ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02384.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:We developed a PCR‐based approach to sequence exons 2 and 3 of HLA‐B44 alleles from genomic DNA. We applied this method to determine the B44 alleles encoded on extended HLA‐A, B, DRB1, DQB1 haplotypes and the degree of mismatching for B44 alleles among marrow transplant patients and their unrelated donors (URD). A total of 81 samples was studied and included 38 patients, 42 donors and the cell “FMB”; the 80 clinical samples were comprised of 8 unpaired patients, 12 unpaired donors, and 30 URD‐recipient pairs. Three alleles encoding B44 were identified, B*4402 (N=51), 4403 (N=32) and a new allele designated B*44KB and named B*4405 (N=4). Of the 27 patients for whom family study was available, there were 13 different B*4402, 7 different B* 4403 and 2 new B*4405 haplotypes. HLA‐A2, Cw*0501, B*4402, DRB1* 0401, DQB1*0301 (n=2); A2, Cw*0501, B*4402, DRB1*1501, DRB5* 0101, DQB1*0602 (n=2); and HLA‐A29, Cw*1601, B*4403, DRB1* 0701, DQB1*0201 (n=5) comprised the most common patient haplotypes. Of 30 URD‐recipient transplant pairs studied, 27 were HLA‐A, B serologically matched and DRB1, DRB3, DRB5, DQB1 allele matched, and 3 pairs were DRB1‐mismatched. All B44 allele mismatching (N=3) occurred among the 27 matched pairs. The novel B*4402‐variant sequence, HLA‐B*4405, was identified in 4 individuals, and in each case was associated with an HLA‐B44, Cw*02022, DRB1*0101, DQB1*0501 haplotype. HLA‐B*4405 and B*4402 are identical in exon 2; in exon 3 however, B*4405 encodes T instead of G at nucleotide position 75 which translates to a substitution of tyrosine for aspartic acid at codon 116. Finally, the published B*4402 sequence derived from cell “FMB” was found to contain an error; the corrected B*4402 sequence encodes G rather

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02385.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Santamaria et al. (Human Immunology 1993 37: 39–50) describe a method of sequence‐based typing (SBT) for HLA‐A, B and C alleles said to give “unambiguous typing of any sample, heterozygous or homozygous, without requiring additional typing information”. From SBT analysis, which involves determination of partial sequences of mixed alleles, these investigators reported that cell lines KT17 (HLA‐B35,62) and OLGA (HLA‐B62) from the reference panel of the 10th International Histocompatibility Workshop express novel variants of HLA‐B15 (B1501‐MN6) and HLA‐B35 (B3501‐MN7) respectively. To study further the novel alleles, we cloned and sequenced full‐length HLA‐B cDNA clones isolated from the KT17 and OLGA cell lines. We find that KT17 expresses B*3501, as assigned by SBT, and B*1501, the common allele encoding the B62 antigen. We were unable to confirm that KT17 expresses the novel B1501‐MN6 variant identified by SBT. For OLGA our analysis confirms the partial sequences obtained by SBT. Thus OLGA expresses B*1501 and a novel HLA‐B allele. The complete sequence of the latter shows it is a hybrid having exons 1 and 2 in common with B*1501 and other B15 subtypes and exons 3–7 in common with B*3501 and related molecules including B*5301 and B*5801. The novel allele has been designated B*1520 because of its sequence similarity with the B15 group; furthermore, serological analysis shows that the B*1520 product does not express epitopes in common with either B35, B53 or B58. The B*1520 heavy chain has a similar isoelectric point to A*3101; B*1520 was undetected by previous applications of isoelectric focusing because B*1520 and A31 are both expressed by OLGA. In conclusion, HLA‐B typing of two cell lines by cDNA cloning and sequencing gives concordant results with SBT for three of the four alleles. The cause of the discrepany for the fourth allele is unknown, however this finding indicates that the novel HLA‐A, B and C sequences emerging from SBT st

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02386.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:HLA‐DR and ‐DQ typings were performed by a combination of RFLP and PCR‐SSP techniques in 234 Danish women with at least three consecutive unexplained fetal losses (recurrent fetal losses) and 360 controls and the DRB1, DQA1 and DQB1 alleles were deduced. In the total group of patients, the frequency of no DRB1‐DQA1‐DQB1 haplo‐type was significantly increased compared with controls. In the subgroup of 97 women with four or more fetal losses (multiple fetal loss group), the frequency of women carrying the DRB1*0101, DQA1*0101, DQB1* 0501; DRB1*0102, DQA1*0101, DQB1*0501 and DRB1*0103, DQA1*0101, DQB1*0501 haplotypes or the DRB1*0301, DQA1*0501, DQB1*0201 haplotype were significantly increased compared with controls (RR=2.1; pc<0.05 with regard to former three haplotypes combined and RR=2.2; pc<0.05 for the latter). The frequency of women with at least one of the four haplotypes was significantly (p<0.002) increased with the number of previous fetal losses in the women's history. Analysis of the DQA1 and DQB1 phenotypes in women with at least four fetal losses showed that DQA1*0501 and DQB1*0501 were increased compared with controls (RR=1.9; pc<0.05 and RR = 2.2; pc<0.025, respectively). Analysis of DRB1‐DQA1‐DQB1/DRB1‐DQA1‐DQB1 genotypes suggested that genotypes comprising both DQA1*0501 and DQB1*0501 alleles (intrans) exhibited a higher RR for experiencing at least four fetal losses (RR=3.4, p=0.002) than each of the alleles did alone. Genotypes comprising a DQB1 allele encoding a β chain negative for aspartate in position 57 were associated with an increased RR (2.3; p<0.01) for multiple fetal losses and the etiologic fraction was high (0.49). The results suggest that genes or gene‐sequences, linked to the DRB1*0101, DQA1*0101, DQB1*0501; DRB1*0102, DQA1*0101, DQB1*0501; DRB1*0103, DQA1*0101, DQB1*0501 and DRB1*0301, DQA1*0501, DQB1*0201 haplotypes, confer susceptibility to multiple fetal losses. These alleles/gene‐sequences are presumably located in the DQA1 and DQB1 loci and may elicit their effect by mediating an autoimmune reaction against one or m

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02387.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Although many studies have established an association between insulin‐dependent diabetes mellitus (IDDM) and the class II region of the human major histocompatibility complex (MHC), it has been difficult to assign susceptibility to a single locus. Recently, two antigen‐processing genes,TAP1andTAP2, have been identified within the region. Previous studies have reached conflicting conclusions as to the role of these genes in IDDM; it is uncertain whether an increased frequency of the alleleTAP2Aand a concomitant decrease inTAP2Bare independent disease associations or secondary to linkage disequilibrium (LD) betweenTAP2AandHLA‐DR3. To further investigate this question, we have characterizedTAP1andTAP2alleles in 129 IDDM patients from Sardinia, a population with limited genetic heterogeneity and a high disease incidence. When compared to 90 random controls, the only significant difference was a decrease in the minor alleleTAP2Cin patients. However, whenHLA‐DRand ‐DQmatched controls were compared, this difference disappeared. Further analysis suggested thatTAP2Cwas in LD with HLA‐DRB1*1401 and subtypes ofHLA‐DRB1*11, alleles which were not observed in the IDDM population. LD was also observed between otherTAPandHLA‐DRalleles, in particular betweenTAP2AandHLA‐DR3in both patients and controls. Our data supports the conclusion that there is no primary association betweenTAP2alleles and IDDM, and that previously reported associations may be due to LD with ot

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02388.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:In spite of the widespread use of rhesus monkeys (Macaca mulatta) in biomedical research, MHC typing of this species is not yet routine. Since suitable antibodies are lacking, serological typing of Mamu‐DQA1 is not feasible. We developed a typing protocol forMhcMamu‐DQA1from published sequences of the second exon ofMamu‐DQA1. This protocol is based on the amplification of the second exon ofMamu‐DQA1with one specific primer pair followed by a “diagnostic” digestion of the PCR products with, at most, 5 different restriction endonucleases. This modified PCR‐RFLP permits the rapid identification of 11 out of 13Mamu‐DQA1alleles in homozygous and heterozygous combinations. The protocol was validated by cloning and sequencing the PCR‐products of several animals of different geographic origin. In addition, an as yet unknown allele was detected by PCR‐RFLP and was subsequently cloned and its nucleotide sequence determined. All examined sequences except the new allele were identical to those previously published. Therefore, we assume that many of theMamu‐DQA1alleles of rhesus monkeys have been identified molecularly and that the typing technique presented here can reliably ide

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02389.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:We performed HLA‐DQA1, ‐DQB1 and ‐DRB1 genotyping using the PCR‐RFLP (polymerase chain reaction‐restriction fragment length polymorphism) method for 32 Japanesepemphigus vulgaris(PV) patients. There was a significant association of either DQB1*0503 or DRB1*1405 with PV, and a negative association of either DQA1*0103 or DQB1*0601 with PV was found. Since the DQB1*0503+ patients had various DR14‐related alleles, we concluded that the association with DQB1 is primary and that the association with DRB1 is simply due to linkage disequilibrium between the DQ and DR genes. These results may indicate that specific HLA class II antigens confer the susceptibility to PV amo

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02390.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Preliminary characterization of an apparently novel bovine leukocyte adhesion protein is described. Two IgG1monoclonal antibodies, UC‐C1 and UC‐H5, raised against established cultures of IL‐2‐dependent bovine peripheral blood lymphocytes (PBL) were found to react with an antigen expressed by the majority of bovine peripheral blood leukocytes. Immunoprecipitation and polyacrylamide gel electrophoresis of the antigen produced a distinct protein band of molecular weight 160 000, and additional diffuse protein bands of approximate molecular weight 180 000, 175 000 and 150 000. Two‐color flow cytometric analyses showed that the antigen was expressed at low density on a small proportion of circulating B lymphocytes, but was highly expressed on all circulating T lymphocytes. The majority of monocytes and all granulocytes expressed the antigen at a density lower than that of T lymphocytes. Peripheral blood lymphocytes stimulated with concanavalin A had an approximately 3‐fold increased expression of the antigen, which was apparent within 18 h and remained stable in long‐term cultures. Expression of the antigen in thymus, analyzed by the immunoperoxidase technique, was predominantly restricted to thymocytes in the immediate subcapsular cortex and medulla; expression in lymph nodes and spleen was predominantly confined to lymphocytes in T‐cell areas. Flow‐cytometric analysis demonstrated that thymocytes and the majority of peripheral and mesenteric lymph node‐derived T cells had relatively low surface density of antigen compared to circulating T cells. Binding of UC‐C1 or UC‐H5 to the antigen on lymphocytes induced homotypic aggregation. UC‐C1 completely blocked binding of FITC‐conjugated UC‐H5 to blood mononuclear cells, suggesting that the antibodies recognize the same

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02391.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02392.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02393.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY