Jen-Chieh Chang

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.

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.

Altered p38 Mitogen-Activated Protein Kinase Expression in Different Leukocytes with Increment of Immunosuppressive Mediators in Patients with Severe Acute Respiratory Syndrome

Severe acute respiratory syndrome (SARS) has spread to a global pandemic, especially in Asia. The transmission route of SARS has been clarified, but the immunopathogenesis of SARS is unclear. In an age-matched case-control design, we studied immune parameters in 15 SARS patients who were previously healthy. Plasma was harvested for detection of virus load, cytokines, and nitrite/nitrate levels, and blood leukocytes were subjected to flow cytometric analysis of intracellular mitogen-activated protein kinases (MAPKs) in different leukocytes. Patients with SARS had significantly higher IL-8 levels (p = 0.016) in early stage, and higher IL-2 levels (p = 0.039) in late stage than normal controls. Blood TNF-α, IL-6, and IL-10, and nitrite/nitrate levels were not significantly elevated. In contrast, TGF-β and PGE2 levels were significantly elevated in SARS patients. Five of the 15 SARS patients had detectable coronaviruses in blood, but patients with detectable and undetectable viremia had no different profiles of immune mediators. Flow cytometric analysis of MAPKs activation by phospho-p38 and phospho-p44/42 (extracellular signal-regulated kinase) expression showed that augmented p38 activation (p = 0.044) of CD14 monocytes associated with suppressed p38 activation (p = 0.033) of CD8 lymphocytes was found in SARS patients. These results suggest that regulation of TGF-β and PGE2 production and MAPKs activation in different leukocytes may be considered while developing therapeutics for the SARS treatment.

Gene–gene and gene–environment interactions on IgE production in prenatal stage

To cite this article: Yang KD, Chang J‐C, Chuang H, Liang H‐M, Kuo H‐C, Lee Y‐S, Hsu T‐Y, Ou C‐Y. Gene–gene and gene–environment interactions on IgE production in prenatal stage. Allergy 2010; 65: 731–739.

Gender‐limited association of cytotoxic T‐lymphocyte antigen‐4 (CTLA‐4) polymorphism with cord blood IgE levels

Allergic mechanism has long been attributed to IgE‐mediated reaction. The relationship between gene polymorphism and cord blood IgE (CB IgE) is unclear. We investigated whether elevation of CB IgE levels was associated with polymorphisms of cytotoxic T‐lymphocyte antigen 4 (CTLA‐4) at (−318) CT and (+49) AG positions in a gender‐limited fashion. CB IgE levels were determined by Pharmacia CAP system and the CTLA‐4 polymorphisms at (−318) and (+49) were determined by restriction fragment length polymorphism (RFLP). A total of 644 consecutive umbilical cord bloods were collected for this study. 32.9% of newborn infants had detectable IgE levels (≥0.35 kU/l). 25.6% of the male newborns had elevated CB IgE levels (≥0.5 kU/l) similar to those of the female newborns (22.7%). The CTLA‐4 polymorphism at (+49) but not (−318) was significantly associated with elevated CB IgE levels (p = 0.004). The association of CTLA‐4 (+49) A allele with elevated CB IgE levels was found only in females. Both male and female infants with different CTLA‐4 (−318) genotypes had no difference in the rates of elevated CB IgE levels. A linkage disequilibrium between CTLA‐4 (+49) G and (−318) C allele was found in this Chinese population. Subjects with the (+49, GG and −318, CC) genotype had a significantly lower rate of elevated CB IgE levels. Association of the CTLA‐4 (+49) polymorphism with elevated CB IgE levels was found only in female infants. Newborn infants with the (+49, GG and −318, CC) genotype tended to have a low rate of elevated CB IgE.

Combination of CTLA-4 and TGFβ1 gene polymorphisms associated with dengue hemorrhagic fever and virus load in a dengue-2 outbreak

The pathogenesis of dengue hemorrhagic fever (DHF) has been considered to be massive immune activation of T cells. Abnormal expression of the immune regulatory molecules, CTLA-4 and TGFbeta1, leads to disturbances of regulatory T cell immune response. We investigate the contribution of CTLA-4 and TGFbeta1 in DHF by analyzing them for association with virus load in blood and polymorphisms of CTLA-4 +49A/G, and TGFbeta1 -509C/T in a DEN-2 outbreak. The increased frequency of the TGFbeta1 -509 CC genotype in patients with DHF was compared to those with dengue fever (OR=1.9, p=0.034). Moreover, the presence of the CTLA-4 +49 G allele and TGFbeta1 -509 CC genotype increased the susceptibility to risk of DHF (OR=2.1, p=0.028) and significantly higher virus load (p=0.013). This finding suggests that a combination of CTLA-4 and TGFbeta1 polymorphisms is associated with the susceptibility of DHF and higher virus load.

Interaction of maternal atopy, CTLA‐4 gene polymorphism and gender on antenatal immunoglobulin E production

Background Genetic heritability and maternal atopy have been correlated to antenatal IgE production, but very few studies have studied gene–maternal atopy interaction on antenatal IgE production. This study investigated the interaction of CTLA‐4 polymorphism with prenatal factors on the elevation of cord blood IgE (CBIgE).

Identification of an association between genomic hypomethylation of FCGR2A and susceptibility to Kawasaki disease and intravenous immunoglobulin resistance by DNA methylation array.

Kawasaki disease (KD) is characterized by systemic vasculitis, and it is the most common acquired heart disease in children. However, the etiology and immunopathogenesis of KD are still unclear. A genome‐wide association study (GWAS) identified polymorphisms in CD40, BLK, and FCGR2A as the susceptibility genes for KD. No epigenetic array studies of KD have previously been published. This study was undertaken to investigate differences in DNA methylation in patients with KD as compared to controls.

Use of proteomic differential displays to assess functional discrepancies and adjustments of human bone marrow- and Wharton jelly-derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) from bone marrow are suitable for the reconstruction of connective tissues and even brain tissue but have limitations in terms of cell expansion and fully specific differentiation. In our current study, we have attempted to adjust and improve the cell expansion and differentiation properties of human MSCs from different tissues. MSCs from normal bone marrow and Wharton jelly were subjected to proteomic differential displays, followed by functional adjustments based on these displays. Bone marrow MSCs expressed more transgelin-2 and differentiated more rapidly into bone nodules but showed a slower growth rate. A knockdown of transgelin-2 expression by specific small interfering RNA (siRNA) significantly increased the growth rate of these cells, the G1/S phase cell cycle transition, and the interaction of cyclin D1 with cdk2. Wharton jelly MSCs expressed the chaperone protein HSP90β at higher levels and differentiated slowly toward an osteogenic lineage. However, the knockdown of HSP90β expression significantly increased bone nodule formation, inhibited cell growth, decreased the number of cells in the G1/S phase of the cell cycle, and decreased the interaction of cyclin D1 with cdk2 and of cyclin E with cdk2. These results were validated by the in vivo repair of segmental bone defects in a mouse model with severe combined immunodeficiency. We thus demonstrate an improvement in the cell expansion and tissue regeneration properties of human MSCs through specific adjustments.

Role of vascular cell adhesion molecules and leukocyte apoptosis in the lymphopenia and thrombocytopenia of patients with severe acute respiratory syndrome (SARS)

Abstract The immunopathogenesis of leukopenia and thrombocytopenia in patients with severe acute respiratory syndrome (SARS) is unclear. In order to explore the leukopenia mechanism, we studied 15 SARS patients who were previously healthy, and 15 age-matched normal controls in a paired design. Soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble Fas ligand (sFasL) in plasma were measured by ELISA, and intracellular activated caspase-3 fragment in different leukocytes was determined by flow cytometry. Patients with SARS had significantly lower lymphocyte and platelet counts and significantly higher sVCAM-1 and sFasL levels compared to healthy controls. sVCAM-1 levels correlated negatively with total leukocytes and platelet counts, but positively with plasma sFasL levels. Intracellular cleaved caspase-3 expression was also significantly higher in lymphocytes from SARS patients in acute phase than in convalescent stage. Lymphopenia and thrombocytopenia in SARS patients may be caused, in part, by enhanced vascular sequestration associated with increased sVCAM-1 levels. However, lymphopenia may be due to enhanced cell death. Inhibition of cell adhesion and caspase-3 activation could, therefore, have prevented SARS patients from developing thrombocytopenia and lymphopenia.