中国胰岛素依赖性糖尿病病人（HLA－DR9）的HLA－DQB1基因顺序分析

用血清学方法研究显示中国人胰岛素依赖性糖尿病（ＩＤＤＭ）与ＨＬＡ－ＤＲ９相关。鉴于白种人中的研究显示ＩＤＤＭ与ＨＬＡ－ＤＱβ链第５７位氨基酸相关，Ａｓｐ－５７对ＩＤＤＭ呈抗性，ｎｏｎ－Ａｓｐ与ＩＤＤＭ易感性相关。我们用ＰＣＲ技术扩增了中国人中血清学ＤＲ９纯合的ＩＤＤＭ患者和正常对照的ＨＬＡ－ＤＱＢ１基因第二外显子并测定了核苷酸顺序，结果未发现ＩＤＤＭ特异ＨＬＡ－ＤＱＢ１等位基因，但发现ＩＤＤＭ病人ＨＬＡ－ＤＱＢ１５７位均为天冬氨酸。表明中国ＩＤＤＭ患者中的ＨＬＡ－ＤＱＢ１５７位天冬氨酸不一定具有保护个体抵抗ＩＤＤＭ的足够能力。ＩＤＤＭ易感性可能涉及多个基因位点的变化，另外还可能与其它遗传因素及环境因素有关。     

胰岛素依赖性糖尿病与HLA的关联研究

胰岛素依赖性糖尿病是自身免疫性疾病,目前认为遗传因素与环境因素相互作用决定,该病的发生,而遗传因素主要是与人类白细胞的抗原系统ＨＬＡ相关,且主要涉及ＨＬＡ－Ⅱ类区域,此外,ＨＬＡ－Ⅰ类基因是ＨＬＡ内非Ⅰ,Ⅱ类基因对该病发生也有一定的作用。     

天花粉蛋白诱发人体免疫抑制的遗传控制：Ⅲ.HLA—DQA1和DQB1…

根据天花粉蛋白（Ｔｋ）作用下淋巴细胞体外增殖的抑制是否由ＣＤ８细胞所介导，健康人群被分成介导型（Ｍ＾＋）和非人导型（Ｍ＾－）两类。我们已有的工作证明，性状Ｍ＾＋／Ｍ＾－由ＨＬＡ连锁的一对孟德尔基因控制，本文采用分子生物学手段，对Ｍ＾＋和Ｍ＾－个体作ＨＬＡⅡ类基因的精细分型，发现编码ＤＱ异二聚体分子的ＤＱＡ和ＤＱＢ两个基因同时决定Ｍ＾＋的表达，而且等位基因ＤＱＡ１＊０５０１和ＤＱＢ１＊０２０１以顺式     

胰岛素依赖性糖尿病与HLA的关联研究

胰鸟素依赖性糖尿病是自身免疫性疾病,目前认为遗传因素与环境因素相互作用决定该病的发生。而遗传因素主要是与人类白细胞抗原系统ＨＬＡ相关,且主要涉及ＨＬＡⅡ类区域。此外,ＨＬＡⅠ类基因及ＨＬＡ内非Ⅰ、Ⅱ类基因对该病发生也有一定的作用。     

雌二醇、睾酮对健康男性外周血单核细胞HLA－DR、HLA－DQ抗原表达的影响

将分离的人单核细胞（Ｍ）在体外分别与雌二醇（Ｅ＿２）、睾酮（Ｔｅ）以及Ｅ＿２＋Ｔｅ共同温育２４小时后，观察Ｍ表面ＨＬＡ－ＤＲ、ＨＬＡ－ＤＱ抗原的表达。结果表明，Ｅ＿２（７．３４×１０￣（－１２）ｍｏｌ／Ｌ）能显著抑制Ｍ表面ＨＬＡ－ＤＲ、ＤＱ抗原的表达，Ｔｅ（３．４７×１０￣（－１１）ｍｏｌ／Ｌ）则能促进Ｍ表面ＨＬＡ－ＤＲ、ＤＱ抗原的表达。Ｅ＿２、Ｔｅ一起作用（浓度分别同上）于Ｍ时，Ｍ表面ＨＬＡ－ＤＲ、ＤＱ抗原表达无明显变化。     

用PCR－RFLP方法研究新疆地区汉族HLA－DQA1和－DQB1基因多态性

应用ＰＣＲ－ＲＦＬＰ技术，对新疆地区汉族健康群体进行了ＨＬＡ－ＤＱＡ１（４９人）和－ＤＱＢ１（４７人）基因分型。在ＤＱＡ１８个等位基因中，ＤＱＡ１０３０１的基因频率最高（３２．５６％），０４０１最低（１．０２％）。在ＤＱＢ１１６个等位基因中，ＤＱＢ１０２０１（２０．２１％）、０３０１（１５．９６％）、０３０３（１４．８９％）为最常见；没有观察到０５０３２、０５０４和０６０５。与河北固安县及江浙沪地区汉族群体进行比较，ＤＱＡ１基因未发现存在差异。而ＤＢ１０６０２（３．１９％）与固安汉族（Ｐｃ＝０．０１４４）和上海汉族（Ｐｃ＝０．０１４０）有显著差异，ＤＱＢ１０５０３（８．５２％）与上海汉族有显著差异（Ｐｃ＝０．０２１６）。     

关于我国HLA－DR、DQ位点抗原多态性血清学研究的分析

就国内外１４家实验室的１５组（包括作者自己）ＨＬＡ－ＤＲ、ＤＱ位点多态性的血清学研究，进行了评析，旨在对今后的发展提出其自己的建议。我国ＨＬＡ研究界的当务之急是团结合作，筛选研制出中国自己成套的Ⅱ类抗原分型血清。     

I型糖尿病的HLA易感基因（文献综述）

Ｉ型糖尿病是一种与ＨＬＡ－Ⅱ类抗原密切相关的疾病，各民族特异的单倍型分布导致了种族间不同的与ＩＤＤＭ的相关格局。这种相关性是由于ＨＬＡ－Ⅱ类抗原作为一个重要角色参与并影响了ＩＤＤＭ的发生，尤其是Ⅱ类抗原中的ＤＱ更为重要，其α链及β链均参与此作用。     

中国南京地区汉族人HLA－DR4DNA的快速分型

应用ＡＲＭＳ（ａｍｐｌｉｆｉｃａｔｉｏｎｒｅｆｒａｃｔｏｒｙｒｎｕｔａｔｉｏｎｓｙｓｔｅｍ）方法对南京地区１００例汉族健康人ＨＬＡ－ＤＲ４进行基因分型，结果表明，本地区汉族人ＨＬＡ－ＤＲ４基因纯合子占６％，杂合子占１５％，非ＤＲ４占７９％。本法较ＰＣＲ／ＳＳＯ法简单、快速，结果可靠，可用于ＨＬＡ配型。检测结果为ＨＬＡ相关联疾病及人类学的研究提供了参考依据。     

胰岛素依赖型糖尿病患者的HLA—DQA1等位基因分析

应用聚合酶链反应与序列特异寡核苷酸探针杂交技术（ＰＣＲ／ＳＳＯＰ），分析了北京地区６１例汉族胰岛素依赖型糖尿病（ＩＤＤＭ）患者和５０例非糖尿病对照者的ＨＬＡ－ＤＱα链第５２位氨基酸的编码基因。结果表明：１．８９％的患者ＨＬＡ－ＤＱα链５２位是精氨酸，其编码基因为ＨＬＡ０ＤＱＡ１０３０１和（或）０５０１，其中１０．８％为０３０１纯合子，７０．３％为０３０１杂合子，１８．９％为０５０１纯合子；２．患者     

HLA-DQB1基因多态性与中国汉族人群系统性红斑狼疮的相关性

目的 探讨HLA-DQB1基因单核苷酸多态性(single nucleotide polymorphisms,SNP)与汉族人群系统性红斑狼疮(systemic lupus erthematosus,SLE)遗传易感性的相关性.方法 通过聚合酶链式反应-连接酶检测反应(polymerase chain reaction-ligase detection reaction,PCR-LDR)技术对908例SLE患者和961例健康对照rs3129716(HLA-DQB1)位点进行基因分型,同时结合临床表现分型,分析该位点与疾病及临床表型的相关性.分型结果用PLINK1.07软件进行统计分析.结果 rs3129716(HLA-DQB1)位点等位基因频率和基因型频率在SLE疾病组和对照组的分布差异无统计学意义(P＞0.05).三种遗传模型下的分析显示,两组间差异无统计学意义(P＞0.05).将SLE患者按血清学指标及临床表现分型,未发现相关性.结论 rs3129716(HLA-DQB1)与中国汉族人群SLE患者遗传易感性不相关.     

HLA-DR,DQ nucleotide sequence polymorphisms in five Melanesian populations

HLA-DRB1 nucleotide sequence polymorphisms have been examined in 304 Melanesians from the Papua New Guinean coast (Madang), islands (Rabaul) and highlands (Goroka), and from New Caledonia and Fiji. A total of 20 HLA-DRB1 alleles were detected by oligonucleotide hybridizations of exon 2 HLA-DRB1 polymerase chain reaction products, in a typing protocol designed to detect all 42 officially-designated HLA-DRB1 alleles. DRB1*1502 and 1101 alleles were the most common alleles in coastal and island Melanesians, while DRB1*1501, 1502 and 1408 predominated in Papua New Guinean highlanders. Undefined mixed lymphocyte reaction determinants in earlier studies of Melanesians could be accounted for in the present study as DRB1*0410, 1407 and 1408 in Papua New Guinean highlanders and as DRB1*1104 and 1602 in coastal people. Nucleotide sequence polymorphisms at HLA-DQA1, -DQB1, -DRB3 and -DRB5 were also determined for estimating HLA-DR,DQ allelic disequilibrium relationships; unusual haplotypes in Melanesians included DBR1*1502, DRB5*0101 and DRB1*0410, DQB1*0402. Previous claims of limited heterogeneity in the HLA-DR allele repertoire in Melanesians are now seen to reflect limitations of early typing reagents rather than any dramatic restriction in HLA-DR allelic diversity.     

2型糖尿病患者C反应蛋白及rs1205多态性研究

观察C反应蛋白（CRP）及其单个核苷酸多态性（SNP）与2型糖尿病（T2DM）的相关性。提取受检者外周血有核细胞DNA,应用荧光标记单碱基延伸分型技术及寡核苷酸微阵列芯片杂交技术检测CRP基因的标签SNP（tag SNP）rs1205;同时采用全自动生化分析仪检测CRP及血脂水平。结果表明,T2DM组血清CRP及甘油三酯（TG）水平明显高于健康对照组（P〈0．05）,两组间的等位基因频率具有统计学差异（P〈0．05）,而两组的基因型频率及每组各基因型间CRP水平未见统计学差异（P〉0．05）。研究结果提示汉族人群CRP和TG水平是T2DM的危险因素,CRP基因rs1205的等位基因可能是T2DM遗传易感基因。     

Identification of a novel HLA-DQB1*06 allele, DQB1*0618

We report here the identification of a novel DQB1*06 allele, DQB1*0618, found in a bone marrow donor. The new allele was detected during routine DNA-based HLA typing by an ambiguous pattern of probe hybridization, obtained by polymerase chain reaction using sequence-specific oligonucleotides (PCR-SSO). Molecular cloning and sequencing confirmed that the new allele is identical to DQB1*0609 at exon 2 except for 3 nucleotide substitutions at positions 353, 356 and 367, also found in other alleles. These nucleotide changes may explain its anomalous reactivity.     

Decreased Secretion of Th2 Cytokines Precedes Up-regulated and Delayed Secretion of Th1 Cytokines in Activated Peripheral Blood Mononuclear Cells from Patients with Insulin-Dependent Diabetes Mellitus

Recent evidence suggests that autoimmune animal diabetes is associated with an imbalance between the Th1 and Th2 arms of the cellular immune system. However, limited data is available regarding the Th1/Th2 imbalance in human Insulin dependent diabetes mellitus (IDDM) patients. Therefore, we examined the peak levels, secretory pattern and total cytokine production (calculated as the area under the curve, AUC) of the Th1 cytokines, IL-2 and IFN-γ, and Th2 cytokines, IL-4 and IL-10, from stimulated peripheral blood mononuclear cells, from 17 IDDM patients and 24 normal controls. In contrast to controls, diabetic patients were characterized by an early, uniformly low secretion of Th2 cytokines, followed by a late increased secretion of Th1 cytokines. This resulted in significant differences in secretory patterns of IFN-γIL-2, IL-4 and IL-10 between the two groups;P<0.001,P<0.005,P<0.005 andP<0.001, respectively. No correlation was found in the diabetic patients between any profiles of the cytokines and their various clinical parameters, including age, gender, disease duration, insulin requirements or glycated hemoglobin levels. In conclusion, our data provides the first comprehensive evidence for an independent and persistent impairment of both Th1 and Th2 cytokine secretory patterns in IDDM patients.     

Complete coding region of the new HLA-DQB1*0612 allele, obtained by RT-PCR

A novel HLA-DQB1 allele was detected by oligotyping in the Bubi population of Equatorial Guinea. In order to characterize the new allelic variant, a RT-PCR method which permitted the cloning of its complete coding region was designed. With this method, we have determined the nucleotide sequence of the new DQB1*0612 allele, related to *0604 and *0609 but differing from them at polymorphic codon 70. A proposal for the improvement of the sequencing strategies of HLA class II alleles is made.     

Increased Numbers ofin vivoActivated T cells in Patients with Recent Onset Insulin-dependent Diabetes Mellitus

Insulin-dependent diabetes mellitus is a T-cell dependent immune mediated disease. Conflicting results regarding the distribution of various T lymphocyte phenotypes in recently diagnosed diabetic patients and in patients with established IDDM compared to controls have been reported. In the present study we evaluated phenotypic characteristics of lymphocytes in IDDM patients. Lymphocytes from 77 newly diagnosed IDDM patients, 58 IDDM patients with disease duration >6 months and 30 non-diabetic controls (including patients with several inflammatory conditions) were analyzed for membrane expression of CD4, CD8, CD45RA, CD45RO and CD27 molecules by FACS analysis. No differences in the percentage of CD8⁺T cells were found between any of the groups. However, the percentage of CD4⁺T cells, and consequently the CD4/CD8 ratio were significantly increased in PBL of recently diagnosed diabetic patients compared to the non-diabetic control group (P<0.005). Interestingly, the fraction of lymphocytes coexpressing CD45RO and CD45RA molecules was significantly increased in recent onset IDDM patients, as well as IDDM patients with disease for a longer duration, compared to controls (P<0.0009 andP<0.007, respectively). IDDM patients had a lower percentage of resting memory T cells (CD45RO⁺CD27⁺) than the non-diabetic controls. The proportion of CD45RO⁺lymphocytes lacking the CD27 molecule ((re)-activated memory cells) was similar in IDDM patients and non-diabetic controls. Our findings confirm and extend previous observations that a disturbance in lymphocyte subset distribution is present in patients with IDDM showing an increase in the percentages of circulating CD4 lymphocytes.