摘要:

Abstract:HLA incompatibility between bone marrow recipients and unrelated donors is one of the main obstacles in bone marrow transplantation. HLA class I and generic class II DR and DQ typing is generally performed by serology. Precise subtyping of HLA class II genes, however, can only be achieved by molecular genetic methods. Here, the final selection of serologically pretyped unrelated bone marrow donors by confirmatory PCR‐SSP (PCR‐sequence‐specific primers) typing and subsequent nucleic acid sequence analysis of the second exon of DRB1, DRB3, DRB4, DRB5, DQB1, and DPB1 alleles is presented. Serologically identical potential marrow donors and their corresponding recipients were analyzed for HLA‐DRB identity by PCR‐SSP analysis. After solid‐phase single‐strand separation, direct sequencing of the allele‐ or group‐specific DRB amplified products was performed by applying fluorophor‐labelled sequencing primers. Electrophoretically separated sequencing products were detected by means of an automated DNA sequencer. Group‐specific amplification and sequencing of DQB1 alleles was carried out for all potential bone marrow donors and recipients, while only the final donor‐recipient pair was analyzed for DPB1 alleles. Thus, the presented amplification strategy in combination with direct sequencing of PCR products allows matching of bone marrow transplant pairs with the highest degree of reliability for the assessment o

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02396.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:PECAM‐1, a member of the immunoglobulin gene superfamily, is widely distributed on cells of the vascular system, and mediates cellular interactions through both homotypic and heterotypic adhesive mechanisms. Previous studies have demonstrated that PECAM‐1 is initially expressed at high levels on CD34+multipotential progenitors in the bone marrow, but is subsequently downregulated in more committed precursors of all lineages. Interestingly, although PECAM‐1 expression is high on circulating monocytes and neutrophils, little is known about the upregulation of PECAM‐1 expression during terminal myelomonocytic differentiation. We have further characterized this process by examining PECAM‐1 expression during chemically‐induced differentiation of the U937, HL‐60 and HEL cell lines. Quantitative Western blot analysis of cellular lysates indicated that PECAM‐1 expression could be upregulated in U937 and HL‐60 cells by phorbol esters or dimethyl sulfoxide. Northern blot analysis showed that PECAM‐1 mRNA levels appeared to increase in parallel with that of PECAM‐1 protein. We also observed a marked difference in the apparent molecular mass of PECAM‐1 that was lineage‐specific, both in differentiated leukemic cell lines and in their corresponding leukocyte population. Immunofluorescence localization indicated that the cellular distribution of PECAM‐1 in U937 and HL‐60 cells was similar to that of their normal circulating counterparts, and that the pattern of distribution again displayed lineage fidelity. The ability to induce the expression of PECAM‐1 molecules having different glycosylation and surface expression patterns may prove useful for further elucidation of the role of PECAM‐1 in hematopoiesis, as well as studies of the cell lineage‐spec

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02397.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:HLA class II DNA typing was conducted for 1335 unrelated Japanese individuals. The study on the linkage disequilibrium revealed a striking conservation of HLA DR13 haplotypes. Among these Japanese, 155 were typed for HLA‐DR13 serologically, and they were correspondent to three DRB1 alleles, DRB1*1301, 1302 and 1307 defined by using the polymerase‐chain reaction and sequence‐specific oligonucleotide probe (PCR‐SSOP) method. The two alleles, DRB1*1301 and 1307 were exclusively associated with each specific DRB3‐DQA1‐DQB1 combination which was DRB1*1301‐DRB3*0101‐DQA1*0103‐DQB1*0603, and DRB1*1307‐DRB3*0202‐DQA1*0501‐DQB1*0301, respectively. DRB1* 1302, the most common DR13 allele in Japanese, had two significant associations with DRB3*0301‐DQA1*0102‐DQB1*0604 (DRB1*1302A) and with DRB3*0301‐DQA1*0102‐DQB1*0605 (DRB1*1302B). In this study, no other DR13 class II combinations were found. Ony the DRB1*1302A halotype was associated with the DPB1*0401 allele while the DRB1*1302B haplotype was not. The complete conservation of these DR13 class II haplotypes was found to extend toward the HLA class I region. They were HLA A3‐B44‐DRB1*1301, A33‐B44‐DRB1*1302A and A33‐B17‐DRB1*1302B. Japanese could be characterized with these three extended haplotypes which were remakrably different from those in Caucasian, Bl

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02398.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:SSP‐PCR (sequence‐specific primer) DNA typing was performed in Terasaki trays using 1.5 μ1 of DNA, and the ethidium‐stained PCR product was measured by direct fluorometric reading. Elimination of the gel electrophoresis step greatly simplified the SSP method. 17 serological DR specificities were discriminated for 239 DNA samples utilizing the new method, standard SSP, sequence‐specific oligonucleotide probe (SSOP), and restriction fragment length polymorphism (PCR‐RFLP). Results showed 98% concordance between the SSP‐PCR assay and conventional methods. DRB1 alleles were determined by PCR‐RFLP in 59 samples, by SSP in 110 samples, and by consensus (all methods) in the rem

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02399.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:TrJ14 is a cytotoxic human IgG1Λ hybridoma mAb that recognized a novel HLA‐A epitope expressed by lymphoblastoid B cells that are homo‐ or heterozygous for A2, A3, A11, A30, A31, A33, A68 and A69. Based on these results, the HLA type of cell line TEM (10w9057) was retyped as A66. When peripheral blood T cells isolated freshly from 265 HLA‐typed normal individuals served as targets, TrJ14 killed cells expressing two TrJ14‐positive HLA‐A alleles, as well as the majority of cells having one TrJ14‐positive and one TrJ14‐negative HLA‐A antigen. However, TrJ14 failed to recognize or reacted weakly with most HLA‐A2 and ‐A3 heterozygous normal T cells when A2 or A3 was coexpressed together with a TrJ14‐negative antigen. The serological reactivity of TrJ14 correlated with the amino acid valine and aspartic acid at positions 76 and 77 of the αl‐domain helix. These amino acids were shared exclusively by all the

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02400.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

Abstract:This study describes the characterization of endogenous peptides associated with the two major subtypes of HLA‐B44. The two subtypes differ for a single amino acid substitution from Asp (HLA‐B*4402) to Leu (HLA‐B*4403) in position 156 of the α2 domain, causing strong alloreactivityin vivo.In order to study the involvement of peptides in this phenomenon, the peptide motifs of the two subtypes were determined from natural peptide pools using Edman degradation. The motif was found to be essentially identical for HLA‐B*4402 and ‐B*4403, with a strong predominance for Glu at position 2, Tyr or Phe at positions 9 and 10 and hydrophobic residues, especially Met, at position 3. Two individual naturally processed ligands of HLA‐B*4403 were sequenced and shown to be derived from intracellularly expressed proteins found in protein sequence databases. The sequence of these natural peptide ligands conform well to the determined motif. These data will allow the prediction of HLA‐B44 restricted peptide epitopes from viral and tumor antigens of known amino acid sequences. Moreover, they indicate that the peptide repertoire presented by HLA‐B*4402 and ‐B*4403 is very similar, suggesting that the strong alloresponse between these two subtypes is not due to presentation of a different se

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1994.tb02401.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.tb02402.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.tb02403.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.tb02404.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.tb02405.x

出版商:Blackwell Publishing Ltd    

年代:1994

数据来源: WILEY