HLA-DQA1 Polymorphism in Idiopathic Dilated Cardiomyopathy

Idiopathic dilated cardiomyopathy (IDC) is a disease ofunknown etiology, for which immune abnormalities, possi bly related to viral infections, are widely suspected butunproven. Many abnormalities in humoral and cellular re sponses have been reported, including the previous sero logic studies of correlation between human leukocyte anti gen DR4 and IDC[1]. However there was little informa tion on the relation between antigens at the DQA1 loci ofthe HLA system and idiopathic dilated c…     

Association of HLA-DQ with idiopathic dilated cardiomyopathy in a northern Chinese Han population

Autoimmune mechanisms are likely involved in the pathogenesis of idiopathic dilated cardiomyopathy (IDC)and components of MHC may serve as markers for the propensity to develop immune-mediated myocardialdamage.This study was conducted to investigate the possible association between HLA-DQA1,-DQB1 allelesand IDC in Han population from northern China by using PCR-based sequence-specific primer (PCR-SSP)technique for HLA genotyping.Among 68 unrelated IDC patients,4 probands of IDC pedigrees and 100healthy controls,we found that the alleles of HLA-DQA1*0501 and HLA-DQB1*0303 conferred susceptibilityto IDC while HLA-DQA1*0201 and HLA-DQB1*0502,*0504 alleles were in negative association with IDC.Theserine at position 57 (SER~(57)) in the exon of HLA-DQB1*0502 and *0504 was confirmed in our experiment as amarker for resistance to IDC.The results suggest that HLA-DQ polymorphism may be involved in thepathogenesis of IDC.Cellular & Molecular Immunology.2004;1 (4):311-314.     

HLA-DQA1和-DQB1等位基因与乙型病毒性肝炎相关性的研究

不同个体对乙型肝炎病毒(Hepatitis B virus, HBV)易感性的差异与个体的免疫特性有关,而后者主要取决于人类主要组织相容性复合体(Major histocompatibility complex, MHC)。人类白细胞抗原(Human leucocyte antigen, HLA)是MHC的基因产物,是首次发现的与疾病有明确关系的遗传系统。HLA复合体作为调节机体免疫应答的重要基因群,与抗HBV免疫反应有着密切的关系,某些特殊的HLA基因型可能影响着HBV感染的慢性化和免疫反应的强弱。作者利用PCR/     

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。     

Association of HLA class II DRB1, DPA1 and DPB1 polymorphism with genetic susceptibility to idiopathic dilated cardiomyopathy in Chinese Han nationality

Although the aetiology of idiopathic dilated cardiomyopathy (IDC) remains unclear, many immunological abnormalities involving changes in cell-mediated and humoral immunity may be associated with cardiac impairment in IDC. Autoimmune mechanisms are likely to participate in the pathogenesis of at least a subgroup of IDC and components of the major histocompatibility complex may serve as markers for the propensity to develop immune-mediated myocardial damage. Human leukocyte antigen (HLA) class II genes, which are highly polymorphic, play an important role in the activating of immune responses and thus control the predisposition for or protection from IDC. This study explores the possible contribution of HLA-DRB and DP polymorphisms to IDC susceptibility. DNA genotyping for HLA-DRB1, DPA1 and DPB1 was performed using polymerase chain reaction-sequencing based typing (PCR-SBT) method in 198 IDC patients and 136 random selected healthy Han ethnic individuals living in Northern China. IDC patients were, sub-grouped into asymptomatics (subgroup A), with arrhythmia (subgroup B) and with overt congestive heart failure (subgroup C) according to the clinical manifestations and electrocardiogram or echocardiographic characteristics. ADP/ATP autoantibody was detected in IDC group by immunoblot analysis. The results revealed that HLA-DR15, -DPB*0601 frequencies were significantly elevated in IDC group compared with normal control. The DPB1*0601 allele in homozygous form or in combination with allele DPB1*2301 or *3901, was found present more often in IDC patients. The predominance of HLA-DR4 allele was observed in subgroup B after stratification. However, the frequency of DPB1*0101 allele increased in the control than in the IDC group. The frequency of HLA-DPB1*0601 allele was significantly higher in IDC patients with positive autoantibody against ADP/ATP carrier of myocardial mitochondria in contrast to those with negative autoantibody. We conclude that HLA-DR4, -DR15, -DPB1*0601 alleles confers susceptibility to, while DPB1*0101 allele confers protection from IDC among individuals of northern Chinese Han nationality. The glutamate at position 69 in the second exon of DPB1*0601, as a key residue for special conformation of HLA-DP, may confer predisposition to IDC. HLA-DR and -DP alleles polymorphisms may serve as genetic markers for IDC and be involved in the regulation of the immune specific response to auto or exterior anti-myocardium antibodies.     

High-resolution sequence typing of HLA-DQA1 and -DQB1 exon 2 DNA with taxonomy-based sequence analysis (TBSA) allele assignment

High-resolution DNA sequencing of exon 2 of DQA1 and DQB1 genes that uses a taxonomy-based sequence analysis (TBSA) method to assign alleles was developed. The system uses fewer primers for polymerase chain reaction (PCR) amplification and sequencing than other methods and yields accurate DQA1 and DQB1 typing when either homozygous or heterozygous DNA samples are tested. The approach was initially corroborated by the correct typing of 10 blinded samples that had been previously typed by PCR using sequence-specific oligonucleotide probes (PCR-SSOP) or serology, and subsequently confirmed by sequencing of cloned PCR products. DNA from peripheral blood cell samples of 130 individuals enrolled in a case-control analysis of HLA determinants of abdominal aortic aneurysm were subsequently evaluated. Overall, 8 different DQA1 and 19 DQB1 alleles were identified. All 21 DQA1 heterozygous combinations and 45 of 49 DQB1 heterozygous combinations were successfully resolved with TBSA. The two pairs of heterozygous DQB1 combinations that were not unambiguously typed required sequence specific PCR amplification for correct allele identification. We conclude that the method provides precise analysis for HLA-DQ typing.     

青海土族、撒拉族HLA-DQA1和-DQB1基因多态性探讨

目的 :检测青海土族和撒拉族健康个体HLA DQA1和 DQB1等位基因频率 ,了解和分析这两个民族的HLA DQA1和 DQB1基因多态性。方法 :用PCR SSO方法检测 132名土族和 80名撒拉族健康个体的HLA DQA1和 DQB1的基因多态性 ,并将所得结果与国内其他民族的同类资料进行比较。结果 :土族和撒拉族在HLA DQA1和 DQB1高频率等位基因分布上有共同点 ,也有一定的独特性。这两种民族HLA DQA1 0 10 2和 0 5 0 1以及DQB1 0 2 0 1、 0 30 1、 0 4 0 2和 0 6 0 2基因分布频率上具有显著性差异。结论 :土族和撒拉族与北方诸民族的等位基因频率接近 ,而与其他南方诸民族的差异相对较大。     

单核苷酸多态性与大肠癌遗传易感性

大肠癌遗传易感性与单核苷酸多态性(SNP)的关系是近年来研究的热点。研究发现COX2,MTHFR等代谢相关基因的某些SNP与大肠癌的发病风险相关,其中携带COX2 9850G-10335A单倍型的个体可显著增加患大肠癌的风险。MMP家族是调控大肠癌侵袭转移的重要基因,MMP7-181G等位型频率可显著增加大肠癌淋巴结转移风险。进一步寻找大肠癌特异性SNP,对筛选大肠癌高危人群,预估发病风险,具有重要意义。     

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.     

Novel associations among HLA-DQA1 and -DQB1 alleles, revealed by high-resolution sequence-based typing (SBT)

Althought it is a valuable tool for the identification of HLA alleles, sequence-based typing (SBT) presents difficulties when used to determine HLA-DQA1 and -DQB1 alleles. Specifically, some HLA-DQA1 alleles have a three-base deletion at codon 56 of exon 2 that interferes with the sequencing read. Moreover, the frequently used primers for HLA-DQB1 may co-amplify the HLA-DQB2 pseudogene. To overcome these problems, we amplified DQA1 exon 2 using five group-specific polymerase chain reactions (PCRs) which allowed separation of deleted from non-deleted DQA1 alleles. DQB1 exon 2 was amplified using two group-specific amplifications. To increase typing resolution, we also analyzed DQA1 exons 1, 3 and 4 and DQB1 exon 3 by PCR using sequence-specific primers (PCR-SSP) or SBT analysis. Using this method we found some important associations between DQA1 and DQB1 alleles: DQA1*05011 and DQB1*0201, DQA1*0505 and DQB1*03011, DQA1*01021 and DQB1*06, DQA1*01022 and DQB1*0502.     

Association of HLA-DRB1*1502-DQB1*0501 haplotype with susceptibility to systemic lupus erythematosus in Thais

In this study, we investigated the association of HLA-DRB1 and DQB1 with Thai patients with SLE. A highly significant increase in the frequency of DRB1*1502 and DQB1*0501 was found in SLE patients compared with normal controls. DRB1*1501 and DRB1*1602 were also slightly increased. In contrast, DRB1*1202 and DQB1*0301 were decreased, and DRB1*0406 and DRB1*1401 were not found in the patients' group. The haplotype analysis revealed that DRB1*1502 - DQB1*0501 was most strongly associated with SLE, and also suggested a primary role for DRB1 rather than DQB1. Taken together with the previous report which demonstrated the association of the same haplotype in Taiwan, our present observations strongly suggested that DRB1*1502 - DQB1*0501 is the major HLA haplotype that confers susceptibility to SLE in the South-east Asian populations.     

Distribution of HLA-DQA1 and -DQB1 alleles and DQA1-DQB1 genotypes among Senegalese patients with insulin-dependent diabetes mellitus

Transracial analysis is one method for distinguishing primary associations between insulin-dependent diabetes mellitus (IDDM) and HLA II alleles from those related to linkage disequilibrium. Black people have different DR-DQ relationships from other races and are a useful group to investigate HLA-D regions associated with IDDM. In this study, we compared the frequencies of HLA-DQA1 and DQB1 alleles in Senegalese IDDM and control subjects. DQA1*0301 was positively associated with insulin-dependent diabetes mellitus (p<10^-9, OR 5.21), as were DQB1*0201 and *0302 (p<10^-7 OR=3.55, p<10^-3 OR=3.20, respectively). The positive associations with DQA1*0301, DQB1*0201 and DQB1*0302 are consistent with all racial groups investigated. However, taken together, the data in Senegalese population show that susceptibility and resistance to IDDM are associated both with particular haplotypes and DQA1-DQB1 heterodimers.     

HLA-DQA13’-UTR序列多态性分析及microRNA的调控

目的 揭示HLA-DQAI基因3’-UTR区多态性的进化机制及其对基因表达的调控功能.方法 利用核苷酸变异率计算、系统发育树构建等群体遗传学分析方法将IMGT/HLA数据库中释放的20条HLA-DQAl全长序列多态性进行系统分析,揭示3’-UTR多态性的进化机制.利用miRanda vl.9软件提供的算法,对HLA-DQA1三种差异较大的3'UTR等位基因序列进行microRNA的靶位点预测,对分值较高的预测利用体外荧光细胞实验进行验证.结果 HLA-DQA1基因的3’-UTR具有极高的核苷酸变异率及跨物种多态性,表明其受到平衡选择的作用.预测出14种分值较高的microRNA-靶标复合 物,且14个microRNA与3种DQA1-3’-UTR等位基因靶标的结合自由能以及分值均有不同程度的差异.通过体外荧光表达实验证明,HLA-DQAl受到hsa-mir-658的负调控作用,且hsa-mir-658对DQAl-3’-UTR 3种等位基因特异性的结合对基因表达产生了影响.结论 HLA-DQAl基因表达水平受到microRNA的负调控,且HLA-DQA1 3’-UTR的多态性可影响与microRNA的结合而引起等位基因特异性表达.     

Association of HLA-DQA1 (rs9272219) with susceptibility to rheumatoid arthritis in a Han Chinese population

HLA-DQA1 (rs9272219) has been previously reported that it is a susceptibility locus in rheumatoid arthritis (RA) of UK Caucasian population and North American; however, it has not reported in RA of Chinese population. Our study was to identify whether or not this relationship is reside between rs9272219 and RA in a Han Chinese population. 207 patients with RA and 199 control subjects were recruited. The single nucleotide polymorphism (SNP) of rs9272219 was tested in alleles and genotype frequencies and the data was analyzed by doing the statistic analysis of odds ratio (OR) and 95% confidence interval (CI) from multivariate unconditional logistic regression analyses after pairwise linkage disequilibrium (LD) was estimated. Finally, the Alleles and genotype frequencies distribution of rs9272219 locus among RA patients and control subjects were in accordance with Hardy-Weinberg equilibrium. We found significant association between rs9272219 and RA of Chinese population (OR 0.494, 95% confidence interval [95% CI] 0.354-0.688, P = 0 and OR 2.541, 95% CI 1.695-3.808, P = 0, respectively). In this study, we found that the SNP of rs9272219 in HLA-DQA1 is a potential susceptibility locus in RA of Han Chinese population; the results suggest that HLA-DQA1 may be related to the development of RA.     

Genetic Relationship Between IL-6 rs1800796 Polymorphism and Susceptibility to Periodontitis

Background: There are accumulating reports for the potential role of Interleukin-6 (IL-6) rs1800796 polymorphism in the risk of periodontitis. However, distinct conclusions are observed. In this study, we have an interest in comprehensively analyzing the genetic relationship between IL-6 rs1800796 and the susceptibility to periodontitis.

Methods: We retrieved the eligible case-control studies from on-line database and conducted a meta-analysis. P-value of association test, OR (odd ratios) and 95% CI (confidence interval) were calculated for the assessment of potential genetic association.

Results: We enrolled a total of 20 case-control studies for pooling analysis. A positive association between periodontitis cases and controls was observed in the overall meta-analysis under all genetic models (all P < 0.05, OR > 1). Similar results were detected in the “population-based, PB” and “China” subgroups (all P < 0.05, OR > 1). In the “Asian” subgroup, there is an increased periodontitis risk under the allele, homozygote, heterozygote, dominant and carrier models (all P < 0.05, OR > 1). Nevertheless, negative results were found in the “Caucasian” subgroup under all models [all P > 0.05]. In addition, a positive association between IL-6 rs1800796 and the risk of chronic periodontitis was detected under the models of allele [G vs. C], GG vs. CC, GG vs. CC+ CG and carrier [G vs. C] (all P < 0.05, OR > 1).

Conclusion: IL-6 rs1800796 may serve as one genetic risk factor for periodontitis patients in the Asian population, especially the Chinese population. G/G genotype of IL-6 rs1800796 appears to be associated with an increased risk of chronic periodontitis.     

内蒙古汉族儿童过敏性紫癜与HLA-DQA1基因的关联性研究

目的 探讨内蒙古汉族儿童HLA－DQA1等位基因与过敏性紫癜（anaphylactoid purpura,AP）的遗传易感性及其与临床特点的关系。方法 用聚合酶链反应－序列特异性引物技术，对70例内蒙古汉族儿童AP和90名健康儿童HLA－DQA1等位基因进行对比分析。结果 （1）病例组DQA1＊0301基因频率为33.4%，明显高于对照组（10.6%）（P＜0.01）；DQA1＊0302基因频率为6.7%，明显低于对照组（19％）（P＜0.01)。（2）单纯皮肤紫癜病例组与对照组DQA1＊0301和0302基因频率比较，差异无显著性（P＞0.05）；伴有胃肠、关节、肾脏损害病例DQA1＊0301基因频率分别为26.7%、28.5%和29.3%，均明显高于对照组（10.6%）（P均＜0.01）；而DQA1＊0302基因频率分别为3.9%、5.7%和9.6%，分别与对照组（19％）比较，肾脏损害组差异无显著性（P＞0.05），胃肠和关节损害组均明显降低（P均＜0.01）。结论 HLA－DQA1＊0301等位基因可能是内蒙古汉族儿童AP发病单体型中一个遗传易感基因，具此基因者可能更易出现胃肠、关节和肾脏损害；而DQA1＊0302等位基因可能为其遗传保护基因，对出现胃肠、关节损害可能有拮抗作用。     

葡萄糖醛酸转移酶1F基因多态性及与肝癌易感性研究

目的 探讨葡萄糖醛酸转移酶1F（UDP－glucuronosyltransferase 1F,UGT1F）基因的多态性及其与肝癌的关系。方法 应用聚合酶链反应、变性梯度凝胶电泳、单链构象多态和DNA测序技术，对84例肝癌患者和144名健康对照的UGT1F基因的多态性进行了研究。结果 在第1外显子和第2内含子发现3个新的单核苷酸多态。即第232位核苷酸T→G颠换，第528位核苷酸A→G转换和第376位核苷酸A→G转移。另外，754位点多态在我们的研究中得到了证实。分析病例组和对照组4个位点等位基因及基因型频率的分布，病例组UGT1F754位点G／G基因型频率（13.10%）和G等位基因频率（29.17%）均显著高于对照组（2.78%和19.44%）。其它位点，两组差异无显著性。结论 UGT1F基因的第2－5外显子高度保守，而第1外显子则呈高度多态。其754位点多态可能与肝癌有关。     

DNA修复基因hOGG1多态与食管癌遗传易感性

目的：研究修复８－羟基鸟嘌呤的ｈＯＧＧ１基因Ｓｅｒ３２６Ｃｙｓ多态与中国人食管癌易感性的关系。方法：采用病例－对照分子流行病学方法,以ＰＣＲ－单链构象多态（ｓｉｎｇｌｅ ｓｔｒａｎｄ ｃｏｎｆｏｒｍａｔｉｏｎ ｐｏｌｙｍｏｒｐｈｉｓｍ,ＳＳＣＰ）技术,分析了２０１名正常对照和１９６例食管癌患者ｈＯＧＧ１基因第３２６位Ｓｅｒ／Ｓｅｒ、Ｓｅｒ／Ｃｙｓ和Ｃｙｓ／Ｃｙｓ基因型分布,并比较不同基因型与食管癌风险的关系。结果：正常人群的Ｓｅｒ／Ｓｅｒ、Ｓｅｒ／Ｃｙｓ和Ｃｙｓ／Ｃｙｓ基因型频率分别为３３．８％、５２．８％和１３．４％,而食管癌病例组则分别为３９．８％、３８．８％和２１．４％。虽然两组人群的Ｃｙｓ等位基因频率基本相同（３９．８％４０．８％）,但食管癌病例组的Ｃｙｓ／Ｃｙｓ基因型频率显著高于对照组（Ｐ＜０．０５）     

HLA-DQA1、-DRB1等位基因与子宫内膜异位症和子宫腺肌病的比较性研究

目的 从免疫遗传学角度比较子宫内膜异位症和子宫腺肌症的异同。方法 采用顺序特异引物聚合酶链反应技术检测51例子宫内膜异位症和45例子宫腺肌病患者的HLA－DQA1和HLA－DRB1等位基因频率，并与44名正常人比较。结果 两患者组HLA－DQA1＊0301等位基因频率（8.8%,5.6%）均明显低于正常对照组，差异有显著性（Pc=0.00,Pc=0.00），而两患者组的HLA－DQA1＊0.0401等位基因频率（7.8％,10.0%）均明显高于对照组，差异有显著性（Pc=0.03,Pc=0.01）；两患者组之间HLA－DQA1、－DRB1各等位基因频率比较，差异无显著性。结论 子宫内膜异位症及子宫腺肌病均与HLA－DQA1＊0301、＊0401相关联，从HLA－DQA1、－DRB1角度分析，子宫内膜异位症与子宫腺肌病的发病机理可能有共同之处。     

Distribution of HLA-DQA1, -DQB1 and DRB1 alleles in black IDDM patients and controls from Zimbabwe

We have used the XI Histocompatibility Workshop sequence-specific oligonucleotide probes to determine the DRB1, DQA1 and DQB1 genotypes by dot-blot hybridization of polymerase chain reaction (pcr)-amplified material from a homogenous black population in Zimbabwe. The DR4 subtype DRB1*0405, the DR3 subtype DRB1*0301, DQB1*0201 and DQB1*0302 and DQA1*0301 and DQA1*0501 were significantly increased in the IDDM group compared to the controls, whereas DRB1*11, DQB1*0602 and DQA1*0102 were significantly decreased. Taken together, the data show that susceptibility and resistance to IDDM are associated both with particular haplotypes and DQA1-DQB1 heterodimers without one or other being overriding.