Association between polymorphisms in the vesicle-associated membrane protein-associated protein A (<Emphasis Type="Italic">VAPA</Emphasis>) gene on chromosome 18p and bipolar disorder

Linkage studies in bipolar disorder (BPD) suggest that a susceptibility locus exists on chromosome 18p11. The vesicle-associated membrane protein-associated protein A (VAPA) gene maps to this region. VAPA interacts with presynaptic proteins and is necessary for vesicular neurotransmission. Dysregulation of synaptic neurotransmission might contribute to the pathophysiology of BPD. In this study, we hypothesize that genetic variations in the VAPA gene contribute to BPD. We tested this hypothesis by genotyping 6 SNPs (rs494015; rs29193; rs29162; rs29145; rs29067; rs29066) in BPD patients (n = 570) and healthy controls (n = 730). Genotype and allele frequencies were compared between groups using Chi square contingency analysis. Linkage disequilibrium (LD) between markers was calculated and estimated haplotype frequencies were compared between groups. Single marker analysis revealed an association of rs29067 and rs29066 with BPD; however, after permutation correction, only rs29066 showed a trend towards an allelic association (P = 0.066). Haplotype analysis did not show any significant association with disease after permutation correction. Our results provide suggestive evidence of an association between SNPs in the 3'UTR of the VAPA gene and BPD. Interestingly, these SNPs are in close proximity to the microsatellite marker D18S464, which showed significant signals in previous linkage studies of BPD. Additional studies are necessary to confirm and elucidate the role of VAPA as a susceptibility gene for BPD on chromosome 18p.     

TGF-β1和ADAM33基因交互作用与儿童哮喘易感性及严重程度相关性

目的:探讨转化生长因子β1(TGF-β1)和解整合素金属蛋白酶33(ADAM33)基因单核苷酸多态性与儿童哮喘易感性及严重程度相关性。方法:采用以医院为基础的病例对照研究(110例哮喘患儿和144例对照)方法,三个多态性位点(V4、T2、T869C)用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)技术进行基因分型,应用MDR软件分析基因各位点之间交互作用。结果:TGF-β1基因的T869C位点基因型及等位基因频率分布在哮喘组和对照组比较差异均无统计学意义(P>0.05)。哮喘组ADAM33基因V4位点C和T2位点A等位基因频率显著高于对照组(P<0.01),而V4位点的CC基因型及T2位点的GA基因型分布在轻中度哮喘组与对照组比较差异无统计学意义(P>0.05),T2位点的AA基因型分布在重度哮喘与对照组比较无显著性差异(P>0.05)。MDR交互作用分析显示,ADAM33基因V4和T2位点构成的2个位点最佳模型;ADAM33基因位点V4、T2和TGF-β1位点T869C构成的3个位点最佳模型两组比较均有统计学差异(P<0.05)。结论:ADAM33基因V4和T2位点与儿童哮喘发生及其严重程度相关,同时TGF-β1基因T869C位点与ADAM33基因V4和T2位点均具有明显交互作用     

Jα33基因工程蛋白与多克隆抗体的制备及初步应用

为了获得同时识别人类和小鼠MAIT细胞的特异性抗体,我们制备了MAIT细胞TCRα链Jα33融合蛋白及其抗体,为MAIT细胞在炎症性肠道疾病(IBD)中作用的研究提供有利的工具。用Jα33的全长序列为引物经PCR扩增Jα33串联体,扩增的串联体片段连接进入T载体,阳性克隆转入pGEX-4T-1表达载体中原核表达GST-Jα33融合蛋白,纯化后的融合蛋白免疫新西兰大白兔制备anti-Jα33多克隆抗体。并分别用Western blot和RT-PCR检测IBD模型小鼠MAIT细胞Jα33的表达。成功获得分子量约为38 kD的融合蛋白,免疫大白兔后得到抗Jα33的多克隆抗体,Western blot结果显示Jα33多克隆抗体能检测到人外周血单个核淋巴细胞表面Jα33的表达,而HeLa中则无Jα33的表达;MAIT细胞Jα33在IBD小鼠肠组织中表达量明显低于正常对照。成功制备了能同时识别人类和小鼠MAIT细胞Jα33的多克隆抗体,为研究MAIT细胞的定位和功能开启了方便之门。