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S Shiokawa

Publications and source records attributed to S Shiokawa.

67 records · Page 4Linked to original sources
PubMed1990

[Antigenic phenotype of the lymphocytic component of regional lymph nodes in patients with cervical cancer and its modulation by lentinan].

Immunohistochemical study with various monoclonal antibodies to the mononuclear cell surface antigens was carried out on the regional lymph nodes in patients with cervical cancer to assess the augmentative effect of lentinan. Zero, 2, 4, or 6 mg of lentinan was administered i.v. one day prior to surgery to patients with cervical cancer (14 cases with FIGO stage 0 and 19 with FIGO stage Ib) and those with benign gynecologic tumors (8 cases with myoma uteri and 6 with ovarian tumor). Frozen sections of fresh pelvic lymph nodes obtained from these patients during surgery were stained by the ABC (avidin-biotin-peroxidase complex) method using several monoclonal antibodies to define the surface phenotype of mononuclear cells. The results were as follows: 1. Pelvic lymph nodes in patients with benign disease: In the absence of lentinan, lymphocytes stained with Leu 3a antibody were more numerous than those stained with Leu 2a, and both were observed mainly in the paracortical area (PC). The number of lymphocytes stained with Leu 4 antibody was practically equal to the sum of those stained with Leu 3a and Leu 2a. HLA-Dr positive lymphocytes were present in moderate numbers in PC and sinus. The above findings were not changed by the administration of lentinan. Cells stained with monoclonal antibodies including Leu 7, 11, M3, and IL-2 receptor (IL-2R) were very few or absent. 2. Pelvic lymph nodes in patients with cervical cancer receiving no lentinan: The findings obtained in these cases were much the same as those in patients with benign tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Differentiation↗
PubMed1990

A novel CD10-positive erythroid cell line, RM10, established from a patient with chronic myelogenous leukemia.

A novel erythroid cell line, RM10, was established from a long-term bone marrow culture of a patient with chronic myelogenous leukemia (CML). RM10 cells were positive for periodic acid Schiff (PAS), but negative for peroxidase and dual esterase. RM10 cells had la, pre B (CD10), myeloid (CD13, CD14, CD33) and erythroid (glycophorin A) markers, but had no other lymphoid, megakaryocytic, or mesenchymal cell markers. RM10 cells spontaneously synthesized hemoglobin, which was markedly enhanced with hemin. Isoelectric focusing of the cell lysates and northern blot analysis of the total cellular RNA revealed hemoglobin synthesis in the cells. Using 125I-labeled recombinant human erythropoietin (Epo), two classes of Epo receptors were demonstrated in the RM10 cells. However, Epo did affect neither growth nor erythroid differentiation of the cells. RM10 cells rapidly differentiated to monocytic cells in the presence of 12-0-tetradecanoylphorbol-13-acetate, and simultaneously expressed glycoprotein IIb/IIIa. RM10 cells had Philadelphia chromosome (Ph), and expressed p210bcr-abl using immunoprecipitation with anti-c-abl and anti-phosphotyrosine antibodies. These results indicate that the RM10 cells have the characteristics of multipotential hemopoietic cells originating from Ph-positive CML and that high affinity Epo receptor class is not a sufficient condition for Epo responsiveness.

Adult↗
PubMed1990

Molecular analysis of the gene of the alpha 1-antitrypsin deficiency variant, Mnichinan.

Mnichinan, a variant of alpha 1-antitrypsin (alpha 1-AT) was detected in a Japanese individual with serum alpha 1-AT deficiency (18 mg/dl), associated with aggregated alpha 1-AT molecules in the hepatocytes. Cloning and sequencing of the 10,627-bp-long region containing the Mnichinan gene and the normal M1(Val213) alpha 1-AT gene revealed all five exons of the Mnichinan gene to be identical with the M1(Val213) alpha 1-AT gene, except for two changes: a TTC trinucleotide deletion in the codon for amino acid Phe52 and a G-A substitution, by which the normal Gly148 (GGG) became Arg148 (AGG). Dot blot analysis of the polymerase chain-reaction-amplified DNA derived from the proband and other family members showed both mutations to be associated with an alpha 1-AT deficiency phenotype. Ninety-eight alpha 1-AT alleles were all negative for both changes. Comparison of the region, except for five exons between the Mnichinan and M1(Val213) genes, demonstrated one base difference in the 5' flanking region and 14 base changes in the introns. All exon-intron junctions were identical, and base changes in the 5' flanking region did not seem significant. The G-A substitution in codon 148 of the Mnichinan gene could not be responsible for the alpha 1-AT deficiency phenotype because Arg- and not Gly- was located at the corresponding position of the protein C inhibitor belonging to the serine protease inhibitor superfamily. The deletion of Phe52 may cause the newly synthesized alpha 1-AT protein to aggregate, resulting in alpha 1-AT deficiency. Comparison of the alpha 1-AT gene sequences available indicated that the C-T substitution at the CpG dinucleotide has an important role in generation of variants and nucleotide changes in the noncoding regions of the alpha 1-AT gene.

Adult↗
PubMed1989

Heterogeneity of the gamma-globin gene sequences in Japanese individuals: implication of gene conversion in generation of polymorphisms.

The linked fetal globin genes (the G gamma- and A gamma-globin genes) were cloned from Japanese individuals with three different haplotypes of the HindIII polymorphisms within the gamma-globin genes. Determination of nucleotide sequences of the segment spanning from IVS2 to the 3' flanking region of each gamma-globin gene revealed that nucleotide differences are located at 43 positions and a stretch of simple GT or GC sequences. Almost half of the nucleotide changes could be accounted for by gene conversion between the G gamma- and A gamma-globin genes. We found that gene conversion had created the SacI polymorphic site just downstream of the A gamma-globin coding region. Association of the SacI polymorphic site with the HindIII polymorphic site suggests that the region containing these two sites was derived from that of the linked G gamma-globin gene through a gene conversion event. The nucleotide sequences obtained here are identical to those of the Caucasoid fetal globin genes of the same haplotypes, with the exception of some sequence changes in the hot spots of mutations. These results indicate that the sequence heterogeneity of the gamma-globin genes can be classified into three major categories according to HindIII haplotypes. The possible mechanisms of generation of the heterogeneity of the gamma-globin gene sequences are discussed.

Base Sequence↗
PubMed1988

Hemoglobin synthesis of both adult and fetal types in a human CML cell line.

We present evidence for the continuous erythroid differentiation of a cell line, KU-812-F, without the addition of an inducer. Erythroid differentiation of these cells was confirmed according to the following findings: 1) erythroid morphology; 2) existence of glycophorin A and carbonic anhydrase I as a membrane marker protein and an enzyme characteristic of erythroid cells, respectively; 3) synthesis of adult type (HbA and HbA2) and fetal type (HbF) hemoglobins, as detected on isoelectric focusing; and 4) transcription of the alpha-, beta-, and gamma-globin genes, as detected on Northern blot analysis.

Cell Differentiation↗
PubMed1988

Molecular analysis of Japanese delta beta-thalassemia.

A DNA fragment containing the deletion junction region from a Japanese individual with homozygous delta beta-thalassemia has been cloned. A clone containing the normal DNA surrounding the 3' breakpoint of this deletion and a clone carrying the G gamma- and A gamma-globin genes of this patient were also isolated. Sequences of the deletion junction and both gamma-globin genes were determined. A comparison of these sequences with previously determined sequences of the normal counterparts revealed that the 5' breakpoint is located between 2,134 and 2,137 base pairs (bp) 3' to the polyA site of the A gamma-globin gene, the 5' breakpoint is located just downstream of the 3' border of the fetal gamma-globin duplication unit, and no molecular defects are evident within the gamma-globin gene region. A comparison between the sequences of the normal DNA surrounding the 3' breakpoint and the normal DNA surrounding the 5' breakpoint shows that deletion is the result of a nonhomologous recombination event. There are A+T-rich stretches near the 5' and 3' breakpoints in the normal DNA, and a portion of an Aly repeat is located in the region 3' to the 3' breakpoint. Southern blot analysis using probes 3' to the beta-globin gene showed that the deletion extends in the 3' direction further than any other deletions associated with delta beta-thalassemia and hereditary persistence of fetal hemoglobin (HPFH) heretofore reported. These results are discussed in terms of the mechanism generating large deletions in mammalian cells and three models for the regulation of gamma-globin and beta-globin gene expression in humans.

Adult↗
PubMed1987

Differential induction of adult and fetal globin gene expression in the human erythremia cell line KMOE.

Induction of globin gene expression in KMOE cells derived from a patient with acute erythremia was studied by Northern blot and S1 analysis. KMOE cells exposed to cytosine arabinofuranoside (Ara-C) synthesized beta-globin gene transcripts, however, in the presence of hemin gamma-globin gene transcripts. An increase in alpha-globin gene transcripts was also detectable in KMOE cells treated with both Ara-C and hemin. Upon exposure to hemin after exposure to Ara-C, or exposure to Ara-C after hemin, there was a 5-10-fold increase in gamma-globin gene transcripts compared to that of cells induced by hemin alone. Neither epsilon nor zeta globin transcripts were detected. The KMOE cell line, therefore, exhibits phenotypic properties of adult and fetal erythroid cells.

Cell Line↗
PubMedDate not supplied

Regulation of the antibody repertoire through control of HCDR3 diversity.

In man, as in mouse, diversification of the antibody repertoire appears to follow a strict developmental program whereby antigen specificities are serially acquired during ontogeny. When compared to the adult repertoire, the fetal antibody repertoire is highly enriched for polyreactive specificities of low affinity. Although the mechanisms governing the development of this fetal repertoire differ between human and mouse, the composition and structure of the fetal antibodies produced by both species are quite homologous. Specifically, both species use similar V gene segments and restrict the sequence and structure of the third complementarity determining region (HCDR3) of the antibody heavy chain. The precise role that this restriction of the HCDR3 might play in the development of immunocompetence in the human remains to be elucidated.

Animals↗