ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01932.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

摘要:

Abstract:A human monoclonal IgM antibody, referred to as TU223, has been produced. The reactivity of TU223 was tested in various cells and cell lines by complement‐dependent microcytotoxicity test and fluorescence‐activated cell sorter analysis. The antigen defined by TU223 was expressed on Epstein‐Barr virus‐transformed B‐cell lines and on some Burkitt's lymphoma cell lines, but was not expressed on normal T cells, B cells or erythrocytes. In addition, expression of the antigen defined by TU223 was also induced on B cells activated by Epstein‐Barr virus or pokeweed mitogen, and on T cells activated by phytohemagglutinin, concanavalin A, pokeweed mitogen or recombinant interleukin‐2. However, no expression of the antigen detected by TU223 was induced at all on recombinant interleukin‐4‐treated B cells or macrophage‐like cell line U937. When the ability of TU223 and various mouse monoclonal antibodies to bind to human differentiation antigens was compared, interestingly, the reactivity of TU223 was found to be very similar to that of mouse monoclonal antibody CD23 (H107), which reacts with Fcs receptor 11. Two‐color analysis revealed that the antigen defined by TU223 is expressed on the cell surface of certain lymphoid cells expressing CD23 antigen. However, it can be concluded that the antigen defined by TU223 is clearly distinct from Fcs receptor 11, based on assay of cross‐blocking between HI07 and TU223. The surface antigen on B85 cells recognized by TU223 had the molecular size of 80–82 kiloDaltons as determined by immunoblotting analysis. In biological function, TU223 inhibited the generation of plaque‐forming cells when added to a pokeweed mitogen‐derived IgG‐secreting system on days 0 to 4. These findings suggest that TU223 recognizes a novel and functional antigen expressed on c

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01933.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

摘要:

Abstract:We modified a previously published PCR‐RFLP for DQA1 typing (1) and examined the predictive value of HEA‐DQA1 in mixed lymphocyte cultures (MLC) among matched (HLA generic types) pairs of unrelated individuals. There were 61/102 (60%) pairs with positive MLC, one‐third of which could be predicted by DQA1* typing alone. DQA1 matching and MLC reactions were classified into 3 groups: 1) DQA1 mismatches showing positive MLC: 19/102 (19%); 2) DQA1 matches showing negative MLC: 41/102 (40%); 3) DQA1 identical showing positive MLC: 42/102 (41%). Five different HLA haplotypes that result from non‐random association of HLA generic types (high delta haplotypes) were overrepresented in the individuals tested. One of these haplotypes carrying HLA‐B7, DR2 was found associated with three different DQA1 alleles (*0201, *0103, *0102). The remaining four high delta haplotypes were associated with one DQA1 allele in all independent examples tested: HLA‐A1, B8, DR3 with DQA1*0501; HLA‐A26, B38, DR4 with DQA 1*0301; HLA‐A2, Bw62, DR4 with DQA1*0301 and HLA‐AI, Bw57, DR7 with DQA1*0201. Forty per cent of the negative MLC were explained in part by the excessive number of individuals carrying two of these four haplotypes, which probably carry determinants in linkage disequilibrium with HLA. Nineteen per cent of HLA‐identical (generic types) unrelated pairs show positive MLC reactions and all of them are DQA1* mismatched, suggesting that DQA1* allele typing should be used to screen samples prio

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01934.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

摘要:

Abstract:We previously introduced HLA‐DQA1,‐DPB1 and DQB1 genotyping with the modified PCR‐RFLP method using some informative restriction enzymes which have either a single cleavage site or alternatively no cleavage site in the amplified DNA region, depending on the HLA alleles, making reading of RFLP band patterns much easier. In this study, 43 HLA‐DRB1 alleles, excluding DRB1*1103 and *1104 for which no restriction enzymes are available to distinguish each from the other, could be defined by this modified PCR‐RFLP method combined with 7 pairs of group‐specific primers. It is impossible to distinguish DRB 1*0701 and DRB1*0702 as they are identical for the second exon of DRB1. For DRI‐DRB1, DR2‐DRB1, DR4‐DRB1, DR7‐DR1, DR9‐DRB1, DRw10‐DRB1 or DRw52 associated antigens (DR3, w11, w12, w13, w14, and DRw8)‐DRB1 gene amplification, the second exon of the DRB1 gene was selectively amplified using each group‐specific primer from genomic DNAs of 70 HLA‐homozygous B‐cell lines and healthy Japanese by PCR. Amplified DNAs were digested with restriction endonucleases and then subjected to electrophoresis assaying simply for cutting, or no cutting, of the DNA, although some alleles can be distinguished only after examination of RFLP band patterns generated and in some cases using double digestion technique with two restriction enzymes. This modified PCR‐RFLP method can be successfully applied to all possible DRB1 heterozygotes, despite the fact that 15 pairs of heterozygotes among them cannot be distinguished theoretically by the PCR‐SSO method, because the PCR‐RFLP method can tell whether two polymorphic sites are linked to each other (cis position) or located on a different chromosome (trans position) by checking the length of RFLP bands generated with double digestion. Thus, the PCR‐RFLP method is technically simple, practical and inexpensive for determination of the HLA

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01935.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

摘要:

Abstract:We have used a PCR‐RFLP method with one generic amplification of HLA‐DPB1 second exon and 6 endonucleases to differentiate the 19 HLA‐DPB1 alleles and 171 heterozygous combinations. The set of primers used in our studies produced fragment sizes different from those published before (1). The HLA‐DPB1 alleles in Caucasians showed a higher frequency of DPB 1*0401 and DPB 1*0402, when compared to a small group of Colombians who showed a higher frequency of DPB 1*0402 and DPB 1*0201. We found three HLA‐DPB1 alleles associated with two HLA haplotypes that result from non‐random association of alleles: DPB1*0401 with HLA‐A26, B38, DR4, DQA1*0301 and DPB1*0101 and DPB1*0401 with HLA‐A1, B8, DR3, DQA1*0501. We also report that 70% of combinations between HLA (generic A,B,C,DR) and DQA1‐identical MLC‐unreactive cell mixtures showed HLA‐DPB1 mismatches, suggesting that HLA‐DPB1 differences are not impo

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01936.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

摘要:

Abstract:A total of 61 DNA sequences from human major histocompatibility class II loci were searched for statistical evidence of past genetic exchange (gene conversion or recombination). Among the 12 A‐locus sequences (derived from DPA1 and DQA1), 4 clusters indicating potential exchange events were found. Among the 49 B‐locus sequences (derived from DOB, DPB1, DPB2, DQB1, DRB1, DRB3, DRB4 and DRB5), 15 clusters were found. The clusters suggested short exchanges (less than 100 bp) within and between loci, and were concentrated in exon 2 (coding for the antigen binding site). The most striking feature of the results was the presence of an approximately 200‐bp region in the middle of B‐locus exon 2 which contained almost no locus‐specific substitutions, which were abundant elsewhere. This suggests either strong selection for locus specificity in the other regions of the gene or a history of frequent between‐locus exchange in this part of exon 2, which is involved in forming the antigen b

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01937.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01938.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1992.tb01939.x

出版商:Blackwell Publishing Ltd    

年代:1992

数据来源: WILEY