摘要:

A nonisotopic oligotyping method using reverse dot blot hybridization was developed for HLA class II DQA1, DQB1, DPB1, DRB1, DRB3, DRB4, DRB5 alleles. The polymorphic second exon of the different genes was amplified by the polymerase chain reaction (PCR). For each gene the amplified DNA was hybridized at stringent conditions to membrane‐bound sequence‐specific oligonucleotides (SSOs) and visualization of positive signals was done by chemiluminescence. A combination of 11, 18, 23 and 31 SSOs was designed to identify 9/13 DQAI, 16/17 DQB1, 23/24 DPB1 and 50/55 DRB1, 4 DRB3, 1 DRB4, 3/4 DRB5 alleles respectively. For the DRB1 locus, an additional DRB1*04 groupspecific PCR was developed to make discrimination between the DR4 alleles possible in different heterozygous combinations. The procedure described here provides rapid and nonisotopic genotyping of heterozygous samples from a variety of sources and can be applied for tissue typing, disease susceptibility studies and forensic medic

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01970.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

The restriction enzyme Pstl showed a two‐allele restriction fragment length polymorphism (RFLP) marker when DNA polymorphism of the gene G8, a novel major histocompatibility complex (MHC) class III gene, was screened. The gene G8 is located ca. 4 kb centromeric of the heat‐shock protein 70 (HSP70) gene cluster. A more detailed mapping indicated that the polymorphic restriction site is actually located in the coding region of the adjacent HSP70–2 gene, where it has been earlier reported to cause a silent mutation. To estimate the frequency of this polymorphism in the normal population, 95 blood donors were analyzed: The gene frequency of the 8.5 kb (designated‘L’) allele was 0.45 and that of the 8.65 kb (‘U’) allele 0.55. However, when 19 families with patients suffering from celiac disease were studied, the gene frequencies in the affected haplotypes (L = 0.76, U = 0.24) significantly deviated from those observed in the normal population and in the non‐affected MHC haplotypes of these families (L = 0.48, U = 0.52). However, the association results from a strong association between the allele ‘L’and the MHC haplotype HLA B8 DR3, a known suspectibility marker of celiac disease. Only one patient, in fact, was negative for the well‐established class II haplotype markers DR3 or DR7. The data therefore confirm the crucial role of MHC class II in suspectibility to celiac disease, but due to a strong linkage disequilibrium within MHC the role of MHC class III genes in disease associations

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01971.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

We have typed 64 Japanese patients with mixed connective tissue disease (MCTD) and 53 Japanese patients with systemic lupus erythematosus (SLE) for HLA‐DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, and DPB1 genes by the HLA‐DNA typing method using the PCR‐SSOP technique. Frequencies of HLA‐DRB1*0401, DRB1*0901, DRB4*0101, and DQA1*03 were increased and those of HLA‐DRB1*0405 and DQB1*0401 were decreased in the patients with MCTD, while the frequencies of HLA‐DRB1*1501, DRB5*0101, and DQB1*0602 were increased in the patients with SLE. The typing results suggest that susceptibility to MCTD is strongly associated with the HLA‐DRB1*0401‐DRB4*0101‐DQA1*03‐DQB1*0301 haplotype, and that to SLE is associated with the HLA‐DRB1*1501‐DRB5*0101‐DQA1*0102‐DQB1*0602 haplotype. The observation that the MCTD‐associated HLA alleles are distinct from the SLE‐associated ones may support the clinical en

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01972.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Rheumatoid arthritis (RA) is associated with the HLA‐DR4 cellular subtypes Dw4 and Dw14 in Caucasians, with Dw15 in Japanese, and possibly with HLA‐DR1 in Israeli Jews. Sequencing studies in Caucasians have shown that these molecules share a common amino acid sequence in the third hypervariable region of the DR molecule (AA 67‐74: LLEQRRAA or LLEQKRAA), suggesting that this sequence is primarily associated with RA. An important argument in favor of this shared‐epitope hypothesis has been the reported association between DR1 and RA in Israeli Jews. However, a later report did not confirm this association, and cellular typing showed that Israeli DR1 consists of three or more subtypes, suggesting that new subtypes might be present. Since no sequencing data on Israeli HLA‐DR1 genes have been reported, we sequenced the first domain (AA 10–91) of the DRB1 gene in 12 DR1‐positive Israeli RA patients, 5 healthy controls and a homozygous typing cell (HTC), defining the major Jewish cellular HLA‐DR1 subtype. DRB1*0102 (DR1 Dw20) was found in 8 RA patients, 3 controls and the HTC “LVA”. DRB1*0101 (DR1 Dw1) was found in 4 RA patients and 2 controls. No other DR1 subtypes were encountered. In all 20 DR1 haplotypes, the DRB1*0101 or 0102 allele was associated with DQA1*0101 and DQB1*0501, being identical to the Caucasian DR1 haplotypes. Thus, at the sequence level, we found no basis for the reported extensive cellular heterogeneity of DR1 in the Israeli population. Both alleles encountered, DRB1*0102 or DRB1*0101, encode a third hypervariable region sequence (AA 67–74) shared with DRB1*0404

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01973.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Multiple Sclerosis (MS) is a demyelinating disease associated with the HLA‐DR2‐related haplotype DRB1*1501, DQB1*0602 in Caucasoids and with DQB1*0602 in DR2‐positive Cantonese. However, many MS patients do not have the high‐risk HLA‐D determinants and alternative genes may contribute to the pathogenesis of MS. One candidate gene is HLA‐DPB1. Our reanalysis of five earlier reports of HLA‐DPB1 antigen distributions in Caucasoid MS patients shows a consistent and highly significant increase (p = 1.5 × 10‐5) in frequency of HLA‐DPw3 in the combined data set. This study tests whether HLA‐DPw3 (DPB1*0301) is also increased in frequency in Australian and Cantonese MS patients and whether any distortion in DPB1 allelic distributions can be attributed to linkage disequilibrium with DQB1*0602. PCR‐RFLPs were used to determine distributions of 20 HLA‐DPB1 alleles in 41 Australian MS patients and 67 controls of known DQB1*0602 status and in 11 Cantonese MS patients and 33 controls positive for HLA‐DR2. HLA‐DP distributions in Australian MS patients and controls positive for DQB1*0602 did not differ, but in those MS patients lacking DQB1*0602, the DPB1*0301 antigen (phenotype) frequency was significantly (p = 0.006) increased (50.0%) when compared with DQB1*0602‐negative controls (9.1%). DPB1*0301 was associated (p = 0.003) with DQB1*0402 (DR8) in Caucasoid MS patients. In Cantonese MS patients positive for DQB1*0602, there was an increase in frequency of DPB1*0201 (66.7%) compared with HLA‐DR2‐positive controls negative for DQB1*0602 (10.0%), possibly due to linkage disequilibrium (p = 0.06) with the DRB1*1501, DQB1*0602 haplotype. In DR2‐positive Cantonese lacking DQB1*0602, DPB1*1301 was increased in MS patients (80%) when compared with controls (13.3%) although the association did not reach statistical significance when the p value was corrected for multiple comparisons; the increase could not be accounted for by linkage disequilibrium with any HLA‐DR2‐related haplotypes. This study suggests that the HLA‐DPB1 locus, or a closely‐linked locus, may contribute to inherited susceptibility for MS in those lac

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01974.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

DQA1, DQB1 and DPB1 alleles were investigated in a large panel of samples carrying HLA‐DQ2‐ and HLA‐DQ9‐bearing extended haplotypes. Every instance of the [HLA‐B8, SCO1, DR3, DQ2a] and [HLA‐B18, F1C3, DR3, DQ2b] extended haplotypes carried the DQA1*‐0501 allele, while every instance of [HLA‐844, F1C31, DR7, DQ2c], (HLA‐B47, FC91,0, DR7, DQ2c] and [HLA‐B57, SC61, DR7, DQ2d] carried the DQA1*0201 allele. All HLA‐DR3, DQ2 and HLA‐DR7, DQ2 extended haplotypes carried the DQB1*0201 allele. Every example of [HLA‐B57, SC61, DR7, DQ9] carried the DQB1*0303 allele. Several associations between the DPB1 alleles and some HLA‐DQ2‐ and DQ9‐carrying extended haplotypes were identified. HLA‐DPw2‐encoding alleles. DPB1*0202 (50%, p<0.001) and 0201 (30%). were found on the [HLA‐B18, F1C30, DR3, DQ2b] extended haplotype. Every instance of the [HLA‐B57, SC61, DR7, DQ9] extended haplotype carried the DPB 1*0401 allele. Also, we have confirmed the associations of the DPB1*0101 and DPB1*0401 alleles with the [HLA‐B8, SC01. DR3, DQ2a] extended haplotype. The analysis of hornozygous typing cells carrying the [HLA‐B44. FC31, DR7, DQ2c] extended haplotype showed that the DPB1*0401 and 0201 alleles were present at similar frequencies (37.5%). while the DPB1*1101 allele was found in only 25% of these haplotypes analyzed. Our results suggest that the constancy of the DNA of the HLA‐B, DR, DQ regions may extend to the DP region and that the extent of such fixity varies, perhaps as the result of

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01975.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

A polymorphic monoclonal antibody (TAL16.1), raised against a mouse L‐cell transfectant expressing the human DRB5*0101 gene from the HLA‐DR15(2) Dw2 DR51 haplotype was shown to have a complex pattern of reactivity to DRB gene products. The antibody bound to a transfectant expressing the DRB5*0101 allele against which it was produced but not to a transfectant expressing the DRB1*1501 allele. These alleles of the DRB1 and DRB5 genes are usually coexpressed on DR15(2) Dw2 DR51 cells. A comparison of the HLA‐DRB amino acid sequences of reactive and non‐reactive cells identified an aspartic acid residue at position 70, conserved in all antibody‐positive cells and absent in antibody‐negative cells, which was postulated as being responsible for conferring the specificity of the antibody. The aspartic acid residue at position 70 is present in DRB5*0101 and DRB5*0102 alleles but absent in DRB5*0201 and DRB5*0202 alleles, allowing the antibody to distinguish between these splits of the DR51 serological specificity. TAL16.1 also binds to the product of the DRB1*0103 allele and discriminates between cells with a DR103 specificity and the other DR1 subtypes, DRB1*0101 and DRB1*0102. In this report the value of transfectants as immunogens for use in the production of monoclonal antibodies of predetermined specificity and as tools for the fine mapping of antibody specificity i

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01976.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01977.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01978.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1993.tb01979.x

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY