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T Akaza

Publications and source records attributed to T Akaza.

At least 55 records · Page 3Linked to original sources
PubMed1997

Identification of HLA-C alleles using PCR-single-strand-conformation polymorphism and direct sequencing.

Alleles encoding HLA-C antigens in Japanese were identified by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) and nucleotide sequencing analyses. The results showed that at least sixteen different alleles code for eight serologically detectable antigen groups and undetectable blanks. Cw1 was mainly encoded by Cw*0102, whereas two split antigens of Cw3, Cw9 and Cw10, were encoded by Cw*0303 and Cw*0304, respectively. Cw4 and Cw6 were encoded by Cw*0401 and Cw*0602, respectively. Seven alleles, Cw*0801, Cw*0803, Cw*1202, Cw*1203, Cw*1402, Cw*1403 and Cw*1502, were found to encode serological HLA-C "blanks" in Japanese. Moreover, errors in the published nucleotide sequences of Cw*0501 and Cw*1201 were corrected. Twenty-one HLA-C alleles were distinguished from each other by means of group-specific PCR amplification followed by the SSCP method developed in the present study. The system using genomic DNAs can be used effectively for identification of new HLA-C alleles.

Alleles↗
PubMed1997

The HLA system in the population of Mongolia.

The frequencies of HLA-A, B, C and DRB1 alleles and haplotype frequencies for HLA-A, B, C and A, B, DR loci were studied in 201 healthy unrelated Khalkha-Mongolians. The most common class I antigens were A24 (25.8%), A2 (23.4%), B61 (12.6%), B51 (8.5%), Cw10 (14.2%), Cw9 (14.1%), Cw7 (13.3%), Cw1 (11.8%) and Cw6 (10.2%). A total of 35 DRB1 alleles were identified in this group of samples. The most frequent DRB1 allele was DRB1*0301 (11.1%), followed by DRB1*0701 (9.7%) and DRB1*1101 (8.5%). One novel DRB1 allele (14MV) and three rare types, DRB1*1111, DRB1*1504 and DRB1*1412, recently described in Jews, the Dai minority of China and Japanese, respectively, were identified in Mongolians. The phylogenetic tree constructed by UPG method put Mongolians in the Northeast Asian cluster. A comparison of three locus haplotype distributions with world populations, revealed that Mongolians share several characteristics in common with other Mongoloids as well as with Caucasoids and Amerindians. The most common A, B and DR haplotype in Mongolians, A33-B58-DR3, was shared with Thai, Thai Chinese and Singapore Chinese. These data support that unique genetic background of Mongolians played a major role in ethnic formation and differentiation of Mongoloid populations.

Alleles↗
PubMed1997

[HLA typing in the Japanese Bone Marrow Program].

The Japan Marrow Donor Program (JMDP) was established in 1992, and managed by the Japan Marrow Donor Foundation and the bone marrow data centers which are affiliated with the Japanese Red Cross. Since 1992, about 82,000 donors and 4,400 patients have been registered. HLA typing of registered donors is performed 2 steps. Serological HLA-A, -B locus typing and low-resolution HLA-DRB1 locus DNA typing are performed at the donor data centers. For the final matching test, serological HLA-A, -B locus typing is performed as reconfirmation and high-resolution HLA-DRB1 DNA typing are performed. The class I DNA typing is performed for A2, A26, B39 and B61. These antigens occur with a high frequency and each can be subtyped into more than three alleles in Japanese. A retrospective study on the effect of matching or mismatching HLA class I and class II alleles between the donor and recipient on the survival rate of 363 transplanted is currently being performed using JMDP samples. In the report of this research project supported by the Ministry of Health and Welfare matching HLA-A, -B locus alleles of donor and recipient decreased the risk of acute GVHD and improved survival after unrelated marrow transplantation.

Bone Marrow Transplantation↗
PubMed1997

[Possible involvement of donor-derived B cells in development of post-transfusion graft-versus-host disease].

Post-transfusion graft-versus-host disease(PT-GVHD) is one of the most severe side effects of blood transfusion. To clarify the mechanism of development PT-GVHD, we characterized possible effector lymphocyte clones, presumably involved in the occurrence of PT-GVHD. By lymphocyte cloning from peripheral blood of a PT-GVHD patient, not only donor-derived cytotoxic T cell clones but also donor-derived B cell clones were established. The B cell clones produce cytotoxic IgG, directed against the patient's class II HLA. It was strongly suggested that donor-derived B cells, through production of cytotoxic IgG directed against the patient's HLA, were involved in the pathogenesis of the PT-GVHD in this case.

B-Lymphocytes↗
PubMed1996

HLA-A null allele with a stop codon, HLA-A*0215N, identified in a homozygous state in a healthy adult.

A healthy adult having no serologically detectable HLA class I A locus antigens was identified. The parents of the individual are consanguineous. Results of a family study indicated that the individual is homozygous for the B46-Cw1-DR8.1 haplotype, which was shown to be positively associated with A*0207 in our previous study. The HLA-A null individual is healthy and exhibits no apparent immunological abnormality. Total RNAs extracted from peripheral blood were converted to cDNAs. The reverse transcriptase-polymerase chain reaction (PCR) product, which is of the same size as the normally expressed gene, was easily obtained from the cDNAs with HLA-A locus-specific primers. The nucleotide sequence of this null allele (A*0215N) was the same as that of A*0207 except for a single nucleotide substitution which resulted in a stop codon in exon 4. From its nucleotide sequence, a truncated molecule was expected to be produced; however, the immunoprecipitation study failed to detect the predicted product. Genomic DNAs from 29 unrelated individuals who expressed only one HLA-A antigen with HLA-B46, were analyzed by a PCR-sequence-specific oligonucleotide method. None of the samples possessed this stop codon. Therefore, A*0215N is likely to be a rare allele generated by a single point mutation from A*0207.

Adult↗
PubMed1996

Identification and DNA typing of two Cw7 alleles (Cw*0702 and Cw*0704) in Japanese, with the corrected sequence of Cw*0702.

Two alleles encoding HLA-Cw7 antigens, tentatively called C7J1 and C7J2, have been identified in Japanese using a PCR-SSCP method. The nucleotide sequence of full-length C7J1 cDNAs showed a high degree of homology to the reported Cw*0702 sequence except in exon 1. We then resequenced the allele carried by the cell line JY in which Cw*0702 was first identified, according to a request from the WHO Nomenclature Committee. The results revealed complete identity between the corrected Cw*0702 sequence and the C7J1 sequence. On the other hand, the C7J2 sequence was completely identical to the reported Cw*0704 sequence. Sequences specific for Cw*0702 and Cw*0704 were confirmed using PCR-RFLP and PCR-SSO methods. Moreover, association analysis with other HLA locus alleles showed positive associations of Cw*0702 with HLA-B7, -B39, and -B67 and of Cw*0704 with HLA-B70 in Japanese.

Alleles↗
PubMed1996

Large-scale DNA typing for human platelet alloantigens by PCR-PHFA (preferential homoduplex formation assay).

Alloimmunization against human platelet alloantigens (HPA) is known to be involved in disorders such as neonatal alloimmune thrombocytopenic purpura, posttransfusion purpura, and refractoriness to platelet transfusion therapy. HPA typing is essential in diagnosis and management of patients. Therefore a reliable and speedy method is necessary for HPA typing. We have successfully applied a new DNA typing method, PCR-preferential homoduplex formation assay (PHFA) method, to typing for the HPA-1, -2, -3, -4, -5 and -6 systems. This method is based on DNA strand competition during hybridization under a precisely controlled temperature gradient between a double-labelled amplicon (standard DNA), prepared from biotin- and DNP-labelled primers, and an unlabelled amplicon (sample DNA). The results obtained by PCR-PHFA typing were in good agreement with the allotypes determined by serological typing and by other DNA typing methods. The PCR-PHFA method can be easily automated, is suitable for typing both small and large numbers of samples, and thus is applicable to routine HPA typing.

Antigens, Human Platelet↗
PubMed1996

Further molecular diversity in the HLA-B15 group.

In order to further clarify the diversity of the HLA-B15 antigens and the correspondence of serological types with alleles in Asians, we screened various B15 serological splits by means of a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method. Subsequently, the genes encoding various B15 variants were sequenced. Two novel alleles, B*1528 and B*1529, were identified: the nucleotide sequence of the former contained a single-base substitution at position 263 in exon 2 as compared to that of the B*1501 allele, which results in an amino acid change at position 64 in the alpha 1 domain, and the nucleotide sequence of the latter differs from that of B*1518 by a single-base substitution at position 272 of exon 2 which results in an amino acid change at position 67 of the alpha 1 domain. One new allele, B*1521, described recently in Australian Aborigines was also identified in Asians in the present study. Moreover, the results of sequencing demonstrated that Asian HLA-B62, B70, and B77 antigens are encoded by B*1501, B*1518, and B*1513, respectively. Two splits of B75 antigens, B75V (TS-1) and B15N, which have been proposed to exist in the Japanese population were encoded by B*1511 and B*1502, respectively. Most of the B15 alleles detected in the present study showed positive associations with other locus antigens. Especially, B*1502 was strongly associated with Cw8, while B*1521 was strongly associated with A34 and Cw6.

Alleles↗
PubMed1996

Routine low and high resolution typing of the HLA-DRB gene using the PCR-MPH (microtitre plate hybridization) method.

We describe HLA-DRB1 typing using polymerase chain reaction-based microtitre plate hybridization (PCR-MPH), which can process large numbers of samples. MPH typing is similar to an enzyme-linked immunosorbent assay (ELISA), in which a tandemly ligated sequence-specific oligonucleotide is immobilized on microtitre wells. The typing procedure consisted of two steps. In the first, PCR-MPH with 16 probes was performed to determine the specificities of the serological levels (DR1, DR2, DR3, DR4, DR11, DR12, DR13, DR14, DR7, DR8, DR9 and DR10) after generic amplification ("low resolution typing'). In the second step, DR1, DR2, DR4, DR12/8 and DR3/11/13/14 were group-specifically amplified based on the results of the first PCR-MPH, and microtitre plate hybridization proceeded in a similar manner to the first step ("high resolution typing'). Low resolution typing was completed within 2 h after generic amplification, and the results of high resolution typing were obtained in another 3.5 h after amplification. The allelic types classified using PCR-MPH were completely concordant with those obtained by PCR-single-strand conformation polymorphism or PCR-restriction fragment length polymorphism.

HLA-DR Antigens↗
PubMed1996

Report of the Japanese Central Bone Marrow Data Center.

The Japan Marrow Donor Program started to recruit donors at the beginning of 1992 with the financial support of the Japanese government, and the first unrelated bone marrow transplantation arranged through this program was performed in 1993. Since 1992, about 82,000 donors and 4,400 patients have been registered. At least one HLA-identical donor has been found for 60% of the patients in the first search after the patients register with the JMDP, and as of the end of September 1996, 880 patients had received bone marrow transplants from unrelated donors. The results of 19.7% of registered patients having received bone marrow transplants and 1.1% of registered donors having donated far exceed our forecasts. This is attributed to the high frequency of several haplotypes in the Japanese. However, no HLA-identical donors have been found for approximately 800 registered patients so far. Thus, it is still necessary to increase the number of donors registered with the JMDP.

Bone Marrow Transplantation↗
PubMed1995

A new B18 sequence (B*1802) from Asian individuals.

A new HLA-B18 allele (B*1802) derived from a Thai individual was sequenced. Comparison of this B18 nucleotide sequence with the published B*1801 sequence indicated that this Asian B18 allele has a nucleotide sequence different from that of B*1801. Three nucleotide changes were observed in exon 3, in which two substitutions at codon 97, AGG in B*1801 to AAT in the B*1802, result in an amino acid change from arginine to asparagine. The residue 97Asn has also been described in some B27 subtypes. A silent mutation was also observed at codon 99, TAC in B*1801 to TAT in the B*1802. This sequence has been reported in many class I alleles published so far. Moreover, 18 HLA-B18-positive samples were examined by the PCR-SSO method using specific probes for B*1801 and B*1802. The results demonstrated that three Asian samples possess B*1802 and share HLA-Cw7, DR12, and DQ7.

Alleles↗
PubMed1995

A new HLA-A9 subtype lacking the Bw4 epitope. Ancestral or revertant allele?

The HLA-A9 group has been subdivided into three serologically defined splits, A23, A24, and A2403. We have found a new HLA-A9 split antigen, tentatively called A24AK, in the Japanese population. Sequence analysis of A24AK (officially assigned A*2404) showed that this new allele was different from HLA-A*2402, which codes for the common A24 antigen, by seven nucleotides, and the two alleles could be discriminated by the PCR-SSCP method. These nucleotide substitutions are predicted to result in substitution of six amino acid residues at positions 76, 79, 80, 81, 82, and 83. In all HLA-A9-group alleles described to date, this region is known to code for the Bw4 epitope, which is usually localized on HLA-B molecules. However, the new allele lacks the Bw4 coding sequence. Sequencing results are supported by results showing that the lymphocytes from A24AK-positive individuals did not react with anti-Bw4 antiserum. The nucleotide sequence of A*2404 in this region was identical to that of A*0101, A*2601, A*2602, and several other alleles. These findings suggest several possible paths of evolution of this new allele.

Alleles↗
PubMed1995

High-resolution HLA-DQB1 typing by combination of group-specific amplification and restriction fragment length polymorphism.

A reliable method for high-resolution HLA-DQB1 typing using the combination of group-specific amplification and RFLP analysis is described. Group-specific amplification was carried out for the alleles of two groups using the two primer pairs under the same PCR conditions. One group contains DQ5 and DQ6 specificities and the other DQ2, DQ3, and DQ4 specificities. Computer analysis on cleavage patterns for 19 alleles of the DQB1 gene showed that the 11 alleles of the former group could be distinguished with five restriction enzymes and the eight alleles of the latter group could be distinguished with four enzymes. We could reduce the number of restriction endonucleases required compared with the number used in previous studies because we selected appropriate restriction enzymes which had at least one recognition site in almost all DQB1 alleles as a form of internal control. Moreover, DQB1*0602 and 0603, which were indistinguishable using the previously reported PCR-RFLP methods, could be distinguished by the present method. The results of typing of 100 samples from Japanese individuals by this method showed no discrepancy with the results obtained by serologic methods. The calculated allele frequencies showed good agreement with those reported at the 11th International Histocompatibility Workshop.

Base Sequence↗
PubMed1995

Sequence-based typing of HLA-A2 alleles using a primer with an extra base mismatch.

A PCR-SBT method using genomic DNA for HLA-A2 alleles was established. To achieve specific amplification for detecting a single base difference between A2 alleles and other HLA-A alleles, a primer having one extra mismatch at the second position from its 3'-end was designed. The primer exhibited a high specificity with annealing temperatures from 64 degrees C to 68 degrees C. Thirty-eight Japanese samples were screened using this method. The majority of Japanese A2 antigens were coded by A*0201. A*0206 and A*0207 were observed at relatively high frequencies. Serologically defined split antigens, A2S and A2AK, which we have recently identified, corresponded to A*0203 and A*0210, respectively. Moreover, A*0207 was strongly associated with B46 and DR8.1.

Alleles↗