positive cells human card8-specific sirna privacy choices/manage cookies nlrp3-gfp formed speck full length nlrp3 nlrp3-speck positive cells wild type nlrp3-flag related subjects authors’ original file interleukin-1β secretion full length card8 leucine-rich repeats single-step method secreted il-1β full access peripheral blood leukocytes card8-knockdown hmdms secreted

Questions {❓}

  • Verma D, Lerm M, Blomgran Julinder R, Eriksson P, Söderkvist P, Särndahl E: Gene polymorphisms in the NALP3 inflammasome are associated with interleukin-1 production and severe inflammation: relation to common inflammatory diseases?

Schema {🗺️}

WebPage:
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         headline:CARD8 is a negative regulator for NLRP3 inflammasome, but mutant NLRP3 in cryopyrin-associated periodic syndromes escapes the restriction
         description:NLRP3 plays a role in sensing various pathogen components or stresses in the innate immune system. Once activated, NLRP3 associates with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and procaspase-1 to form a large protein complex termed inflammasome. Although some investigators have proposed a model of NLRP3-inflammasome containing an adaptor protein caspase recruitment domain-containing protein 8 (CARD8), the role of this molecule remains obscure. This study aimed to clarify the interaction between CARD8 and wild-type NLRP3 as well as mutant forms of NLRP3 linked with cryopyrin-associated periodic syndromes (CAPS). In here HEK293 expression system, cells were transfected with the cDNAs for inflammasome components. Also used were peripheral blood mononuclear cells (PBMCs) and human monocyte-derived macrophages (HMDMs) from healthy volunteers. The interaction of CARD8 and NLRP3 was studied by immunoprecipitation. The effect of CARD8 expression on IL-1β secretion was assessed by ELISA. CARD8 knockdown experiments were carried out by transfection of the specific siRNA into HMDMs. In HEK293 cells, CARD8 interacted with wild-type NLRP3, but not with CAPS-associated mutant NLRP3. CARD8 significantly reduced IL-1β secretion from cells transfected with wild-type NLRP3, but not if they were transfected with mutant NLRP3. In addition, association of endogenously expressed CARD8 with NLRP3 was confirmed in resting PBMCs, and CARD8 knockdown resulted in higher amount of IL-1β secretion from HMDMs. Until specific stimuli activate NLRP3, CARD8 holds NLRP3, and is supposed to prevent activation by subtle stimuli. However, CAPS-associated mutant NLRP3 is unable to bind with CARD8, which might be relevant to the pathogenesis of CAPS.
         datePublished:2014-02-12T00:00:00Z
         dateModified:2014-02-12T00:00:00Z
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      headline:CARD8 is a negative regulator for NLRP3 inflammasome, but mutant NLRP3 in cryopyrin-associated periodic syndromes escapes the restriction
      description:NLRP3 plays a role in sensing various pathogen components or stresses in the innate immune system. Once activated, NLRP3 associates with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and procaspase-1 to form a large protein complex termed inflammasome. Although some investigators have proposed a model of NLRP3-inflammasome containing an adaptor protein caspase recruitment domain-containing protein 8 (CARD8), the role of this molecule remains obscure. This study aimed to clarify the interaction between CARD8 and wild-type NLRP3 as well as mutant forms of NLRP3 linked with cryopyrin-associated periodic syndromes (CAPS). In here HEK293 expression system, cells were transfected with the cDNAs for inflammasome components. Also used were peripheral blood mononuclear cells (PBMCs) and human monocyte-derived macrophages (HMDMs) from healthy volunteers. The interaction of CARD8 and NLRP3 was studied by immunoprecipitation. The effect of CARD8 expression on IL-1β secretion was assessed by ELISA. CARD8 knockdown experiments were carried out by transfection of the specific siRNA into HMDMs. In HEK293 cells, CARD8 interacted with wild-type NLRP3, but not with CAPS-associated mutant NLRP3. CARD8 significantly reduced IL-1β secretion from cells transfected with wild-type NLRP3, but not if they were transfected with mutant NLRP3. In addition, association of endogenously expressed CARD8 with NLRP3 was confirmed in resting PBMCs, and CARD8 knockdown resulted in higher amount of IL-1β secretion from HMDMs. Until specific stimuli activate NLRP3, CARD8 holds NLRP3, and is supposed to prevent activation by subtle stimuli. However, CAPS-associated mutant NLRP3 is unable to bind with CARD8, which might be relevant to the pathogenesis of CAPS.
      datePublished:2014-02-12T00:00:00Z
      dateModified:2014-02-12T00:00:00Z
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         Rheumatology
         Orthopedics
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               name:Division of Biomedical Laboratory Sciences, Tokyo Medical and Dental University Graduate School of Health Care Sciences, Tokyo, Japan
               type:PostalAddress
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      name:Division of Biomedical Laboratory Sciences, Tokyo Medical and Dental University Graduate School of Health Care Sciences, Tokyo, Japan
      name:Division of Biomedical Laboratory Sciences, Tokyo Medical and Dental University Graduate School of Health Care Sciences, Tokyo, Japan

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