摘要:

With the use of polymerase chain reaction (PCR) and sequence‐specific oligonucleotide probe (SSOP), we established a DNA typing method of the HLA ‐ A locus. A pair of primers to amplify the highly polymorphic region of HLA‐A gene including exon 2 and exon 3 was designed and the amplified DNAs were hybridized with 91 types of32P labeled SSOPs. This method allowed discrimination of all known HLA‐A alleles except for two combinations, A*0201 or A*0209 and A*0207 or A*0215N, which have identical sequences in exon 2 and exon 3. Another pair of primers was designed for amplification of exon 4 and the PCR products were hybridized with 5 SSOPs to distinguish A*0201 and A*0207 from A*0209 and A*0215N, respectively. In this study, 81 B‐lymphoblastoid cell lines (BLCL) homozygous for HLA and 553 unrelated healthy Japanese individuals were determined for their HLA‐A genotypes. Based on the genotyping results, frequency of HLA‐A alleles and linkage disequilibrium between HLA‐A and HLA‐B in the Japanese population were investigated. In addition, four new HLA‐A alleles were identified and their nucleotide sequences in exon 2 and exon 3 were determined to confirm

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02520.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We here present a sequencing strategy for the HLA‐A locus which is generally applicable for all HLA class I genes. The typing strategy is based on a group‐specific amplification according to the serologically defined antigens. The PCR products carry the typing‐relevant polymorphic regions of the 2nd and 3rd exon including the 2nd intron. The sequencing primers were designed to match conserved sequence motifs in the 2nd intron allowing a nested sequencing approach in 3′ and 5′ direction. These conserved regions were identified after sequence compilation of the 2nd intron of 143 clinical samples and 48 cell lines mostly from the 9th and 10th IHWC representing all serologically defined groups of alleles. This strategy allowed the use of only one 5′ and one 3′ sequencing primer regardless of the amplified allele. Therefore, it was possible to use dye terminator as well as dye primer sequencing chemistry. The amplification strategy allowed the separation of the haplotypes in almost all cases. Thus, an assignment of heterozygous positions requiring high sequencing quality was not necessary, allowing the application of Sequenase as well as TaqPolymerase as sequencing enzyme. Concerning the resolution of heterozygosity it is obvious that this approach is superior to a typing system using a single pair of generic primers followed by direct sequencing, since the latter technique is not capable of defining the cis/trans linkage of polymorphic sequences and, hence, cannot exclude the presence of un

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02521.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We studied C4A and C4B polymorphisms and HLA‐B and ‐DR associations in the San, Khoi and Xhosa. C4A and C4B alleles were determined using conventional protein allotyping methods. The C4A*3, C4B*1 haplotype had a high frequency (30–55%) in all populations. The frequency of C4A*3, C4B*Q0 was 7–19%. The C4A*Q0, C4B*1 haplotype was frequent (15%) in the Khoi but very rare in the San (P0.001). C4A*12 A*91, C4B*Q0 was frequent in the Xhosa (15%) but rare in the San and Khoi (P0.001). Alleles C4A*5 and C4A*6, and the C4B*2 B*92 duplication were only found in the Xhosa. C4A alleles A*4, A*45, A*58, A*12, A*14, A*19 and the C4A*3 A*91 duplication were only found in the San/Khoi population group. In the San, fourteen extended haplotypes were found in a relatively high frequency (2–7%). In the Xhosa, one extended haplotype (B42, C4A*12 A*91, C4B*Q0, DR18) was found in a very high frequency (13%) and was characteristic for this group; five other extended haplotypes were found with a low frequ

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02522.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Abstract:A single human leukocyte antigen (HLA) class II allele,DQB1*0301, is strongly associated with melanoma, and theHLA‐DRlocus provides the telomeric boundary for melanoma susceptibility in the HLA class II region of chromosome 6. However, the centromeric boundary is unknown. This study was designed to determine whether the adjacent upstream transporter associated with antigen processing (TAP) locus,TAP2, constitutes the centromeric boundary of disease susceptibility in melanoma. Molecular oligotyping ofTAP2genes was performed for 36 Caucasian patients with melanoma and for 32 Caucasian control individuals by both amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) and PCR‐sequence‐specific oligonucleotide (SSO) typing.TAP2allele frequencies in the melanoma patients were compared to those in non‐melanoma Caucasian control populations, and toHLA‐DQallele frequencies determined by molecular oligotyping. WhileHLA‐DQB1*0301was more common in this group of 36 melanoma patients compared to a group of 200 controls (56 percent vs. 27 percent, Bonferoni‐corrected chi‐square p=0.01), no significant differences were observed inTAP2allele frequencies between melanoma patients and controls. TheTAP2locus represents the centromeric boundary of disease susceptibility for melanoma in the class II region of chromosome 6p. These results support an etiologic role forHLA‐DQB1*0301in melano

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02523.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We describe for the first time the use of PCR based techniques to analyze the MHC class II polymorphism of the Bulgarian population. The present study provides the HLA‐DRB, DQB1 allele frequencies in 116 Bulgarian individuals and DQA1 alleles frequencies in 100 subjects. DNA from these individuals was typed for DRB and DQB1 typed by the PCR‐ Allele Specific Amplification (PCR‐ASA) method and DQA1 by PCR followed by hybridization using Sequence Specific Oligonucleotides (PCR‐SSO). Allele and haplo‐type frequencies and linkage disequilibria are computed by the standard methods used for the XIth International Histocompatibility Workshop. The highest frequencies are 0.159, 0.109 and 0.085 for DRB1*1101, DRB1*1601 and DRB1*1301 respectively. Among the eight DQA1 alleles detected, DQA1*0501 (0.344) is found to be much more frequent than the two most frequent alleles DQA1*0102 (0.225) and DQA1*0101 (0.151). Twelve DQB1 alleles are found and three of them, DQB1*0301 (0.280), DQB1*0502 (0.153) and DQB1*0201 (0.133) showed the highest frequencies. The haplo‐type DRB1*1101‐DQA1*0501‐DQB1*0301 (0.079) predominate clearly, followed by DRB1*1601‐DQA1*0102‐DDQB1*0502 (0.055) and DRB1*0101‐DQA1*0101‐DQB1*0501. These results indicate that the Bulgarian population is characterized by features representative of the European anthropological type with a substantial contribution from the

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02524.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Celiac disease (CD) has one of the strongest class II HLA associations of any human illness. We used DNA‐RFLP typing to study the class II HLA genotypes of celiac disease patients from the West of Ireland, the geographic area with the highest rate of celiac disease in the world. We confirmed the high frequency of HLA‐DR3 in this population, and we were also able to demonstrate the additional risk of developing celiac disease imparted by HLA‐DR7. This was done by clearly distinguishing DR7, DQ2 haplotypes from DR7, DQ9 haplotypes, and by “subtraction analysis” of haplotype frequencies. As reported in other populations, most of the patients without DR3 were heterozygous for DR7 and DR11 or 12 (DR5), or had DR4. We used PCR‐RFLP and direct sequencing of amplified DNA to examine HLA‐DR4 subtypes. The frequency of HLA‐DR4 was markedly decreased in patients compared with controls (p=0.000001) and there was a significant alteration of DR4 subtypes of the patients compared with controls (p=0.0227). Moreover, all of the CD patients (5 of 5) with DR4 had a haplotype associated with the DQB1*0302 allele compared with only 11 of 23 control subjects with DR4. Our results in this population with exceptionally high risk of CD strongly support the DQ heterodimer hypothesis and suggest that the recently described sequence difference between the DQB1*02 alleles of DR3 and DR7 may contribute to a synergistic increased risk when these haplotypes are inherited together. In addition, our findings suggest a role for HLA‐DQ in

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02525.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02526.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02527.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

NewHLA‐Blocus alleles have been found in South American Amerindian populations but were largely absent in North American Amerindian tribes also descended from this first Paleo‐Indian migration. We have now extended these studies to the Navajo, descendants of the second Nadene migration. No new functional alleles were found at theBlocus of this tribe. This limited study supports the notion that while newBlocus variants are common in South American Amerindians, it is more difficult to find newBlocus alleles in North American native peoples. Whether this dichotomy is due to differences in pathogen environment and/or population structures between North and South America remains a subject of speculat

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02528.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02529.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY