上海汉族人HLA-DR、DQ抗原分布调查

用第一届国际红十字HLA会议的全套定型血清调查了上海汉族人HLA-DR和DQ的分布频率.HLA-DR的分解物和HLA-DQ的分型在中国是首次.结果:HLA-DR的基因频率为DR1 0.0054,DR4 0.0906,DR7 0.0847,DRw80.0499,DR9 0.1553,DRw10 0.0109,DRw11 0.0847,DRw12 0.1086,DRw130.0906,DRw14 0.0219,DRw15 0.1332,DRw16 0.0191,DRw17 0.0556,DR空白0.0895;HLA-DQ的基因频率为DQw2 0.1426,DQw4 0.0527,DQw5 0.0642,D Qw6 0.2393,DQw 7 0.2324,DQw 8 0.0219,DQw 9 0.1521,DQ空白0.0946.     

HLA class II alpha chain gene polymorphisms in patients with insulin-dependent diabetes mellitus, dermatitis herpetiformis, and celiac disease

We have investigated DNA polymorphism of the class II alpha chain genes in HLA typed patients with insulin dependent diabetes mellitus (IDDM; n = 79), celiac disease (CD; n = 46), dermatitis herpetiformis (DH; n = 53), and controls (n = 86). Preferential allelic associations of HLA genes and gene products have thus been constructed for susceptibility to these diseases. DR alpha and DQ alpha gene polymorphisms indicated heterogeneity of HLA DR3, DRw6, and DR7, and HLA DR2 and DRw6, respectively. In DR7 positive CD patients a 3.8-kilobase (kb) DR alpha fragment, which correlated with DQw3, was found in only 11% of patients compared with 45% of corresponding controls (P less than 0.05). An increased frequency of a DX alpha genotype UU in all three diseases was found (IDDM 59%, DH 45%, CD 48%, compared to 21% in controls, P less than 0.001), which is not explained solely by the increased frequencies of DR3-DX alpha U. We therefore conclude part of the genetic susceptibility for these three conditions is encoded by genes within the DQ-DX subregion.     

Association of I and II class HLA antigens with Cushing's syndrome]

Incidence of class I and II HLA antigens was studied in 69 patients suffering from Icenko--Cushing syndrome. Positive association was reported for antigens DRw53, DQw3, DR4, DR5, negative for antigen DR2. Antigen combinations DR5, DR7 and DR4, DR5 proved more prevalent. The risk to develop the disease rose 2-3-fold in case of HLA-DR5 phenotype occurrence with antigen DR4 or DR7. Association with DRw53 and DQw3 antigens showing wide specificity seemed most significant.     

HLA antigens in Japanese patients with myasthenia gravis.

HLA antigens in 104 Japanese patients and 41 families with myasthenia gravis (MG) were investigated. The frequencies of DR9 and DRw13 were significantly increased in the patients who developed MG before 3 yr of age. The DQw3 antigen was positive for all the patients that developed MG before 15 yr with only one exception. All the examined cases that developed MG before 3 yr (including this DQw3 negative patient) had the same DQA and DQB DNA restriction fragments. These HLA frequencies decreased as the age of onset increased, and no significant association was observed in adult-onset MG. No patients had B8, DR3, and DQw2. The relative risk was higher for the DR9/DRw13 heterozygotes (37.4) than for DR9 (16.4) or DRw13 (7.1) in the childhood-onset MG. Statistical analysis suggested that DR9 and DRw13 (or DQw1 and DQw3) act synergistically in the disease development. Family study revealed diverse DR9 haplotypes. The most frequent DRw13 haplotype was Bw44-BFF-C4A3B1-DRw13-DQw1, which may be evolutionarily related to the caucasian B8-DR3-DQw2 haplotype. These results showed that MG in early childhood in Japanese individuals is genetically different from that in adulthood and that in caucasians.     

P52Characteristics of HLA Class I and Class II Antigens of the Somali Population

HLA antigens of the Somali population are not categorised as well as those of other international ethnic groups. We analysed the HLA antigens of 76 unrelated Somalis who lived in the west of England. HLA ‐A, ‐B, ‐C and DRB1 typing was performed by polymerase chain reaction using sequence‐specific oligonucleotide probes (PCR‐SSOP) at a low‐intermediate resolution level. Phenotype frequency, gene frequency and haplotype frequency were used to study the relationship between Somalis and other relevant populations. The antigens with highest frequencies were HLA ‐A1, A2, and A30; B7, B51 and B39; Cw7, Cw16, Cw17, Cw15 and Cw18; DR 13, DR17, DR8 and DR1. HLA haplotypes with high significance and characteristics of the Somali population are B7‐Cw7, B39‐Cw12, B51‐Cw16, B57‐Cw18. The result of HLA class I and class II antigen frequencies show that the Somali population appear more similar to Arab or Caucasoid than to African populations. The results are consistent with hypothesis, supported by cultural and historical evidence, of common origin of the Somali population. This study will serve as a reference for further anthropological studies, as well as studies of associations between HLA and disease.     

Familial Atrioventricular Heart Block of Adult Onset: Electrocardiogram and HLA Typing Analysis

EBISAWA, K., et.al .: Familial Atrioventricular Heart Block of Adult Onset: Electrocardiogram and HLA Typing Analysis . A family was investigated because of heart block and sinus arrhythmia. The electrical characteristics were: (1) adult onset in all members; (2) complete heart block with atrial fibrillation in 2, and first- or second-degree heart block in 7 members; and (3) sinus arrhythmia in 3 members. Human leukocyte antigen (HLA) typing was performed. HLA A2, B39, Cw7, and DR12 were positive in 4 of 5 members in the heart block group. In the sinus arrhythmia group, HLA DR12 was positive in all members. In the normal group, none of these HLA typings was positive except one. These findings indicate a tighter relationship between heart block and the HLA locus than previously thought.     

HLA-Cw基因常规检测区外第1、5、6、7外显子核苷酸序列测定的研究及临床应用

目的 研究中国人群HLA-Cw基因第1、5、6、7外显子的分子遗传多态性,探讨增加第1、5 6 7外显子核苷酸序列测定在临床组织配型工作中的重要性及意义.方法 应用PCR-SBT法,对324份样本的HLA-Cw基因第2、3、4外显子作常规测序分型.对检出的模棱两可结果,设计HLACw第1、5 6 7外显子序列测序引物并优化测序反应条件,增加第1、5、6、7外显子核苷酸序列分析.结果 对HLA-Cw基冈第2、3、4外显子常规检测,一次性获得等位基因前4位数分型结果 的样本占23.8%(77/324);出现模棱两可结果的样本数占76.2%(247/324),检出的模棱两可等位基因组合有73种;增加HLA-Cw基因的第1、5、6、7外显子多态性检测,可解决Cw* 030201/030202、030301/0320N、Cw* 040101/0409N/0430、Cw* 070201/0750、Cw* 0403/0409N/0430和Cw* 080101/0822等10种常见的模棱两可等位基因组合.结论 在临床HLA-Cw基因配型中增加第1、5、6、7外显子多态性检测,有助于解决测序分型中的模棱两可的结果 和提高HLA-Cw基因分型精确度,对临床组织配型工作具有重要意义.     

HLA-Cw基因常规检测区外第1、5、6、7外显子核苷酸序列测定的研究及临床应用

目的 研究中国人群HLA-Cw基因第1、5、6、7外显子的分子遗传多态性,探讨增加第1、5 6 7外显子核苷酸序列测定在临床组织配型工作中的重要性及意义.方法 应用PCR-SBT法,对324份样本的HLA-Cw基因第2、3、4外显子作常规测序分型.对检出的模棱两可结果,设计HLACw第1、5 6 7外显子序列测序引物并优化测序反应条件,增加第1、5、6、7外显子核苷酸序列分析.结果 对HLA-Cw基冈第2、3、4外显子常规检测,一次性获得等位基因前4位数分型结果 的样本占23.8%(77/324);出现模棱两可结果的样本数占76.2%(247/324),检出的模棱两可等位基因组合有73种;增加HLA-Cw基因的第1、5、6、7外显子多态性检测,可解决Cw* 030201/030202、030301/0320N、Cw* 040101/0409N/0430、Cw* 070201/0750、Cw* 0403/0409N/0430和Cw* 080101/0822等10种常见的模棱两可等位基因组合.结论 在临床HLA-Cw基因配型中增加第1、5、6、7外显子多态性检测,有助于解决测序分型中的模棱两可的结果 和提高HLA-Cw基因分型精确度,对临床组织配型工作具有重要意义.     

HLA-Cw基因常规检测区外第1、5、6、7外显子核苷酸序列测定的研究及临床应用

目的 研究中国人群HLA-Cw基因第1、5、6、7外显子的分子遗传多态性,探讨增加第1、5 6 7外显子核苷酸序列测定在临床组织配型工作中的重要性及意义.方法 应用PCR-SBT法,对324份样本的HLA-Cw基因第2、3、4外显子作常规测序分型.对检出的模棱两可结果,设计HLACw第1、5 6 7外显子序列测序引物并优化测序反应条件,增加第1、5、6、7外显子核苷酸序列分析.结果 对HLA-Cw基冈第2、3、4外显子常规检测,一次性获得等位基因前4位数分型结果 的样本占23.8%(77/324);出现模棱两可结果的样本数占76.2%(247/324),检出的模棱两可等位基因组合有73种;增加HLA-Cw基因的第1、5、6、7外显子多态性检测,可解决Cw* 030201/030202、030301/0320N、Cw* 040101/0409N/0430、Cw* 070201/0750、Cw* 0403/0409N/0430和Cw* 080101/0822等10种常见的模棱两可等位基因组合.结论 在临床HLA-Cw基因配型中增加第1、5、6、7外显子多态性检测,有助于解决测序分型中的模棱两可的结果 和提高HLA-Cw基因分型精确度,对临床组织配型工作具有重要意义.     

HLA分子三维结构与GVHD相关性研究

目的 进一步探索GVHD的发病机制,提高临床移植疗效。方法 HLA分型方法包括血清-细胞学、基因和基因亚型分型。血清-细胞学分型采用国际通用的微量淋巴细胞毒试验和混合淋巴细胞培养;基因分型采用PCR-SSP方法,对基因亚型进行DNA测序,再进行HLA三维结构模拟。结果 (1)供-受体间HLA-Ⅰ、Ⅱ类血清学相合,组基因相合,基因亚型不同,但HLA三维结构模拟两者差别不明显,骨髓移植后仅发生了Ⅱ度GVHD;(2)供-受体间,基因亚型不同,两者仅差1个氨基酸,供-受体间抗原三维结构差别明显,骨髓移植后发生Ⅳ度GVHD;(3)受体为HLA-A~*0201,供体为A~*6801,二者血清学呈强交叉反应,其它位点均相合,移植后发生Ⅳ度GVHD,后经HLA三维结构模拟,两者差别明显。结论 当异基因骨髓移植供-受体间HLA基因亚型不同时,移植后GVHD的发生程度与HLA分子三维结构差别大小有密切关系;GVHD与抗原氨基酸差别的数量没有直接关系。     

P53Electron Microscopical Localisation of CD61 (Beta3 Integrin) on Placental Syncytiotrophoblast Microvilli

HLA antigens of the Somali population are not categorised as well as those of other international ethnic groups. We analysed the HLA antigens of 76 unrelated Somalis who lived in the west of England. HLA -A, -B, -C and DRB1 typing was performed by polymerase chain reaction using sequence-specific oligonucleotide probes (PCR-SSOP) at a low-intermediate resolution level. Phenotype frequency, gene frequency and haplotype frequency were used to study the relationship between Somalis and other relevant populations. The antigens with highest frequencies were HLA -A1, A2, and A30; B7, B51 and B39; Cw7, Cw16, Cw17, Cw15 and Cw18; DR 13, DR17, DR8 and DR1. HLA haplotypes with high significance and characteristics of the Somali population are B7-Cw7, B39-Cw12, B51-Cw16, B57-Cw18. The result of HLA class I and class II antigen frequencies show that the Somali population appear more similar to Arab or Caucasoid than to African populations. The results are consistent with hypothesis, supported by cultural and historical evidence, of common origin of the Somali population. This study will serve as a reference for further anthropological studies, as well as studies of associations between HLA and disease.     

A Study of Human Leukocyte D Locus Related Antigens in Graves'' Disease

An association between Graves' disease and the human leukocyte antigen (HLA) system has previously been reported. The disease was more strongly associated with the HLA D locus antigen Dw3 than with HLA B8. Products of the HLA D locus are determined by the interaction of test cells with standard typing lymphocytes, a technically difficult procedure. Recently, it has been possible to type serologically for D locus related (DRw) specificities on peripheral bone marrow-derived (B) lymphocytes. Blood B lymphocytes from 50 unrelated controls and 41 patients with Graves' disease were typed for seven HLA DRw specificities. 28 patients with Graves' disease (68%) were positive for DRw3, in contrast to 14 controls (28%); whereas only 21 patients (50%) were HLA B8 positive, compared with 13 (26%) controls. Thus, positivity for DRw3 afforded a relative risk for Graves' disease of 5.5, whereas that for HLA B8 amounted to 3.0. Additionally, a family with multiple cases of Graves' disease in which the disease was previously shown to be inherited with the haplotype, was linked to DRw2, which suggests that the susceptibility to the disease was inherited in association with that antigen. Two HLA B/glyoxalase recombination events were observed in this family; in both instances HLA DRw followed HLA B. This study thus demonstrates that the disease susceptibility gene for Graves' disease is in strong linkage disequilibrium with DRw3; however, it may be associated with other DRw specificities and inherited within family units in association with them.     

DNA序列分析鉴定HLA新等位基因B~*0740

目的确认一个中国人群中的新HLA等位基因。方法应用单倍型特异性分离(haplotype-specific ex-traction,HSE)技术和DNA序列分析技术鉴定HLA-B~*的新等位基因。结果被检标本HLA-B位点有一个等位基因的核苷酸序列与任何已知的HLA等位基因均不同,与同源性最高的B~*0705相比,在exon3区域中的605位碱基由A>T,引起编码178位的氨基酸由赖氨酸(K)变成甲硫氨酸(M)。血清学分型表明新等位基因的免疫学表型仍为HLA-B7。家系分析提示,HLA-B~*0740基因来源于其母亲。结论该等位基因序列为HLA-B位点的一个新等位基因,于2005年2月由WHO HLA因子命名委员会正式命名为HLA-B~*0740。     

Association of polar amino acids at position 26 of the HLA-DQB1 first domain with the anticentromere autoantibody response in systemic sclerosis (scleroderma).

HLA class II alleles (detected by DNA typing) were determined in 116 Caucasians with systemic sclerosis positive and negative for anticentromere autoantibodies (ACA). Significantly increased frequencies of HLA-DR5(DRw11) (P = 0.009) and the Dw13(DRB1*0403, *0407) subtypes of DR4 (probability corrected, Pc = 0.005) were seen in ACA positive patients, and HLA-DR1 and DRw8 were also increased. These findings appeared to reflect linkage disequilibrium of DR5(DRw11) and many DR4(Dw13) haplotypes with HLA-DQw7 and DR1 with DQw5. In fact, the presence of a DQB1 allele having a polar glycine or tyrosine at position 26 of the DQB1 first domain versus a hydrophobic leucine accounted for 100% of ACA positive Caucasian systemic sclerosis patients compared to 69% of the ACA negative SS patients (P = 0.0008) and 71% of Caucasian controls (P = 0.0003) as well as all 7 ACA patients of non-Caucasian background. Furthermore, the genotype frequency of DQB1 alleles lacking leucine at position 26 was 73% in ACA positive SS patients, compared to 42% of ACA negative patients (P = 1.2 x 10(-5)) and 38% of controls (P = 5.8 x 10(-7)). These data, then, suggest that the second hypervariable region of the HLA-DQB1 chain may form the candidate epitope associated with the ACA response.     

The importance of class I and class II HLA in cadaveric renal transplantation

The importance of avoiding mismatches (MM) at Class I and Class II HLA antigens in cyclosporine-treated renal allograft patients is controversial. In order to assess the role of HLA, 200 consecutive cadaveric renal allografts over a 4-year period were analysed. All patients received cyclosporine/predinisone immunosuppression and 75% were induced with ALG. Minimum follow-up period was one year. HLA A, B, DR, DQ, and DRw52/53 typing were available on 77-100% of allografts. A beneficial effect was noted at the HLA A locus. One-year survival was 87.2% in the 0 and 1 HLA A MM group combined vs 73.8% in the 2 HLA A MM group (p less than 0.05). The mean creatinine level at one year was also lower in the 0 plus 1 MM vs 2 MM group: 152.8 mumol/L vs 184.8 mumol/L, respectively (p less than 0.05). Significantly fewer rejection episodes occurred in the 0 and 1 HLA DQ MM group combined vs the 2 MM group. Steroid-resistant rejection episodes (SRRE) were not associated with the number of HLA MM. Patients who had an SRRE had significantly higher mean current and historical peak panel reactive antibodies (PRA) than patients who did not have SRRE. These results indicate that avoiding mismatches at the HLA A locus may improve renal allograft survival, and matching at HLA DQ may predispose patients to a more quiescent post-transplant course. The degree of preoperative sensitization may be an important etiologic factor in SRRE.     

Functional polymorphism of each of the two HLA-DR beta chain loci demonstrated with antigen-specific DR3- and DRw52-restricted T cell clones

HLA-DR3- and HLA-DRw52-associated functional polymorphism was investigated with selected tetanus toxoid (TT)-specific T cell clones. We have shown earlier that HLA-DR antigens are encoded by two distinct loci, DR beta I and DR beta III. The alloantigenic determinant(s) defined by the serological HLA-DR3 specificity map to the former, while the supratypic HLA-DRw52 determinants map to DR beta III. Furthermore, we have recently recognized by DNA sequencing three alleles of HLA-DRw52 at locus DR beta III, referred to as 52 a, b, and c. Our objective was to correlate the pattern of T cell restriction with the gene products of individual DR beta chain loci and with the three newly described alleles of locus DR beta III. Among the selected T cell clones, 5 reacted exclusively when TT was presented by HLA-DR3+ APCs (TT-DR3-APC). In contrast, two T cell clones were stimulated by TT-DRw52-APC. More specifically, these two T cell clones (Clones 10 and 16) were stimulated by different subsets of TT-DRw52-APC. Clone 16 responded to some DR3 and TT-DRw6-APC, while clone 10 was stimulated by other TT-DR3 and TT-DRw6, and all TT-DR5-APC. This same pattern of DRw52 restriction was found in panel, as well as in family studies. Because this suggested a correlation with the pattern of DRw52 polymorphism observed earlier by DNA sequencing and oligonucleotide hybridization, the APC used in these experiments were typed for the 52 a, b, and c alleles of locus DR beta III by allele-specific oligonucleotide probes. This distribution overlapped exactly with the stimulation pattern defined by the T cell clones. Clone 16 responded to TT-52a-APC, clone 10 to TT-52b-APC, and both clones to a TT-52c-APC. The response of the T cell clones was inhibited differentially by mAbs to DR. Raising TT concentration, or increasing HLA-class II expression with INF-gamma both affected the magnitude of response of the TT-specific clones but did not modify their specificities. These results demonstrate that a restriction specificity can be attributed to the DR beta III locus and illustrate the functional relevance of the polymorphism observed at this locus. This is of special interest in view of the striking difference in the pattern of structural diversity among alleles of DR beta I and DR beta III.     

HLA-B40交叉反应组血清学与高分辨度DNA分型的比较研究

目的 比较研究中国汉族人群ＨＬＡＢ４０ 交叉反应组血清学分型与高分辨度ＤＮＡ 分型结果。方法研究样本１９９ 份,包括血清学分型Ｂ４０ 组阳性样本１７７ 份、可疑阳性１２ 份和阴性对照１０ 份。同时对血清学分型和高分辨度顺序特异引物聚合酶链反应（ＰＣＲＳＳＰ） ＤＮＡ 分型进行比较。结果 ＤＮＡ分型成功率９９ ．５ ％ ,总耗时５ 小时,特异性和敏感性好,可准确分辨出Ｂ４０ 组１０ 个等位基因。血清学分型成功率９９ ．４ ％ ,总耗时２ 小时,总误差２０ 个,误差率１１ ．２ ％ 。检出中国汉族人群Ｂ４０ 组各等位基因所占比例：Ｂ６０ 为６７ ．４ ％ 、Ｂ６１ 为２０ ．８ ％ 、Ｂ４８０１ 为９ ．６ ％ 、Ｂ４００５ 为２ ．２ ％ 。结论 高分辨度ＰＣＲＳＳＰ 方法用于ＨＬＡＢ４０ 组分型比血清学分型更加精确,适合于临床应用。     

GVHD病因学的新视野:HLA分子三维结构研究

目的 :进一步探索GVHD的发病机理 ,提高临床移植疗效。方法 :采用国际通用的微量淋巴细胞毒试验和混合淋巴细胞培养方法 ;基因分型采用PCR SSP方法 ,对基因亚型进行DNA测序 ,再进行HLA三维结构模拟。结果 :① 4名骨髓移植患者中 ,例 1供 受体间HLA Ⅰ、Ⅱ类血清学分型相合 ,DQB1基因亚型不同 ,例 2、3、4均有 1个Ⅰ类抗原 (分别为HLA A、B和A)血清学分型不合和 1个基因亚型不同。HLA三维结构模拟例 1、3、4供 受者差别明显 ,骨髓移植后发生了Ⅲ～Ⅳ度GVHD ;例 2三维结构模拟差别不明显 ,仅发生Ⅰ度GVHD ;② 4个病例供 受体间HLA分子分别相差 14、17、2 0、12个氨基酸 ,相差数量与GVHD无相关性 ;3例重度GVHD供 受体的HLA分子三维结构均在抗原结合区的β转角处出现明显差异 ,而 1例轻度GVHD则无明显差异。 结论 :当异基因骨髓移植供 受体间HLA基因亚型不同时 ,移植后发生GVHD的程度与HLA分子三维结构差别大小有密切关系 ;三维结构的差别又与差异氨基酸所处的位置有关 ;GVHD与抗原氨基酸差别的数量多少没有直接关系。     

Successful immunosuppressive therapy in a patient with autoantibodies to insulin receptors and immune complex glomerulonephritis

Altered mental status, acanthosis nigricans, immune complex glomerulonephritis with nephrotic syndrome, fasting hypoglycemia, and postprandial hyperglycemia associated with anti-insulin receptor antibodies (type B insulin resistance) developed in a 43-year-old black woman who initially was treated for diabetes mellitus. Her HLA phenotype was A2, A29, Bw45(w6), B13(w4), Cw3, DR1 (DQw1). Her serum contained immune complexes, low complement levels, and antibodies that bound to the glomerular mesangium of mouse kidney. All clinical and serologic abnormalities resolved with combination cyclophosphamide and glucocorticoid treatment, and low doses of these agents have maintained the remission for more than a year.     

应用中高分辨率分型方法分析中国人群HLA-B15组的多态性

目的　了解人类白细胞抗原(HLA)系统中最常见、最复杂且最难分型的B15组的多态性情况，并探讨其是否影响移植时供的选择。方法　对194名随机健康人群用反向PCR—SSO方法做HLA—AB分型，同时，对部分样品作HLA—AB血清学分型，以期得到与等位基因对应的血清学特异性。结果　本研究共得到7种属于B15组的等位基因，包含了4种血清学特异性。在中国人群HLA—B15组中，B^*1501占绝大多数，B^*1502，B^*15ll，B^*1518，B^*1527的分布频率相似。结论 B^*1527较常见，其与B^*1501(两均为B62)在供受间错配、是否能导致移植物抗宿主反应(GVHD)等方面，需要作进一步的研究。结果提示在移植时对于HLA—B15组如仅做低分辨率分型是不够的。