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Biomedical subjects

A Takashima

Publications and source records attributed to A Takashima.

At least 127 records · Page 7Linked to original sources
PubMed1995

Papulonodular mucinosis associated with systemic lupus erythematosus: possible mechanisms of increased glycosaminoglycan accumulation.

BACKGROUND: The mechanism for the production of papulonodular mucinosis in patients with lupus erythematosus (LE) is not known. OBJECTIVE: Our purpose was to determine whether fibroblasts in a patient with LE and papulonodular mucinosis produced more mucin than normal fibroblasts and whether this mucin production could be stimulated by the patient's serum. METHODS: Skin fibroblasts from a patient with systemic LE and massive papulonodular mucin deposition, as well as normal fibroblasts, were incubated in the presence of serum from the patient or from a healthy volunteer. The production of glycosaminoglycan by fibroblasts was analyzed. RESULTS: Fibroblasts from the patient produced more glycosaminoglycan than did normal fibroblasts. Glycosaminoglycan production was increased in all cells when incubated in the presence of the patient's serum. CONCLUSION: Cutaneous mucin deposition in patients with papulonodular LE skin lesions is associated with increased glycosaminoglycan production by dermal fibroblasts. Our preliminary observations suggest glycosaminoglycan production by these fibroblasts appears to be stimulated by a factor, (or factors) in the patient's serum that is yet to be identified.

Adult↗
PubMed1995

Different localization of tau protein kinase I/glycogen synthase kinase-3 beta from glycogen synthase kinase-3 alpha in cerebellum mitochondria.

We examined the subcellular distribution of two glycogen synthase kinase-3 (GSK-3) isoforms in rat cerebellum. Results from immunoelectron microscopy and subcellular fractionation revealed that one isoform, tau protein kinase I/GSK-3 beta (TPKI/GSK-3 beta), was present in mitochondria, but GSK-3 alpha was not. Although the two GSK-3 isoforms seem to have similar properties, the difference of subcellular localization observed here suggests that TPKI/GSK-3 beta fulfills some specific function in mitochondria.

Animals↗
PubMed1995

Correlation among secondary structure, amyloid precursor protein accumulation, and neurotoxicity of amyloid beta(25-35) peptide as analyzed by single alanine substitution.

Structure-neurotoxicity relationships of amyloid beta (25-35) peptide were studied by replacing each amino acid with Ala. In contrast to the general tendency in hydrophobicity-toxicity relationships, replacement of Asn27 yielded a more hydrophobic but less toxic analog and that of Met35 gave a less hydrophobic but more toxic one. Sedimentation profiles and CD spectra indicated that peptide aggregation via intermolecular beta-sheet formation is essential for the neurotoxicity of amyloid beta (25-35) peptide. The correlation between neurotoxicity and amyloid precursor protein accumulation suggested that the latter is one of the pathways of the neuronal death caused by amyloid beta protein.

Alanine↗
PubMed1995

Interleukin-7-dependent interaction of dendritic epidermal T cells with keratinocytes.

Dendritic epidermal T cells (DETC), a member of the epithelial tissue-type gamma delta T-cell family, are characterized by their exclusive residence within mouse epidermis, their dendritic morphology, and their monoclonal nature in the T-cell-receptor configuration. Here we review our recent studies on the interleukin (IL)-7-dependent interaction of DETC with neighboring keratinocytes. Keratinocytes express constitutively the mRNAs for IL-7 and secrete biologically relevant amounts of IL-7. This cytokine, in turn, serves as a growth factor for DETC, as evidenced by the proliferative responses to recombinant or keratinocyte-derived IL-7 of the 7-17 DETC line and of DETC freshly purified from mouse skin. The 7-17 DETC line undergoes apoptotic cell death in response to external stimuli known to deplete DETC in situ (e.g., ultraviolet B radiation or corticosteroid treatment), and IL-7 prevents this apoptosis, thereby promoting long-term survival. These results document the crucial role played by IL-7 in maintaining the survival and growth of DETC in epidermis. IL-7 mRNA expression in keratinocytes is abrogated by ultraviolet B radiation, whereas it is up-regulated by interferon-gamma, which is secreted by DETC upon activation. More specifically, interferon-gamma induces the preferential expression of truncated forms (2.6 and 1.5 kb) of IL-7 transcripts, in addition to the 2.9- and 1.7-kb transcripts that are expressed constitutively, and this regulation occurs through the usage of alternative transcription initiation sites. These results suggest unique pathways through which IL-7 production is regulated in keratinocytes by external stimuli (e.g., ultraviolet B) as well as T-cell-derived cytokines (e.g., interferon-gamma). We propose that keratinocyte-derived IL-7 is an essential component of the epidermal cytokine milieu.

Animals↗
PubMed1995

Identity and functional properties of novel skin-derived fibroblast lines (NS series) that support the growth of epidermal-derived dendritic cell lines.

We have established recently a series of unique cell lines (NS series) from dispase-separated mouse epidermis that promote the growth of epidermal-derived antigen-presenting cell lines (XS series). The purposes of this study were to determine the identity of NS cells and to characterize their functional properties. NS cells were distinguishable from leukocytes by the lack of typical surface markers and by the failure to respond to leukocyte growth factors. Despite their epidermal derivation, NS cells were distinct from keratinocytes by the absence of cytokeratins. On the other hand, NS cells were indistinguishable from lines of dermal fibroblasts by their a) morphology, b) surface phenotype, and c) intracellular deposits of type I collagen. Growth of the XS antigen-presenting lines has been promoted by co-culturing with gamma-irradiated NS cells, and this activity could be replaced with NS cell-conditioned media alone, but not with paraformaldehyde-fixed NS cells. Each clone derived from the NS01 line secreted XS cell-growth-promoting activity, and this activity was blocked by monoclonal antibodies against colony-stimulating factor-1 receptors. Dermal fibroblasts also promoted the growth of XS cells in a colony-stimulating factor-1-dependent manner. By contrast, culture supernatants from other cell lines derived from skin (e.g., Pam 212 keratinocytes, 7-17 dendritic epidermal T cells, or XS lines) failed to promote XS cell growth. These results indicate that NS cells belong to the fibroblast lineage and that they share the intrinsic property to secrete large amounts of colony-stimulating factor-1 with dermal fibroblasts. Dermal cells may support the growth of skin-associated antigen-presenting cells in vivo.

Animals↗
PubMed1995

2B4, a new member of the immunoglobulin gene superfamily, is expressed on murine dendritic epidermal T cells and plays a functional role in their killing of skin tumors.

Dendritic epidermal T cells (DETC), which are skin-specific members of the tissue-resident gamma delta T-cell family, are characterized by their potential to kill selected tumor targets by a non-major histocompatibility complex (MHC)-restricted mechanism. We have recently identified a new receptor molecule, 2B4, that appears to be associated with non-MHC-restricted recognition of tumor targets by natural killer cells. The purpose of this study was to determine whether DETC express 2B4 molecules, and, if so, to assess their functional roles in DETC-mediated killing of tumor targets. Short-term DETC lines as well as DETC freshly procured from skin expressed surface 2B4, as detected with a specific monoclonal antibody. Removal of interleukin (IL)-2 from DETC cultures caused substantial reduction in 2B4 expression levels as well as a reduction in cytotoxic capacity against YAC-1 targets in a standard 51Cr-release assay. Conversely, exposure to IL-2, but not to IL-7, elevated both 2B4 expression and cytotoxicity. To assess the functional roles played by surface 2B4, we pretreated DETC lines with anti-2B4 antibody and then tested for their killing potential. Anti-2B4, but not the control antibody, augmented their capacity to lyse YAC-1 targets (51Cr-release assays) and to disrupt the monolayers of Pam-212-transformed keratinocytes (visual assessment). Thus, we conclude that DETC express, in an IL-2-dependent manner, 2B4 molecules, which may play a unique role in the killing of skin-derived tumors.

Animals↗
PubMed1995

Phenotypic and functional heterogeneity among murine epidermal-derived dendritic cell clones.

We have established recently long-term dendritic cell lines from the epidermis of newborn BALB/c mice. These lines, termed XS series, resembled epidermal resident Langerhans cells or their progenitors in terms of surface phenotype, antigen-presenting capacity, and growth factor requirement. We examined in this study the degree of clonal heterogeneity among XS cells with respect to each of these features. Twelve stable clones were established by limiting dilution microculture from 8-10-week-old cultures of the XS52 or XS20 line. Despite the uniform expression of CD45, these clones varied substantially in their expression of Ia, B7-1, and B7-2 molecules. They also varied significantly in their relative efficiency in activating T cells. Finally, remarkable clone-to-clone heterogeneity was also observed in their growth factor responsiveness; some clones responded equally well to granulocyte macrophage-colony-stimulating factor-1, whereas others responded preferentially to one or the other of these factors. We propose that the observed clonal heterogeneity in XS cells reflects possible heterogeneity in the state of maturation and mitotic potential among the starting populations, i.e., skin-associated dendritic cells in newborn mice.

Animals↗
PubMed1995

Interleukin (IL)-15 promotes the growth of murine epidermal gamma delta T cells by a mechanism involving the beta- and gamma c-chains of the IL-2 receptor.

Dendritic epidermal T cells (DETC) are skin-specific members of the epithelial gamma delta T-cell family in mice. We have reported previously that the growth of DETC is promoted by interleukin (IL)-2 in an autocrine fashion, or by IL-7, which is secreted by neighboring keratinocytes. Here we report that DETC growth is promoted by IL-15, a newly discovered T-cell growth factor that is produced in lymphoid as well as nonlymphoid tissues. Recombinant IL-15 promoted the growth of the 7-17 DETC line in a time- and dose-dependent fashion. Using monoclonal antibodies against alpha-, beta-, or gamma c-chains of the IL-2 receptor complex, we observed that the combination of anti-beta chain and anti-gamma c chain antibodies blocked IL-15 responsiveness completely, whereas anti-alpha chain had no effect. These results indicate that this gamma delta T-cell line uses the beta/gamma c heterodimer for proliferative responses to IL-15. Antibodies against IL-2 or IL-7 did not block IL-15-driven proliferation of 7-17 DETC, indicating that IL-15 promotes their growth in an IL-2- and IL-7-independent manner. Both the surface expression of beta/gamma c heterodimers and the IL-15 responsiveness of 7-17 DETC were highest 1 to 8 days after concanavalin A stimulation, and both declined substantially 21 days after stimulation, illustrating regulation by the state of cell activation. Working with epidermal cells that were freshly procured from CBA mice, we noted that IL-15 promoted conavalin-A-triggered growth of Thy-1+ cells (i.e., DETC), but not of the Thy-1- cells. The gamma c-chain was not expressed by freshly procured DETC, becoming detectable within 48 h after concanavalin A stimulation. We propose that IL-15 facilitates the growth of epithelial gamma delta T cells by a beta/gamma c receptor-dependent mechanism.

Animals↗
PubMed1995

UVB-dependent modulation of epidermal cytokine network: roles in UVB-induced depletion of Langerhans cells and dendritic epidermal T cells.

The epidermis of mice consists of three cellular components, i.e., keratinocytes, Langerhans cells (LC), and dendritic epidermal T cells (DETC). Each epidermal subpopulation produces a different set of cytokines, thereby forming a unique cytokine milieu. These cytokines, in turn, support the survival and growth of LC and DETC and regulate their immunological functions. LC and DETC play important, but distinct, effector roles in protective immunity against antigens that are generated in or penetrate into the epidermis. Acute or chronic exposure of mice to ultraviolet B (UVB) radiation is known to impair this cutaneous immunity, as evidenced by the failure to induce T cell-mediated immune reactions, by the generation of antigen-specific immunological unresponsiveness, and by the development of skin cancers. Importantly, these changes are associated with reduced densities of LC and DETC in UVB-exposed skin, suggesting that the deficiency in these epidermal leukocytes may account for some of the deleterious influences of UVB radiation on skin. Here I will review the recent advance in our understanding of the mechanisms by which UVB radiation may deplete LC and DETC from epidermis. More specifically, I will discuss the following possibilities: a) UVB-mediated suppression of the production of relevant growth factors for LC and DETC, b) UVB-induced abrogation of surface expression of growth factor receptors, and c) UVB-triggered apoptotic cell death in epidermal leukocytes.

Animals↗
PubMed1995

Hydrogen peroxide mediates UV-induced impairment of antigen presentation in a murine epidermal-derived dendritic cell line.

Ultraviolet-B (290-320 nm) radiation is known to impair the antigen-presenting cell (APC) function of Langerhans cells (LC), skin-specific members of the dendritic cell (DC) family. We sought to address mechanisms of this effect, focusing on the role played by hydrogen peroxide. For this purpose, we used a newly established murine DC line, XS52, which resembles epidermal LC in several respects. The APC capacity of XS52 cells, using two different CD4+ T cell clones as responders, was inhibited significantly (> 50%) by exposure to UV radiation (unfiltered FS20 sunlamps) at relatively small fluences (50-100 J/m2). Ultraviolet radiation also inhibited growth factor-dependent proliferation of XS52 cells. On the other hand, cell surface phenotype was relatively well preserved after irradiation; expression levels of B7-1 and B7-2 were reduced slightly, while other molecules (e.g. Ia, CD54, CD11a and CD18) were not affected. With respect to the role played by hydrogen peroxide, pretreatment with purified catalase (900 U/ mL) prevented UV-induced inhibition of APC function. Short-term exposure to 3 mM H2O2 or t-butyl H2O2 mimicked UV radiation by inhibiting APC function. Finally, intrinsic catalase activity was substantially lower in XS52 cells compared with Pam 212 keratinocytes. These results indicate that the generation of hydrogen peroxide alone is sufficient to produce some, but not all, of the deleterious effects of UV radiation on DC derived from the skin.

Animals↗
PubMed1995

Regional characteristics of the membrane response of an identified crayfish nonspiking interneuron to intracellularly injected current.

1. We investigated the membrane response of the local directionally selective (LDS) interneuron, a nonspiking cell identified in the terminal abdominal ganglion of crayfish, to intracellularly injected current in different regions within the cell by single-electrode, discrete current-clamp experiments. The site of electrode impalement into the cell was visualized in situ together with the cell structure under a dissecting microscope. 2. The LDS interneuron has dendritic branches on both hemiganglia connected by a thick segment crossing the midline. Irrespective of the site of electrode impalement, the interneuron showed outward rectification upon depolarization from the resting potential level. When hyperpolarizing current was injected, a linear relationship was observed between the voltage response of the interneuron and the amount of injected current. Upon large hyperpolarization, however, the interneuron showed inward rectification. 3. The input resistance of the interneuron measured within the linear range of the membrane response was significantly lower in the transverse segment than in lateral dendrites (0.001 < P < 0.01). The time constant of the transient voltage response to step current injection was also significantly shorter in the transverse segment than in the lateral dendrites (0.001 < P < 0.01). 4. Although the regional difference in the input resistance could be accounted for, at least partly, by different geometric conditions of each dendritic branch into which current was injected, the regional difference in the time constant of the membrane response cannot be accounted for by structural differences because the time constant is independent of the membrane area. It is thus suggested that the passive properties of the interneuron membrane that are related to its response time constant show regional variability within the cell.

Animals↗
PubMed1995

Effects of neuromedins and related peptides on the body temperature of rats.

Neuromedin B (NMB) and neuromedin C (NMC) are peptides found in the mammalian central nervous system, and their concentrations are particularly high in the hypothalamus. The amino acid sequences of these peptides are similar to that of bombesin, which is known to induce marked hypothermia not only in amphibians but also in mammals. The effect of neuromedins and related peptides on the body temperature regulation was examined in the rat under a thermoneutral environment. The findings indicated that NMC caused moderate hypothermia following intracerebroventricular administration, while the effect of NMB was meager, suggesting that these two neuropeptides possess different physiological functions in the brain.

Amino Acid Sequence↗
PubMed1994

Cloning and expression of eel natriuretic-peptide receptor B and comparison with its mammalian counterparts.

A comparative study of the natriuretic-peptide receptor NPR-B was performed by cloning and expressing, in COS-1 cells, the NPR-B receptor subtype from the eel gill which exhibited a strong C-type-natriuretic-peptide (CNP)-induced guanylate cyclase activity. Like other mammalian NPR-B receptors, the eel NPR-B receptor consisted of a ligand-binding extracellular domain, a hydrophobic transmembrane domain, a kinase-like domain and a guanylate cyclase domain. Sequence comparison among the eel and mammalian receptors revealed a relatively low similarity (approximately 44%) in the extracellular domain compared to a very high similarity (approximately 84%) in the cytoplasmic regulatory and catalytic domains. This low similarity allowed identification of the amino acid residues or candidate regions important for the ligand-binding activity. RNase protection analysis of the eel NPR-B mRNA demonstrated that the message was predominantly expressed in the liver and atrium as well as in the gill with moderate-to-small amounts in the brain, ventricle, esophageal sphincter, stomach, posterior intestine and kidney. The high NPR-B mRNA levels in the liver, atrium and gill were found to decrease markedly when eels were transferred from fresh water to seawater and kept there for 2 weeks. Since similar changes are known to occur in the ligand CNP levels when eels are facing osmotic challenges, the CNP/NPR-B system appears to play an important role in their successful adaptation to salinity changes.

Amino Acid Sequence↗
PubMed1994

Contact sensitization induces proliferation of heterogeneous populations of hapten-specific T cells.

To characterize the T cells that are activated during the induction of contact hypersensitivity (CH), two sets of studies were conducted: 1) dinitrophenol (DNP)-specific proliferative responses of T cells in draining lymph nodes of BALB/c mice sensitized epicutaneously to dinitrofluorobenzene (DNFB) were examined, and 2) from these lymph node cells, DNP-specific T cells were cloned by limiting dilution microculture and analyzed by FACS for surface markers, by RT-PCR, HT2 bioassay and ELISA for cytokine expression at mRNA and protein levels respectively, and by proliferation assay for cytokine and antigen-presenting cell (APC) requirements. Our results show that alpha beta TCR-bearing T cells of both the CD4+ and CD8+ subtypes from lymph nodes of DNFB-skin-painted mice proliferate specifically to dinitrobenzene sulfonate (DNBS) in vitro. Four DNP-specific, CD4+ T-cell clones were characterized: clone 5S4 secreted IL-4 and required Il-4 for optimal growth; clone 5S10 secreted IL-2 and required IL-2 for optimal growth; clone 5S2 secreted IL-4 but required IL-2 for optimal growth; and clone 5S8 secreted IL-2 predominantly at 5 months, but switched to production of IL-4 at 7 months. All four clones secreted IL-10, and proliferated to DNBS when Langerhans cell (LC)-enriched epidermal cells were used as APC. These findings indicate that heterogeneous populations of DNP-specific T cells are activated in draining lymph nodes during the induction of CH to DNFB in BALB/c mice.

Administration, Cutaneous↗
PubMed1994

Impaired spatial learning by vasoactive intestinal peptide in Morris water maze task in the rat.

Intracerebroventricular administration of vasoactive intestinal peptide (VIP) disturbed the learning by rats of the location of a platform submerged in a water pool. When the platform was removed from the pool, VIP injection produced marked impairment of the ability to find a previously learned location in the pool. This spatial memory impairment caused by VIP was restored by peripheral pre-administration of cerulein.

Animals↗
PubMed1994

Open-field behavior of rats following intracerebroventricular administration of neuromedin B, neuromedin C, and related amphibian peptides.

Neuromedin B (NMB) and neuromedin C (NMC) have amino acid sequences similar to those of various amphibian skin peptides and thus may elicit behavioral effects similar to those of bombesin. The present study was performed to determine the central effects of neuromedins (including pyro-NMB and pyro-NMC) in comparison with those of amphibian peptides using an open-field test situation. At 1 min after intracerebroventricular administration of the peptides, there was a marked decrease in the duration of locomotion, the distance moved, and rearing behavior, but a marked increase of excessive grooming and violent scratching. Pronounced behavioral changes were still observed 30 min after injection of the peptides, except for NMB. Thus, NMB seemed to be rapidly degraded by endogenous peptidases in the brain, suggesting that it might be the essential form of this neuropeptide family in the mammalian brain.

Amino Acid Sequence↗
PubMed1993

Keratinocyte-derived IL-7 serves as a growth factor for dendritic epidermal T cells in mice.

Dendritic epidermal T cells (DETC) are CD3+, CD45+, CD4-, CD8-, TCR-V gamma 3/V delta 1+ T lymphocytes that reside in symbiosis with keratinocytes in mouse epidermis. To address mechanisms by which these cells survive and proliferate within the epidermal environment, we have tested the hypothesis that cytokines secreted by neighboring keratinocytes play relevant roles. The present study was conducted to determine whether keratinocytes produce biologically relevant amounts of IL-7, and, if so, to study its effects on DETC. The long term cultured DETC line, 7-17, and freshly isolated DETC exhibited dose- and time-dependent proliferative responses to rIL-7. These responses were blocked completely by anti-IL-7 antibodies, whereas anti-IL-2 had no effect, indicating that DETC respond to IL-7 by an IL-2-independent mechanism. Proliferative responses depended on the state of cell activation; DETC stimulated 2 to 5 days earlier with Con A proliferated optimally to added IL-7, whereas cells stimulated 10 days earlier did not proliferate. DETC that failed to proliferate responded to IL-7 by showing prolonged survival; cells maintained for 7 days with IL-7 alone retained their capacity to proliferate in response to Con A. Mouse epidermal cells and Pam 212 keratinocyte line both expressed IL-7 mRNA constitutively, as demonstrated by reverse transcription-polymerase chain reaction analyses. The production of IL-7 by mouse keratinocytes was also confirmed; Pam 212 culture supernatants supported DETC proliferation, and this activity was diminished by 50% with added anti-IL-7 antibodies. These results indicate that keratinocytes produce IL-7 in biologically relevant amounts, which, in turn, serve to promote the survival and growth of DETC. IL-7-mediated communication between epithelial cells and gamma delta T cells may represent one mechanism to sustain the indefinite residence of gamma delta T cells in epithelial tissues of mice.

Animals↗