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Deregulation of the IL-1β axis in chronic recurrent multifocal osteomyelitis.

Scianaro R, Insalaco A, Bracci Laudiero L, De Vito R, Pezzullo M, Teti A, De Benedetti F, Prencipe G - Pediatr Rheumatol Online J (2014)

Bottom Line: The Interleukin (IL)-1β released in the medium of PBMC cultures after treatment with lipopolysaccharides (LPS) alone or LPS and ATP was measured by ELISA.CASP-1 and IL-1β transcript levels were significantly higher also in PBMCs from CRMO patients in remission compared to healthy controls.PBMCs from CRMO patients in active disease stimulated in vitro with LPS showed a significant increase in IL-1β release compared to healthy control cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Rheumatology Unit, Bambino Gesù Children's Hospital, Rome, Italy.

ABSTRACT

Background: This study aims to investigate the inflammasome response in peripheral blood mononuclear cells (PBMCs) and the expression of inflammasome components in bone biopsies from patients with chronic recurrent multifocal osteomyelitis (CRMO).

Methods: The expression of inflammasome components mRNAs was evaluated in PBMCs isolated from 15 CRMO patients and 13 healthy controls by quantitative real-time PCR. The Interleukin (IL)-1β released in the medium of PBMC cultures after treatment with lipopolysaccharides (LPS) alone or LPS and ATP was measured by ELISA. Immunohistochemical staining for Apoptosis-associated Speck-like protein (ASC), caspase-1 (CASP-1), Nod-like receptor protein-3 (NLRP3) and IL-1β expression was performed in bone biopsies from CRMO patients.

Results: mRNA levels of ASC, CASP-1 and IL-1β were significantly higher in freshly isolated PBMCs from CRMO patients in active disease than in healthy controls. CASP-1 and IL-1β transcript levels were significantly higher also in PBMCs from CRMO patients in remission compared to healthy controls. PBMCs from CRMO patients in active disease stimulated in vitro with LPS showed a significant increase in IL-1β release compared to healthy control cells. Immunohistochemistry staining of bone tissue revealed the expression of inflammasome components in CRMO osteoclasts.

Conclusions: Our data suggest that an abnormal regulation of IL-1β axis may be involved in CRMO pathogenesis.

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Characterization of PBMCs from CRMO patients. (A-E) Real Time-PCR analysis of gene products involved in the regulation of IL-1β, including ASC, NLRP3, CASP-1, IL-1β and of the pro-inflammatory cytokine TNF-α in freshly isolated PBMCs obtained from CRMO patients in remission (gray dots) or with active disease (black dots) and healthy controls (white dots). Values are presented as arbitrary unit (AU). (F-G) IL-1β released in supernatants by PBMCs isolated from CRMO patients in remission or in active disease and from healthy controls, after stimulation with 10 ng/ml LPS for 3 hours (F) or 10 ng/ml LPS for 2 hours followed by treatment with 2 mM ATP for 1 hour (G). IL-1β was measured by ELISA. Black lines represent the mean value. *p < 0.05, **p < 0.01 vs healthy controls.
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Figure 1: Characterization of PBMCs from CRMO patients. (A-E) Real Time-PCR analysis of gene products involved in the regulation of IL-1β, including ASC, NLRP3, CASP-1, IL-1β and of the pro-inflammatory cytokine TNF-α in freshly isolated PBMCs obtained from CRMO patients in remission (gray dots) or with active disease (black dots) and healthy controls (white dots). Values are presented as arbitrary unit (AU). (F-G) IL-1β released in supernatants by PBMCs isolated from CRMO patients in remission or in active disease and from healthy controls, after stimulation with 10 ng/ml LPS for 3 hours (F) or 10 ng/ml LPS for 2 hours followed by treatment with 2 mM ATP for 1 hour (G). IL-1β was measured by ELISA. Black lines represent the mean value. *p < 0.05, **p < 0.01 vs healthy controls.

Mentions: ASC, CASP-1 and IL-1β mRNA levels were significantly higher in PBMCs freshly isolated from CRMO patients during active disease compared to PBMCs from healthy controls. The mRNA expression of CASP-1 and IL-1β was also significantly higher in PBMCs from patients in remission compared to healthy controls (Figure 1A-D).


Deregulation of the IL-1β axis in chronic recurrent multifocal osteomyelitis.

Scianaro R, Insalaco A, Bracci Laudiero L, De Vito R, Pezzullo M, Teti A, De Benedetti F, Prencipe G - Pediatr Rheumatol Online J (2014)

Characterization of PBMCs from CRMO patients. (A-E) Real Time-PCR analysis of gene products involved in the regulation of IL-1β, including ASC, NLRP3, CASP-1, IL-1β and of the pro-inflammatory cytokine TNF-α in freshly isolated PBMCs obtained from CRMO patients in remission (gray dots) or with active disease (black dots) and healthy controls (white dots). Values are presented as arbitrary unit (AU). (F-G) IL-1β released in supernatants by PBMCs isolated from CRMO patients in remission or in active disease and from healthy controls, after stimulation with 10 ng/ml LPS for 3 hours (F) or 10 ng/ml LPS for 2 hours followed by treatment with 2 mM ATP for 1 hour (G). IL-1β was measured by ELISA. Black lines represent the mean value. *p < 0.05, **p < 0.01 vs healthy controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4109750&req=5

Figure 1: Characterization of PBMCs from CRMO patients. (A-E) Real Time-PCR analysis of gene products involved in the regulation of IL-1β, including ASC, NLRP3, CASP-1, IL-1β and of the pro-inflammatory cytokine TNF-α in freshly isolated PBMCs obtained from CRMO patients in remission (gray dots) or with active disease (black dots) and healthy controls (white dots). Values are presented as arbitrary unit (AU). (F-G) IL-1β released in supernatants by PBMCs isolated from CRMO patients in remission or in active disease and from healthy controls, after stimulation with 10 ng/ml LPS for 3 hours (F) or 10 ng/ml LPS for 2 hours followed by treatment with 2 mM ATP for 1 hour (G). IL-1β was measured by ELISA. Black lines represent the mean value. *p < 0.05, **p < 0.01 vs healthy controls.
Mentions: ASC, CASP-1 and IL-1β mRNA levels were significantly higher in PBMCs freshly isolated from CRMO patients during active disease compared to PBMCs from healthy controls. The mRNA expression of CASP-1 and IL-1β was also significantly higher in PBMCs from patients in remission compared to healthy controls (Figure 1A-D).

Bottom Line: The Interleukin (IL)-1β released in the medium of PBMC cultures after treatment with lipopolysaccharides (LPS) alone or LPS and ATP was measured by ELISA.CASP-1 and IL-1β transcript levels were significantly higher also in PBMCs from CRMO patients in remission compared to healthy controls.PBMCs from CRMO patients in active disease stimulated in vitro with LPS showed a significant increase in IL-1β release compared to healthy control cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Rheumatology Unit, Bambino Gesù Children's Hospital, Rome, Italy.

ABSTRACT

Background: This study aims to investigate the inflammasome response in peripheral blood mononuclear cells (PBMCs) and the expression of inflammasome components in bone biopsies from patients with chronic recurrent multifocal osteomyelitis (CRMO).

Methods: The expression of inflammasome components mRNAs was evaluated in PBMCs isolated from 15 CRMO patients and 13 healthy controls by quantitative real-time PCR. The Interleukin (IL)-1β released in the medium of PBMC cultures after treatment with lipopolysaccharides (LPS) alone or LPS and ATP was measured by ELISA. Immunohistochemical staining for Apoptosis-associated Speck-like protein (ASC), caspase-1 (CASP-1), Nod-like receptor protein-3 (NLRP3) and IL-1β expression was performed in bone biopsies from CRMO patients.

Results: mRNA levels of ASC, CASP-1 and IL-1β were significantly higher in freshly isolated PBMCs from CRMO patients in active disease than in healthy controls. CASP-1 and IL-1β transcript levels were significantly higher also in PBMCs from CRMO patients in remission compared to healthy controls. PBMCs from CRMO patients in active disease stimulated in vitro with LPS showed a significant increase in IL-1β release compared to healthy control cells. Immunohistochemistry staining of bone tissue revealed the expression of inflammasome components in CRMO osteoclasts.

Conclusions: Our data suggest that an abnormal regulation of IL-1β axis may be involved in CRMO pathogenesis.

Show MeSH
Related in: MedlinePlus