IDDM与HLA-DQA1及其启动子QAP 的相关性研究

目的 研究胰岛素依赖性糖尿病（IDDM）的易感性与HLA－DQA1及其启动子QAP基因多态性是否存在关联。方法 采用PCR－RFLP分析DQA1等位基因多态性;采用PCR－SSO检测QAP多态性。结果 等位基因分析发现,患者中DQA1＊0301、QAP3．2等位基因频率显著高于正常人（RR＝2．80、2．43,P＜0．01）;而DQA1＊0601、QAP4．2在患者中分布频率较正常人显著降低（RR＝0．10,P＜0．025）。启动子与结构基因连锁分析表明：QAP3．2－DQA1＊0301单元型与IDDM关联,QAP3．1－DQA1＊0301与IDDM无关联;QAP4．20－DQA1＊0601则具有保护作用。结论 1．IDDM与HLA的关联不仅存在于结构基因,而且也存在于启动子;2．同一结构基因与不同的启动子连锁可呈现不同的关联格局。     

上海地区汉族人HLA-DQB1启动子多态性以及QBP-DQB1连锁分析

为分析上海地区汉族人HLA DQB1启动子 (QBP )多态性 ,以及QBP与DQB1的连锁不平衡 ,我们选用 2 4个序列特异性寡核苷酸探针 (SSO )检测QBP多态性。在目前已知的 12种QBP等位基因中 ,我们仅检测到 10种 (未发现新的QBP类型 ) ,并且发现QBP与DQB1的某些等位基因之间存在一定的连锁格局。我们的结果与高加索人群研究结果比较发现 ,无论是QBP基因频率还是QBP DQB1单元型频率 ,在两种不同人群中都存在明显差异。结果提示 ,由于不同的QBP等位基因功能不同 ,因此QBP DQB1之间不同的单倍型组合可能是HLA与疾病关联的另一个因素。     

上海地区汉族人HLA—DQB1启动子多态性以及QBP—DQB1连锁分析

为分析上海地区汉族人HLA－DQB1自动子（QBP）多态性,以及QBP与DQB1的连锁不平衡,我们选用24个序列特异性寡核苷酸探针（SS0）检测QBP多态性。在目前已知的12种QBP等位基因中,我们仅检测到10种（未发现新的QBP类型）,并且发现QBP与DQB1的某些等位基因之间存在一定的连锁格局。我们的结果与高加索人群研究结果比较发现,无论是QBP基因频率还是QBP－DQB1单元型频率,在两种不同人群中都存在明显差异。结果提示,由于不同的QBP等位基因功能不同,因此QBP－DQB1之间不同的单倍型组合可能是HLA与疾病关联的另一个因素。     

上海地区汉族人HLA—DQA1启动子多态性以及QAP,DQA1单元型 …

目的：分析上海地区汉族人ＨＬＡ－ＤＱＡ１启动子（ＱＡＰ）多态性,以及ＱＡＰ与ＤＱＡ１的连锁关系。方法：采用ＰＣＲＲＦＬＰ分析ＤＱＡ１等位基因多态性;采用ＰＣＲ－ＳＳＯ检测ＱＡＰ多态性。结果：在９６个个体中检测到９种ＤＱＡ１启动子（ＱＡＰ）等位基因。ＱＡＰ与ＤＱＡ１之间有３种连锁格局：①ＱＡＰ与ＤＱＡ１存在一对一关系;②一种ＱＡＰ可与不同的ＤＱＡ１组成多种单元型;③一种ＤＱＡ１等位基因可受到多种类型     

中国汉族人HLA-DQAl~*0601基因转录启动子区显示新的核苷酸序列

本文测定了中国汉族人HLA纯合细胞SMY-43A的DQA1~*0601基因启动子区(QAP)核苷酸序列,发现顺式作用元件W box中-215和-216位核苷酸组成与已报道的白种人DQA1~*0601的QAP序列不同,由于SMY-43A的Ⅱ类基因单倍型组成为中国人特有,提示相应的QAP多态性可能具有人种差异.     

应用PCR/SSO方法进行西安汉族人群HLA-DR,DQ位点的DNA分型

对80例西安汉族人HLA第Ⅱ类抗原的DRB、DQA1和DQB1的等位基因多态性进行分析,发现的特异性包括HLA-DRB1 27种,DRB3 3种,DRB5 3种,DQA1 8种,DQB1 14种,共55种。按血清学所定的特异性将DRB1位点的等位基因对应分类,出现频率由高到低排列依次为DR5,DR2,DR6,DR9,DR4,DR7,DR8,DR3,DR1,DR10。在DQA1和DQB1等位基因中,DQA1*0301和DQB1*0301、DQB1*0303的频率是最高的。和以前所报道的北方人群中DRB1*1301与DQB1*0604相连锁的结论不同的是,该人群中DRB1*1301与DQB1*0603相连锁,且未发现北方人群中所报道的DRB1*1403。这一结论与地处南方     

湖北地区汉族健康人群白细胞介素-1α基因启动子-889C/T位点多态性的研究

目的 :了解湖北地区汉族健康人群白细胞介素 1α基因 (IL 1A )启动子 -889位点的单核苷酸多态性 (SNP) ,比较其在不同种族间的分布特点。方法 :采用聚合酶链反应 限制性片段长度多态性 (PCR RFLP)分析的方法 ,检测 184名健康者IL 1A启动子 -889位点的SNP ,分析其基因型和等位基因频率。结果 :CC、CT、TT三种基因型 ,以C等位基因发生频率最高 ( 92 .4% ) ,T等位基因次之 ( 7.6% ) ;与挪威人群比较 ,其基因型和等位基因频率均存在极显著性差异 (P <0 .0 0 1) ,而与日本人群相比也有显著差异 (P <0 .0 5)。结论 :IL 1A启动子 -889位点存在SNP ,其在不同种族间的分布存在明显的差异 ,这种差异有可能是导致一些疾病在不同种族间的发病率和临床表现显著不同的因素之一     

P-选择素启动子区C-2123G、T-1817C基因多态性的研究

目的研究P-选择素(P-selectin)基因启动子区C-2123G、T-1817C多态性在中国湖北地区健康汉族人群中的分布,同时比较不同种族间基因型及等位基因频率分布差异.方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)的分析方法,检测200名健康者P-selectin基因启动子区C-2123G、T-1817C的基因型并计算其基因型频率及等位基因频率.结果中国湖北地区健康人群P-selectin C-2123G基因各基因型频率:CC型8.0%,CG型40.5%,GG型51.5%;C,G各等位基因频率分别为28.2%,71.8%,这种基因多态性分布在男女间无显著性差异(P＞0.05).与德国和英国比较,发现不同种族间P-selectin C-2123G基因型分布及等位基因频率均存在显著差异(P＜0.001,P＜0.001).P-selectin T-1817C基因各基因型频率:TT型65.5%,TC型28.5%,CC型6.0%;T,C各等位频率分别为79.75%,20.25%,这种基因多态性分布在男女间无显著性差异(P＞0.05).与英国比较,发现不同种族间P-selectin T-2123基因型分布及等位基因频率均存在显著差异(P＜0.001).结论中国湖北地区汉族人群中存在P-selectin启动子区C-2123G、T-1817C基因多态性,这种多态性在同种族男女间无差异,在各族间存在着较大的差异.     

广西瑶族与汉族人群TGF-β1基因启动子-509 C/T多态性的研究

目的研究TGF-β1(转化生长因子-β1)基因多态性在广西瑶族与汉族人群中的分布.方法应用聚合酶链反应-限制性片段长度多态性技术检测150名瑶族人和190名汉族人的TGF-β1基因多态性,比较两组人群TGF-β1基因型和等位基因的分布频率.结果在瑶族人中CC基因型占46.0%、CT基因型占43.3%、TT基因型占10.7%;在汉族人中CC基因型占30.0%、CT基因型占51.1%、TT基因型占19.9%.两组人群TGF-β1基因型的分布频率差异有显著性(P＜0.05).结论瑶族与汉族人群TGF-β1基因多态性分布频率差异有显著性.     

浙江汉族人群HLA-DPA1和-DPB1基因多态性分析

目的 检测浙江地区汉族人群HLA-DPA1和HLA-DPB1等位基因及单倍型频率.方法 应用PCR-直接测序分型法对100名健康、无血缘关系的浙江汉族人外周血标本进行HLA-DPA1和HLADPB1等位基因分析.结果 在100份标本中检出8个HLA-DPA1等位基因和19个HLA-DPB1等位基因.HLA-DPA1等位基因频率较高的依次为DPA1*020202(47.0%)、DPA1*010301(38.5%)和DPA1*020101(10.5%).HLA-DPB1等位基因中,频率较高的依次为DPB1*0501(39.5%)、DPB1*020102(13.5%)和DPB1*040101(13.0%).连锁分析发现共有44个HLA-DPA1-DPB1单倍型,单倍型频率最高为DPA1*020202-DPB1*0501(29.5%).结论 浙江地区汉族人群HLA-DPA1和DPB1基因座等位基因具有丰富的多态性,2个基因座呈现连锁不平衡.     

用聚合酶链反应-直接测序分型法研究江苏汉族人群HLA-DQA1和DQB1基因多态性

目的检测江苏地区汉族人群HLA-DQA1和DQB1等位基因及单倍型的频率，分析该人群DQA1、DQB1基因多态性和DQA1-DQB1单倍型特点。方法应用聚合酶链反应-直接测序分型法（PCR-sequence-based typing ，PCR-SBT）方法对100名健康、无血缘关系的江苏汉族人群的HLA-DQA1和DQB1进行基因分型。结果共检出7个DQA1等位基因和13个DQB1等位基因。DQA1等位基因中，DQA1＊0301／02／03的基因频率最高（29．5％），其次为DQA1＊0501（18．5％）、DQA1＊0102（17．0％）、DQA1＊0201（12．5％）；DQB1等位基因中，DQB1＊0201／02（21．5％）、DQB1＊0301／09（14．5％）、DQB1＊0303（13．5％）和DQB1＊0603（11．5％）最为常见。分析得出30种DQA1-DQB1单倍型，DQA1＊0301／02／03-DQB1＊0303（12．5％）、DQA1＊0201．DOB1＊0201／02（10．5％）、DQA1＊0501-DQB1＊0201／02（9．5％）、DQA1＊0501-DQB1＊0301／09（7．0％）为常见的单倍型。结论江苏汉族人群HLA-DQA1和DQB1基因具有较为丰富的多态性，基因频率分布具有中国北方群体的特征且具有一定的独特性。     

Polymorphism of DQA1 promoter in Italian population

We have investigated polymorphism in the 5′-URR of the DQA1 gene by PCR-SSO method in a group of 55 Italian healthy individuals olygotyped for DRB1, DQA1, DQB1 genes and in 20 10th IHWS cell lines as controls. We used primers and oligos (X and Y box) supplied by 12th IHWS and a DIG-11-ddUTP/AMPPD method. We have detected eight QAP variants (1.1,1.2,1.3,1.4,2.1,3.1,4.1,4.2) in our samples. As far as the association of DR/DQ haplotype and QAP sequences, we observed cases of one to one relationship (DQA1^*0201 and QAP2.1, DQA1^*0301 and QAP3.1, DQA1^*0401 and QAP4.2, DQA1^*0501 and QAP4.1); cases in which the same QAP allele was present in different DQA1-DRB1 haplotypes (QAP1.2 with DQA1^*0102 in DRB1^*15-DQB1^*0602 and DRB1^*16-DQB1^*0502 haplotypes or with DQA1^*0103 in the DRB1^*15-DQB1^*0601 haplotypes; QAP1.3 linked to DQA1^*0102, DQA1^*0103 or DQA1^*0104 in different haplotypes; QAP4.1 linked to DQA1^*0501 in DRB1^*11-DQB1^*0301, DRB1^*0301-DQB1^*0201, DRB1^*1303-DQB1^*0301 haplotypes or to DQA1^*0601 in DRB1^*0803-DQB1^*0301); cases where the same DQA1 allele is associated with different QAP sequences according to the DRB1 specificity (DQA1^*0102 allele with QAP1.2 or QAP1.4 in DRB1^*1302). Besides, we have observed that the QAP1.3, previously reported associated with DQA1^*0101-DRB1^*1401 haplotype, is really linked to DQA1^*0104-DRB1^*1401 haplotype. An intriguing data is that sometimes the same QAP is linked to different DQA1 alleles but to the same generic DRB1 allele: DRB1^*02 haplotype includes always the QAP1.2 variant but can bring different DQA1 alleles (^*0102 or ^*0103) and DRB1^*08 haplotype has always the QAP4.2 variant with different DQA1 alleles (^*0401 or ^*0601). The variability of linkage QAP-DQA1 can give further informations about HLA susceptibility in autoimmune diseases and in regulation of immune response in transplantation and oncology.     

Polymorphism in the upstream regulatory region of DQA1 gene in the Italian population

Abstract: Polymorphism in the 5'-upstream regulatory region of the DQA1 gene has been recently described. Using PCR-SSO method and SSCP analysis we have investigated this polymorphism in a group of 111 Italian blood donors which had been oligotyped for DRB1, DQA1 and DQB1 genes. Eight allelic variants were detected. Looking at the relationships among QAP sequences and DQA1 and DRB1 genes, three alternative situations were found: 1. a one-to-one relation between QAP and DQA1 alleles, independently of the other class II genes; 2. the same QAP allele in association with different DQA1-DRB1 haplotypes; 3. the same DQA1 allele with different QAP sequences according to the DRB1 specificity. No unexpected associations with DQB1 gene were found. These results must be interpreted considering that DQA1 and DRB1 genes are transcribed in opposite directions so that the promoter region of DQA1 gene lies between DQA1 and DRB1, close to the former but several hundreds kb away from the latter.     

Polymorphism of the DQA1 promoter region (QAP) and DRB1, QAP, DQA1, DQB1 haplotypes in the Dai minority population in South-Western China

We have investigated the polymorphism of the DQA1 promoter region (QAP) and we have deduced four point (DRB1, QAP, DQA1, DQB1) haplotypes of 60 unrelated healthy Dai minority individuals using the polymerase chain reaction and Dig-ddUTP labeled oligonucleotides. A total of eight QAP alleles (QAP1.1, 1.2, 1.3, 1.4, 3.1, 3.2, 4.1 and 4.2) were detected and two QAP alleles, QAP1.5 and QAP2.1 were absent in this population. The most predominant allele was QAP1.2 with 80% allele frequency. We also found that QAP alleles are in strong linkage disequilibrium with certain alleles of the neighboring loci DQA1 and DQB1. Complete positive association was found for QAP4.1-DQA1^*05, QAP4.2-DQA1^*0601, QAP1.2-DR2 group, QAP3.2-DRB1^*09, QAP4.1-DRB1^*03. A total of 28 different four point (DRB1-QAP-DQA1-DQB1) haplotypes were deduced and the most frequent haplotypes were DRB1^*1602-QAP1.2-DQA1^*0102-DQB1^*0502 (N = 18, H.f. = 15%) and DRB1^*09-QAP3.2-DQA1^*03-DQB1^*03032 (N = 18, H.f. = 15%) followed by the haplotypes DRB1^*1401-QAP1.3-DQA1^*01-DQB1^*0502, DRB1^*1202-QAP4.2-DQA1^*0601-DQB1^*0301 and DRB1^*1502-QAP1.2-DQA1^*0101-DQB1^*0501 with H.f. 9.1%, 6.7% and 5.0% respectively. The other 23 haplotypes were all less than 5% (H.f. 0.8%-5%). The relationship between the QAP alleles and DQA1 in the Dai minority is the same as that in the Chinese and the Caucasoid population.     

应用PCR-SSP方法研究越南中部京族HLA-DQA1等位基因的多态性

目的：检测越南中部京族HLA-DQA1等位基因的多态性。方法：应用PCR-SSP方法对105名健康，无血缘关系的越南中部京族儿童、青年进行HLA-DQA1分型。结果：检出10个HLA-DQA1等位基因，频率最高为DQA1*0104(21．3％)和最低为DQA1*0601(1．5％)。结论：结果表明越南中部京族人的HLA-DQA1等位基因分布与中国各民族的HLA-DQA1等位基因分布有差异。     

High-resolution genotyping of HLA-DQA1 in the GoKinD study and identification of novel alleles HLA-DQA1*040102, HLA-DQA1*0402 and HLA-DQA1*0404

In order to achieve high-resolution HLA-DQA1 genotyping, it is necessary to identify polymorphisms in exons 1, 2 and 3. We present a high-resolution sequence-based typing (SBT) strategy for genotyping exons 1, 2 and 3 of the polymorphic HLA-DQA1 locus. This method is an improvement upon previously presented methods, because it utilizes the minimum number of SSP-PCR assays to obtain clear DNA sequence in both the forward and reverse directions of all three exons. All known HLA-DQA1 alleles are resolved with the exception of HLA-DQA1*010101 and HLA-DQA1*010102 for which the distinguishing polymorphism is located in exon 4 and does not result in an amino acid change. This method has enabled our laboratory to identify three new HLA-DQA1 alleles - HLA-DQA1*040102, HLA- DQA1*0402 and HLA-DQA1*0404 - in the Genetics of Kidneys in Diabetes (GoKinD) study population. Additionally, we present single-allele amplification methods, which identify the coding sequences of HLA-DQA1 exons 1, 2, 3, intron 2 and 300 bp of the HLA-DQA1 promoter (QAP). This study, also describes the QAP for most of the known HLA-DQA1 alleles, three HLA-DQA2 promoter sequences and the intron 2 sequences for HLA-DQA1*040101, HLA-DQA1*040102, HLA-DQA1*0402 and HLA-DQA1*0404.     

中国海南黎族群体HLA-DRB1、DQA1、DQB1基因多态性研究

[摘要] 目的 了解海南黎族群体HLA-DRB1、DQA1、DQB1基因的遗传多态性。方法 应用PCR-SSP方法对随机抽取的94名海南黎族无血缘关系的健康个体进行HLA-DRB1、DQA1、DQB1基因分型。结果 鉴定了海南黎族群体HLA-DRB1位点的16种等位基因,DQA1位点的10种等位基因,DQB1位点的19种等位基因,包括了DR、DQ位点目前已知的全部血清学特异性,三个位点的基因型分布均符合Hardy-Weinberg平衡（P>0.5）。结论 提供了一套比较完整准确的海南黎族群体DRB1、DQA1、DQB1等位基因的基因频率和连锁不平衡参数。对群体遗传和疾病关联研究具有参考意义。     

越南中部京族HLA-DQA1等位基因多态性分析

目的　检测和分析越南中部京族HLA DQA1等位基因的多态性。方法　应用PCR SSP方法对 2 14名健康、无血缘关系的越南中部京族儿童、青年进行HLA DQA1分型。结果　检出 10个HLA DQA1等位基因 ,频率最高的为DQA1 0 10 4 (2 5 .8% ) ,最低的是DQA1 0 6 0 1(1% )。结论　越南中部京族人的HLA DQA1等位基因分布具有民族特征 ,与中国各民族及泰国人的HLA DQA1等位基因分布有差异。     

HLA—DQA1,DQB1基因与特发性膜性肾病关联研究

为探讨上海地区特发性膜性肾病（ＩＭＮ）与ＨＬＡ－Ⅱ类基因的关联，本研究用聚合酶链反应（ＰＣＲ）及序列特异性寡核苷酸（ＳＳO）探针杂交方法，对３３例上海地区汉族ＩＭＮ患者及７０例同一地区 正常人作了ＨＬＡ－ＤＱＡ１、－ＤＱＢ１位点的等位基因分型，并比较其基因频率。结果表明：ＩＭＮ组ＤＯＡ１０１０１等位基因频率高于对照组，统计学处理有显著性差异（ＲＲ＝３．０４３，Ｐ＝０．００２，Ｐｃ－０．０１８）。ＤＱＢ１０６０４等位基因频率较对照组高，而ＤＱＢ１０３０１等位基因频率较对照组低，但统计学处理后差异均不显著（Ｐ＜０．０５，Ｐｃ＞０．０５），以上结果提示，１ＭＮ的发病机制可能与免疫遗传因素有关。