Construction of mouse interleukin-10 recombinant adenovirus vector and gene modification for dendritic cells / 中国组织工程研究

BACKGROUND: Few reports concern the effects of dendritic cells-a kind of antigen presenting cells, and interleukin-10 (IL-10) on airway hyperreactivity or inflammation. OBJECTIVE: To construct mice IL-10 recombinant adenovirus vector Ad-mIL-10 to acquire the dendritic cells modified by mIL-10, which can provide a foundation for the further study. METHODS: Mouse IL-10 (mIL-10) gene comprise of enzyme cutting spot was synthesized according to the mIL-10 gene sequence and multiclone spot of adenovirus vector, connected to pMD18-T vector and sequenced. MIL-10 was subcloned to BD Adeno-X~(TM) vector, packed and augmented in HEK 293 cells, following determine the protein expression, and the vector was transfected to mice bone marrow-derived dendritic cells. RESULTS AND CONCLUSION: Recombinant adenovirus vector Ad-mIL-10 was successfully synthesized, packed and augmented, which could highly express protein IL-10. Bone marrow-derived dendritic cells were successfully cultured and transduced in vitro. It suggested that it is feasible to transfect mice dendritic cells by Ad-mIL-10 adenovirus vector. The study can provide more sufficient theoretic evidence for the possibility of correlative gene therapy.

Association between HLA-DQA1, -DQB1 gene polymorphisms and susceptibility to asthma in northern Chinese subjects / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>Human leukocyte antigen (HLA) class II genes, especially HLA-DQ genes, which are highly polymorphic, have been thought to be candidate loci for the etiology of asthma, and shown to be involved in antigenic presentation. This study was conducted to investigate whether susceptibility or resistance to asthma is associated with HLA-DQA1 and DQB1 genes polymorphism.</p><p><b>METHODS</b>Venous blood samples were collected from northern Chinese population with Han ethnic. (1) One hundred and twenty-five unrelated asthmatic individuals and 52 subjects from 12 asthmatic pedigrees. (2) Ninety-six healthy controls without asthma and atopy with the same ethnic. Genomic DNA was extracted using standard phenol-chloroform method. The second exon of HLA-DQA1 and DQB1 genes were amplified by sequence-specific primer-polymerase chain reaction (SSP-PCR) method. All asthmatics had their serum IgE (total and specific) antibody or skin-prick test measured, bronchial reactivity to methacholine (Mch) and bronchial reversibility by beta(2)-agonist evaluated.</p><p><b>RESULTS</b>HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were significantly higher in asthmatics than those in healthy controls (0.408 vs 0.177, P < 0.01; 0.568 vs 0.198, P < 0.01). Odds ratios (ORs) were 3.203 (95% CI 1.699 - 6.037), 5.328 (95% CI 2.883 - 9.849) respectively. Conversely, HLA-DQA1 * 0301 allele and HLA-DQB1 * 0301 were significantly decreased in asthmatics compared to healthy controls (0.296 vs 0.50, P < 0.01; 0.4 vs 0.563, P < 0.05); Logistic regression analysis showed that HLA-DQA1 * 0104 allele was associated independently with asthma etiology, OR [represented by Exp(B)] was 5.0942 with 95% CI 2.2520 - 21.1813; Spearman's analysis showed that HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were positively associated with atopy, the correlation coefficient were 0.183 and 0.289 respectively, P < 0.01. By contrast, HLA-DQA1 * 0301 allele was negatively related to atopy, the correlation coefficient was -0.168, P < 0.05; linkage analysis did not support the view that HLA-DQA1/DQB1 genes were linked to asthma with LOD value being 0.72.</p><p><b>CONCLUSIONS</b>HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were implicated in susceptibility to asthma and atopy, HLA-DQA1 * 0301 allele and HLA-DQB1 * 0301 might be protective factor against asthma. Asthma and atopy are multifactorial disorders, HLA-DQA1 and DQB1 genes are involved in the regulation of immune specific response to common allergen.</p>

Role of endothelin and nitric oxide in the pathogenesis of asthma / 中国病理生理杂志

AIM:To explore whether the balance between endothelin(ET) and nitric oxide(NO) plays an important role in airway hyperresponsiveness of asthmatic rats. METHODS:The tension of isolated perfused rat tracheal rings was measured after ET-1 stimulation and incubation of JKC 302 and L-NAME. RESULTS:ET-1 constricted isolated rat tracheal ring, produced slowly developing and long-lasting contraction. The ET-1-induced contraction response of asthmatic rat tracheal ring was higher than that of normal control group (P＜0.01). JKC 302, a selective ETA receptor antagonist, partly blocked ET-1-induced contraction in asthmatic rat trachea ring. L-NAME significantly augmented the constriction caused by ET-1. CONCLUSION:The effects of ET on bronchomotor tone may be modified by NO as this is a bronchomotor, and the imbalance between ET and NO may play an important role in asthma pathogenesis.