人类白细胞Ⅱ类抗原DR、DQ基因型与妊娠期糖尿病的相关性研究

目的探讨人类白细胞II类抗原DR、DQ基因型与妊娠期糖尿病的相关性。方法对26例GDM孕妇及同期入院的42例正常健康孕妇,采用序列特异性引物聚合酶链反应技术(PCR-SSP)检测HLA-II类抗原DR和DQ的等位基因。结果研究中发现,DQA1*0101、DQA1*0201、DQB1*0609、DRB l*07-DQA1*0201-DQB1*0201基因频率在GDM中显著高于正常对照组,两组比较,统计学有差异(Ρ<0.05)。DQB1*0301基因频率在GDM中显著低于正常对照组,两组比较,有统计学差异(Ρ<0.05)。结论人类白细胞II类抗原DR、DQ基因型与GDM的易感性和保护性存在关联。DQA1*0101、DQA1*0201、DQB1*0609、DRB l*07-DQA1*0201-DQB1*0201基因是GDM的易感基因。DQB1*0301基因是GDM的保护基因。     

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     

采用PCR-RFLP技术检测中国人HLA纯合细胞DQ基因多态性

本文采用PCR-RFLP方法对7株中国人HLA纯合细胞的DQA1、DQB1基因作基因分型,检出了这些细胞株的DQ基因类型,证实了中国人群存在着一种DRw12与DQA1*0601-DQB1*0301连锁的新单倍型。     

云南汉族系统性红斑狼疮的易感及抵抗单体型研究

目的：探讨云南汉族系统性红斑狼疮（SLE）在HLA－DRB1、DQA1、DQB1等座位的易感抵抗单体型,方法：采用多聚酶链反应－序列特异性引物（PCR－SSP）技术对63例动态汉族SLE患者及54名同民族健康对照进行DRB1、DQA1、DQB1基因分型。结果：与正常对照组比较,SLE病人中有5个单体型频率显著升高;11个单体型频率在病例组中明显降低。结论：云南汉族SLE的易感单体型为DQA1^*0102-DQB1^*0601,DR15-DQA1^*0102-DQB1^*0601,DR15-DQA1^*0102-DQB1^*0602,DR15-DQA1^*0101-DQB1^*0601,DR15-DQA1^*0103-DQB1^*0601;其余均为低抗单体型。     

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。     

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。     

用PCR-SSO检测IgA肾炎HLA-DRB1、DQA1、DQB1等位基因及其临床意义

利用聚合酶链区应（PCR）和序列特异性寡核苷酸（SSO）探针技术对47例经临床及免疫荧光证实的IgA肾病（IgAN）患者HLA-DRB1、DQA1、DQB1等位基因频率进行了检测。结果显示IgAN患者组DR4基因频率明显高于正常人组,相反DQB1*0602基因频率与对照组相比呈显著下降。IgAN患者中蛋白尿组DR4基因频率显著高于对照组,而肉眼血尿组与对照组无显著差异。约 1/4 DR4基因阳性的IgAN病理表现为局灶节段硬化性肾小球肾炎。 IgAN肾衰组DR4阳性的发生率显著高于非肾衰组。由此可见,IgAN中HLA-DR4基因频率而著增高, DR4阳性IgAN临床多表现蛋白尿,易发生肾衰,病理多呈局灶节段硬化型;DQB1*0602等位基因对IgAN可能有一定抵抗性。这些研究结果提示IgAN有免疫遗传的背景。     

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。     

天花粉蛋白引起个体间CD8细胞负调节功能差异性表达的免疫学机制

在天花粉蛋白 (Tk )作用下 ,正常人群可因CD8细胞是否显示负向调节作用而分为介导型 (M+)和非介导型 (M-)两类 ,并作为一对遗传性状受HLA DQ基因调控。本研究表明 ,M-个体CD8细胞丧失抑制活性是因为激活了一群CD8VV+反抑制T细胞 (Tcs) ,而Tcs的激活可能依赖于CD4阳性诱导细胞的存在。由于Tk可以在M-个体中却不能在M+个体中激活这一诱导细胞 ,造成只有前者出现CD8Tcs及特定的应答格局。由此推论 ,DQ基因对Tk相关免疫抑制的调节点在Tcs的诱导水平。     

中国天疱疮患者的免疫遗传易感性研究

目的探讨中国天疱疮患者的免疫易感基因位点.方法本实验应用序列特异性引物聚合酶链式反应(SSP-PCR)技术对HLA-II类等位基因进行特异性体外扩增,分析了天疱疮患者HLA基因的DR位点和DRB1、DQB1多态性,并将天疱疮患者的基因频率与正常人结果进行χ2检验,以探讨中国天疱疮易感性基因位点.结果天疱疮患者中DR4的频率为93.3%(28/30),DR14的频率为63.3%(19/30),天疱疮患者中的DRB1*04和DRB1*14的频率分别为40%(12/30)和53.3%(16/30),经χ2分析,与正常对照组相比具有显著的差异性;在28个DR4+的天疱疮患者中有13个DRB1*0402,在19个DR14+天疱疮患者中DRB1*1401频率显著增高(P<0.01).所有DRB1*04+患者均为DQB1*0302,所有DRB1*14患者均为DQB1*0503.结论中国人天疱疮易感性与HLA的DR4、DR14抗原以及DRB1*04、DRB1*14、DRB1*0402、DRB1*1401基因位点与高度相关.     

HLA-DQ1 haplotype ruin the effect of cis complementation of DQA10501-DQB10201 in association with trichosanthin-induced immunosuppression

CD8 cell-mediator (M+) or non-mediator (M−) are distinguishable for healthy subjects according to whether their CD8 T cells keep the down-regulatory function in Trichosanthin (Tk)-induced immunosuppression. Tk is a plant protein of 247 amino acid residues purified from a Chinese medicinal herb. The M+ phenotype has been shown in our previous work to be strongly associated with HLA-DQ2. By genotyping with PCR-based techniques, the essential alleles of the DQ2 were identified as DQA1^*0501 and DQB1^*0201, which were either in cis (DR3) or in trans (DR5, DR7) position. A more detailed examination of the HLA association pattern with M+/M− in 42 Chinese candidates, however, revealed another two points of interest. 1) The cis complementation did not work if another DQA1^*01- or DQA1^*02-related haplotype (e.g. DRB1^*0101-DQA1^*0101-DQB1^*0501) were combined. The later seemed to behave like a ‘negative’ factor superimposed on the ‘positive’ role of DQA1^*0501-DQB1^*0201 haplotype in heterozygous condition. 2) In addition to DQA1^*0501, the DQB1^*0201 was actually able to combine all available DQA1 alleles except DQA1^*01 family to form the trans complementation. Again, the DQ1 haplotype acted negatively. It is thus likely that the cis and trans complementary association of DQA1^*0501-DQB1^*0201 could only be detected conditionally or only appeared as a special case in the Tk-induced immunosuppression.     

Cis-trans complementation of DQA1-DQB1 genes are modulated by DQ alleles: an immunogenetics analysis of DQ association with the down-regulatory function of CD8 cells in trichosanthin-induced immunosuppression

The initiation of a CD8 cell-mediated pathway (M⁺) was adopted as a phenotypic trait to analyse genetic predisposition in trichosanthin (Tk)-induced immunosuppression. Tk is a natural protein antigen with 247 amino acid residues. Based on DNA typing for DR, DQ, DP and TAP genes, data in this paper indicate that only DQ genes were primarily involved and that the alleles DQA1*0501 and DQB1*0201 were strongly associated with the M⁺ phenotype in cis (on DR3 haplotype) or trans (on DR5/DR7 heterozygotes) complementation. This is consistent with our observation that only the DQ-positive cells were capable of expanding after being co-cultured with Tk for 96h. Two points of interest were noted. (1) The susceptible haplotype DRB1*0301-DQA1*0501-DQB1*0201 showed an association with the M⁺ phenotype only if combined with DRB1*04-, DRB1*08-, or DRB1*09-related haplotypes. When co-presented with DRB1*11-, DRB1*15-, DRB1*07-related haplotypes, however, no cis complementation could be detected. A detailed analysis of the association patterns indicated that the DQB1 locus of the non-susceptible haplotypes was the main factor for up- or down-modulation. (2) For M⁺ phenotype-related trans complementation in Tk-induced suppression, it was found that not only DQA1*0501-DQB1*0201 (DR5/7) alleles, but also associated DQA1*0301-DQB1*0201 (DR4/7, 9/7) alleles, were involved. The allele DQB1*0201 was not associated with the DQA1 alleles in DRB1*01-, DRB1*15-, DRB1*13-, DRB1*07-related haplotypes. The results obtained indicate that there are some additional genetic factors involved in the functional expression of cis and trans complementation of DQA1 and DQB1 genes, among which the DQ alleles play a critical role as self-regulators.     

Asp57-negative HLA DQ beta chain and DQA1*0501 allele are essential for the onset of DQw2-positive and DQw2-negative coeliac disease.

The genetic predisposition to coeliac disease is associated with the HLA DQw2 allele. Coeliac patients lacking the DQw2 allele are very rare and always exhibit the DR4-DQw3 haplotype. We performed oligotyping of polymerase chain reaction (PCR)-amplified DQA1 and DQB1 genes in six DQw2-negative and 30 DQw2-positive coeliac patients. The DQB analysis showed that all six DQw2-negative patients possessed the DQB1*0302 allele. The other DQB alleles found in five of these patients were DQB1*0501, DQB1*0604 and DQB1*0302. The DQ beta chains encoded from all these alleles have the replacement of aspartic acid residue at position 57 (Asp57), as well as the DQB1*0201 allele which was found in all 30 DQw2-positive coeliac patients. The DQw2-negative proband who lacked the homozygous Asp57 replacement exhibited the DQA1*0501 allele in the DQA1 gene. The DQA1*0501 allele was also found in 27 of the 30 DQw2-positive coeliac patients. Among this group of coeliacs, the four cases lacking the DQA1*0501 allele exhibited the homozygous Asp57 replacement in the DQ beta chain. Our results indicate that Asp57-negative DQ beta alleles are involved in both DQw2-positive and -negative coeliac patients. Moreover, when the Asp57-negative DQ beta chain is encoded from only one of the two DQB1 genes the DQA1*0501 allele is always present.     

The HLA DQB 1*0502 allele is neutrally associated with insulin-dependent diabetes mellitus in the Sardinian population

In the Sardinian population a very high incidence of insulin-dependent diabetes mellitus (IDDM) and the lack of HLA-DR2 protective effect due to the high frequency of the A2, Cw7, B17, 3F31, DR2, DQw1 extended haplotype has been reported. This haplotype, carrying a Serine at position 57 of the DQB1*0502 allele, has been previously reported to be underrepresented in patients when compared to controls. In order to provide an explanation for this finding, we defined by RFLP analysis the HLA haplotype of 45 Sardinian IDDM patients and 49 controls. All DR-2DQw1 subjects were molecularly characterized at the HLA DQA and DQB loci. All DR2-positive patients and the vast majority of the DR2-positive controls had the DQB1*0502 allele at the DR2-linked DQB1 locus, with no statistically significant difference between the two groups. All DQA1 genes were the ones expected, with only two exceptions. Nine out of 10 of the DR2-positive patients were compound heterozygotes for DQB1*0201/DQB1*0502 alleles; only this allele combination was significantly increased (p less than 0.0003). Our data suggests that a) the DQB1*0502 allele is neutral for IDDM development and b) the susceptibility to IDDM in our DR2-positive patients is related to the compound heterozygous state between the neutral DQA1*0102/DQB1*0502 and the susceptibility DQA1*0501/DQB1*0201 alleles.     

原发性肥厚型心肌病与HLA-DQ的相关性研究

本文用PCR/SSO方法对41例原发性肥厚型心肌病患者和52例正常人HLA-DQA1和DQB1基因的多态性进行分析.发现,在肥厚型心肌病患者中,DQA1＊0201,DQB1＊0504,0502等位基因频率明显较低,其相对风险值分别为9.51、5.87和11.60;而DQA1*0501,DQB1＊03031的频率明显地高,相对风险值分别为2.93和6.65.初步认为,原发性肥厚型心肌病与某些HLA-DQ基因相关联.     

A study of Italian pediatric celiac disease patients confirms that the primary HLA association is to the DQ(alpha 1*0501, beta 1*0201) heterodimer.

Celiac disease (CD) has been recently reported to be primarily associated with the DQ(alpha 1*0501, beta 1*0201) heterodimer encoded in cis on DR3 haplotype and in trans in DR5,7 heterozygous individuals. The high incidence of DR5,7 heterozygotes, reflecting the high frequency of the DR5 allele in Italy, makes the analysis of the Italian CD patients critical. Polymerase chain reaction-amplified DNA from 50 CD patients and 50 controls, serologically typed for DR and DQw antigens, was hybridized with five DQA1-specific oligonucleotide probes detecting DQA1*0101 + 0102 + 0103, DQA1*0201, DQA1*0301 + 0302, DQA1*0401 + 0501 + 0601, and DQA1*0501 and a DQB1-sequence-specific oligonucleotide probe recognizing DQB1*0201 allele. As expected by the DR-DQ disequilibria, DQA1*0201 [62% in patients versus 26% in controls, relative risk (RR) = 5] and DQA1*0501 (96% versus 56%, RR = 19) show positive association with the disease. Of CD patients, 92% (50% DR3 and 42% DR5,7) compared to 18% of the controls carry both DQA1*0501 and DQB1*0201 alleles, so that the combination confers an RR of 52, higher than both the risks of the single alleles (DQA1*0501 RR = 19, DQB1*0201 RR = 30), confirming the primary role of the dimer in determining genetic predisposition to CD both in DR3 and in DR5,7 subjects.     

Genetic differences in HLA-DQA1* and DQB1* allelic distributions between celiac and control children in Santiago, Chile.

Celiac disease is a permanent gluten intolerance strongly associated with HLA class II antigens. The over presentation of particular HLA alleles and haplotypes has been described in several populations. Different lines of evidence obtained during the last years suggest that a particular HLA-DQ heterodimer, encoded by the DQA1*0501 and DQB1*0201 genes in cis or trans conformation, confers the primary disease susceptibility. We report the HLA class II allelic distribution and DQA1/ DQB1 genotypes in 62 Chilean celiac patients compared with 124 control subjects in Santiago, Chile. We found a pronounced increase of the "susceptible" alleles :DQA1*0501 (0.480 vs 0.169, Pc < 0.0005), DQB1*0302 (0.430 vs 0.242, Pc = 0.002) and DQB1*0201 (0.250 vs 0.125, Pc = 0.037) in celiac patients in comparison with control children. As for "protective" alleles, we detected a high frequency of DQA1*0101 (0.310 vs 0.160, Pc = 0.01), DQA1*0201 (0.105 vs 0.010, Pc < 0.0075) and DQB1*0301 (0.250 vs 0.100, Pc = 0.010) in controls. In relation to risk haplotypes, the main combination observed was the conformation DQ8 (DQB1*0302/DQA1*0301) over DQ2 (DQB1*0201/DQA1*0501). In conclusion, results show that celiac disease in Chilean patients is primarily associated with DQ8 conformation. This is concordant with the high frequency of DR4 alleles (in linkage disequilibrium with DQB1*0302) detected in Amerind groups in Chile, where DQB1*0302 is more frequent than DQB1*0201.     

DRB1-DQA1-DQB1 loci and multiple sclerosis predisposition in the Sardinian population

Multiple sclerosis (MS) is a common neurological disease caused by genetic and environmental factors. Previous genetic analyses have suggested that theMHC/HLA region on chromosome 6p21 contains an MS- predisposing component. Which of the many genes present in this region is primarily responsible for disease susceptibility is still an open issue. In this study, we evaluated, in a large cohort of MS families from the Mediterranean island of Sardinia, the role of allelic variation at the HLA-DRB1, DQA1 and DQB1 candidate loci in MS predisposition. Using the transmission disequilibrium test (TDT), we found significant evidence of association with MS in both the Sardinian- specific DRB1*0405(DR4)- DQA1*0501-DQB1*0301 haplotype and the DRB1* 0301(DR3)-DQA1*0501-DQB1*0201 haplotype. Detailed comparative analysis of the DRB1-DQA1- DQB1 haplotypes present in this data set did not identify an individual locus that could explain MS susceptibility. The predisposing effect is haplotype specific, in that it is confined to specific combinations of alleles at the DRB1, DQA1 and DQB1 loci. Cross- ethnic comparison between the two HLA haplotypes associated with MS in Sardinians and the DRB1*1501 (DR2)-DQA1*0102-DQB1* 0602 haplotype, associated with MS in other Caucasian populations, failed to identify any shared epitopes in the DR and DQ molecules that segregated with disease susceptibility. These results suggest that another MHC gene(s), in linkage disequilibrium with specific HLA-DRB1, DQA1, DQB1 haploypes, might be primarily responsible for genetic susceptibility to MS. Alternatively, the presence of complex interactions between different HLA haplotypes, other non-HLA predisposing genes and environmental factors may explain different associations in different populations.      

HLA-DR and -DQ genotypes of celiac disease patients serologically typed to be non-DR3 or non-DR5/7.

The susceptibility to develop celiac disease (CD) seems to be primarily associated to a particular HLA-DQ alpha/beta heterodimer encoded by the DQA1*0501 and DQB1*0201 alleles, in cis position on the DR3-DQ2 haplotype or in trans position by DR5-DQ7/DR7-DQ2 heterozygotes. However, exceptional patients exist who are neither DR3 nor DR5/DR7, particularly among Southern European populations. We therefore examined the DRB1, DQA1, and DQB1 alleles of 13 Spanish CD patients who were serologically typed to be neither DR3 nor DR5/DR7. Five patients were found to carry the DQA1*0501 and DQB1*0201 alleles either in cis or in trans position, three of them had previously been serologically mistyped. However, two of these patients carried DQA1*0501 and DQB1*0201 on haplotypes other than DR3 or DR5 in combination with DR7. One of the latter patients carried an unusual DR4-DQ2 haplotype, while another had an unusual DR8-DQ2 haplotype. Four of the remaining eight patients carried DR4-DQ8 haplotypes. Taken together, our findings provide further evidence that the DQ alpha/beta heterodimer encoded by the DQA1*0501 and the DQB1*0201 alleles confers the primary HLA-associated susceptibility to develop CD. However, our studies also corroborate that a second (and "weaker") HLA-associated CD susceptibility gene may be present on some DR4-carrying haplotypes.     

The distribution of DQ genes in the Saharawi population provides only a partial explanation for the high celiac disease prevalence

Celiac disease (CD) is a multifactorial disorder of the small intestine caused by a permanent dietary intolerance to gluten. The combined presence of the HLA class II DQA1*0501 and DQB1*0201 alleles represents the major genetic component for disease predisposition. It has been shown that the Saharawi refugees living in northern Africa have a very high frequency of CD. In the present study we analysed this population to evaluate the degree of association with CD of the haplotypes and genotypes at the main HLA-DQB1 and DQA1 disease loci. We found a strong association of the DR3, DQB1*0201-DQA1*0501-positive haplotypes and genotypes. A very high frequency of DR3, DQB1*0201-DQA1*0501 was also observed in the general Saharawi population. These results indicate that there is a good correlation between disease prevalence and frequency of the main predisposing haplotype in the background population. However, the correlation is incomplete because similar frequencies of DR3 are also observed in populations such as the Sardinians showing a much lower prevalence of CD. We can conclude that the distribution of DQ genes in the Saharawi population only provides a partial explanation for the high prevalence of CD. Other factors, such as rapidly changing dietary habits and/or non-DQ genes, may also play some role.