CARD9S12N facilitates the production of IL-5 by alveolar macrophages for the induction of type 2 immune responses.

The adaptor CARD9 functions downstream of C-type lectin receptors (CLRs) for the sensing of microbial infection, which leads to responses by the TH1 and TH17 subsets of helper T cells. The single-nucleotide polymorphism rs4077515 at CARD9 in the human genome, which results in the substitution S12N (CARD9S12N), is associated with several autoimmune diseases. However, the function of CARD9S12N has remained unknown. Here we generated CARD9S12N knock-in mice and found that CARD9S12N facilitated the induction of type 2 immune responses after engagement of CLRs. Mechanistically, CARD9S12N mediated CLR-induced activation of the non-canonical transcription factor NF-κB subunit RelB, which initiated production of the cytokine IL-5 in alveolar macrophages for the recruitment of eosinophils to drive TH2 cell-mediated allergic responses. We identified the homozygous CARD9 mutation encoding S12N in patients with allergic bronchopulmonary aspergillosis and revealed activation of RelB and production of IL-5 in peripheral blood mononuclear cells from these patients. Our study provides genetic and functional evidence demonstrating that CARD9S12N can turn alveolar macrophages into IL-5-producing cells and facilitates TH2 cell-mediated pathologic responses.

Single-nucleotide polymorphism and haplotypes of TNIP1 associated with systemic lupus erythematosus in a Chinese Han population.

OBJECTIVE: To determine the association of systemic lupus erythematosus (SLE) with single-nucleotide polymorphisms (SNP) in the TNIP1 gene and compare the expression of this gene in cases and controls from a Chinese Han population in this replication study. METHODS: Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was used to genotype 19 SNP in TNIP1 in Chinese Han patients with SLE (n = 341) and controls (n = 356). Genotypes were analyzed by codominant, dominant, and recessive models. Analysis of allele frequencies and linkage disequilibrium was also performed. Western blotting and qRT-PCR were used to measure the expression of these genes in peripheral blood mononuclear cells of SLE cases and controls. RESULTS: Seven SNP loci were significantly associated with SLE in our population (p < 0.05 for all comparisons). Two TNIP1 gene haplotypes (ATTGCGC and GTCCTAT) were associated with SLE (p = 0.0246 and p = 0.0024, respectively). Western blotting and qRT-PCR results provide evidence that patients with SLE had significantly reduced expression of TNIP1/ABIN-1 relative to controls. CONCLUSION: Analysis of SNP in the TNIP1 gene and expression of this gene in peripheral blood lymphocytes indicated these SNP were associated with the occurrence of SLE in Han Chinese patients. Future studies should examine the roles of these SNP in the pathogenesis of SLE.

PGC-1α prevents apoptosis in adipose-derived stem cells by reducing reactive oxygen species production in a diabetic microenvironment.

AIMS: To examine whether overexpression of peroxisome proliferator activated receptor-gamma coactivator-1 alpha (PGC-1α) can prevent apoptosis in adipose-derived stem cells (ASCs) by reducing reactive oxygen species (ROS) production and enhancing mitochondrial function in a diabetic environment. METHODS: After the isolation, expansion and characterisation of rat ASCs, we overexpressed PGC-1α in ASCs using an adenoviral vector encoding green fluorescent protein (GFP) or PGC-1α and tested the apoptotic effect under conditions of high glucose, hypoxia and serum deprivation. The production of intracellular ROS and mitochondrial ROS was evaluated using dihydroethidium and CM-H2XRos fluorescent probes. RESULTS: Under conditions of high glucose, hypoxia and serum deprivation, the overexpression of PGC-1α in ASCs decreased apoptosis and led to an increased survival rate. The ASCs modified with PGC-1α produced lower intracellular and mitochondrial ROS. The mitochondrial morphology and structure in the PGC-1α-ASC group remained relatively complete compared with the control group. CONCLUSIONS: These results reveal a crucial protective role for PGC-1α in the treatment of diabetes mellitus and its complications using stem cells therapy.

Recombinant VirB5 protein as a potential serological marker for the diagnosis of bovine brucellosis.

The molecular tag vaccine against Brucella abortus and serological testing are the main methods of prevention of brucellosis used currently. They can discriminate vaccinated animals and humans from those naturally infected. In this study, we constructed a gene deletion mutant strain, B. abortus S19 virB5 with a molecular tag. Recombinant VirB5 was expressed and purified for evaluation as a diagnostic reagent for bovine brucellosis. In total, 400 sera samples were tested using a VirB5 antigen-based enzyme-linked immunosorbent assay (ELISA) and the results were compared with those of the standard tube agglutination test (SAT). This showed that the sensitivity was 88.2%, specificity was 97.8% and accuracy was 94.8%. Recombinant VirB5 could also be used to discriminate B. abortus-infected mice from mice infected with the B. abortus S19 virB5 mutant strain. It was concluded that recombinant VirB5 could be used as a potential antigen and serological marker for the diagnosis of bovine brucellosis.

Simple and rapid spectrophotometric determination of trace titanium (IV) enriched by nanometer size zirconium dioxide in natural water.

A novel method for preconcentration of Ti(IV) with nanometer size ZrO(2) and determination by spectrophotometry has been developed. Ti(IV) was selectively adsorbed on 300 mg ZrO(2) from 500 mL solution at pH 6.0, then eluted by 5 mL 11.3 mol L(-1) HF. The eluent added was diantipyrylmethane (DAPM, as chromogenic reagent) and ascorbic acid (as masking agent), used for the analysis of Ti(IV) by measuring the absorbance at 390 nm with spectrophotometry, based on the chromogenic reaction between the Ti(IV) and DAPM. This method gave a concentration enhancement of 100 for 500 mL sample, eliminated the sizable interferences on direct determination with spectrophotometry. Detection limit (3 sigma, n=11) of 0.1 microg L(-1) was obtained. The method was applied to determine the concentration of Ti(IV) in river water and seawater and the analytical recoveries of Ti(IV) added to samples were 97.6-101.3%.