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Biomedical subjects

T Juji

Publications and source records attributed to T Juji.

At least 181 records · Page 10Linked to original sources
PubMed1995

Suppressed mixed lymphocyte reaction in couples with intrauterine growth retardation pregnancies.

OBJECTIVE: The objective of this study was to gain insight into the pathogenesis of intrauterine growth retardation (IUGR). METHOD: Immune reactivity against paternal MHC class-II antigens was determined in 18 women with IUGR pregnancies and 18 normal pregnant women, employing one-way mixed lymphocyte reaction (MLR). RESULTS: One-way MLR between the women with IUGR pregnancies and their husbands was significantly suppressed compared with that between the women with IUGR pregnancies and males unrelated to them. However the MLR of normal pregnant women was essentially similar regardless of whether stimulating cells were obtained from their husbands or not. The degree of HLA class-II antigen shared by IUGR couples was not related to the degree of specific suppression of MLR towards the husbands. CONCLUSION: IUGR may be associated with specifically suppressed patients' MLR towards their husbands, which may provide insight into the underlying pathophysiology of IUGR.

Adult↗
PubMed1995

HLA and anti-GQ1b IgG antibody in Miller Fisher syndrome and Guillain-Barré syndrome.

We investigated serological human leukocyte antigen (HLA) types in patients with histories of Miller Fisher syndrome (MFS) and Guillain-Barré syndrome (GBS) with ophthalmoplegia, in whom serum anti-GQ1b IgG antibody was present during the acute phase. We examined class I antigens (A, B and C) in 32 patients and class II antigens (DR and DQ) in 30, but found no association. We conclude that particular serologically defined HLA types are not preferred for the immunoresponse of anti-GQ1b IgG antibody in MFS and GBS.

Autoantibodies↗
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↗
PubMed1995

Lack of primary association between transporter associated with antigen processing genes and atopic dermatitis.

We examined polymorphisms of transporter associated with antigen processing (TAP) genes in 37 Japanese patients with atopic dermatitis and 52 control subjects. We have evaluated, again, the specificity polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method since our previous report and have also developed a mismatch PCR-RFLP method for discriminating two dimorphic sites of TAP1 gene and four dimorphic sites of TAP2 gene. Amplified products from genomic DNA were digested with restriction endonucleases: Sau3A1 for TAP1 codon 333 (Ile-Val), AccI for TAP1 codon 637 (Asp-Gly), and BfaI for TAP2 codon 687 (Stop-Gln). The other sites of TAP2 gene were analyzed by mismatch PCR-RFLP: AccII for TAP2 codon 379 (Val-Ile), RsaI for codon 565 (Ala-Thr), and MspI for codon 665 (Thr-Ala). We observed three TAP1 alleles and six TAP2 alleles. Infrequent allele TAP1 C was observed in one patient and one control subject. We did not identify any differences in TAP allele frequencies between those patients with atopic dermatitis and Japanese control subjects. Analysis of TAP gene polymorphisms will provide better understanding of susceptibility loci in HLA class II-associated disease because TAP genes are located between HLA-DQB1 and HLA-DPB1 loci.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1995

Analysis of disease-associated amino acid epitopes on HLA class II molecules in atopic dermatitis.

We investigated the association between HLA class II alleles and severe atopic dermatitis with high serum IgE levels (greater than 8000 U/ml). The frequencies of HLA-DRB1*1302 and DQB1*0604 were increased, whereas the frequency of HLA-DQB1*0302 was decreased. A strong haplotype, HLA-DRB1*1302-DQB1*0604, has been reported in the Japanese population, and this haplotype is conserved in patients with AD. Further analysis of the amino acid epitopes on the HLA-DR beta 1 and DQ beta 1 domains revealed that DR beta 1 71Glu (RR = 5.71, p < 0.05) and DQ beta 1 30His (RR = 3.25 p < 0.01), and 57Val (RR = 3.13, p < 0.05) were increased in frequency. DR beta 1 71Glu was exclusively unique to DRB1*1302. DQ beta 1 30His was shared by the following alleles: DQ beta 1*0501, *0502, *0503, and *0604, which were all increased in patients with AD. DQ beta 1 57Val was shared by DQB1*0501 and *0604. A well-known haplotype, HLA-DRB1*1302-DQB1*0604, contains DR beta 1 71Glu and DQ beta 1 30His and 57Val. Therefore HLA-DR beta 1 71Glu and/or DQ beta 1 30His/57Val are considered to play the most important role in the development of atopic dermatitis.

Adolescent↗
PubMed1995

DNA-based typing of human platelet antigen systems by polymerase chain reaction-single-strand conformation polymorphism method.

Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method has been established to discriminate genotypes for the human platelet antigen (HPA) systems HPA-1, HPA-2, HPA-3, HPA-4, and HPA-5. Gene fragments which contain polymorphic sequences corresponding to the HPA-1, HPA-2, HPA-3, HPA-4, and HPA-5 systems were PCR-amplified with specific primers. The amplified DNA was denatured and subjected to non-denaturing polyacrylamide gel electrophoresis followed by silver staining. The results obtained by the PCR-SSCP method were in good agreement with those of the allotypes determined by serological typing. Furthermore, the results agreed with those obtained by other DNA-based typing methods such as PCR-allele-specific restriction enzyme analysis and PCR-sequence-specific primer. These results indicate that PCR-SSCP is a simple and sensitive method for determining HPA genotypes and identifying unknown polymorphisms.

Antigens, Human Platelet↗
PubMed1995

Establishment of a T-cell line from lymphocytes presumably implicated in posttransfusion graft-versus-host disease.

Posttransfusion graft-versus-host disease (PTGVHD) is known to develop in immunocompetent patients exhibiting clinical symptoms such as erythroderma, fever, liver dysfunction, diarrhea and pancytopenia. It is speculated that transfused blood donors' lymphocytes might recognize the recipients' HLAs as alloantigens. The thus stimulated lymphocytes might proliferate, expand and finally attack the host's immune system or tissues. However, details regarding these expanded donor cells such as: (1) whether they represent one clone or more, (2) the composition of lymphocyte subsets, and (3) the target HLA antigens of recipients, are not clear, since T-cell lines derived from PTGVHD patients have not yet been obtained. The aim of this study is to characterize T-cells responsible for PTGVHD and to identify their target molecules. For that purpose, we attempted to establish T-cell lines derived from a PTGVHD patient. We show that the established T-cell line, proven to be derived from donor lymphocytes, showed a CD4+ phenotype and had cytotoxic activities. Furthermore, we describe that the target of the cytotoxic T-cell line (CTL) is an HLA-DRB1*0405-related molecule of the patient.

Base Sequence↗
PubMed1995

Multiplex ARMS-PCR-RFLP method for high-resolution typing of HLA-DRB1.

A reliable method for high-resolution HLA-DRB1 typing using the combination of group-specific amplification and RFLP analysis is described. Group-specific PCR amplification (multiplex ARMS-PCR) was carried out under the same conditions for all groups using seven different primer pairs divided into four groups: (1) DR1 and DR10; (2) DR2, DR7 and DR9; (3) DR3 DR5, DR6 and DR8, and (4) DR4. The subsequent polyacrylamide gel electrophoresis was used to determine the group(s) contained in each sample. DR1, DR2/7, DR3/5/6/8, DR4, DRB1*0901 and DRB1*1001 could be distinguished easily using this system. Computer analysis of the various restriction enzyme cleavage sites was carried out on 105 DRB1 allele sequences. It was shown that all DRB1 alleles, except for five allele pairs and some alleles possessing silent mutations, could be distinguished with commonly available restriction endonucleases. Computer analyses on the discrimination of the heterozygous and homozygous combinations were also carried out. Although some heterozygous combinations could no be distinguished with single digestion, double digestion using two restriction enzymes could distinguish most of such heterozygotes. The results of the typing of 100 Japanese individuals using this method showed good agreement with those obtained by other methods.

Alleles↗
PubMed1994

A simple method for detecting single base substitutions and its application to HLA-DPB1 typing.

We have developed a simple and reliable method, PCR-PHFA (polymerase chain reaction dependent preferential homoduplex formation assay), for detection of single base substitutions within PCR amplicons. This technique is based upon strand competition during hybridization between a double labeled amplicon, prepared from biotin and DNP labeled primers, and an unlabeled amplicon. Under the precisely controlled temperature gradient, the preferential formation of a homoduplex over a heteroduplex occurs. After annealing, the identical sequence of the double labeled and unlabeled amplicon resulted in a low population of regenerated double labeled dsDNA due to strand exchange between them. Even when the two differed by only a single base substitution, double labeled molecule was regenerated efficiently because of preferential homoduplex formation. The regenerated double labeled molecule was captured onto a streptavidin coated microtiter plate and quantified enzymatically with a chromogenic substrate. The technique has been successfully applied in HLA-DPB1 typing. Furthermore, we detected a mutated gene even in the presence of a large excess of the corresponding normal gene.

Base Sequence↗
PubMed1994

Analysis of risk factors for post-transfusion graft-versus-host disease in Japan. Japanese Red Cross PT-GVHD Study Group.

We distributed a questionnaire to highlight the effects of post transfusion graft-versus-host disease (PT-GVHD) and to elucidate the risk factors that would predispose people to the disease. The questionnaire described the pathogenesis and the clinical profiles of PT-GVHD and was distributed to doctors in Japan whose hospital conducted more than 1000 units of blood transfusions a year. Doctors were asked about their awareness and experience of PT-GVHD cases. Those who had seen cases of PT-GVHD were then asked to detail clinical course and laboratory data. Of the 14,083 doctors who replied to the first question, 47.4% did not realise that PT-GVHD could occur in immunocompetent hosts. From 304 cases where detailed information was supplied, 171 were considered clinically compatible with PT-GVHD. From these it seems that the risk factors linked to PT-GVHD are cardiovascular surgery, cancer, fresh and consanguineous blood transfusion, and being male. Patients with any of these factors should have some prophylactic therapy such as irradiation of blood before transfusion.

Adolescent↗
PubMed1994

Alloreactive feature of HLA-DP-specific cytotoxic T-cell clone.

The alloreactive feature of CD4+ cytotoxic T-cell clone that could specifically lyse the cells bearing DPB1*0202 sequence was described. The clone was generated from a mixed culture of peripheral blood lymphocytes derived from siblings who were HLA-A, B, C, DR, and DQ identical by serological typing and whose DNA sequence of the second exon of DPB1 was one-allele mismatched by oligonucleotide typing (responder, PDB1*0201/0402; stimulator, DPB1*0202/0402). Specific cytotoxic activity of the clone was strictly limited against the cells bearing DPB1*0202 and was not able to lyse the other tested cells bearing DPB1*0201, 0301, 0401, 0501, 0601, 0901, 1301, and 1601. The cytotoxic activity of the clone was blocked by treatment of target cells with anti-DP monoclonal antibodies (B7/21). On the other hand, treatment of target cells with blocking agents of endogenous or exogenous antigen transport pathway (brefeldin A (BFA), endogenous; chloroquine, exogenous) had no effect on the cytotoxic activity of the clone. These results strongly favor the view that the DP epitopes recognized by the clone are conformational epitopes conferred by specific amino acids in hypervariable regions of the HLA-DP second DPB1 exon and the contribution of peptides in the HLA grooves to the conformational epitope motif is less likely.

Acyclovir↗