Search PubMed⌕ Search

Biomedical subjects

N Tanigaki

Publications and source records attributed to N Tanigaki.

At least 37 records · Page 2Linked to original sources
PubMed1989

Serological detection and molecular localization of allelic HLA-DP supertypic epitopes.

A DP serological allospecificity was identified using 125I-labeled preparations of HLA class II molecules isolated from cells of HLA homozygous typing cell lines, SLE (DRw6, DQw1, DPw3) and WT46 (DRw6, DQw1, DPw2), and depleted of DR molecules by absorption with an anti-DR monoclonal antibody. The specificity, provisionally called WT, was carried by class II molecules possessing the characteristics of DP molecules on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and detected primarily in DPw1, DPw3 and DPw5 cells and exceptionally in some DPw2 cells including WT46 and DPw4 cells on a large panel of DP-pretyped B cell lines mostly derived from the 10th International Histocompatibility Workshop B cell reference panel. It was apparently allelic to another DP serological specificity BUT previously defined on DP molecules isolated from cells of DPw2+ HLA deletion mutant cell line LCL 721.82. On the same cell panel; the BUT specificity was negative in all DPw1, DPw3 and DPw5 cells, and positive in all DPw2 and DPw4 cells and also in DPw2B and DPw4B cells except the cells typed WT+. This DP association pattern was similar to that of the known allelic sequences, GGPM and DEAV, in DP beta F segment, one of the six variable segments in the second exon of DP beta gene. Thus, genomic DNA from 22 B cell lines pretyped for BUT and WT specificities were enzymatically amplified for the second exon of DP beta gene by the use of locus-specific oligonucleotide primers and hybridized with 32P-labeled oligonucleotide probes corresponding to the F segment sequence variations. This oligonucleotide typing showed perfect one-to-one correlation with the BUT and WT serological typing. The typing also revealed that WT46 cells, although typed DPw2, have the DEAV sequence common to DPw1, DPw3 and DPw5.

Alleles↗
PubMed1989

DNA typing of an HLA-DR bilocus specificity by gene amplification and oligonucleotide hybridization.

The structure of the DRB1*03 gene has been interpreted as the product of a gene conversion event involving a DRB3 gene as donor and resulting in the introduction of two short segments of the DRB3 sequence into the DRB1 locus. The serological counterpart of this double insertion is the TR81 specificity. Consequently, the TR81-specifying sequences can reside on either DRB1 or DRB3, or on both loci. Within each of the two sequence stretches a single nucleotide may be responsible for the generation of the TR81 alloantigen. Oligonucleotide probes corresponding to these stretches and to their allelic variants were constructed. They were used, under stringent hybridization conditions, to detect TR81-specifying sequences in the DNA of HLA-homozygous cell lines carrying different haplotypes of the DRw52 family. Prior to hybridization the DNA was amplified with either DRB1-specific or DRB3-specific primers. Using this approach it was possible to perform a "DNA typing" of the TR81-specifying sites separately on both the DRB1 locus and the DRB3 locus.

Alleles↗
PubMed1989

Human anti-HLA-DQw2 monoclonal antibody secreted by an Epstein-Barr-virus--transformed lymphoblastoid cell line: assessment of the monoclonality, allospecificity, and target.

IgM molecules were purified by the use of anti-IgM antibody-coupled Sepharose from the culture supernatant of an Epstein-Barr-virus-transformed lymphoblastoid cell line, MP1, that secretes alloantibodies possessing HLA-DQw2 specificity as defined by the cytotoxicity assay. The obtained IgM preparation was labeled with radioactive iodine-125I and fractionated by gel filtration. It contained pentameric IgM and smaller oligomeric IgMs. When tested by the direct cellular binding assay against a panel of HLA-typed cell lines, they all showed the DR3 and DR7 association pattern characteristic of DQw2. A weak but significant binding was detected for DR1, DR6, and DR9. On isoelectrofocusing, MP1 pentameric IgM gave a restricted banding pattern comparable to monoclonal IgM obtained from a patient with Waldenström's syndrome. Moreover, the pattern was identical to that of IgM purified from the culture supernatant of a defined hybrid clone, 162, that was generated by fusing MP1 cells with heteromyeloma D33 cells. The target class II molecules showed the dimeric structure that conforms to DQw2 molecules.

Antibodies, Monoclonal↗
PubMed1989

[Methods for optimized cultivation of hair cells from C3H mice].

Methods for the optimized culturing the hair cells from 4-day-old C3H mice are described. The skin was obtained and soaked in 500 unit/ml dispase at 4 degrees C for 24 hr. Then the epidermis was removed and the dermis was further treated with 0.25% collagenase for 1 hr at 37 degrees C. The hair roots isolated by collagenase digestion of the dermis were dispersed by the treatment with the mixture of trypsin and EDTA into a cell suspension and plated on a substrate. Attachment of hair cells was dependent on serum concentrations and enhanced by collagen coating of the surface of dishes. MCDB 153 was beneficial for the growth of hair cells and discouraged for the growth of dermal fibroblasts. The cells could not grow at inoculum size of less than 3 x 10(4) cells/cm2. Addition of a crude bovine pituitary extract in MCDB 153 had the greatest impact on hair cell growth. Functional integrity of the cultured hair cells were maintained, since the hair-specific cytoskeletal proteins were expressed in these cells under the present experimental conditions. These desirable conditions could allow selective growth of the hair cells and provide the model system which could be used for the study of hair growth.

Animals↗
PubMed1988

Serological recognition of HLA-DR allodeterminant corresponding to DNA sequence involved in gene conversion.

HLA class II molecules were isolated from mouse L cells transfected with a DR alpha gene and an allele, 52a, of locus DR beta III from an HLA-homozygous cell line, AVL, of the DR3 haplotype. The isolated molecules were found to possess a new allospecificity, named TR81. This specificity behaved allelic to the previously described specificity TR22 encoded by another allele, 52b, of the DR beta III locus. The TR81 specificity was also present on the DR beta I gene product of the DR3 haplotype. The nucleotide sequence of the gene encoding TR81 differs from TR81-negative DR beta genes of the DRw52 family in only two codons, both located in the regions known to be involved in a gene conversion event. Consequently, the following conclusions can be formulated. (a) TR81 is a bi-locus specificity and allelic to TR22 only in its DR beta III locus localization. (b) The TR81 specificity is the phenotypic counterpart of the gene conversion event which led to the generation of the DR beta I gene of the DR3 haplotype. (c) One or both individual amino acid substitutions in the first domain of the DR beta chain are responsible for the TR81 allospecificity. (d) Since TR81 is expressed on the DR beta I chain of the DR3 haplotype, it is possible that TR81 and DR3 represent the same serological specificity.

Alleles↗
PubMed1988

A high proportion of sera of heroin addicts possesses anti-HLA class I and class II reactivity.

The anti-HLA reactivity of sera from 210 heroin addicts was tested by the direct binding with 125I-labeled preparations of HLA class I and class II molecules purified from human B-cell lines of various HLA haplotypes. A high proportion (81.7%) of the sera tested possessed anti-HLA class I and II reactivity. The reactivity did not show any allospecificity and was apparently mediated by antibodies. The control included 100 healthy blood donors, 25 male homosexuals positive for anti-HIV (human immunodeficiency virus) antibodies, and 25 patients positive for HBsAg (hepatitis B surface antigen). Of these controls, only one of the healthy blood donors was positive for anti-HLA reactivity (P much less than 0.001). Among heroin addicts, the reactivity was independent of the presence of either HBsAg or anti-HIV antibodies in the serum.

HLA Antigens↗
PubMed1988

Correlations between polymorphisms at the DNA and at the protein level of DRw52 haplotypes, revealed with a variety of techniques.

LB-Q1 and LB-Q4 are two subtypes of DRw52, defined by proliferative T-cell clones. These subtypes represent a polymorphism of the DR beta III gene. Similar subtypes of DRw52 can be defined by oligonucleotide typing, serology and RFLP analysis. In the present study we compared these typing techniques on a panel of 22 HLA-D homozygous, DRw52-positive typing cells. All typing techniques correlated very well. Three subtypes of DRw52 could be identified. Our results show that typing for cellularly defined structures can be done with a variety of noncellular techniques. This observation has important implications for matching in unrelated bone marrow transplantation and for disease association studies.

DNA↗
PubMed1987

Detection of HLA-DP serological allodeterminants by the use of radioiodinated DP molecules.

HLA class II molecules were partially purified from cells of an HLA deletion mutant cell line, LCL721.82, that lost DR and DQ expression but retained DPw2 specificity and labeled with radioactive 125I. The radioiodinated preparation bound to DP-specific monoclonal antibody B7/21 as well as rabbit anti-HLA class II antiserum. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the component involved in these bindings gave, unlike known HLA class II molecules, a sharp and dense band of approximately 60 kDa under nonreducing conditions and a single but diffuse band of approximately 30 kDa under reducing conditions. By screening 401 anti-HLA class II alloantisera, including those distributed in the 9th International Histocompatibility Workshop and also those locally available, eight were found to possess significant binding activity. Specificity analysis of these eight binding-positive antisera on a panel of DP-pretyped HLA homozygous typing cells revealed the presence of two clusters, one corresponding to an allodeterminant associated with DPw1, 2 and 3 and the other to that associated with DPw2 and 4. These two determinants were shown by the sequential binding test to reside on the B7/21-defined HLA class II molecules. Thus, two major conclusions were drawn: two distinct allodeterminants are carried by a single DP molecule; and these serologically detected DP allodeterminants are supertypic to cellularly defined DP allospecificities.

Antibodies, Monoclonal↗
PubMed1987

Human lymphoblastoid cell lines secreting antibodies with restricted HLA specificity.

Peripheral B lymphocytes obtained from three healthy individuals who had been immunized against peripheral blood lymphocytes from appropriate HLA-incompatible donors were transformed by the use of Epstein-Barr virus. The transformed blastoid B cells were repeatedly subcultured by means of "cluster picking," and the HLA antibody-producing cultures were identified by testing the culture supernatants by means of the cytotoxicity assay, using the corresponding donor cells. Thus far, four cell lines that secrete cytotoxic HLA antibodies (MP1, 3, 4, and 5) have been established. Specific immunoabsorption experiments revealed that the antibody activity is carried by lambda-type IgM for MP1, by kappa-type IgM for MP3 and MP5, and by both for MP4. Specificity analysis of a panel of HLA-pretyped cells indicated that MP1 detects DQw2, whereas MP5 recognizes B7. The specificity of MP3 was similar to a DQ specificity termed DC5 (probably equivalent to TA10) but not the same. In the case of MP4, both of the lambda-type and kappa-type antibodies appeared to be directed toward new HLA class II determinants.

Antibody Specificity↗
PubMed1987

Immunochemistry of the HLA class II molecules isolated from a mouse cell transfected with DQ alpha and beta genes from a DR4 haplotype.

Cells from a mouse B lymphoma were transfected by DQ alpha and DQ beta genes derived from a DR4 haplotype. Quantitatively, the resulting expression of human class II molecules was similar to that of human B lymphoblastoid cell lines. Qualitatively, the transformant class II molecules differed from "normal" class II molecules in their carbohydrate moiety. As for their antigenic specificity, they were shown to carry two determinants previously identified on DQ molecules controlled by DR4 haplotypes, i.e., DQw3 and DCHON. The transformant molecules did not carry a third DR4-associated specificity, DC5 (equivalent to TA10), and must possess a structure allelic to DC5. However, no corresponding alloantigenic specificity was detected by a screening of relevant alloantisera.

Animals↗
PubMed1987

[Specific inhibitory action of a novel antidepressant paroxetine on 5-HT uptake].

Effect of a novel antidepressant, paroxetine, on the uptake of serotonin (5-HT), noradrenaline (NA) and dopamine (DA) as well as on various neuro-receptors were investigated in comparison with those of the tricyclic antidepressants amitriptyline, chlorimipramine and imipramine. Paroxetine showed a potent 5-HT uptake inhibitory action, giving the NA/5-HT ratio of 886 in comparison with the ratios of 1.7, 15 and 1.5 for amitriptyline, chlorimipramine and imipramine, respectively. On the other hand, paroxetine showed almost no inhibitory action on the binding of the [3H]-labeled ligands examined in this study [( 3H]quinuclidinyl benzilate, [3H]5-HT, [3H]ketanserine, [3H]pyrilamine, [3H]dihydroalprenolol, [3H]prazosin, [3H]clonidine and [3H]spiroperidol). In contrast, the tricyclic antidepressants showed inhibitory action on a number of bindings and also revealed comparatively high affinities especially for muscarine, histamine-1 and alpha 1-adrenaline receptors responsible for the side effects. From the above findings, it can be concluded that paroxetine has only a weak affinity for various neuro-receptors and inhibits specifically 5-HT uptake.

Animals↗
PubMed1986

Immunochemical analysis of a cell transfected with an HLA-DR gene reveals a new alloantigenic specificity within HLA-DRw52.

The HLA-DR antigen has been prepared from the surface of a mouse fibroblast cell line transfected with a single HLA-DR beta-chain gene as well as single HLA-DR alpha and invariant chain gene. Since the HLA-DR beta chain gene studied corresponds to the DR beta III locus, the DR serological specificities detected on the transformed cells can be assigned to this locus. The use of the HLA-DR-producing mouse cell line has led to the identification of a new serological specificity included within DRw52 and associated with some DR3, some DRw6 and all DR5 haplotypes studied. Most likely this new specificity corresponds to an allelic polymorphism at the DR beta III locus of DRw52 individuals and can serve as a new serological marker for this subset of DR3, DR5 and DRw6 haplotypes.

Alleles↗