All Phenotypes Abraxas2<tm1Nju> MGI Mouse

immune system phenotype ( J:277205 )

N	• isolated bone marrow-derived macrophages (BMDMs) exhibit normal proliferation, differentiation, and LPS-induced secretion of IL-6 and TNF-alpha in condition medium relative to wild-type BMDMs

abnormal macrophage physiology ( J:277205 , J:363478 )

• LPS-primed BMDMs show impaired pro-caspase-1 and pro-IL-1beta processing in response to ATP, nigericin, and MSU stimulation, consistent with defective activation of the NLRP3 inflammasome (J:277205)

• LPS-primed BMDMs show a significant reduction in active caspase-1 levels, PYCARD/ASC speck formation, and PYCARD/ASC oligomerization in response to nigericin stimulation, and a weaker interaction between NLRP3 and PYCARD/ASC in response to ATP treatment, indicating impaired NLRP3-PYCARD/ASC complex formation (J:277205)

• LPS-primed BMDMs show no obvious deubiquitination of NLRP3 in response to stimulation with ATP or nigericin, unlike similarly treated wild-type BMDMs; notably, the poly-ubiquitination level of the endogenous NLRP3 protein is normal after LPS priming (J:277205)

• however, LPS-primed BMDMs show normal IL-1beta and caspase-1 maturation in response to flagellin and poly(dA:dT) treatment, indicating normal NLRC4 and AIM2 inflammasome activation (J:277205)

• moreover, LPS-primed BMDMs show normal PYCARD/ASC speck formation after poly(dA:dT) transfection, normal K+ efflux, Ca2+ flux, and membrane potential after ATP treatment, and normal mitochondrial ROS after nigericin treatment (J:277205)

• both untreated and MG132-treated BMDMs show increased ubiquitination of endogenous BRCC3 (BRCA1/BRCA2-containing complex, subunit 3) (J:363478)

decreased circulating interleukin-1 beta level ( J:277205 )

• in an LPS-induced sepsis model, mice i.p. injected with LPS (15 mg/kg) show a significant reduction in serum IL-1beta levels at 3 h post-injection

decreased interleukin-1 beta secretion ( J:277205 )

• in vitro, LPS-primed BMDMs produce significantly less IL-1beta in response to ATP stimulation; re-expression of exogenous ABRAXAS2/ABRO1 protein restores IL-1beta secretion to a level seen in wild-type BMDMs in response to inflammasome activation

• LPS-primed BMDMs produce significantly less IL-1beta in response to other NLRP3 inflammasome stimuli, including nigericin, silica, monosodium urate crystals (MSU), muramyl dipeptide (MDP), and aluminum (Alum), indicating defective activation of the NLRP3 inflammasome

• after LPS + ATP or LPS + nigericin treatment, BMDMs show a ~70% decrease in IL-1beta release relative to wild-type BMDMs

• in vivo, mice i.p injected with MSU show a ~60% decrease in IL-1beta secretion in the peritoneal lavage fluid, whereas inflammasome-independent TNF-alpha secretion is normal

• similarly, mice i.p. injected with Alum show a significantly lower production of IL-1beta in the lavage fluid

• however, LPS-primed BMDMs show normal IL-1beta secretion in response to anthrax lethal toxin, Salmonella flagellin, and poly(dA:dT), indicating normal activation of the respective NLRP1, NLRC4, and AIM2 inflammasomes

decreased interleukin-18 secretion ( J:277205 )

• LPS-primed BMDMs generate significantly less IL-18 in response to ATP stimulation

decreased inflammatory response ( J:277205 )

• in NLRP3-dependent peritonitis models, mice i.p. injected with MSU or Alum show a significantly lower IL-1beta secretion in the lavage fluid and impaired recruitment of neutrophils and inflammatory monocytes to the peritoneal cavity relative to wild-type controls

decreased susceptibility to endotoxin shock ( J:277205 )

• in an LPS-induced sepsis model, mice i.p. injected with LPS (15 mg/kg) show significantly lower serum IL-1beta levels and markedly increased survival, with ~90% surviving to 9 days whereas all wild-type controls die by 6 days after LPS injection

decreased protein level ( J:363478 )

• untreated bone marrow cells and BMDMs show a severe reduction in the protein levels of BRCC3 (BRCA1/BRCA2-containing complex, subunit 3), with no change in Brcc3 mRNA levels seen in BMDMs relative to wild-type cells

• BMDMs treated with cycloheximide (an inhibitor of protein synthesis) show a severe reduction in the half-life of BRCC3 protein, indicating that ABRAXAS2/ABRO1 increases the protein stability of BRCC3

• BMDMs treated with MG132 (a proteasome inhibitor) show a time-dependent rescue of BRCC3 protein levels, whereas BMDMs treated with E-64 (a lysosome inhibitor) do not, suggesting that BRCC3 undergoes proteasome-mediated degradation