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Biomedical subjects

R Ueda

Publications and source records attributed to R Ueda.

At least 289 records · Page 16Linked to original sources
PubMed1991

Molecular cloning of the gene coding for the human T cell differentiation antigen CD7.

The CD7 molecule is a differentiation antigen found on the surface of T lymphocytes and also on a very minor fraction of acute nonlymphocytic leukemia (ANLL). To study the genomic structure of the CD7 gene, two clones (SY4 and SY22) were isolated by screening a genomic library with a CD7 cDNA probe. Restriction mapping of these two phage clones showed that both overlapped each other, covering a total length of 23 kilobases (kb). Transfection of mouse L cells demonstrated that SY22 contains the gene expressing the CD7 antigen reactive with monoclonal CD7 antibody (Tp40), while SY4 does not. Subcloning of a 10.5 kb fragment from a 14.4 kb insert of SY22 contained the structural gene for the CD7 antigen. Detailed restriction mapping and partial sequence analysis revealed the CD7 gene to consist of four exons. By RNase protection assay, multiple initiation sites -122 base pairs (bp) to -38 bp from ATG translation initiation site were demonstrated. The promoter region had high G + C content and contained two SP1 binding sites (CCGCCC) and an AP2 binding site (CCCCAGGC), but lacked CAAT and TATA motifs.

Amino Acid Sequence↗
PubMed1991

bcl-2 translocation in Japanese B cell lymphoma: novel bcl-2 translocation with immunoglobulin heavy chain diversity segment.

Breakpoints of a lymphoma case with bcl-2 gene rearrangement that did not show comigration of immunoglobulin (Ig) heavy chain joining (JH) fragment were cloned. Sequence analysis revealed that the translocation broke the 3' side of the Ig heavy chain diversity (DH) segment at the heptamer recombination signal and each end was ligated to the bcl-2 locus. Since Southern blot demonstrated that both alleles of JH were rearranged, this translocation was suggested to have occurred at the step of VH-DH, or DH-DHJH recombination, one step later than that of DH-JH recombination where the common pattern of bcl-2 rearrangement generally occurs. Cases that showed comigration with JH fragment were also studied by polymerase chain reaction with 5' bcl-2 oligomer and 3' JH consensus anti-sense oligomer since it has been demonstrated that bcl-2 translocation at the major breakpoint clustering region (mbr) in American cases clusters within an about 150 bp region in the mbr. The results demonstrated that four out of five cases studied were amplified, indicating that the same clustering mechanism exists for Japanese cases. The present study, together with our previous report on Ig kappa-bcl-2, indicated that bcl-2 translocation in Japanese B cell lymphomas might occur at a later stage of B cell development, as compared with that in American cases. Less involvement of bcl-2 in Japanese B cell lymphoma may also be in part explainable by low susceptibility to bcl-2 rearrangement at the step of DH-JH recombination.

Base Sequence↗
PubMed1991

The p53 gene is very frequently mutated in small-cell lung cancer with a distinct nucleotide substitution pattern.

The p53 gene has been implicated as a tumor-suppressor gene whose disruption is involved in the pathogenesis of common human cancers. The results of extensive analysis of p53 mutations in non-small cell lung cancers (NSCLCs) have revealed that p53 is mutated in 45% of NSCLC with base changes different from those of colon cancer. In this study, we examined 17 SCLC tumor samples taken directly from 15 patients as well as the corresponding nine tumor cell lines. Mutations changing the p53 coding sequence were found in 11 of 15 patients (73.3%) and showed a similar but distinct nucleotide substitution pattern compared with NSCLC, suggesting that a different mutagenic process is involved. In addition, a strong correlation was seen between the presence of p53 mutations in tumors and the successful establishment of the corresponding cell lines, suggesting that p53 mutations can confer a selective growth advantage in vitro (and probably also in vivo).

Amino Acid Sequence↗
PubMed1991

Epitope analysis of cluster 1 and NK cell-related monoclonal antibodies.

By flow cytometric assays, we tested the antibodies of the Second International Workshop on Small Cell Lung Cancer Antigens against 20 normal peripheral leukocytes, four small cell lung cancer (SCLC) cell lines (one classic type, and three variant type) and one gastric cancer line (KATO 3). Thirteen antibodies (Code # 4, 12, 21, 31, 34, 41, 48, 58, 60, 61, 74, 77, 82) among 98 registered antibodies showed a very similar pattern to antibody NE150, which was previously characterised as SCLC cluster 1. Since NE150 showed a positive reaction to the natural killer (NK) cell population, the serological specificity was compared with NK cell-associated antibodies, NKH1 (CD56), Leu7 (CD57) and Leu11 (CD16). Only NKH1 antibody showed a similar pattern to NE150, when tested against various target cells including SCLC lines and peripheral leukocytes, suggesting that NKH1 is a cluster 1 antibody, although it was already classified as CD56 of hematopoietic cells. By sequential immunoprecipitation, the antigen detected by NE150 antibody was depleted by preincubation with NKH1 antibody, but the reactivity of NE150 was not inhibited by NKH1 antibody, suggesting that NE150 and NKH1 detect different epitopes on the same antigen molecule. Epitope analysis was also conducted with 13 antibodies of cluster 1. Ten were found to detect the same epitope as NE150. The other three did not inhibit the binding of NE150 or NKH1, suggesting that there are at least three epitopes. Since the cluster 1 antibodies were demonstrated to detect NCAM, the present results suggest the presence of at least three epitopes on this molecule.

Antibodies, Monoclonal↗
PubMed1991

Coexpression of the stem cell factor and the c-kit genes in small-cell lung cancer.

Stem cell factor (SCF) is a pluripotent growth factor which is suggested to play an important role in proliferation and differentiation in various types of fetal and adult tissues as the ligand of the c-kit proto-oncogene product. However, very little is known about expression of the SCF gene in human malignancies. We analysed DNA and RNA extracted from 28 cell lines and 16 fresh tumor specimens of lung cancer as well as 24 cancer cell lines of various origin for SCF expression. Now we report that the SCF gene is expressed in a wide variety of human cancers including lung cancer, in marked contrast to c-kit, which is expressed in very few types of cancers. As a consequence, coexpression of both the ligand and the receptor is seen only in small-cell lung cancer, suggesting possible involvement of autocrine stimulation via this ligand-receptor system in the pathogenesis of this aggressive cancer. In addition, this study revealed that the human SCF gene is transcribed into two major forms of alternatively spliced mRNAs with different molar ratio in fetal, adult and malignant tissues.

Base Sequence↗
PubMed1991

[BCL-2 gene in lymphocytic malignancy].

In t(14;18) (q32;q21) lymphomas, bcl-2 gene is activated by the juxtaposition of immunoglobulin (Ig) gene. The fused bcl-2-Ig gene generates chimeric mRNAs which consist of bcl-2 at 5' portion and Ig at 3' portion. Chimeric mRNA does not disrupt the bcl-2 coding frame of 239 amino acid polypeptide. Bcl-2-Ig transgenic mice demonstrated the extended B cell survival and the follicular lymphoproliferation, but they did not develop a malignancy until 25 weeks. Ten percent of them, however, developed malignant diffuse large-cell lymphomas after a long latency. Forty percent of these malignancies demonstrated the c-myc rearrangement, indicating that multiple step changes are required for malignant transformation in bcl-2 activated cells. Study on the bcl-2 gene rearrangement in Japanese B cell lymphoma and B-CLL revealed that 10 out of 32 cases of follicular lymphoma (31%), 5 out of 56 cases of diffuse lymphoma (9%) and 2 out of 30 cases of B-CLL (7%) were rearranged. Less frequency of B cell lymphoma, particularly follicular lymphoma in Japan might be partly due to the less bcl-2 involvement than in American cases. The ratio of bcl-2 involvement in B-CLL is not significantly different between Japan and U.S.A.. bcl-2 rearrangement at 5' promoter region is noted for Japanese B-CLL which was demonstrated for American cases. The clinical application of polymerase chain reaction for bcl-2 translocation was also discussed.

Animals↗
PubMed1990

Single clonal origin of neoplastic B-cells with different immunoglobulin light chains in a patient with Richter's syndrome.

A 71-year-old man was found to have chronic lymphocytic leukemia (CLL) and diffuse large cell lymphoma (DLC) simultaneously and was diagnosed as Richter's syndrome. The CLL had mu lambda surface immunoglobulin (sIg) whereas the DLC had mu kappa sIg. However, the immunoglobulin (Ig) gene rearrangement and surface marker analysis demonstrated that both CLL and DLC had identical rearrangement patterns of the Ig heavy chain (IgH) and identical surface markers CD5+, CD19+, and CD20+. These facts imply that in this case the two malignancies are of single clonal origin initially, and that different sIg of CLL and DLC do not, therefore, necessarily indicate the biclonality of these malignancies. The origin of DLC in Richter's syndrome remains controversial. This case suggests difficulty in concluding the biclonality of these malignancies. For a conclusion on clonality to be definitive, there is a need for cloning and nucleotide sequencing of rearranged Ig genes in more patients with Richter's syndrome.

Aged↗
PubMed1990

A case of infantile acute monocytic leukemia caused by vertical transmission of the mother's leukemic cells.

A case of infantile acute monocytic leukemia (AMoL), which was probably transmitted from a pregnant woman with leukemia to her unborn infant, is presented. A woman had AMoL when her third infant was born. This infant, who was a boy, also suffered from AMoL when he was 20 months of age. The infant's leukemic cells had the same cytochemical and immunophenotypic patterns as the mother's leukemic cells. By cytogenetic analysis, the majority of the infant's leukemic marrow cells had the 46,XX karyotype and showed no Y body by quinacrine staining. Moreover, the phenotype for human major histocompatibility system, HLA-A, HLA-B, and HLA-DR of the infant's leukemic cells was consistent with that of the mother's lymphocytes. Thus, the infant's leukemic clone was found to be identical to the mother's leukemic clone. His lymphocytes could not react with the mother's leukemic cells or his own leukemic cells in mixed lymphocyte culture, suggesting that the HLA homozygosity of the mother may have played a role in inducing immunologic tolerance to the immigrated leukemic cells in the infant. This is the first report of an infantile leukemia transmitted from a mother with leukemia, supposedly through the placenta.

Adult↗
PubMed1990

Genuine CD7 expression in acute leukemia and lymphoblastic lymphoma.

CD7 has been used as a valuable marker for normal and malignant T cells and also for a proportion of acute nonlymphocytic leukemia (ANLL) cells. Difference in reactivity was noticed among CD7 antibodies, however, when tested against ANLL cells and myeloid/monocytoid cell lines; Tp40 antibody produced in our laboratories was not reactive with the HL-60 promyelocytic line, whereas 4A antibody was reactive, even though both detected a quite similar or an identical epitope on CD7 molecule. Preincubation of HL-60 cells with human immunoglobulin preparation clearly negated the reactivity by 4A, suggesting that 4A antibody is not reactive to CD7 itself, but it probably binds with immunoglobulin G Fc receptors expressed on HL-60 cells. Five cases of ANLL which were positive with 4A antibody were selected and tested with Tp40 antibody, and only two were found to be positive. Expression of CD7 mRNA in these two cases (but not in other cases) was also demonstrated by Northern blotting with a cDNA probe for CD7 recently cloned in our laboratories, indicating that CD7 is expressed on a certain fraction of ANLL, although the positive cases may be smaller than the reports so far appeared. A Northern blot study was also conducted with two acute lymphocytic leukemia cases and one lymphoblastic lymphoma case with CD7+, CD2-/+/-, CD5-/+/- phenotype and germline T cell receptor beta genes. CD7 mRNA is expressed in all three cases and CD3 mRNA is also observed in two cases, suggesting that these tumor cells are of T precursor origin.

Acute Disease↗
PubMed1990

Analysis of the fine specificities of 11 mouse monoclonal antibodies reactive with type 2 blood group determinants.

The specificity of 11 mouse monoclonal antibodies reacting selectively with type 2 blood group structures was analyzed in detail by studying their reactivities with a panel of standard glycolipids, glycolipids from erythrocytes and blood group glycoproteins. The antibodies reacted with monofucosyl type 2 H, difucosyl type 2 structures (Le gamma) or both; none of the antibodies reacted with type 1 (H, Lea, or Leb) structures. Only a small proportion of the antibodies were completely specific for either type 2H or Le gamma structures. None of the antibodies had identical patterns of reactivity and their specificities were individually distinct. Seven antibodies preferentially agglutinated O and A2 erythrocytes. Anti-Le gamma-specific antibodies, except mAb101, did not agglutinate erythrocytes or react with glycolipids from erythrocytes, indicating the absence of Le gamma structures in erythrocyte glycolipids. The ability of some antibodies to react with A erythrocytes was shown to be due to cross-reactivity of the antibodies with type 3 (repetitive) A structures. The study demonstrates that monoclonal anti-carbohydrate antibodies tend to react with a range of related, and even distantly related, structure in a pattern characteristic of each antibody and that very few antibodies have extremely restricted specificities.

ABO Blood-Group System↗
PubMed1990

Nuclear localization of antigens detected by a monoclonal antibody against a synthetic peptide of rfp finger protein.

A monoclonal antibody (MAb) was generated against a synthetic peptide corresponding to amino acids 148 to 163 of the rfp protein with zinc finger domains. The MAb, designated RFP-1 (IgM), which was positive with the immunizing peptide in enzyme-linked immunosorbent assay, was reactive in immunoblotting with an in vitro translated rfp product as well as with native proteins in cell extracts made from mouse testis and HL-60 human leukemia cell line, both of which were previously shown to express high levels of rfp mRNA. When HL-60 cells were fractionated into nuclear and cytoplasmic components, the protein reactive with RFP-1 MAb was detectable only in the nuclear fraction. By the avidin-biotin complex immunoperoxidase method, this MAb strongly stained over 90% of the nuclei of human and mouse spermatogenic cells, except mature spermatozoon, and of human testicular tumor cells. In other human adult tissues, up to 60% of positive cells were observed. These antibody activities were clearly absorbed by pre-incubation of RFP-1 MAb with the immunizing peptide. These results thus indicated that RFP-1 MAb recognizes a nuclear protein which is expressed at high levels in male germ cells.

Amino Acid Sequence↗
PubMed1990

Successful treatment of acute leukemia with t(4;11) in an infant with congenital hypothyroidism.

We describe a case of acute leukemia with t(4;11) (q21;q23) in a 3-month-old girl suffering from congenital hypothyroidism. The blast cells were cytochemically and immunologically classifiable as acute lymphoblastic leukemia of an early B-cell lineage (HLA-DR +, B4 +, CALLA-). but we treated this patient with a protocol designed mainly for acute nonlymphocytic leukemia, employing Adriamycin, vincristine, and cytosine arabinoside. Although the prognosis of this type of leukemia is known to be extremely poor, our patient is currently alive and in continuous complete remission lasting 35 months to date. This case may demonstrate a potential alternative therapeutic strategy for acute leukemia with t(4;11) as well as clinical evidence for the mixed-lineage characteristics of this condition. However, the pathogenetic role of congenital hypothyroidism in the development of acute leukemia is still uncertain.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1990

Point mutations of c-ras genes in human bladder cancer and kidney cancer.

Point mutations of c-ras genes at codons 12, 13 and 61 were analyzed in 26 cases of bladder cancer and 16 cases of kidney cancer. DNA prepared from either frozen tissues or 10% formalin-fixed, paraffin-embedded tissues were amplified by means of polymerase chain reaction methods, and mutations were analyzed by dot blot hybridization assays with oligonucleotide probes. In three cases of bladder cancer c-ras mutations were found, at codons 13 and 61 of c-Ha-ras and at codon 61 of c-Ki-ras, while no mutation was found in kidney cancer. No mutation was found in normal bladder epithelial tissues from the same patients. Our findings, taken together, may indicate relative scarcity of c-ras mutations in these types of human cancer. The results of dot blot hybridization assays and DNA sequencing showed a G-to-C transition of the first nucleotide at codon 13 c-Ha-ras. This is the first time that such a point mutation has been detected in human cancer tissues.

Aged↗
PubMed1990

Long-term follow-up study of cerebral palsy children and coping behaviour of parents.

The purpose of this study was to understand: (a) the feelings, thoughts and actions of parents at the time their children were diagnosed as having cerebral palsy; (b) the crisis periods in raising their children; (c) the important persons who supported the parents during the 'acceptance' phase; and (d) the roles of mothers and fathers in raising the children. The subjects were 28 mothers and fathers who had sons or daughters with cerebral palsy. The latter offspring were aged 22-29 years at the time of this study. They were interviewed at their homes with a semi-structured method retrospectively. The results showed that most parents became aware of their children's disability in infancy and most of their children were diagnosed as having cerebral palsy by around 2 years of age. The mothers' reactions to the diagnosis were emotional and those of the fathers were realistic in coping with the problem. Although crisis periods arose throughout the periods of growth and development of the children, the infancy period was the more critical for mothers and the toddlerhood, school-age and adolescence periods were more critical for fathers. The important support people were the spouses. The mothers took care of the children while the fathers provided an income; however, some fathers withdrew from competing for achievement in their jobs.

Adaptation, Psychological↗
PubMed1990

Assay for the soluble interleukin-2 receptor by sandwich enzyme linked immunosorbent assay.

A system for the detection of soluble IL-2R by sandwich enzyme linked immunosorbent assay (ELISA) was established. This assay system has good reproducibility, and was found to be specific for soluble IL-2R by examination of the binding of anti-IgM antibody or anti-IL-2R monoclonal antibody (mAb) with IgM or soluble IL-2R and inhibition by IL-2 of the binding of anti-IL-2R mAb to soluble IL-2R. The IL-2R molecules in supernatant of phytohemagglutinin-stimulated peripheral blood mononuclear cells were of 44,000-61,000 molecular weight as estimated by size of exclusion high performance liquid chromatography. The results of determinations for soluble IL-2R with a Eurogenetics kit (sandwich enzyme linked immunosorbent assay) were significantly correlated with those by the our method.

Enzyme-Linked Immunosorbent Assay↗