ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02610.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Seven samples with irregular PCR‐SSO hybridization patterns, observed during routine HLA‐DRB typing, were studied in more detail. Group‐specific amplification, followed by hybridization with relevant SSOs strengthened the suggestion that these samples contained new DRB alleles. DRB exon 2 segments were amplified, cloned and sequenced and revealed: DRB1*1121 [MUL] is similar to DRB 1*1102 in which codon 85 changed from GTT(V) into GTC(A); DRB1*1419 [AKKAL]is similar to DRB1*1402 with codon 71 changed from AGG(R) into AAG(K); DRB1*1420 [OND‐52971] is a DRB1*1406 with codon 37 changed from AAC(N) into TTC(F); DRB1*1421 [TGI]is similar to DRB1*1417 with codon 71 changed from AGG(R) into AAG(K); DRB3*0203 [POS] is similar to DRB3*0202 in which codons 37–38 are changed from TAC GCG(YA) into TCC GTC(SV); DRB5*0103 was found in two unrelated individuals of Oriental origin [IND‐24 and IND‐59]and is similar to DRB5*0102 in which codon 71 AGG(R) changed into ACG(T). This particular sequence variation at position 71 has not yet been described. The new DRB sequences were confirmed using the sequencing based typing technique. Low resolution PCR‐SSP typing failed to amplify two of the DRB1*14 variants, whereas high resolution PCR‐SSP resulted in aberrant patterns. Class II alloantisera identify the codon 71 changes in DRB1*1419 and *1421 with respect to the MC1(

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02611.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

TAP, LMP and DM genes map within the major histocompatibility complex (MHC) class II region between the DQB1 and DPB1 loci, and are involved in the processing of peptides bound to HLA class I or class II molecules. In order to determine the various linkage disequilibria existing between these genes and HLA class II genes, we have analyzed TAP1, TAP2, LMP2, DMA, DMB, DRB1, DQA1, DQB1 and DPB1 polymorphisms in 162 unrelated healthy Caucasian individuals. Many positive or negative associations were observed between alleles at these loci, such as between DR/DQ and TAP2, DM or LMP, between DP and DMB, and between TAP2 and DM, TAP2 and LMP. Conversely, no linkage disequilibrium was detected between some closely related genes (DR/DQ and TAP1, TAP1 and TAP2, LMP2 and DM), in agreement with the existence of recombination hot spots in this region. Other weak linkage disequilibria are likely to exist in this region. These data allow to define some conserved MHC class II haplotypes including HLA class II and TAP, LMP and DM alleles. Furthermore, the knowledge of such linkage disequilibria is of outstanding importance in order to avoid misinterpretation of the data when studying MHC class II associations with autoimmune diseases.

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02612.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

This study reports the first data on gene frequencies of platelet alloantigens HPA‐1, HPA‐2, HPA‐3, HPA‐4 and HPA‐5 in a population of unrelated Danish blood donors using PCR‐techniques. The observed gene frequencies fit the Hardy‐Weinberg equilibrium, and the calculated phenotype frequencies are similar to those obtained in other Caucasian populations: HPA‐la and ‐lb occur in 96.6% and 30.3% of 557 unrelated respectively. HPA‐2a and ‐2b in 99.4% and 15.9% of 163 tested, HPA‐3a and ‐3b in 88.3% and 63.2% of 163 tested, HPA‐4a and ‐4b in 100% and 0% of 131 tested, and finally HPA‐5a and ‐5b in 100% and 15.7% of 427 tested. It is a major technical improvement to use PCR techniques for genomic typing of HPA. Not only is it possible to perform HPA typings in severely thrombocytopenic patient and on amniotic fluid cells of the fetus of alloimunized mothers, but it must be expected that accuracy of the HPA typing will increase considerably, as has been the case with genomic HLA class II typing. Finally, use of PCR technique combined with allele‐specific primers is suitable for accurate large scale typing of platelet donors, which may be

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02613.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Molecular typing of HLA DQB1 alleles, employing sequence‐specific primers (SSP) for PCR amplification, was used to test a novel method that eliminates the requirement for subsequent gel electrophoresis or additional hybridization steps by directly detecting positive reactions. We have evaluated the performance of this fluorescence‐based oligonucleotide probe assay to assign the most common DQB1 alleles on DNA from 14 homozygous cell lines and in a blind study of 50 diabetic patient samples that had been previously typed at the DQB1 locus using SSOP and conventional SSP‐based approaches. We used a panel of 14 DQB1 SSP primer pairs, internal control primers, and a combination of 4 fluorescent oligonucleotide probes to detect 14 alleles or groups of alleles and controls. We can reliably detect single‐base allelic differences, observe 100% concordance with the results obtained using both of the standard methods, and are able to further subtype several alleles that are not easily distinguished using SSOP (e.g. DQB1 *0401/0402 and DQB1 *0302/ 0303). Sequence‐specific priming and exonuclease‐released fluorescence (SSPERF) detection is technically simple and can be performed in less than 2 hours, including DNA extraction, PCR amplification, data analysis and allele identification. This method is particularly useful for the analysis of large numbers of samples, for which high throughput is critical and for which gel‐based approaches are difficult to perform. This technique may also be useful for small‐scale class I and class II molecular typing in clinically orient

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02614.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

HLA‐C polymorphism of 11 individuals from Papua New Guinea was studied by serology and DNA typing (SSP ARMS‐PCR). To resolve certain discrepancies HLA‐C alleles were cloned and sequenced. Five alleles were identified by sequencing, four of which; Cw*0304, Cw*0401, Cw*12022 and Cw*1502 have been identified previously in other populations. The fifth allele, which was found in four individuals is a novel HLA‐C allele. The new allele, called HLA‐Cw*0403 is most similar to HLA‐Cw*0401, differing by 10 nucleotides, 9 of which are located in the region from nucleotide 98 to 218. This region of Cw*0403 is identical to both HLA‐Cw*0201 and Cw*02022. The 9 nucleotide differences between Cw*0401 and Cw*0403 result in 6 amino acid differences in the α1 domain. These amino acids in Cw*0403 may contribute to the serological typing of some, but not all Cw*0403 expressing cells. The Final difference between Cw*0401 and Cw*0403 is a coding substitution at nucleotide 979 in exon 5. The guanine found in Cw*0403 is identical to all HLA‐C alleles except HLA‐Cw*0401, which has an adenine. The Cw*0403 allele was most likely formed by a gene conversion event between Cw*02 and Cw*04, involving a minimum of 121 to a maximum

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02615.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We have sequenced DNA from six new DR52‐associated DRB1 alleles initially detected by PCR/SSOP analysis. Three DR8 associated alleles differed from previously known alleles by single nucleotide substitutions. DRB1*0807 and DRB1*0811 both vary from DRB1*08021 at codon 57 resulting in two different amino acids at this residue. DRB1 *0807 was identified in samples of Brazilian origin while *0811 was identified among samples from the Tlingit Native American population of Southeast Alaska. DRB1*0814, identified in a family of Chinese origin, differed from DRB1*08032 at codon 12 at both the nucleotide and the amino acid level. In addition, two alleles of DR11, DRB1*1113 and *1119, were each detected in Caucasian individuals. DRB1*1113 differs from other DR11 alleles at codons 37, 67, 70 and 74, while DRB1*1119 differs from *1101 by a single nucleotide substitution at codon 67. Finally, DRB1*1418 was detected in a sample from an Asian or Pacific Islander and shares sequences with several other DR52‐associated DRB 1 alleles. These six DRB 1 alleles appear to have been generated by either gene conversion events, DRB1*1113 and * 1418, or by point mutations, DRB1*0814, *0807, *0811 and *1119, although the single nucleotide substitutions found in the latter three alleles are also present in at least one other DRB1 allele and, therefore, could have been the product of gene conversi

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02616.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Following a successful immune response against invading microorganisms, the majority of activated T cells is eliminated, while a minor fraction survives as memory T cells. A decline in T lymphocyte growth factors such as interleukin‐2 (IL‐2) appears to play a role in the elimination of previously activated T cells. Thus, removal of IL‐2 from proliferating T cells not only induces growth arrest, but triggers a massive cell death due to apoptosis. While the apoptotic response involves a series of well‐described events, it remains less clear how apoptosis is regulated following IL‐2 withdrawal. Here, we provide evidence that CD 18 molecules (β2‐integrins) play a regulatory role in the apoptotic response following removal of IL‐2 from previously activated, antigen specific CD4+ T cell lines. Thus, CD 18 mAb inhibited the apoptotic response to IL‐2 deprivation, whereas mAb against other adhesion molecules (CD28, CD29, CD49d, CD80, CD86) did not. Secondly, IL‐2 withdrawal resulted in a retarded apoptotic response in LFA‐1 (CD11a/CD 18) negative T cells obtained from a leukocyte adhesion deficiency (LAD) patient, as compared to LFA‐1 positive T cell lines. Thirdly, co‐culture of LFA‐1 positive‐ and negative‐ T cells at different ratios induced apoptotic responses that were higher than expected, had the two lymphocyte populations not been interacting and significantly higher than that seen in pure LFA‐1 negative T cells. Supema‐tants from LFA‐1 positive T cell cultures undergoing apoptosis did not induce an enhanced apoptotic responses in LFA‐1 negative T cells, and, reversely, culture supernatants from LFA‐1 negative T cells did not rescue LFA‐1 positive cells from undergoing apoptosis. The apoptotic response was partly blocked by IL‐15, a newly identified T cell growth factor. Taken together, these findings suggest that CD 18 molecules (β2‐integrins) play a regulatory role in the apoptotic response following cytokine withdrawal, and that the regulation is mediated, at least partly, through T‐T cell interactions. Thus, apoptotic death following IL‐2 deprivation appears

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02617.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02618.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

The DQB1*06011 allele first classified and registered with the codon ACC at position 51(1) was recently corrected to ACG by Dr. Akinori Kimura (2) and in independently confirmed in our laboratory (3). The correct nucleotide sequence for this allele is shown below. The DQB1*06011 allele was found in two sisters of Turkish nationality who had been serologically typed for class I as HLA‐A11, A33, B44, B52, Cw4. Nucleotide sequencing based typing of HLA class II alleles disclosed DRB1*0701, *15021, DRB4*01011/*0103, DRB5*0102, DQA1*0103, *0201, DQB1*02, *06011, DPB1*0401,*1101

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02619.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY