摘要:

Abstract:Naturally‐processed self peptides bound to HLA class I molecules of a T‐cell leukemia (HLA‐A1, A31, B38, B58) were isolated for sequence analysis. Acid‐eluted peptides were subjected to reversed‐phase HPLC separation and single‐fraction sequencing was performed by Edman degradation. The peptides were found to be mostly nonamers and could be grouped into three distinct structural motifs. One of the peptide groups consistently displayed histidine at position 2 and a bulky hydrophobic residue at the C‐terminus (XHXPXXXXY/F). The only HLA class I structure expressed by this T‐cell leukemia which is consistent with the binding of peptides carrying this sequence motif is HLA‐B38. A peptide binding assay confirmed this assignment. HLA‐B38 is present in 10–12% of the Jewish population and is associated with several autoimmune disorders. The HLA‐B38 motif may be an important tool for identifying potential T‐cell epitopes involved in these diseases and for d

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02361.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:A murine monoclonal antibody (mAb) UN1 was produced on the basis of selective reactivity with human thymocytes. Characterization of UN1 by immunofluorescence gave a high intensity of labeling with the majority of human thymocytes. Expression was preferentially associated with immature thymocytes (CD3dim) compared to mature cells, whereas only a subpopulation of peripheral blood lymphocytes was weakly stained. No specific binding to monocytes or granulocytes was detected. The T‐cell lines HPB‐ALL, H9 and MOLT‐4 were all positively bound by UN1. Immunohistological staining of thymic tissues showed that mAb UN1 detected cells in both the cortex and medulla of fetal thymus, whereas the reaction in thymus samples from young children was mainly with medullar cells. By western blotting analysis, the antigen recognized by mAb UN1 corresponds to a membrane polypeptide with a molecular weight of approximately 120 kDa present on thymocytes and HPB‐ALL cells. The mAb UN1 was submitted to the 5th International Workshop and Conference on Human Leukocyte Differentiation Antigens, Boston, 1993. UN1 did not cluster in any of the old or new clusters of differentiation discussed at the conference, indicating its unique reactivity. Together with the data presented in this paper, this suggests that the UN1 antibody defines a previously undescribed molecule present on the cell surface of thymocytes and a minority of peripheral blood lymp

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02362.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:The utility of the MLC assay as a test of HLA‐D region matching and predictor of acute graft‐versus‐host disease (GvHD) was evaluated in 157 patients receiving marrow grafts from HLA‐A, B identical related haploidentical donors. All donors and recipients were tested by HLA‐DR serology, by Dw phenotyping with homozygous typing cells (HTC) and by standard MLC. Ninety‐nine of the donor‐recipient pairs were mismatched for a serologically defined HLA‐DR antigen while 109 pairs were mismatched for the HLA‐D region by HTC typing. Donor anti‐recipient relative responses (RR) in MLC, corresponding to the GvHD vector in marrow transplantation, ranged from ‐4% to 100%, with a median of 25%. A comparison of reactivity in MLC with presence or absence of matching by Dw phenotyping, however, showed a significant overlap in the distribution of RRs from HLA‐Dw matched versus Dw mismatched pairs, suggesting that the MLC was not a reliable predictor of HLA‐Dw matching. Using an optimally‐defined cutoff of 3% RR, the MLC was correlated with risk of developing clinically significant grades II–IV acute GvHD (p = 0.03) but not with risk of developing severe grades III‐IV GvHD (p = 0.18). In contrast, matching by Dw phenotype was a significant predictor of GvHD, with Dw‐compatible transplant recipients less likely to develop either grades II‐IV (p = 0.004) or III‐IV (p = 0.036) GvHD than Dw‐incompatible transplant recipients. Overall, these results underscore the difficulty in using the MLC to measure HLA‐D region compatibility and predict the risk of severe graft‐versus‐host disease among patients receiving related haploidentical marrow grafts. HLA‐D (HTC) typing results correlate primarily with DRB compatibility, and with the advent of DRB1 allele matching by sequence‐specific oligonucleotide probes (SSOP) or by direct sequencing, the precision in donor matching achievable with these methods is far greater th

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02363.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:The diversity of HLA‐C exon‐2 alleles in 56 HLA‐A, B, DRB and DQB1‐matched patient‐unrelated marrow donor pairs was examined by non‐cloning polymerase chain reaction‐based sequencing of genomic DNA. This method allows simultaneous analysis of both alleles in heterozygous samples. All Cw5‐positive individuals encoded a sequence which differed from the published Cw*0501 sequence at position 61. Among 82 samples assigned a single antigen by serologic testing, 64 (78%) were heterozygous for two distinct alleles when tested by sequencing. Cw*1202, 1601 and 15 were identified in samples for which no phenotype could be assigned (C “blank”). Finally, 7 of the 56 HLA‐A, B, DRB, DQB1‐matched pairs (12.5%) were mismatched for one or both HLA‐C alleles. We conclude that sequence‐based methods constitute the optimal strategies for typing HLA‐C alleles in the unrelated

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02364.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:Previous studies on expressed bovine MHC class II polymorphism using one‐dimensional isoelectric focusing (1D‐IEF) allowed the identification of at least 12 allelic variants of the DRB3 gene. So far, only limited data have been available on the expression of other class II genes. The present study involved biochemical analysis of bovine MHC class II molecules using a set of monoclonal antibodies presupposed to be bovine DR and DQ reactive. After essential modification of the standard electrophoresis conditions used for 1D‐IEF typing of bovine DR products, biochemical polymorphism was observed for non‐DR molecules, revealing polymorphic sets of basic and acidic focusing bands. Because of the extensive DNA polymorphism described for bovine DQA and DQB genes, and the apparent similarity with the focusing pattern of human DQ products, these molecules were considered to be the bovine DQ homologues. The definition of the DQ‐associated banding patterns was made possible by using two half‐sib sire families. Four different DQA‐like patterns and nine DQB‐like patterns were detected. Segregation of the DQ types was supported by serological class I and class II typing. These results show that it is now possible to discriminate between expressed bovine DR and

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02365.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:HLA‐DR2 is negatively associated with insulin‐dependent diabetes mellitus (IDDM). The aim of the present study was to analyze DR2‐positive patients among 425 consecutively diagnosed unrelated Swedish children with IDDM and in 367 matched controls. HLA‐DRB, ‐DQA and ‐DQB were determined by Taq I restriction fragment length polymorphism analysis. Amplification by polymerase chain reaction (PCR) and hybridization with sequence‐specific oligonucleotide probes was done for DQA1, DQB1 and DRB1 and DRB5. DR2 was positive in 11/425 patients (3%) and 101/367 (28%) controls (OR 0.07, p<0.0001). Of the 11 DR2‐positive patients, PCR was done in 10, of whom 8 were positive for DRB1*1601‐DRB5*0201 compared to 4/96 (4%) controls (OR 92.0; p<0.001) while the remaining 2 were positive for DRB1*1501‐DRB5*0101 compared to 92/96 (96%, OR 0.01; p<0.001). In 2 patients, a recombination between the haplotypes DQB1*0502‐DQA1*0102 (DQ5)‐DRB1*1601‐DRB5*0201 (DR16 Dw21) and DQB1*0301‐DQA1*0501 (DQ7)‐DRB1*1602‐DRB5*0202 (DR16 Dw22) was observed resulting in the DQB1*0301‐DQA1*0501 (DQ7) DRB1*1601‐DRB5*0201 (DR16 Dw22) haplotypes. The second haplotype was DR3 DQ2 in 6/11 and DR4 DQ8 in 2/11 DR2‐positive patients. In all 3 DQB1*0602‐DQA1*0102‐DR15‐positive patients the second haplotype was DR4‐positive. In order to test whether physicochemical properties of the DR2 molecules were associated with IDDM, we constructed three‐dimentional models of the peptide binding and T‐cell recognition sites (α1 and β1 domains) of five subtypes of DR2‐DRB1, based on the published DR1 crystal structure. No correlations were observed for DR molecule physicochemical properties and diabetes susceptibility. Islet cell antibodies, insulin autoantibodies and GAD65 antibodies, were measured in DR2‐positive patients (n = 11) and controls (n = 101). Despite the presence of the DR2 haplotype the antibody markers were significantly elevated in the patients compared to the controls (GAD65 3/10 patients and 2/101 controls; ICA 7/11 patients and 1/101 controls and IAA 3/11 patients and 0/101 controls). In conclusion, of the five subtypes of DR2, only one, the DRB1*1501, DRB5*0101, DQB1*0602‐DQA1*0102 haplotype, was negatively associated with IDDM. DQ may

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02366.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.tb02367.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.tb02368.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.tb02369.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.tb02370.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY