摘要:

A novel DQ6 allele (DQB1*0611) was identified via direct DNA sequencing in an African‐American donor for bone marrow transplantation. The allele was not suspected on the basis of a sequence specific PCR assay which instead indicated the presence of DQB1*0602. DQB1*0602 and DQB1*0611 differ in exon 2 only at codon 9 resulting in a tyrosine substitution for phenylalanine. A modification of current DQB1 sequence specific PCR assays was devised which allows distinction between the closely related DQB1*0602 and DQB1*0611 alleles. Preliminary allele frequency studies suggest that DQB1*0611 is rare both in a non‐African American sample and in Americans of African descent carrying DR11, DQ6 haplotypes. The selection of various DQB1*0611 detection methods is discus

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02621.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We have established a ligation based typing method to detect HLA‐B*27 alleles at the DNA level. The method requires amplification of exon 2 of the HLA‐B locus from genomic DNA by the polymerase catalyzed chain reaction (PCR) using group specific primers. An aliquot of the PCR amplification product, heat stable ligase and a pair of oligonucleotide probes, designed to hybridize adjacently to HLA‐B*27 specific sequences of the amplified DNA are subsequently thermocycled. If the probes are perfectly complementary they become ligated otherwise they stay separated. The ligation of probes can be detected through their different labels by an enzyme linked immunosorbent assay (ELISA). Ligation based detection of beta‐actin sequences which have been co‐amplified serves as positive control for each PCR reaction. We observed complete concordance when typing 76 HLA‐B*27 positive and 107 HLA‐B*27 negative individuals either by serology or by the ligation based approach. We conclude that ligation based typing is a reliable tool for the DNA based detection of HLA‐B*27 alleles. The procedure allows automation to a large extent and should be easily applicable to the typing of other HLA

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02622.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

In the present study PCR primers were designed for detecting all knownDPA1variability, i.e. the presently recognized sixDPA1alleles 0103 to 0401, and also for separation of the fourDPA1*02alleles, by PCR amplification with sequence‐specific primers (PCR‐SSP). For each sample seven different PCR reactions were performed which allowed the identification of all DPAl alleles and the resolution of allDPA1genotypes. Forty‐eight cell lines and 100 donor spleen cells were investigated by theDPA1PCR‐SSP technique. In the forty‐eight known workshop cell‐lines no false positive or false negative results were obtained. The 100 donor spleen cells were only typed by the PCR‐SSP technique and in their DNAs only one or twoDPA1alleles were found. Twenty cell lines and twenty donor spleen cells were typed on two separate occasions and interpreted blindly. The reproducibility between the repeated typings was 100%. The length of the specific products ranged from 103 to 258 base pairs and the amplification patterns obtained were easy to interpret. In conclusion,DPA1typing by the PCR‐SSP method is an accurate typing technique with high sensitivity, specificity and reproducibility. Analysis of the distribution ofDPA1alleles was performed in 100 Caucasian samples, 100 African samples and 80 Oriental samples, including separation of the fourDPA1*02alleles. The population study showed a characteristic distribution ofHLA‐DPA1alleles. Each ethnic group appeared to have one (Caucasians), or two (Africans and Orientals), frequentDPA1allele(s) and a high frequency ofDPA1homozygotes, suggesting that, like for theDPB1locus, balancing selection does not appear to be affecting the evolutio

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02623.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

A monoclonal antibody (mAb), designated mNI‐58A, was produced by immunizing mice with the lipopolysaccharide (LPS)‐stimulated monocyte‐like cell line, U937. The antigen defined by mNI‐58A was widely expressed on various lymphoid cells and all cell lines examined except the erythroid cell line, K562. When the reactive patterns between mNI‐58A and the mAbs to various human differentiation antigens (CD11a, CD11b, CD11c, CD14, CD16, CD18, CD23, CD28, CD29, CD31, CD43, CD44, CD45RA, CD50, CD54, CD58, CD80, CD 102, CD 106, HLA‐class I and ‐class II antigen) were compared, that of mNI‐58A was found to be similar to those of the leukocyte function‐associated antigen‐1 (LFA‐1) mAbs. Using a competitive immunofluorescence binding assay it was found that the preincubation with one of the CD11a mAbs, 2F12 completely blocked the subsequent binding of mNI‐58A. mNI‐58A prevented the homotypic cell aggregation of the phorbol myristate acetate (PMA)‐activated U937 cells (referred to as PMA‐U937) and PMA‐activated Epstein‐Barr virus (EBV)‐transformed B cell lines, B‐85 and Mann. mNI‐58A markedly induced the spread formation of the PMA‐U937 cells following this blocking of the homotypic cell aggregation, whereas 2F12 did not under the same condition. The spread formation induced by mNI‐58A was completely blocked by cytochalasin B (CyB), cytochalasin D (CyD), cycloheximide (CHX) or protein kinase C inhibitors, sphingosine and H‐7. The U937 cells markedly adhered to the tumor necrosis factor‐α (TNF‐α)‐stimulated human umbilical vein endothelial cells (HUVECs) and also to the extracellular matrix protein, fibronectin, but mNI‐58 A did not enhance or block these adhesion processes. mNI‐58A precipitated two glycoproteins with molecular weight 180 kDa and 95 kDa as determined by SDS‐PAGE analysis, which were identical to the LFA‐α (CD 11 a) and β (CD 18) chains of leukocyte integrin precipitated by the CD 11 a mAbs, respectively. Sequential immunoprecipitation studies using the CD1 la mAb (2F12) also indicate that mNI‐58 A recognizes an epitope on the α‐chain of the LFA‐1 motecule. The ability of mNI‐58 A to block the PMA‐U937 cells and to induce the spread formation of these cells suggests that mNI‐58A is a novel mAb reacting with an epitope on the α

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02624.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

HLA class II variation was analyzed in nine Native American populations of Colombia using PCR/SSOP typing methods. Under the auspices of the Expedition Humana, approximately 30 unrelated native Colombian Indian samples each from the Tule (NW Pacific Coast), Kogui (Sierra Nevada), Ijka (Sierra Nevada), Ingano (Amazonas), Coreguaje (Amazonas), Nukak (Amazonas), Waunana (Pacific), Embera (Pacific) and Sikuani (Northeastern Plains) were collected and analyzed at the DRB1, DQA1, DQB1 and DPB1 loci. The number of different DRB1, DQA1, DQB1 and DPB1 alleles in the Colombian Indians is markedly reduced in comparison with neighboring African Colombian populations, which exhibit a very high degree of class II variability, as discussed in an accompanying paper. In the Colombian Amerindian groups, DR2 (DRB1*1602), DR4 (DRB1*0407, *0404, *0403 and *0411), DR6 (DRB1*1402) and DR8 (DRB1*0802) comprise>95% of all DRB1 alleles. We also found an absence of DR3 in all populations, and DR1, DR7 and DR9 allelic groups were either very rare or absent. Each Colombian Amerindian population has a predominant DRB1 allele (f=˜0.22–0.65) and DRB1‐DQA1‐DQB1 haplotype. Several novel DR‐DQ haplotypes were also found. At the DPB1 locus, DPB1*0402 (f=0.28‐0.82), *1401 (f=0.03–0.45), and *3501 (f=0.03–0.27), were the three most prevalent alleles, each population maintaining one of these three alleles as the predominant (f>0.26) DPB1 allele. The reduction of diversity for the HLA class II alleles in the Colombian Indians is suggestive of a population bottleneck during the colonization of the Americans, with little to no subsequent admixture with neighboring African Colombian populations in the

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02625.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Psoriasis vulgaris is associated with the HLA‐Cw6 and Cw7 antigens. However, it has not yet been clarified if the HLA‐Cw6 and Cw7 genes themselves are the susceptible gene related to this disease or if it is some other non‐HLA gene in a linkage disequilibrium with these HLA‐C alleles. The S gene, recently identified in the HLA class I region 160 kb telomeric of HLA‐C, encodes a keratin‐like protein and is expressed specifically in the granular layer of the epidermis. Therefore, it is tempting to speculate that the S gene is one of the strong candidate genes responsible for the pathogenesis of psoriasis vulgaris. Direct sequencing of the first and second exon of the S gene after polymerase chain reaction (PCR) amplification has allowed the identification of two diallelic polymorphic sites in exon 1 and seven diallelic polymorphic sites in exon 2, three among which result in amino acid exchanges, a Ser‐Phe substitution at amino acid position 186, a Gly‐Val substitution at position 393 and a Ser‐Leu substitution at position 394. No significant difference in the dimorphic distributions of the S gene was observed between the patients with psoriasis vulgaris and healthy controls, suggesting that the susceptible gene for psoriasis is not

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02626.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

A serological family study identified an HLA‐A “blank” segregating through three generations of apparently healthy individuals. The HLA‐A*0301 allele was assigned by DNA genotyping in each of the three individuals. Complete absence of cellular expression of the HLA‐A3 antigen was associated with a 6 nucleotide deletion in exon 3 of the A*0301 gene. The in‐frame deletion of nucleotides 373–378 results in the absence of residues C101 and D102 from the mature HLA‐A heavy chain. Cysteine 101 is involved in the formation of the highly conserved disulfide bridge in the α2 domain of the class I molecule, and deletion of this residue is believed to be highly disruptive to proper folding and function of the

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02627.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

PCR/SSOP typing methods were used to analyze the HLA Class II DRB1, DQA1, DQB1 and DPB1 loci of samples from three African American populations of Colombia. Forty samples from the Cauca (Pacific), and twenty samples each from the Choco (North Pacific Coast) and the Providencia (Caribbean island) populations, were collected and the Class II loci analyzed under the auspices of the Expedicion Humana. Despite the limited number of samples analyzed, the African Colombian populations exhibit a very high degree of class II polymorphism. A great diversity of DRB1 alleles was found, with representatives from all serological classes, including 19 DRB1 alleles in the Providencia, 16 in the Cauca and 14 in the Choco groups. In addition, a novel DQB1*02 allele (*0203) was found in two individuals from the Cauca population of the Pacific Coast. The sequence of the DQB1*0203 allele, associated with DR3, differs from DQB 1*0201 by only one nucleotide substitution (C←A) in the second position of codon 57, resulting in an Ala to Asp change. The addition of DQB1*0203 brings the total number of DQB1 alleles identified to date to 26. HLA class II diversity is much greater in these African Colombian populations than that seen in nearby Amerindian populations. Analysis of regional Colombian African American HLA population genetics is discussed with respect to the Colombian Amerindian HLA genetics described in an accompanying pape

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02628.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

A full‐length cDNA clone and two overlapping genomic clones corresponding to the canine DQA class II gene were isolated and sequenced. Restriction mapping and sequence data allow identification and orientation of the five exons corresponding to the alpha (α) chain. Sequence analysis of exon 2 amplified from 17 unrelated dogs of various breeds identified seven alleles. The structure of the canine DQA gene is similar to HLA‐DQA1 and other mammalian DQA genes. This study will serve as a reference for developing a typing system for the DLA‐DQA gene for donor and recipient matching in the canine model for organ and bone marrow transplan

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02629.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

We have identified two new A*74 alleles (A*7402 and 7403) in two unrelated individuals. A*7402 differs from A*7401 by a single amino acid substitution in the signal peptide and may be the result of a gene conversion event at the 3′ end of exon 1. A*7403 differs from A*7401 by a single amino acid exchange in the α1 domain and is most likely due to a point mutation in exon 2, since no HLA class I donor allele has been found. Since A*7402 appears to be the ancestor of the other two A*74 alleles, it is possible that A*7401 and 7403 have been created by successive point mutations. The sequences of the expressed proteins of A*7401 and 7402 are identical. The heavy chain sequence of A*7403 differs from these alleles at the crucial residue 79 which is located in the sequence stretch of the al α‐Helix where the Bw4/Bw6 determinants have been identified and which probably affects TCR interaction. This variation can therefore be expected to stimulate alloreactive T cells, graft rejection and graft versus host disease emphasizing the relevance for matching in bone marrow transplantation with alternative d

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1996.tb02630.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY