Biomedical subjects

Y Tanigawa

Publications and source records attributed to Y Tanigawa.

At least 37 records · Page 2Linked to original sources

PubMed1997

Large-scale immunohistological staining using polyethylene glycol-embedded sections mounted on 96-well plates for monoclonal antibody screening.

We have developed a rapid, large-scale immunohistological staining method for monoclonal antibody screening. Sections embedded in polyethylene glycol are mounted on 96-well plates and then subjected to immunological procedures using a 96-well format. With this method, we can prepare 10 plates (960 samples) in 1 day and then perform immunohistological procedures in 1 day by using an enzyme-linked immunosorbent assay-like 96-well format. Thus, we can avoid difficulties seen in conventional immunohistological methods that use sections mounted on glass slides. This method offers high-throughput screening of monoclonal antibodies. In addition, it has the potential of being automated.

Animals↗

PubMed1997

[An autopsy case of chronic granulomatous disease diagnosed by biopsy].

This report concerns a male patient aged 25 years, diagnosed at the age of 12 years as suffering from chronic granulomatous disease. This patient had p47-phox deficiency. He was admitted to this hospital because of fever and dyspnea accompanied by right spontaneous pneumothorax. He failed to respond to medical treatment. He died from respiratory failure four months after admission. Autopsy demonstrated pigmented lipid histiocytes characteristic of CGD. These characteristic pigmented cells were distributed in the spleen, liver, lymph nodes and in the small intestine. As for the nature of the pigment, lipofuschin-like compound were identified. Granulomatous component was seen in the mucosa of the stomach obtained by operation. The presence and characteristic distribution of such pigmented macrophages in tissue in young adults may suggest the diagnosis of CGD.

Adult↗

PubMed1996

Ubiquitin-like moiety of the monoclonal nonspecific suppressor factor beta is responsible for its activity.

Monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by murine T cell hybridoma, possesses pleiotrophic Ag-nonspecific suppressive functions. Most recently, a cDNA clone encoding MNSF beta (a subunit of MNSF) was isolated and characterized. The MNSF beta cDNA encodes a 14.5-kDa fusion protein with MNSF-like activity consisting of an 8-kDa ubiquitin-like segment (ubi-L) and the ribosomal protein S30 (6.5 kDa). To determine whether ubi-L itself has biologic activity, cDNA encoding the ubi-L region was expressed in bacteria and the recombinant product was tested for the activity. Ubi-L showed MNSF-like biologic activity without any cytotoxic action. Interestingly, the addition of ubiquitin to the assay inhibited ubi-L-induced suppression. IFN-gamma, which is known to enhance the expression of MNSF receptor, induced splenocytes to secrete ubi-L by increasing mRNA. Ubi-L has species-specific action and the ability to selectively inhibit the B cell proliferative response stimulated by soluble but not by Sepharose-bound anti-Ig Ab. In addition, okadaic acid, a serine/threonine phosphatase 1 and 2A inhibitor, showed a synergistic inhibitory action with ubi-L, indicative of the possible involvement of phosphatase(s) in the regulation of ubi-L action.

Amino Acid Sequence↗

PubMed1996

Monoclonal nonspecific suppressor factor beta (MNSF beta) inhibits the production of TNF-alpha by lipopolysaccharide-activated macrophages.

The monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by a murine T cell hybridoma, shows a pleiotropic antigen-nonspecific suppressive function. Most recently, a cDNA encoding a subunit of MNSF (MNSF beta) has been isolated and characterized. Recombinant form of MNSF beta (rMNSF beta) inhibits lymphokine functions, as does native MNSF. In this study, we investigated whether rMNSF beta also affects macrophage function in terms of LPS-induced TNF-alpha production by a mouse macrophage cell line, J774. rMNSF beta suppressed the TNF-alpha production in a dose-dependent manner. This suppressive effect was remarkably reduced when rMNSF beta was added after 6 h of LPS stimulation. In addition, enhancement of TNF-alpha production by IFN-gamma was also suppressed by rMNSF beta. The suppressive effect was partly neutralized by the addition of the serine/threonine phosphatase inhibitor, okadaic acid. This finding suggests that serine/threonine protein phosphatases type 1 and/or 2A may be implicated in the mechanism of action of MNSF.

Animals↗

PubMed1996

Ubiquitin-like polypeptide inhibits the IgE response of lipopolysaccharide-activated B cells.

Monoclonal non-specific suppressor factor (MNSF), a lymphokine produced by a murine hybridoma, was originally found to inhibit the generation of lipopolysaccharide (LPS)-induced Ig-secreting cells. Most recently, we demonstrated the recombinant form of the ubiquitin-like segment (rUbi-L) or MNSF beta, an isoform of MNSF, has a MNSF activity. To investigate the possible role of rUbi-L, a 8.5 kDa ubiquitin-like polypeptide, in the regulation of Ig isotype secretion, rUbi-L was added to purified B cell cultures stimulated with LPS plus IL-4. rUbi-L notable suppressed the IgE and IgG1 responses when added at culture initiation. In addition, rUbi-L had a strong effect on IgG3 production and a little effect on IgM production by LPS-stimulated B cells, whereas the level of other isotypes (IgG2a, IgG2b and IgA) was not affected. These findings demonstrate the isotype-specific suppression, but not pan-suppression, of Ubi-L. IFN-alpha and IFN-gamma, which are also known to inhibit the IgE response, showed a synergistic effect with Ubi-L, albeit the effects of IFN-alpha were smaller. The action was reversed by the addition of neutralizing antibodies of these cytokines. Therefore, Ubi-L, a ubiquitin-like protein, may have an important immunoregulatory role on the IgE response.

Animals↗

PubMed1995

Molecular cloning and characterization of a cDNA encoding monoclonal nonspecific suppressor factor.

The monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T-cell hybridoma that inhibits the generation of lipopolysaccharide-induced immunoglobulin-secreting cells in an antigen-nonspecific manner. A cDNA clone encoding MNSF beta (an isoform of MNSF) was isolated and expressed in bacteria. The sequence obtained is virtually identical to the Fau protein, a product of the ubiquitously expressed fau gene with unknown function. Northern blot analysis demonstrated a single, 0.6-kb transcript. Specific polyclonal antibodies against synthetic peptides corresponding to the deduced amino acid sequences were elicited in rabbits. Immunoprecipitation experiments with these antibodies showed that MNSF beta is released extracellularly in an aggregate form, albeit it lacks a signal peptide sequence. The anti-MNSF beta affinity eluate from the MNSF-producing murine hybridoma (E17) and concanavalin A-activated splenocyte culture supernatants inhibited the immunoglobulin production by lipopolysaccharide-activated splenocytes. Recombinant MNSF beta also showed a similar biologic activity. Thus, ubiquitin-like protein(s) may be involved in the regulation of the immune responses.

Amino Acid Sequence↗

PubMed1995

The role of cyclic AMP in the lipopolysaccharide-induced suppression of thymidine kinase activity in macrophage.

We observed that lipopolysaccharide (LPS, 1 micrograms/ml) can suppress [3H]thymidine incorporation into acid-insoluble fraction in a mouse macrophage cell line J774 (over 70% at 6 h) without affecting the uptake of [3H]thymidine or DNA polymerase activity. Paralleling this suppression, a decrease in the thymidine kinase (TK) activity, but not of thymidine monophosphate (TMP) kinase and thymidine diphosphate (TDP) kinase, was observed. LPS dose-dependently increased intracellular cAMP levels to about 3.5-times basal at 6 h, proportionally to the decrease of the TK activity. Elevation of intracellular cAMP by several reagents also decreased TK activity. Apparently LPS treatment elevates cAMP concentration by decreasing the low Km cAMP phosphodiesterase activity (58% at 6 h). The time course of cAMP-dependent protein kinase (PK-A) activity during the first 6 h after LPS treatment correlated with that of cAMP concentration. Treatment with a PK-A inhibitor restored about 63% of LPS-induced reduction of TK activity at 6 h. At longer times, however, there was a discrepancy between the change of cAMP concentration or PK-A activity and the reduction of TK activity. Therefore, protein kinase activation caused by the accumulation of intracellular cAMP probably triggers some mechanism responsible for the reduction of the TK activity.

Cells, Cultured↗

PubMed1995

Antinuclear antibodies in healthy aging people: a prospective study.

In order to evaluate the expression of antinuclear antibodies (ANA) in normal elderly individuals over time and clinical significance, a cross-sectional ANA testing in healthy Japanese was performed, followed by annual evaluations of ANA positive aged (> or = 65 years) and a control group. ANA was more prevalent in the aged (11.4% vs. 3.8%) and most were persistent after 4 years. Anti-ssDNA and anti-histone antibodies were increased in aged ANA positive as compared to ANA negative controls. Except for a history of spontaneous abortion, there was no differences in clinical findings. HLA DRB1*0901 and the DQB1*0602 + 0302 + 0303 set of alleles were increased in ANA positive. Therefore, ANA in the aged were persistent, apparently directed toward chromatin elements, and shared MHC associations with autoimmune diseases. Longer follow-up may be necessary to improve the evaluation of clinical significance of ANA in the aged.

Adult↗

PubMed1995

Cloning of a gene of the DEAD box protein family which is specifically expressed in germ cells in rats.

The cloning of a rat cDNA, RVLG, which encodes a novel protein of 713 amino acid residues with a molecular mass of 77,953, is described. The putative RVLG protein shares amino acid domains that are conserved in the DEAD protein family. It is approximately 52% identical and 85% similar in amino acid sequence to the XVLG1, a product which we previously cloned from Xenopus ovaries, suggesting it is the homologue of XVLG1. The size of the mRNA for RVLG is 3.1kb and can be detected specifically in the gonads of male and female adult rats.

Adenosine Triphosphatases↗

PubMed1995

Monoclonal nonspecific suppressor factor beta inhibits interleukin-4 secretion by a type-2 helper T cell clone.

Monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T cell hybridoma that inhibits the generation of lipopolysaccharide-induced immunoglobulin secreting cells in an antigen-nonspecific manner. Most recently, a cDNA clone encoding MNSF beta, a subunit of MNSF, was isolated and sequenced (Nakamura et al., Proc. Natl. Acad. Sci. USA 1995. 92: 3463). The MNSF beta cDNA encodes a 14.5-kDa ubiquitin-like protein showing several MNSF activities. We have previously observed that hybridoma-derived native MNSF can inhibit interleukin (IL)-4 secretion by bone marrow-derived mast cells. Therefore, we studied whether MNSF beta is also able to inhibit IL-4 secretion by the type-2 helper T cell clone D10. G4.1. MNSF beta inhibited both the IL-4 secretion and the proliferation of concanavalin A-activated D10 cells, although neither effect was complete.

Animals↗

PubMed1995

Human nonspecific suppressor factor (hNSF): cell source and effects on T and B lymphocytes.

The human nonspecific suppressor factor (hNSF), the probable counterpart of the murine monoclonal nonspecific suppressor factor (MNSF), has been isolated from the ascitic fluid of a patient with systemic lupus erythematosus and characterized. hNSF presents an inhibitory activity on the proliferation and IgG production of mitogen stimulated human PBMC. In the present study, we demonstrate that hNSF can be isolated from the supernatants of ConA-activated T cells, but not from CD8-depleted T cells, indicating the CD8+ T cells are the major source of the factor. We also studied the effects of hNSF on purified human B and T cells; hNSF strongly inhibited the proliferation and Ig secretion by highly purified B cells induced by SAC plus IL-2, as well as the proliferation of T cells activated by Con A plus IL-2. These results indicate that hNSF is a CD8+ T cell product with strong antigen-nonspecific immunoregulatory action on both lymphocyte populations.

B-Lymphocytes↗

PubMed1995

Effect of lipopolysaccharide on thymidine salvage as related to macrophage activation.

Lipopolysaccharide (LPS), known as one of the potent activators of macrophages, has inhibitory effects on the proliferation of normal macrophages and macrophage-like cell lines. We report here that LPS dose- and time-dependently suppressed the tritiated thymidine ([3H]TdR) incorporation into the acid-insoluble fraction with a significant inverse correlation to the tumour necrosis factor-alpha (TNF) production in the J774.1 macrophage cell line. Among the three tested enzymes involved in DNA synthesis, only thymidine kinase (TK) activity decreased progressively in parallel with the decline in [3H]TdR incorporation, reaching 97% inhibition within 12 hr of LPS treatment, while changes in the activities of other two enzymes, DNA polymerase alpha and thymidylate synthase (TS), were less significant. On the other hand, LPS inhibited the cell proliferation only incompletely, as judged by 62% inhibition of cell growth at 36 hr. Even in the experiments done in a TdR-free medium, cell growth was inhibited by LPS to the same extent, suggesting that TK was not directly involved in the proliferation of J774 cells. LPS also inhibited the conversion of TdR to thymidine monophosphate (TMP) in murine peritoneal exudate macrophages (PEM). Thus LPS-induced suppression of TdR salvage related to TNF production is common in both normal and neoplastic macrophages, and therefore may be of potential importance in the process of macrophage activation.

Animals↗

PubMed1994

Monoclonal non-specific suppressor factor (MNSF) inhibits the IL4 secretion by bone marrow-derived mast cell (BMMC).

We investigated whether monoclonal non-specific suppressor factor (MNSF) has an effect on the IL4 secretion by BMMC sensitized with anti-DNP IgE monoclonal antibody. The sensitized BMMC showed a maximum increase of IL4 secretion at 10 ng/ml antigen activation. When MNSF (10 U) was added, it remarkably suppressed the IL4 secretion (82 +/- 3%). MNSF also suppressed the IL4 secretion by the mast cell line, CFTL-15, but not CFTL-12. Northern blot analysis showed that, when the same amounts of MNSF were added, IL4 mRNA expression was decreased, suggesting that the suppression results from the inhibition of a transcript.

Animals↗

PubMed1994

[Alpha fetoprotein-producing primary lung cancer].

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Animals↗

PubMed1993

Loss of poly(ADP-ribose) synthetase activity without change in arginine-specific ADP-ribosyltransferase activity by acid-treatment; application of the treatment to simple assay for transferase in the presence of poly(ADP-ribose) synthetase.

We investigated the effects of acid treatment (pH 3.5) on the activities of arginine-specific ADP-ribosyltransferase (mono(ADP-ribosyl)transferase) (EC 2.4.2.31) and poly(ADP-ribose) synthetase (EC 2.4.2.30) purified from chicken liver, and we observed that the former enzyme retained completely its activity while there was no evidence for activity of the latter enzyme. Kinetic parameters, including Km values for NAD and whole histones in the reaction catalyzed by acid-treated ADP-ribosyltransferase, were the same as those in the reaction catalyzed by non-treated enzyme. The stereospecificities of the reaction forming ADP-ribosylarginine by acid-treated and non-treated ADP-ribosyltransferases were indistinguishable. We made use of this acid treatment to determine ADP-ribosyltransferase activity in chicken and chick-embryo liver extracts, with a filter assay. The enzyme activities (means +/- S.D. for three separate experiments) of 1-year-old chicken and 13-day-old chick embryo livers were 372.7 +/- 80.20 and 110.6 +/- 27.22 nmol ADP-ribose/g wet liver per h, respectively. This acid treatment is useful for filter assay with labelled NAD of arginine-specific ADP-ribosyltransferase in the fraction containing poly(ADP-ribose) synthetase.

ADP Ribose Transferases↗

PubMed1992

Species dependency of in vitro macrophage activation by bacterial peptidoglycans.

The effect of various bacterial cell wall components on in vitro biological function of murine peritoneal exudate macrophages was evaluated. We examined four different parameters of metabolic activity and monokine secretion. Peritoneal exudate macrophages from rats and guinea pigs, all of the strains tested, were stimulated by whole bacterial cell wall preparations, purified bacterial cell wall peptidoglucans, its water-soluble peptidolglycan fragments, muramyl dipeptides and amphipathic substances. Murine peritoneal exudate macrophages were activated by amphipathic substances of gram-positive bacteria. However, macrophages from mice, irrespective of strains, were not stimulated in the in vitro assay systems by purified bacterial cell wall peptidoglycan, water-soluble bacterial peptidoglycan fragments or muramyl dipeptides. These results suggest that macrophage activation by bacterial peptidoglycan in vitro is animal species specific.

Acetylmuramyl-Alanyl-Isoglutamine↗

PubMed1992

Changes in cAMP-dependent phosphorylation of specific cytosol protein in the peri- and postmenopausal ovary.

Changes in cAMP-dependent phosphorylation of 53 kDa protein in a human ovary with respect to the aging process were studied. The phosphorylation of endogenous proteins in ovarian cytosol decreased with aging. However, ovarian aging was accompanied by an increase in the cAMP-dependent phosphorylation of 53 kDa specific protein while the amount of this protein decreased with aging. The plasma gonadotropin levels of women with regular menstrual periods were significantly different from those of women whose menstrual periods had in the recent past become irregular (FSH: p less than 0.01; LH: p less than 0.05); however, there was no significant difference in the plasma estradiol levels between these two groups. Our data show the possible correlations between the non-steroidal ovarian factor (53 kDa protein) and/or its phosphorylation by cAMP-dependent protein kinase and the ovarian aging process.

Adult↗

PubMed1991

Localization of an endogenous ADP-ribose acceptor, p33, in polymorphonuclear cell granules in chicken liver interlobular connective tissue.

We investigated immunohistochemically the localization of p33, an endogenous substrate protein for an arginine-specific ADP-ribosyltransferase in chicken liver. Polymorphonuclear-pseudo-eosinophilic granulocytes (heterophils) in interlobular connective tissues of the liver were exclusively and strongly stained with the antibody against p33. Strong reactivity was associated with granules in cytoplasm of the heterophils. When the chicken liver nuclear fraction was washed, the transferase activity was released into the 600 x g supernatant fraction while a nuclear enzyme poly(ADP-ribose) synthetase was retained in the pellet fraction. These results indicate that p33 and probably also ADP-ribosyltransferase, found in the liver nuclear fraction [Tanigawa et al. (1984) J. Biol. Chem. 259, 2022-2029, Mishima et al. (1988) Eur. J. Biochem. 179, 267-273], originate from interlobular heterophils of the chicken liver.

Animals↗