Ping-Chang Yang

To detect the percentage of Th17 cells in spleen and IL-17 level in bronchoalveolar lavage fluid in Dermatophagoides farinae allergic asthma mice. Twenty BALB/c mice were randomly divided into control group (n=10) and asthma group (n=10). Mice in control group were treated with PBS plus 2 mg Al(OH)3 and those in asthma group were sensitized with 200 µl solution [50 µg Dermatophagoides farinae crude extracts plus 2 mg Al (OH)3] on day 0, 7 and 14. One week after the last sensitization, all mice were intranasally challenged with 50 µg Dermatophagoides farinae crude extracts daily for 7 days. Twenty-four hours after the last challenge, mice were sacrificed. The sera, bronchoalveolar lavage fluid (BALF) and spleens were collected. The serum levels of IgE and IgG1, and IL-17 level in BALF were determined by ELISA. The percentage of Th17 cells in spleen was tested by flow cytometry. The serum levels of IgG, and IgE in asthma group were (0.10 ± 0.01) pg/ml and (1.15 ± 0.10) pg/ml, respectively, which were higher than that of the control [(0.06 ± 0.01) pg/ml and (0.04 ± 0.01) pg/ml] (P < 0.05). IL-17 level in asthma group (85.13 ± 2.36) pg/ml was higher than that of the control [(48.27 ± 4.14) pg/ml] (P < 0.01). The percentage of Th17 cells in asthma group [(5.19 ± 0.68)%] was also higher than that of the control [(0.95 ± 0.19)%] (P < 0.01). Meanwhile, the percentage of Thl7 cells in spleen was positively correlated with IL-17 level in BALF (r = 0.851, P < 0.01). Compared with healthy mice, both the percentage of Th17 cells in spleen and IL-17 level in BALF have increased significantly in Dermatophagoides farinae allergic asthma mice.

The DNA fragment encoding β-hexosaminidase was synthesized, and cloned into pET-28a vector. The constructed plasmid pMD18-T-β-hexosaminidase was transformed into E. coli Top 10 and followed by expression of the protein induced by IPTG. SDS-PAGE result showed that the relative molecular mass of the recombinant protein was about M, 55 000. The full length of β-hexosaminidase gene was 1 410 bp. Bioinformatics analysis revealed that β-hexosaminidase was composed with 469 amino acid residues with a calculated molecular weight of Mr 55,000, and its secondary structure was composed of strand (14.71%), helix (30.70%), and loop (54.58%). β-hexosaminidase was a hydrophilic protein without signal peptide, and located in the extracellular space.

The epithelial barrier dysfunction plays a critical role in the pathogenesis of a broad array of immune diseases. Alix protein is involved in the endolysosome system. This study aims to elucidate the role of Alix in the maintenance of epithelial barrier function. The results showed that Alix was detected in T84 cells at both mRNA and protein levels. Exposure to Staphylococcal enterotoxin B (SEB) markedly suppressed the expression of Alix in T84 cells, which could be blocked by knocking down the Toll like receptor 2. The exposure to SEB did not affect the TER, but markedly increased the permeability of T84 monolayers to OVA; the OVA passing through T84 monolayers still preserved the antigenicity manifesting inducing antigen specific T cells proliferation. Alix protein plays a critical role in the maintenance of the barrier function of T84 monolayers.

North Am J Med Sci, Official publication of ,Canada

Allergic rhinitis is a skewed immune reaction to common antigens in the nasal mucosa; current therapy is not satisfactory and can cause a variety of complications. In recent decades, the incidence of allergic rhinitis is increasing every year. Published studies indicate that probiotics are beneficial in treating allergic rhinitis. This review aims to help in understanding the role of probiotics in the treatment of allergic rhinitis. We referred to the PubMed database as data source. This review focuses on the following aspects: The types of probiotics using in the treatment of allergic rhinitis, approaches of administration, its safety, mechanisms of action, treating results, and the perspectives to improve effectiveness of probiotics in the treatment of allergic rhinitis. This review reports the recent findings regarding the role of probiotics in the treatment of allergic rhinitis. Probiotics are a useful therapeutic remedy in the treatment of allergic rhinitis, but its underlying mechanisms remain to be further investigated.

The pathogenesis of inflammatory bowel disease (IBD) has not been fully understood yet. Eosinophils (Eo) are one type of the major proinflammatory cells of the chronic inflammation in the intestine. CD98 is involved in the pathogenesis of a number of inflammations. This study aims to elucidate the role of CD98⁺ Eos in the initiation of intestinal inflammation. The colon biopsies were collected from 60 patients with IBD. The expression of CD98 in the biopsies was examined by immunohistochemistry. The serum levels of the flagellin (FGN) antibody and Eo-derived mediators in the culture supernatants were assessed by enzyme-linked immunosorbent assay. The role of FGN on Eo activation was examined in a cell culture model. The role of FGN in the induction of colitis was observed in a mouse model. Compared to normal controls, the frequency of CD98⁺ Eos was markedly increased in the IBD colon mucosa. FGN were detected in the colon biopsies and in the sera of IBD patients. Exposure to FGN induced the expression of galectin 3 (the ligand of CD98) in dendritic cells. The exposure to galectin 3 activated the CD98⁺ Eos. After treatment with FGN intrarectally, mice with eosinophilia showed severe inflammation in the colon. The interaction of galectin 3 and CD98 can induce Eos to release chemical mediators that contributes to the initiation of the intestinal inflammation.

The enterocytes have the potential to absorb noxious substances, such as microbial products, from the gut lumen. How the enterocytes process the substances to harmless materials is not fully understood. This study aims to elucidate the role of ubiquitin E3 ligase TNFAIP3 (TNFAIP3) in facilitating the degradation of endocytic microbial products in enterocytes. Human intestinal epithelial cell line, HT-29 cells, was cultured to monolayers using as an in vitro model to observe the endocytosis and degradation of microbial products, Staphylococcal enterotoxin B (SEB) in epithelial cells. The RNA interference was employed to knock down the TNFAIP3 gene in HT-29 cells to observe the role of TNFAIP3 in the degradation of endocytic SEB. The role of TNFAIP3 in facilitating the endosome/lysosome fusion was observed by immunocytochemistry. Upon the absorption of SEB, the expression of TNFAIP3 was increased in HT-29 cells. Silencing the TNFAIP3 gene in HT-29 cells resulted in a large quantity of SEB to be transported across the HT-29 monolayers to the transwell basal chambers; the transportation was via the intracellular pathway. TNFAIP3 was required in the fusion of SEB-carrying endosomes and lysosomes. TNFAIP3 plays a critical role in the degradation of endocytic SEB in enterocytes.