Hong-Ren Yu

The relationship of eosinophilia to intravenous immunoglobulin treatment failure in Kawasaki disease

To investigate the role of eosinophils in Kawasaki disease (KD) and the relationship to initial intravenous immunoglobulin (IVIG) treatment failure. A retrospective analysis of all children who were admitted and met the criteria of KD between 1999 and 2005. The patients were divided into IVIG‐responsive and IVIG‐resistant groups. A total of 185 patients were enrolled during the study period. A series of blood eosinophils and biochemistry studies were correlated to the effectiveness of IVIG. The neutrophils percentage before IVIG treatment (pre‐IVIG), leukocyte counts within 3 days after IVIG treatment (post‐IVIG), liver enzyme, albumin levels, and post‐IVIG eosinophils percentage were all significantly different between the two groups in univariate analysis. Under multivariate analysis with logistic regression, post‐IVIG eosinophilia [peripheral blood (PB) eosinophils ≥4%] had an inverse correlation to KD patients with IVIG‐resistance (p = 0.003). Also, pre‐IVIG hypoalbuminemia (albumin ≤3.0 g/dl) was positively correlated to IVIG‐resistance (p = 0.018). Further analysis showed that the PB eosinophils was markedly increased in the acute stage and returned to normal 3 weeks after IVIG treatment (p < 0.001). Eosinophil levels are highly elevated in the acute stage of KD both before and after the IVIG treatment. Post‐IVIG treatment eosinophilia has an inverse correlation to KD patients with IVIG‐resistance and may indicate IVIG‐responsive. This may be a valuable factor to survey for the necessity of a second dose IVIG treatment.

Serum albumin level predicts initial intravenous immunoglobulin treatment failure in Kawasaki disease

Objectives:  Kawasaki disease (KD) is a systemic vasculitis primarily affecting children who are <5 years old. Intravenous immunoglobulin (IVIG) is the standard therapy for KD. However, many patients with KD still show poor response to initial IVIG treatment. This study was conducted to investigate the risk factors for initial IVIG treatment failure in KD.

ITPKC single nucleotide polymorphism associated with the Kawasaki disease in a Taiwanese population.

Kawasaki disease (KD) is characterized by systemic vasculitis with unknown etiology. Previous studies from Japan indicated that a gene polymorphism of ITPKC (rs28493229) is responsible for susceptibility to KD. We collected DNA samples from 1,531 Taiwanese subjects (341 KD patients and 1,190 controls) for genotyping ITPKC. In this study, no significant association was noted for the ITPKC polymorphism (rs28493229) between the controls and KD patients, although the CC genotype was overrepresented. We further combined our data with previously published case/control KD studies in the Taiwanese population and performed a meta-analysis. A significant association between rs28493229 and KD was found (Odds Ratio:1.36, 95% Confidence Interval 1.12–1.66). Importantly, a significant association was obtained between rs28493229 and KD patients with aneurysm formation (P = 0.001, under the recessive model). Taken together, our results indicated that C-allele of ITPKC SNP rs28493229 is associated with the susceptibility and aneurysm formation in KD patients in a Taiwanese population.

CASP3 gene single-nucleotide polymorphism (rs72689236) and Kawasaki disease in Taiwanese children

Kawasaki disease (KD) is characterized by systemic vasculitis of unknown etiology. A study from Japan reported that G to A substitution of a single-nucleotide polymorphism (SNP) located in the 5′-untranslated region of caspase 3 (CASP3) (rs72689236), which was associated with nuclear factor of activated T cell-mediated T-cell activation, is responsible for susceptibility to KD. This study was conducted to investigate whether the polymorphism of CASP3 is responsible for susceptibility and coronary artery lesion (CAL) formation in KD in the Taiwanese population. A total of 1092 subjects (341 KD patients and 751 controls) were investigated to identify an SNP of rs72689236 using Invader assays (Third Wave Technologies). Our data provided a borderline significant association between the genotypes and allele frequency of rs72689236 in control subjects and KD patients (P=0.0535 under the dominant model; P=0.0575 under the allelic model). The A allele of rs72689236 in KD patients and in patients with CAL and intravenous immunoglobulin resistance was seen in a higher frequency. Importantly, a significant association was obtained between rs72689236 and KD patients with aneurysm formation (P=0.009, under the recessive model). The A allele of rs72689236 is very likely to be a risk allele in the development of aneurysm in patients with KD.

Association of lower eosinophil-related T helper 2 (Th2) cytokines with coronary artery lesions in Kawasaki disease.

Kawasaki disease (KD) is a systemic febrile vasculitis particular coronary artery involvement. Eosinophilia has been found in our and other studies in KD. This study further investigates whether eosinophil‐related T helper 2 (Th2) cytokines or the activation marker (eosinophil cationic protein – ECP) is involved in KD with coronary artery lesions (CAL). A total of 95 KD patients were enrolled for this study. Plasma samples were subjected to the measurement of interleukin (IL)‐4, IL‐5, and eotaxin by Luminex‐Bedalyte multiplex beadmates system and to the measurement of ECP by fluoroimmunoassay. Patients with KD had higher eosinophils than controls. Eosinophil‐related mediators: IL‐4, IL‐5, eotaxin, and ECP levels were also higher in KD patients than controls before intravenous immunoglobulin (IVIG) treatment. After IVIG treatment, ECP decreased but IL‐4, IL‐5, and eotaxin increased significantly. The higher the IL‐5 and eosinophil levels after IVIG treatment, the lower rate of CAL was found. Changes of eosinophils after IVIG treatment were positively correlated to changes of IL‐5 levels but not ECP levels. An increase of eosinophils and IL‐5, but not ECP levels after IVIG treatment, was inversely correlated with CAL formation in KD.

miRNA-125b regulates TNF-α production in CD14+ neonatal monocytes via post-transcriptional regulation

Neonates, although deficient in cell immunity, frequently reveal sepsis with augmented proinflammatory reactions. Here, we found that neonatal monocytes produced significantly higher TNF‐α mRNA and protein than adult monocytes. Assessment of the transcriptional factor found no significant difference of NF‐κB p65 level between neonatal and adult monocytes. Addition of Act D to access the half‐life of TNF‐α mRNA revealed no significant difference of the LPS‐induced TNF‐α mRNA half‐life between them, whereas CHX increased neonatal TNF‐α mRNA significantly. This suggests that a post‐transcriptional mechanism involves the augmentation of TNF‐α production by neonatal monocytes. To examine whether miRNA was involved in the post‐transcriptional regulation, differential displays of miRNA array between neonatal and adult MNCs were performed, along with the discovery of hsa‐miR‐103, hsa‐miR‐125b, hsa‐miR‐130a, hsa‐miR‐454‐3p, and hsa‐miR‐542‐3p, which were greater than a twofold decrease or increase after LPS treatment for 4 h. The functional validation identified that miR‐125b decreased significantly in association with higher TNF‐α expression by neonatal monocytes after LPS stimulation. Transfection of the miR‐125b precursor into neonatal monocytes significantly repressed the TNF‐α mRNA and protein expression, suggesting that miR‐125b negatively regulates TNF‐α expression in neonatal monocytes. Modulation of miRNA expression may be used to regulate TNF‐α production in newborns with altered proinflammatory reactions.

Prenatal and postnatal probiotics reduces maternal but not childhood allergic diseases: a randomized, double‐blind, placebo‐controlled trial

The prevalence of atopic diseases has increased rapidly in recent decades globally. The administration of probiotics to reduce gastrointestinal inflammation has been popular, but its role in the prevention or treatment of allergic disease remains controversial. This study evaluated the effectiveness of prenatal and postnatal probiotics in the prevention of early childhood and maternal allergic diseases.

A unique plasma proteomic profiling with imbalanced fibrinogen cascade in patients with Kawasaki disease.

Kawasaki disease (KD) is the leading cause of acquired heart disease during childhood in the developed countries. The mechanism and biomarkers of KD remain to be determined. In this study, we sought to elucidate potential plasma proteomic markers in KD patients in comparison to that in febrile controls. Plasma samples from KD patients and febrile controls were subjected to two‐dimensional polyacrylamide gel electrophoresis analysis. Differential protein displays between KD patients and febrile controls were determined. Fibrinogen beta and gamma chains, alpha‐1‐antitrypsin (A1AT), CD5 antigen‐like precursor (CD5L), and clusterin were increased in KD patients, whereas immunoglobulin free light chains were decreased, as compared with controls. The differential protein displays were validated with enzyme‐linked immunosorbent assay tests. We found higher fibrinogen‐related proteins (fibrinogen, A1AT, clusterin, and CD5L), along with a lower level of the immunoglobulin free light chains that involve fibrin degradation in KD. Results from this study showing a unique proteomic profiling with abnormal fibrinogen cascade may afford a good biomarker of KD and a better strategy to prevent cardiovascular complications of KD by correcting abnormal fibrin deposition or degradation.

Different antigens trigger different Th1/Th2 reactions in neonatal mononuclear cells (MNCs) relating to T-bet/GATA-3 expression

Neonates are known to have poor cellular immunity, especially poor Th1 response. We investigated how neonatal mononuclear cells raised different Th1/Th2 reactions in response to different antigens. Employing Dermatophagoides pteronyssinus (Der p) extract and varicella zoster virus (VZV) as antigens, we assessed Th1/Th2 reactions as demonstrated by IL‐4/IFNγ production and mRNA expression, and transcriptional factors T‐bet/GATA‐3 mRNA expression in mononuclear cells from human umbilical cord blood (CBMC). Results showed that VZV induced a dramatic increase of IFNγ production by adult peripheral blood mononuclear cells (PBMC), whereas VZV did not drive CBMC to release significant IFNγ production (1614.7±362.0 vs. 49.0±29.3,p<0.005). However, Der p induced higher IFNγ production by CBMC than VZV (298.1±171.8 vs. 49.0±29.3, P=0.047). In contrast, VZV did not induce significant IL‐4 production either by CBMC or by PBMC. Der p induced a comparative IL‐4 production by CBMC and PBMC (2.58±0.84 vs. 2.04±0.37, p>0.05). A real‐time RT‐PCR analysis of IL‐4 and IFNγ mRNA expression showed that VZV induced a significantly higher IFNγ, but not IL‐4, mRNA expression in PBMC than CBMC. Der p did not induce significant difference of IFNγ or IL‐4 mRNA expression in PBMC and CBMC. VZV enhanced Th1‐related transcription factor T‐bet mRNA expression, in association with later down‐regulation of Th2‐related GATA‐3 mRNA expression in PBMC. However, VZV did not up‐regulate T‐bet or down‐regulate GATA‐3 expression significantly in CBMC. In contrast, Der p induced an early GATA‐3 expression and later T‐bet expression in CBMC. These results suggest that different antigens trigger various Th1/Th2 reactions in PBMC and CBMC resulting from kinetic changes of T‐bet/GATA‐3 expression.

Homocysteine induces smooth muscle cell proliferation through differential regulation of cyclins A and D1 expression

The mechanism of homocysteine‐induced cell proliferation in human vascular smooth muscle cells (SMCs) remains unclear. We investigated the molecular mechanisms by which homocysteine affects the expression of cyclins A and D1 in human umbilical artery SMCs (HUASMCs). Homocysteine treatment induced proliferation of HUASMCs and increased the expression levels of cyclins A and D1. Knocking down either cyclin A or cyclin D1 by small interfering RNA (siRNA) inhibited homocysteine‐induced cell proliferation. Furthermore, treatment with extracellular signal‐related kinase (ERK) inhibitor (PD98059) and dominant negative Ras (RasN17) abolished homocysteine‐induced cyclin A expression; and treatment with phosphatidylinositol 3‐kinase (PI3K) inhibitor (LY294002) and mammalian target of rapamycin (mTOR) inhibitor (rapamycin) attenuated the homocysteine‐induced cyclin D1 expression. Homocysteine also induced transient phosphorylation of ERK, Akt, and p70 ribosomal S6 kinase (p70S6K). Neutralizing antibody and siRNA for β1 integrin blocked cell proliferation, expression of cyclins A and D1, and phosphorylation of ERK and Akt. In conclusion, homocysteine‐induced differential activation of Ras/ERK and PI3K/Akt/p70S6K signaling pathways and consequent expression of cyclins A and D1 are dependent on β1 integrin. Homocysteine may accelerate progression of atherosclerotic lesions by promoting SMC proliferation. J. Cell. Physiol. 226: 1017–1026, 2011.