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Immunoregulatory protein profiles of necrotizing enterocolitis versus spontaneous intestinal perforation in preterm infants.

Chan KY, Leung FW, Lam HS, Tam YH, To KF, Cheung HM, Leung KT, Poon TC, Lee KH, Li K, Fok TF, Ng PC - PLoS ONE (2012)

Bottom Line: In FHs-74 Int cells, Ang-2, IL1-RII and uPAR mRNA expressions were significantly induced by the combined treatment with lipopolysaccharide (LPS) and platelet activating factor (PAF).The magnitude of changes in these proteins was significantly more extensive in NEC infants, reflecting the different nature of injury and/or severity of inflammation.Such exaggerated immunologic responses may account for the high morbidity and mortality in NEC compared with SIP patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.

ABSTRACT
Necrotizing enterocolitis (NEC) and spontaneous intestinal perforation (SIP) are the most common acute surgical emergencies associated with high morbidity and mortality in preterm infants. We aimed to compare the profiles of immunoregulatory proteins and identify novel mediators in plasma of NEC and SIP infants. We also investigated the expression of target genes in resected intestinal tissues and an enterocyte cell line. Using Cytokine Antibody Array assay, we reported the first comparative profiles of immunoregulatory proteins in plasma of NEC and SIP infants, and showed that dysregulated proteins belonged to functionally diversified categories, including pro- and anti-inflammation, angiogenesis, cell growth, wound healing, anti-apoptosis, cell adhesion and extracellular matrix reorganization. Validation by ELISA confirmed significantly higher concentrations of interleukin (IL)-6, angiopoietin (Ang)-2, soluble type II interleukin-1 receptor (sIL-1RII), and soluble urokinase-type plasminogen activator receptor (suPAR) in NEC infants compared with gestational age-matched control, and a lower level of an epidermal growth factor receptor, secreted form of receptor tyrosine-protein kinase ErbB3 (sErbB3), compared with SIP infants. mRNA expressions of IL1-RII and uPAR were up-regulated in resected bowel tissues from NEC infants, indicating that immunoregulation also occurred at the cellular level. In FHs-74 Int cells, Ang-2, IL1-RII and uPAR mRNA expressions were significantly induced by the combined treatment with lipopolysaccharide (LPS) and platelet activating factor (PAF). Our study provided plasmatic signatures of immunoregulatory proteins in NEC and SIP infants, and demonstrated involvement of multiple functional pathways. The magnitude of changes in these proteins was significantly more extensive in NEC infants, reflecting the different nature of injury and/or severity of inflammation. We speculate that dysregulation of IL-6, Ang-2, IL-1RII and uPAR occurred at both systemic and cellular levels, and probably mediated via LPS and endogeneous PAF signals. Such exaggerated immunologic responses may account for the high morbidity and mortality in NEC compared with SIP patients.

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mRNA expression levels of target genes in FHs-74 Int cell line in response to LPS and PAF.FHs-74 Int cells were cultured in the presence of LPS (100 ng/mL) or PAF (25 mM) for 6 h individually or in combination. qPCR analysis of target mRNA expressions showed that combined treatment with LPS and PAF significantly increased levels of Ang-2, IL1-RII and uPAR (n = 4; P<0.05), whereas single stimulant, LPS or PAF, did not alter the expression levels. Results are presented as mean and SEM of expression levels relative to β-actin.
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pone-0036977-g004: mRNA expression levels of target genes in FHs-74 Int cell line in response to LPS and PAF.FHs-74 Int cells were cultured in the presence of LPS (100 ng/mL) or PAF (25 mM) for 6 h individually or in combination. qPCR analysis of target mRNA expressions showed that combined treatment with LPS and PAF significantly increased levels of Ang-2, IL1-RII and uPAR (n = 4; P<0.05), whereas single stimulant, LPS or PAF, did not alter the expression levels. Results are presented as mean and SEM of expression levels relative to β-actin.

Mentions: Combined treatments with LPS and PAF significantly increased expression levels of Ang-2, IL1-RII and uPAR in FHs-74 Int cells upon culture at a reduced serum concentration (P<0.05; Figure 4B, C and D). Treatment with single stimulant, either LPS or PAF, did not significantly alter the expression levels, though a non-significant trend of increase was observed in the PAF cultures.


Immunoregulatory protein profiles of necrotizing enterocolitis versus spontaneous intestinal perforation in preterm infants.

Chan KY, Leung FW, Lam HS, Tam YH, To KF, Cheung HM, Leung KT, Poon TC, Lee KH, Li K, Fok TF, Ng PC - PLoS ONE (2012)

mRNA expression levels of target genes in FHs-74 Int cell line in response to LPS and PAF.FHs-74 Int cells were cultured in the presence of LPS (100 ng/mL) or PAF (25 mM) for 6 h individually or in combination. qPCR analysis of target mRNA expressions showed that combined treatment with LPS and PAF significantly increased levels of Ang-2, IL1-RII and uPAR (n = 4; P<0.05), whereas single stimulant, LPS or PAF, did not alter the expression levels. Results are presented as mean and SEM of expression levels relative to β-actin.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351425&req=5

pone-0036977-g004: mRNA expression levels of target genes in FHs-74 Int cell line in response to LPS and PAF.FHs-74 Int cells were cultured in the presence of LPS (100 ng/mL) or PAF (25 mM) for 6 h individually or in combination. qPCR analysis of target mRNA expressions showed that combined treatment with LPS and PAF significantly increased levels of Ang-2, IL1-RII and uPAR (n = 4; P<0.05), whereas single stimulant, LPS or PAF, did not alter the expression levels. Results are presented as mean and SEM of expression levels relative to β-actin.
Mentions: Combined treatments with LPS and PAF significantly increased expression levels of Ang-2, IL1-RII and uPAR in FHs-74 Int cells upon culture at a reduced serum concentration (P<0.05; Figure 4B, C and D). Treatment with single stimulant, either LPS or PAF, did not significantly alter the expression levels, though a non-significant trend of increase was observed in the PAF cultures.

Bottom Line: In FHs-74 Int cells, Ang-2, IL1-RII and uPAR mRNA expressions were significantly induced by the combined treatment with lipopolysaccharide (LPS) and platelet activating factor (PAF).The magnitude of changes in these proteins was significantly more extensive in NEC infants, reflecting the different nature of injury and/or severity of inflammation.Such exaggerated immunologic responses may account for the high morbidity and mortality in NEC compared with SIP patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.

ABSTRACT
Necrotizing enterocolitis (NEC) and spontaneous intestinal perforation (SIP) are the most common acute surgical emergencies associated with high morbidity and mortality in preterm infants. We aimed to compare the profiles of immunoregulatory proteins and identify novel mediators in plasma of NEC and SIP infants. We also investigated the expression of target genes in resected intestinal tissues and an enterocyte cell line. Using Cytokine Antibody Array assay, we reported the first comparative profiles of immunoregulatory proteins in plasma of NEC and SIP infants, and showed that dysregulated proteins belonged to functionally diversified categories, including pro- and anti-inflammation, angiogenesis, cell growth, wound healing, anti-apoptosis, cell adhesion and extracellular matrix reorganization. Validation by ELISA confirmed significantly higher concentrations of interleukin (IL)-6, angiopoietin (Ang)-2, soluble type II interleukin-1 receptor (sIL-1RII), and soluble urokinase-type plasminogen activator receptor (suPAR) in NEC infants compared with gestational age-matched control, and a lower level of an epidermal growth factor receptor, secreted form of receptor tyrosine-protein kinase ErbB3 (sErbB3), compared with SIP infants. mRNA expressions of IL1-RII and uPAR were up-regulated in resected bowel tissues from NEC infants, indicating that immunoregulation also occurred at the cellular level. In FHs-74 Int cells, Ang-2, IL1-RII and uPAR mRNA expressions were significantly induced by the combined treatment with lipopolysaccharide (LPS) and platelet activating factor (PAF). Our study provided plasmatic signatures of immunoregulatory proteins in NEC and SIP infants, and demonstrated involvement of multiple functional pathways. The magnitude of changes in these proteins was significantly more extensive in NEC infants, reflecting the different nature of injury and/or severity of inflammation. We speculate that dysregulation of IL-6, Ang-2, IL-1RII and uPAR occurred at both systemic and cellular levels, and probably mediated via LPS and endogeneous PAF signals. Such exaggerated immunologic responses may account for the high morbidity and mortality in NEC compared with SIP patients.

Show MeSH
Related in: MedlinePlus