ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01799.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:The restriction fragment length polymorphism (RFLP) of the two human HLA‐B‐associated transcripts (BATs) genes, BATl and BAT2, was investigated using 5 different restriction enzymes and two human BATl and BAT2 cDNA probes. Two of the enzymes,NcoI andRsaI, revealed polymorphic patterns which were investigated in healthy Danes. The cDNA/restriction enzyme combination BATl/NcoI identifies polymorphic bands at 12 kb, 8 kb, 2.5 kb, and 1.1 kb, while the BAT2/RsaI combination identifies polymorphic bands at 3.3 kb, 2.7 kb, 2.3 kb, and 0.9 kb. The frequencies of these markers were determined in 90 unrelated Danes. Co‐dominant segregation and allelic behavior was seen for the BATl/NcoI 12 kb and 8 kb bands and the BAT2/RsaI 2.7 kb and 2.3 kb bands, respectively. It is possible that the BAT2IRsaI 3.3 kb band represents a rare allele of the BAT2/RsaI system. The BAT2/RsaI 2.3 kb marker was strongly negatively associated with HLA‐B8 and HLA‐DR3 while there were no strong associations between the BATl and BAT2 markers, but the BAT2/RsaI 2.7 kb marker was strongly positively associated with the TNFα/NcoI 5.5 kb marker, which in turn is positively associated with HLA‐B8 and HLA‐DR3. The negative associations between the BAT2/Rsa1 2.3 kb marker and HLA‐B8 and HLA‐DR3 raise the question as to whether this BAT allele may play a role in protecting against certain a

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01800.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:Among the DR specificities undefined by serology, DR‐BON is peculiar because RFLP cannot distinguish it from the well‐defined allele DR1 even if the two specificities are very different functionally. The occurrence of two DR‐BON‐like alleles in the same family, one associated to the DQw5 split of DQwl and the other associated to the DQw7 split of DQw3, enabled us to compare the properties of these alleles. The RFLP analysis showed a typical DR1‐like picture for both alleles when probed with DR beta, but for one of the alleles the DQ beta probe gave a DQw7 pattern. Primary mixed lymphocyte cultures showed weak to moderate stimulation between cells from individuals identical for one haplotype and differing for the DR‐blank haplotypes, but by test with cloned reagents we were not able to define differences between the two DR‐blank molecules. Two‐dimensional gel electrophoresis and spot‐test with a probe covering the third hypervariable region of the DRBl gene showed no difference between the two alleles. We thus think that the two DR alleles are identical and that the stimulation observed in primary cultures probably is caused by incompatibility for DQ

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01801.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:We have previously reported that in nonresponders to hepatitis‐B (HB) vaccine there was an HLA‐linked immune suppression gene for hepatitis‐B surface antigen (Is‐HBsAg) controlling the nonresponsiveness to HBsAg, through HBsAg‐specific suppressor T cells, and that the Is‐HBsAg was in strong linkage disequilibrium with the HLA‐Bw54‐DR4‐DRw53 haplotype (1). We have now done the HLA typing on an additional 6 nonresponders, and using the system of T‐cell proliferative response to HBsAg we found that the Is‐HBsAg controlled the nonresponsiveness to HBsAg through HBsAg‐specific suppressor T cells in nonresponders to HB vaccine who have HLA‐Bw54‐DR4‐DRw53‐DQw4 haplotype. T‐and B‐cell recognition of HB vaccines might play an important role at 3 to 5 weeks after the last immunization. Use of an anti‐HLA monoclonal antibody has shown that the HLA‐DR molecule plays an important role in helper T‐cell proliferation in nonresponders, although the role of HLA

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01802.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:Modifications of the standard microcytotoxicity assay make it possible to use this technique to screen both alloantisera and monoclonal antibodies with mouse L cells transfected with Class II genes. It is necessary to maintain a protein‐rich environment in order to prevent nonspecific complement lysis. Selection of the complement itself is also an important factor, the best results being acheived using a commercially available complement that had previously been absorbed with mouse cells and used at a dilution of 1/8. Using this modified method with transfectants of Dw2 origin we could show that alloantisera against DRw15 recognize the DRB1*1501 gene product, whereas broad DR2 sera react only with the DRB5*0101 product. This technique can be applied succesfully to study the fine specificity of polymorphic monoclonal antibodies, as shown by the reactivity of HU‐30 which binds to the LDR2b transfectant and not to the LDR2a, indicating that the antibody recognizes an epitope present on the DRBl chain and not the DRB5 chain of DR2 cell li

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01803.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:The HLA‐DR4 specificity revealed a relative risk of 8.5 (x2= 99.6; p<0.0001) when 193 type I diabetics were compared to 305 controls. Prevalence of the HLA‐DR4‐associated DQ types, i.e. DQw7 and DQw8, were determined, using a restriction fragment length polymorphism (RFLP) typing that combines the probe/enzyme combinations DQB/TaqI and DQB/BamHI. The HLA‐DQw8 specificity was confined to HLA‐DR3/DR4 heterozygous patients when compared to controls (x2= 4.9; p<0.025) or to all other DR4‐heterozygous patients (x2= 6.7; p<0.01). No association with HLA‐DQw8 was seen in HLA‐DRl/DR4 or HLA‐DR“X”/DR4 (X ≠ 1,3,4) heterozygous patients. Due to the excess of HLA‐DR3/DR4 patients the DQw8 allele is a risk factor in type I diabetics, but in HLA‐DRl/DR4 and DRX/DR4 heterozygotes one might suggest that DOB1 and DRB combinations confer HL

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01804.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

摘要:

Abstract:Recently, a new technique for HLA class II genotyping has been introduced, the polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method, claimed to be a practical alternative to conventional serological and cellular HLA class II typing (1‐3). The PCR‐RFLP technique is ingenious, relatively rapid and does not require hybridization with sequence‐specific oligonucleotide probes. However, analysis of whether homozygous and heterozygous combinations of PCR‐RFLP patterns for the various investigated HLA class If loci are unique or not unfortunately shows that only 19% ofDRBhomozygous and heterozygous combinations are unique. The figures for theDQA1,DQBandDPBloci are 56%, 29% and 65%, respectively. As not all nucleotide sequences analyzed in the above‐mentioned studies (1‐3) gave rise to unique PCR‐RFLPs and as more sequences now are known, for theDRB1, DRB3, DRB5andDPB1loci (4), the frequencies of unique PCR‐RFLP patterns for the different HLA class II loci will be reduced even further. Thus, the present analysis demonstrates that the PCR‐RFLP technique, performed as described in references 1‐3, is not yet ready to be used for routine HLA class II genotyping. The resolution of the PCR‐RFLP method can be improved by various modifications. However, the role of a modified PCR‐RFLP technique in HLA class II typin

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01805.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1990.tb01806.x

出版商:Blackwell Publishing Ltd    

年代:1990

数据来源: WILEY