PubMed Health⌕ Search

Biomedical subjects

M Imada

Publications and source records attributed to M Imada.

At least 37 records · Page 2Linked to original sources
PubMed1996

Fel d 1 peptides: effect on skin tests and cytokine synthesis in cat-allergic human subjects.

We tested peptide immunotherapy in cat-allergic humans, using a formation of two synthetic peptides, IPC-1 and IPC-2, each of which is 27 amino acids long and contains T cell-reactive regions of Fel d 1, the major cat allergen. In this exploratory, randomized, double-blind, parallel-group study, 42 subjects received s.c. injections of treatment peptides 250 micrograms or placebo weekly for four consecutive weeks. Changes in immediate- and late-phase skin test reactivity, and in antigen-driven cytokine synthesis were assessed. Epicutaneous (end-point titration) and intradermal tests were performed with cat extract (ALK SQ Cat Hair) containing Fel d 1, before the first injection, then 2, 6 and 24 weeks after the fourth and last injection of peptides or placebo. IL-4, IL-10 and IFN-gamma expression by circulating peripheral blood mononuclear cells (PBMC) in response to cat extract was measured using short-term bulk culture of PBMC and short-term limiting dilution analysis. Subjects who received peptide immunotherapy did not tolerate significantly more cat extract containing Fel d 1 in the skin tests 2, 6 or 24 weeks after the last injection than they did at baseline, and their late-phase responses did not decrease significantly compared to baseline. Substantial IL-4, IL-10 and IFN-gamma responses were observed following primary culture of cat antigen-stimulated PBMC; however, the intensity of cytokine synthesis and the IFN-gamma: IL-4 ratio were unchanged in peptide- and placebo-treated groups 6 and 24 weeks after the last injection. A few hours after the injections, subjects receiving peptides reported more allergic rhinitis and asthma symptoms and more pruritus than those receiving placebo. We conclude that under the conditions tested, peptide immunotherapy did not reduce immediate- or late-phase skin reactivity to cat extract containing Fel d 1 or modify cat antigen-specific cytokine production significantly.

Adult↗
PubMed1996

Measurement of nitric oxide in human nasal airway.

The nasal output of nitric oxide (NO) is known to be high, but there have been varying reports of the exact level. We attempted to establish a quantitative measurement of nasal NO, and looked for a possible relationship with nasal resistance, at rest and during exercise. Nasal airway ventilation was performed by using an air pump at a constant flow rate, whilst the soft palate was elevated voluntarily. In a preliminary study, the flow rate for sampling was changed and concentrations of NO were measured. After determination of flow rate, rhinomanometry for nasal resistance and measurement of nasal NO by chemiluminescence were carried out before and after moderate exercise. The concentration of NO ([NO]) exhibited a hyperbolic relationship with flow rate of ventilation (V'E), indicating [NO] x V'E = net nitric oxide output (V'NO) = constant. Hence, nasal NO was expressed quantitatively as V'NO (nL.min-1). For 1 L.min-1 of ventilation, [NO] varied between 2 and 500 parts per billion (ppb) (mean = 323 ppb). The average nasal V'NO in 12 healthy male subjects was 323 +/- 91 nL.min-1 (mean +/- SD). After exercise on a treadmill (10 degrees, 5 km.h-1) for 4 min, nasal V'NO decreased to 229 +/- 63 nL.min-1. At rest, expiratory and inspiratory nasal resistance was 0.27 +/- 0.04 and 0.27 +/- 0.06 Pa.s.cm-3, respectively. After exercise, expiratory and inspiratory nasal resistance decreased to 0.17 +/- 0.04 and 0.16 +/- 0.04 Pa.s.cm-3, respectively. These data indicate that nasal NO can be measured quantitatively as V'NO and might be involved in the control of nasal resistance.

Adult↗
PubMed1995

Antigen mediated and polyclonal stimulation of human cytokine production elicit qualitatively different patterns of cytokine gene expression.

Polyclonal activators are widely used as surrogate antigens in analysis of human cytokine gene expression. An implicit assumption is that the T cell activation and cytokine production observed in response to polyclonal activation provides a more intense, but qualitatively identical, reflection of results that would be obtained with antigen. Here we demonstrate that stimulation using accessory cell independent (immobilized anti-CD3 mAb) or dependent [phytohemagglutinin (PHA) or soluble anti-CD3 mAb] polyclonal activators yields different conclusions from those that are obtained in response to antigen-specific T cell activation. Cytokine synthesis in 1-5 day bulk cultures of fresh peripheral blood mononuclear cells (PBMC) from 52 subjects evenly divided between grass pollen sensitive allergic rhinitis subjects and normal, non-atopic controls were examined. Antigen-specific re-stimulation elicited elevated IL-4 and IL-10 production and lower IFN-gamma synthesis among allergic subjects than normal non-atopic control subjects. This commitment of fresh PBMC towards a Th2-like response in atopics and the dominance of the IFN-gamma response seen in non-allergic subjects was reinforced when the ratio of IFN-gamma:IL-4 production in bulk culture was examined. Atopic individuals exhibited median IFN-gamma:IL-4 values of 0.07, whereas grass pollen stimulated cytokine production by normal subjects yielded a ratio of 4.8. In marked contrast, IL-2, IL-4, IL-10 and IFN-gamma production elicited using polyclonal activators, though much more intense, did not differ between allergic and non-allergic subjects (Wilcoxon rank sum test P > > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

Allergen-stimulated interleukin-4 and interferon-gamma production in primary culture: responses of subjects with allergic rhinitis and normal controls.

The balance of interleukin-4 (IL-4) to interferon-gamma (IFN-gamma) production that is induced following exposure to common environmental antigens is believed to be instrumental in determining whether hypersensitivity or clinical unresponsiveness results to that antigen. To date, evaluation of cytokine (protein) production has been based predominantly on allergen-reactive CD4 T-cell clones or activation of fresh unselected peripheral blood mononuclear cell (PBMC) populations with non-physiological stimuli such as phorbal myristate acetate (PMA) and calcium ionophore, phytohaemagglutinin (PHA), anti-CD3 or anti-CD2/anti-CD28 monoclonal antibodies (mAb). Here, ultrasensitive IL-4 and IFN-gamma assays were optimized to allow direct analysis of antigen-stimulated cytokine production by fresh human PBMC. Primary cultures of cells from grass pollen-sensitive allergic rhinitis subjects and non-atopic controls were stimulated using a range of grass pollen allergen concentrations in the absence of exogenous cytokines or polyclonal activators. The majority of subjects (45 of 52) exhibited chloroquine-sensitive, CD4-dependent cytokine production in allergen-stimulated, short-term primary culture. Median IL-4 production was substantially greater among atopics (13.0 pg/ml versus < 1 pg/ml, Mann-Whitney U test, P < 0.0000001) and IFN-gamma was lower (P = 0.008), providing direct evidence for an imbalance in both IL-4 and IFN-gamma production among circulating, pollen-reactive cells in individuals with seasonal allergic rhinitis. The distinction in the allergen-driven cytokine responses elicited from normal and atopic donors was underscored by examination of the ratios of IFN-gamma:IL-4 synthesis. Non-atopic individuals exhibited intense IFN-gamma dominance of the T-cell response, in marked contrast to that observed among grass pollen-sensitive individuals (median IFN-gamma:IL-4 ratios of 14.0 versus 0.096, P = 0.000002). The observation that essentially all individuals produced IFN-gamma (+/- IL-4) following antigen stimulation in vitro argues that the most relevant consideration in determining susceptibility to immediate hypersensitivity versus clinical tolerance to environmental allergens is not a genetically defined capacity to recognize the antigen (i.e. if allergen-reactive T cells are present in that individual) but the nature of the cytokine response.

Adolescent↗
PubMed1993

Cyclic AMP-mediated augmentation of thrombomodulin gene expression: cell type-dependent usage of control regions.

We reported that a cell surface thrombin receptor, thrombomodulin (TM), was regulated by cyclic AMP in fibroblasts and in parietal endoderm-like cells derived from F9 embryonal carcinoma cells. In this paper, the genetic basis for augmentation of TM expression by cyclic AMP was studied in F9 and BALB/3T3 cells. Transient expression assays were performed with plasmid constructs containing various 5' flanking sequences of the TM gene and a reporter gene, chloramphenicol acetyltransferase (CAT). Two regulatory DNA regions, the proximal (-411 to -50) and the distal (-1026 to -850), were located. Interplay of the two regions was suggested using a heterologous thymidine kinase promoter in differentiated F9 cells. Both proximal and distal regions contributed to cyclic AMP-dependent augmentation of CAT expression in differentiated F9 cells, whereas only the proximal region was functional in BALB/3T3 cells. The two cell types responded differently also to a protein synthesis inhibitor, cycloheximide, with respect to TM message accumulation. In BALB/3T3 cells TM message accumulation was refractory to the inhibitor in contrast to that of differentiated F9 cells, which was only partially so. We propose that there are at least two separate genomic DNA regions that regulate cyclic AMP-dependent TM gene expression and that their functions are cell type dependent.

3T3 Cells↗
PubMed1993

Human tissue factor pathway inhibitor (TFPI) gene: complete genomic structure and localization on the genetic map of chromosome 2q.

Tissue factor pathway inhibitor (TFPI), a protease inhibitor that circulates in association with plasma lipoproteins (VLDL, LDL and HDL), helps to regulate the extrinsic blood coagulation cascade. We have cloned a 125-kb genomic region containing the entire human TFPI gene on six overlapping cosmids and prepared a restriction map of this contig to clarify gene structure. More than half (45 kb) of the 85-kb gene is occupied with 5' noncoding elements: coding begins at exon 3. A HindIII RFLP identified with one cosmid was genotyped in the CEPH panel of 59 reference families. Linkage analysis using markers on human chromosome 2 located the TFPI gene on 2q, 36 cM proximal to D2S43(pYNZ15) and 13 cM distal to the crystalline gamma-polypeptide locus CRYGP1(p5G1).

Base Sequence↗
PubMed1993

T cell response to embryonal carcinoma F9 cells: induction and characterization of T cell receptor alpha beta+ double-negative cytotoxic T lymphocytes.

We investigated the mechanism of T cell response to murine embryonal carcinoma F9 cells. Thy-1+, CD4-, CD8- (double-negative) cytotoxic effector cells were induced in spleen cells obtained from immune A.BY mice to F9 cells, and the cytotoxic activity was major histocompatibility complex (MHC)-unrestricted. Furthermore, CD4+ T cells were essential for the induction of double-negative cytotoxic T lymphocytes directed to F9 cells. Most of the double-negative cytotoxic T lymphocyte lines obtained by long-term culture of the effector cells had CD3 molecule and T-cell receptor beta chain on their cell surface, and the CD3 molecule was found to be involved in target cell recognition. The T cell receptor alpha beta+ double-negative cytotoxic T lymphocyte line (2A5) also lysed various tumor cells in a non-MHC-restricted manner, but did not lyse concanavalin A-stimulated blasts of 129 strain, from which F9 cells had originated. These results indicate that T cell receptor alpha beta+ double-negative cytotoxic T lymphocytes induced by F9 cells recognize a common antigen(s) expressed on F9 cells and other tumor cells but not minor histocompatibility antigens.

Animals↗
PubMed1991

Anatomical analysis of the human lacrimal drainage pathway under an operating microscope.

Mesoscopic structures of the lacrimal drainage pathway were observed under an operating microscope in six cases of 5 adult cadavers. In all these cases the upper and lower canaliculi fused to form the common canaliculus. Two cases with a large common canaliculus showed large projections of mucosa which reduced the inner space of the common canaliculus. The remaining 4 cases with a small common canaliculus did not show any projections. These findings indicate that the human lacrimal system can function physiologically through a small narrow space of the common canaliculus.

Aged↗
PubMed1991

[Morphological and functional features of endodermal cells in rat yolk sac, with special reference to the fetal macrophage differentiation].

Morphological and functional features of the yolk sac endodermal cells with special reference to the fetal macrophage differentiation were investigated morphologically under the light and electron microscopes and immunologically with the antigen phenotypic analysis and the phagocytic activity-test, using the syngeneic DA rat-embryos from 8 to 16 days of gestation. Based on the staining property with toluidin blue and the ultrastructural features, the endodermal layer from day 8 to 16-yolk sacs has been known to consist of two kinds of cell type; 10% "clear" cells with clear cytoplasm and 90% "dark" cells with dark cytoplasm. Numerous primary lysosomes, phagolysosomes, lipid droplets and coated vesicles distributed preferentially in the supranuclear portion of endodermal cells. A broad intercellular space was found between "clear" cells and "dark" cells, indicating the loose intercellular binding. It was often found that "clear" cells tend to migrate from the endodermal layer into the mesenchymal layer, where the poor development of basement membrane was seen between them. Cells phagocytosing red blood cells, that resemble morphologically "clear" cells, were also observed in the fetal liver. At ten hours after latex-injection into the yolk sac cavity of 14 days embryos, some cells which phagocytosed latex beads in their cytoplasm were found in the endodermal layer, and also in the liver tissue and loose connective tissue of fetus. These cells were stained positively with monoclonal antibody Mar3 which recognizes preferentially rat-mononuclear phagocyte system. In vitro-latex uptake of separated endodermal cells was also demonstrated by the culture-study of endodermal cell suspension. The present findings indicate that the yolk sac-endodermal layer derived from the proximal endoderm consists of at least two kinds of cell-population with a great similarity to tissue macrophages in morphological and functional senses, and support the concept that some cell-populations of endodermal layer may migrate into fetal tissue and are closely related to the differentiation of fetal macrophages and their precursors.

Animals↗
PubMed1990

Identification of fetomodulin, a surface marker protein of fetal development, as thrombomodulin by gene cloning and functional assays.

Fetomodulin (FM) was previously shown to be a surface marker protein of parietal endoderm by in vitro differentiation of F9 embryonal carcinoma cells and by immunohistochemistry of in vivo embryos. BALB/3T3 and sarcoma S180 cells of the mouse were also shown to possess a protein which was indistinguishable from FM by immunological and structural criteria. We now show by protein and DNA sequencing and by functional assays that FM is identical to thrombomodulin, an anticoagulant endothelial thrombin receptor. Partial amino acid sequences of FM from S180 cells suggested homology between FM and thrombomodulin. An FM cDNA fragment was obtained by screening an expression library, which was constructed with restricted BALB/3T3 cDNA, with polyclonal anti-FM antibody. Several longer cDNA clones were than isolated using this fragment as a probe. They elucidated a 3369-bp partial sequence which encompassed 93% of the coding sequence. The remaining structure was determined from a genomic DNA clone. The deduced FM structure proved to be identical to that of thrombomodulin of mouse lung. Affinity-purified FM of BALB/3T3 and differentiated F9 cells was as active as thrombomodulin of the lung in binding thrombin and also as an anticoagulant. Structural and functional identity of the two proteins was thus confirmed. During embryonic development, FM immunoreactivity is localized not only in vasculatures but also at sites of cell-to-cell contact, including lung bud and neural epithelium, which were not expected a priori to possess this endothelial surface protein. FM may be a multifunctional protein with unique roles in embryonic development.

Amino Acid Sequence↗
PubMed1990

Differential expression of fetomodulin and tissue plasminogen activator to characterize parietal endoderm differentiation of F9 embryonal carcinoma cells.

Fetomodulin is a surface marker protein of differentiated F9 embryonal carcinoma cells. Gene cloning has recently identified it as thrombomodulin which binds thrombin and proteolytically activates protein C. Activity assays and RNA blotting were adopted to analyze F9 cell differentiation with specific reference to another well-characterized marker, tissue plasminogen activator. Retinoic acid induced primitive endoderm differentiation of F9 cells and simultaneously activated tissue plasminogen activator synthesis. This differentiation, however, did not result in fetomodulin expression. When primitive endoderm cells were exposed to 1 mM dibutyryl cyclic AMP, the tissue plasminogen activator level rose further within 6 hr. In contrast, the cofactor activity of fetomodulin stayed below a detectable level for as long as 15 hr and then increased with time. Expression of the two marker proteins appeared to be regulated differently.

Animals↗
PubMed1990

Epithelial cell nodules developing in the cecum of irradiated mice. Some comparisons with jejunal nodules.

A method for producing macroscopic epithelial nodules was developed in order to investigate the properties of epithelial stem cells in the large intestine of mice. The cecum was exteriorized and exposed to various doses of X-ray radiation. Numbers of nodules subsequently developing were counted and plotted against radiation dose. From the logarithmic regression line, the susceptibility to irradiation of nodule-forming stem cells (NFSC) was determined. The susceptibility of cecal NFSC was comparable to the value reported for jejunal NFSC. Mice of (C57BL/6 x DS) F1-Pgk-1b/Pgk-1a that carried X-chromosome inactivation mosaicism for the phosphoglycerate kinase gene were used for examination of nodule clonality. Most cecal nodules contained only 1 type of phosphoglycerate kinase, suggesting a monoclonal origin of the nodules. Histochemical studies showed the presence of absorptive epithelial, goblet and entero-endocrine cells in 17-day-old nodules, implying multipotentiality of the NFSC. In spite of these similarities between cecal and jejunal NFSC, the macroscopic appearance of cecal nodules was quite different from that of jejunal nodules. Only crypt-like structures were observed in the former, whereas both crypt-like and villus-like structures were present in the latter. A comparison between cecal and jejunal nodules may be useful for understanding the morphogenesis of the intestinal mucosa.

Animals↗