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Increased expression of the NLRP3 inflammasome components in patients with Behçet's disease.

Kim EH, Park MJ, Park S, Lee ES - J Inflamm (Lond) (2015)

Bottom Line: Also, increased expression of NLRP3 and ASC was observed in 25 BD skin lesions compared to 25 erythema nodosum patients.Compatible with this, secretion of IL-1β by PBMCs stimulated with LPS alone or LPS plus ATP was increased in BD compared to healthy controls, which was suppressed by caspase-1 inhibitor.Our findings suggest the possible link between increased IL-1β secretion and increased expression of NLRP3 inflammasome components in BD patients with skin manifestations.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Ajou University School of Medicine, 164 Worldcup-ro, Yeongtong-Gu, Suwon, 443-380 South Korea ; Present address: Department of Dermatology, Cheil General Hospital and Women's Healthcare Center, Dankook University College of Medicine, Cheonan, South Korea.

ABSTRACT

Background: Behçet's disease (BD) is a systemic inflammatory disease with manifestations including recurrent oral and genital ulcerations, and vasculitis involving the skin, mucosa, joints, eyes, veins, arteries, nervous and gastrointestinal systems. BD is seen as a disease at the crossroad between autoimmune and autoinflammatory syndromes, possibly triggered by an aberrant response to infectious stimuli. The relevance of Gram negative bacteria-mediated oral inflammation with the increased expression of NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), leading to systemic inflammation, prompted us to investigate the expression of NLRP3 inflammasome components and its link with IL-1β hypersecretion.

Findings: When peripheral blood mononuclear cells (PBMCs) from 15 active, 15 stable BD patients and 15 healthy volunteers were stimulated, the basal and LPS-induced expressions of NLRP3 inflammasome components were significantly increased at both mRNA and protein levels in BD patients compared to healthy controls. Also, increased expression of NLRP3 and ASC was observed in 25 BD skin lesions compared to 25 erythema nodosum patients. Compatible with this, secretion of IL-1β by PBMCs stimulated with LPS alone or LPS plus ATP was increased in BD compared to healthy controls, which was suppressed by caspase-1 inhibitor.

Conclusion: Our findings suggest the possible link between increased IL-1β secretion and increased expression of NLRP3 inflammasome components in BD patients with skin manifestations.

No MeSH data available.


Related in: MedlinePlus

The induced expression of NLRP3, ASC and caspase-1 is increased in Behçet’s disease (BD). PBMCs were initially stimulated for 4 h with LPS (100 ng/ml). After 4 h, ATP (1 mM) was added to the cells for another 15 min (LPS/ATP). a Representative western blot analysis and quantitation of NLRP3, ASC and caspase-1 from cell lysates of stimulated PBMCs. β-actin was used as loading control. (lane 1,4,7: no treatment, lane 2,5,8: LPS, lane 3,6,9: LPS/ATP) (n = 5 per group). b The mRNA expression of NLRP3, ASC and caspase-1 was measured by real time quantitative RT-PCR and normalized against the expression levels of glyceraldehyde 3-phosphate-dehydrogenase. The relative values are shown as a fold change to HC with no treatment (n = 8 per group). Data are represented as mean ± S.D. (*p < 0.05), (−) no treatment. NLRP3: NACHT, LRR, and PYD domains-containing protein 3; PBMCs: peripheral blood mononuclear cells; LPS: lipopolysaccharide; ATP: adenosine 5-triphosphate; HC: healthy volunteers
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Fig2: The induced expression of NLRP3, ASC and caspase-1 is increased in Behçet’s disease (BD). PBMCs were initially stimulated for 4 h with LPS (100 ng/ml). After 4 h, ATP (1 mM) was added to the cells for another 15 min (LPS/ATP). a Representative western blot analysis and quantitation of NLRP3, ASC and caspase-1 from cell lysates of stimulated PBMCs. β-actin was used as loading control. (lane 1,4,7: no treatment, lane 2,5,8: LPS, lane 3,6,9: LPS/ATP) (n = 5 per group). b The mRNA expression of NLRP3, ASC and caspase-1 was measured by real time quantitative RT-PCR and normalized against the expression levels of glyceraldehyde 3-phosphate-dehydrogenase. The relative values are shown as a fold change to HC with no treatment (n = 8 per group). Data are represented as mean ± S.D. (*p < 0.05), (−) no treatment. NLRP3: NACHT, LRR, and PYD domains-containing protein 3; PBMCs: peripheral blood mononuclear cells; LPS: lipopolysaccharide; ATP: adenosine 5-triphosphate; HC: healthy volunteers

Mentions: Toll-like receptor signaling induces transcription of NLRP3 and IL-1β [6, 7]. NLRP3 inflammasome is activated by canonical stimuli like ATP or Nigericin and noncanonical stimuli like live gram negative bacteria [8]. Therefore, we checked whether LPS alone, or ATP stimulation after LPS priming (LPS/ATP), affected the expression of NLRP3 inflammasome components in PBMCs of BD patients. The protein levels of, NLRP3, ASC and caspase-1 were higher following LPS stimulation compared to no stimulation in all the groups and the levels increased significantly in active and stable BD compared to HC (Fig. 2a). Following LPS/ATP stimulation, NLRP3 and ASC protein levels were significantly up-regulated only in active BD compared to HC (Fig. 2a). The mRNA levels of, NLRP3, ASC and caspase-1 were higher after LPS/ATP stimulation compared to single stimulus of LPS in all the groups. Furthermore, they were significantly increased in the presence of LPS or LPS/ATP in active and stable BD compared to HC (Fig. 2b). These findings show that LPS/ATP stimulation resulted in significantly higher expression of NLRP3 inflammasome component at protein and mRNA levels in PBMCs of BD patients.Fig. 2


Increased expression of the NLRP3 inflammasome components in patients with Behçet's disease.

Kim EH, Park MJ, Park S, Lee ES - J Inflamm (Lond) (2015)

The induced expression of NLRP3, ASC and caspase-1 is increased in Behçet’s disease (BD). PBMCs were initially stimulated for 4 h with LPS (100 ng/ml). After 4 h, ATP (1 mM) was added to the cells for another 15 min (LPS/ATP). a Representative western blot analysis and quantitation of NLRP3, ASC and caspase-1 from cell lysates of stimulated PBMCs. β-actin was used as loading control. (lane 1,4,7: no treatment, lane 2,5,8: LPS, lane 3,6,9: LPS/ATP) (n = 5 per group). b The mRNA expression of NLRP3, ASC and caspase-1 was measured by real time quantitative RT-PCR and normalized against the expression levels of glyceraldehyde 3-phosphate-dehydrogenase. The relative values are shown as a fold change to HC with no treatment (n = 8 per group). Data are represented as mean ± S.D. (*p < 0.05), (−) no treatment. NLRP3: NACHT, LRR, and PYD domains-containing protein 3; PBMCs: peripheral blood mononuclear cells; LPS: lipopolysaccharide; ATP: adenosine 5-triphosphate; HC: healthy volunteers
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4487834&req=5

Fig2: The induced expression of NLRP3, ASC and caspase-1 is increased in Behçet’s disease (BD). PBMCs were initially stimulated for 4 h with LPS (100 ng/ml). After 4 h, ATP (1 mM) was added to the cells for another 15 min (LPS/ATP). a Representative western blot analysis and quantitation of NLRP3, ASC and caspase-1 from cell lysates of stimulated PBMCs. β-actin was used as loading control. (lane 1,4,7: no treatment, lane 2,5,8: LPS, lane 3,6,9: LPS/ATP) (n = 5 per group). b The mRNA expression of NLRP3, ASC and caspase-1 was measured by real time quantitative RT-PCR and normalized against the expression levels of glyceraldehyde 3-phosphate-dehydrogenase. The relative values are shown as a fold change to HC with no treatment (n = 8 per group). Data are represented as mean ± S.D. (*p < 0.05), (−) no treatment. NLRP3: NACHT, LRR, and PYD domains-containing protein 3; PBMCs: peripheral blood mononuclear cells; LPS: lipopolysaccharide; ATP: adenosine 5-triphosphate; HC: healthy volunteers
Mentions: Toll-like receptor signaling induces transcription of NLRP3 and IL-1β [6, 7]. NLRP3 inflammasome is activated by canonical stimuli like ATP or Nigericin and noncanonical stimuli like live gram negative bacteria [8]. Therefore, we checked whether LPS alone, or ATP stimulation after LPS priming (LPS/ATP), affected the expression of NLRP3 inflammasome components in PBMCs of BD patients. The protein levels of, NLRP3, ASC and caspase-1 were higher following LPS stimulation compared to no stimulation in all the groups and the levels increased significantly in active and stable BD compared to HC (Fig. 2a). Following LPS/ATP stimulation, NLRP3 and ASC protein levels were significantly up-regulated only in active BD compared to HC (Fig. 2a). The mRNA levels of, NLRP3, ASC and caspase-1 were higher after LPS/ATP stimulation compared to single stimulus of LPS in all the groups. Furthermore, they were significantly increased in the presence of LPS or LPS/ATP in active and stable BD compared to HC (Fig. 2b). These findings show that LPS/ATP stimulation resulted in significantly higher expression of NLRP3 inflammasome component at protein and mRNA levels in PBMCs of BD patients.Fig. 2

Bottom Line: Also, increased expression of NLRP3 and ASC was observed in 25 BD skin lesions compared to 25 erythema nodosum patients.Compatible with this, secretion of IL-1β by PBMCs stimulated with LPS alone or LPS plus ATP was increased in BD compared to healthy controls, which was suppressed by caspase-1 inhibitor.Our findings suggest the possible link between increased IL-1β secretion and increased expression of NLRP3 inflammasome components in BD patients with skin manifestations.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Ajou University School of Medicine, 164 Worldcup-ro, Yeongtong-Gu, Suwon, 443-380 South Korea ; Present address: Department of Dermatology, Cheil General Hospital and Women's Healthcare Center, Dankook University College of Medicine, Cheonan, South Korea.

ABSTRACT

Background: Behçet's disease (BD) is a systemic inflammatory disease with manifestations including recurrent oral and genital ulcerations, and vasculitis involving the skin, mucosa, joints, eyes, veins, arteries, nervous and gastrointestinal systems. BD is seen as a disease at the crossroad between autoimmune and autoinflammatory syndromes, possibly triggered by an aberrant response to infectious stimuli. The relevance of Gram negative bacteria-mediated oral inflammation with the increased expression of NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), leading to systemic inflammation, prompted us to investigate the expression of NLRP3 inflammasome components and its link with IL-1β hypersecretion.

Findings: When peripheral blood mononuclear cells (PBMCs) from 15 active, 15 stable BD patients and 15 healthy volunteers were stimulated, the basal and LPS-induced expressions of NLRP3 inflammasome components were significantly increased at both mRNA and protein levels in BD patients compared to healthy controls. Also, increased expression of NLRP3 and ASC was observed in 25 BD skin lesions compared to 25 erythema nodosum patients. Compatible with this, secretion of IL-1β by PBMCs stimulated with LPS alone or LPS plus ATP was increased in BD compared to healthy controls, which was suppressed by caspase-1 inhibitor.

Conclusion: Our findings suggest the possible link between increased IL-1β secretion and increased expression of NLRP3 inflammasome components in BD patients with skin manifestations.

No MeSH data available.


Related in: MedlinePlus