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1.
Kyobu Geka ; 62(13): 1122-5, 2009 Dec.
Article in Japanese | MEDLINE | ID: mdl-19999087

ABSTRACT

The aim of this study was to confirm that the emphysematous changes had increased in the remaining lung on the operation side after lobectomy. Fourteen patients underwent quantitative analysis by computed tomography (CT) and respiratory function testing before and after the operation for upper or lower lobectomy of the lung between March 2005 and May 2007. The degree of emphysema was calculated by dividing the volume of the emphysematous region (CT values: -1,024 to -950 HU) by the volume of the entire lung (CT values: -1,024 to -600 HU) using a 1 mm thickness high resolution CT slice. Comparison by paired t-test showed significant differences between the emphysema rates pre and post operation in the operation side lung (15.3 +/- 7.9% and 21.7 +/- 10.0%, p = 0.02), but there were no significant differences in the contralateral lung (15.8 +/- 7.5% and 17.7 +/- 8.4%, p = 0.25). On the other hand, there was no significant change in the percent forced expiratory volume in one second (FEV1.0%) between pre and post operation (74.4 +/- 10.5% and 75.5 +/- 7.5%, p = 0.60). We consider that it is necessary to note that the emphysematous changes increased in the remaining lung on the operation side after lobectomy even though the FEV1.0% did not decrease at present.


Subject(s)
Pneumonectomy , Pulmonary Emphysema/diagnosis , Humans , Postoperative Complications , Respiratory Function Tests
2.
Vet J ; 172(1): 141-6, 2006 Jul.
Article in English | MEDLINE | ID: mdl-15927493

ABSTRACT

The outermost layer of skin, the epidermis, is cornified epithelial tissue composed of keratinocytes. To maintain the structure and function of the epidermis, the regulation of proliferation, differentiation, and cornification of keratinocytes is crucial, and various soluble factors secreted by keratinocytes are involved in these regulations. Previously, work has shown that keratinocytes secreted the protein Kdap (keratinocyte differentiation-associated protein) associated with the formation of cornified cell envelopes, a specialized protective barrier structure on the periphery of terminally differentiating keratinocytes. In the present report, the canine counterpart of human Kdap is identified and an attempt has been made to define its physiological role in canine keratinization. Canine Kdap (cKdap) showed structural features commonly observed in other counterparts and is secreted from transfected cells. The expression profile of cKdap mRNA, which was restrictively expressed in cornified epithelial tissues besides skin has also been determined. These findings indicate that there is a strong association between cKdap expression and cornification, which supports previous observations that Kdap is involved in the synthesis and/or degradation of cornified cell envelopes in humans and mice.


Subject(s)
Aspartic Acid Endopeptidases , Epidermal Cells , Keratinocytes/physiology , Amino Acid Sequence , Animals , Aspartic Acid Endopeptidases/chemistry , Aspartic Acid Endopeptidases/genetics , Base Sequence , Cell Differentiation , Cell Division , Cloning, Molecular , Dogs , Epidermis/metabolism , Epidermis/physiology , Gene Expression , Keratinocytes/metabolism , RNA, Messenger/analysis , Species Specificity
3.
Vet Immunol Immunopathol ; 110(1-2): 179-86, 2006 Mar 15.
Article in English | MEDLINE | ID: mdl-16213030

ABSTRACT

The C-type lectin receptor has been shown to recognize carbohydrate moieties of self and non-self antigens, thus serving as an innate immune receptor. Using bioinformatics and molecular cloning techniques, we isolated a bovine gene that encodes a polypeptide of 206 amino acids with structural features shared by mouse and human dectin-2, including a high homology with mouse dectin-2 (66%), a type II configuration, a short cytoplasmic domain without tyrosine-based signal motifs, a carbohydrate recognition domain, a putative N-glycosylation site, and an EPN motif involved in the Ca(2+)-dependent binding of hexose carbohydrates. These results reveal this bovine gene to be a counterpart of mouse dectin-2. Moreover, the bovine dectin-2 gene showed heterogeneity in mRNA (the generation of alternatively spliced transcript) and segmentation into six exons, which are also observed in mouse dectin-2. Inconsistent with mouse dectin-2 mRNA, the bovine counterpart is abundantly expressed by Langerhans cells compared to macrophages; however, lymph nodes showed the highest expression level of bovine dectin-2, while spleen and lung showed the highest expression levels of mouse and human dectin-2. In cattle, dectin-2 expressed by dendritic cells may be clinically involved in the recognition of invading antigens in lymph nodes.


Subject(s)
Cattle/genetics , Lectins, C-Type/genetics , Amino Acid Sequence , Animals , Base Sequence , Cattle/immunology , Cloning, Molecular , Female , Gene Expression Regulation , Langerhans Cells/immunology , Lectins, C-Type/biosynthesis , Lectins, C-Type/immunology , Molecular Sequence Data , RNA/chemistry , RNA/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid
4.
Vet Res Commun ; 29(6): 499-505, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16215840

ABSTRACT

Epidermal Langerhans cells (LC) express a high-affinity receptor for IgE (FcepsilonRI), consisting of two chains (alpha and gamma chains) in humans that allows LC to perform Fc receptor-mediated uptake of allergens. We found that canine LC express alpha and gamma chains but not beta chain of FcepsilonRI, identical to human but not to mouse LC, which do not express functional FcepsilonRI (only gamma chain is expressed). This finding indicates that canine LC have FcepsilonRI-mediated function similar to or identical to human LC, raising the possibility that canine species provides a better model than mouse to understand the pathogenesis of human atopic dermatitis and investigate the therapeutic effect of drugs.


Subject(s)
Dogs/physiology , Langerhans Cells/metabolism , Receptors, IgE/biosynthesis , Animals , Epidermal Cells , Epidermis/metabolism , Gene Expression , RNA, Messenger/metabolism
5.
Br J Dermatol ; 148(4): 654-64, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12752121

ABSTRACT

BACKGROUND: Both intercellular and intracellular signals are transduced primarily by interactions of secreted and/or membrane-anchored polypeptides, and they play a pivotal role in regulating proliferation, differentiation and apoptosis of keratinocytes within the epidermis. Despite recent identification of these polypeptides, it is likely that several important molecules remain undisclosed. OBJECTIVES: To identify novel genes encoding secreted or membrane-anchored polypeptides expressed by human keratinocytes. METHODS: We employed a signal sequence (SS) trap of a 5'-end-enriched cDNA library prepared from primary cultured human keratinocytes. Gene expression analysis was performed using Northern blotting. R Screening of 4018 cDNA clones yielded 82 positive clones (57 independent genes), most of which encoded SSs in their N-termini. Most of the positive clones were known genes registered in the GenBank database. Seven genes were identified in the EST database, four of which encoded novel membrane-anchored polypeptides with features of type I transmembrane proteins; the other three genes encoded novel non-type I transmembrane polypeptides. These EST genes were expressed differentially by keratinocytes subjected to low vs. high calcium concentrations and by basal vs. squamous cell carcinomas. CONCLUSIONS: Using the SS trap, we isolated many genes known to be involved in constituting epidermal structures and others that had not previously been associated with keratinocytes. In addition, we identified novel genes (EST genes) that differ in kinetics of gene expression in keratinocyte differentiation. Our results validate the effective use of this SS trap method for identifying secreted and membrane-anchored polypeptides expressed by human keratinocytes. The identification will better illuminate the molecular mechanisms responsible for co-ordinated regulation of epidermal homeostasis.


Subject(s)
Keratinocytes/metabolism , Membrane Proteins/genetics , Amino Acid Sequence , Cell Differentiation/genetics , Cells, Cultured , DNA, Complementary/genetics , Gene Expression , Gene Expression Profiling , Gene Library , Humans , Infant, Newborn , Keratinocytes/cytology , Molecular Sequence Data , RNA, Messenger/genetics , Signal Transduction/genetics
6.
Br J Dermatol ; 148(4): 680-8, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12752124

ABSTRACT

BACKGROUND: Nuclear factor-kappaB (NF-kappaB) is a transcription factor involved in a number of signalling pathways in many cell types. NF-kappaB in mice has been implicated as an important regulator of keratinocyte proliferation and differentiation. OBJECTIVES: To evaluate the role of NF-kappaB in keratinocyte growth in human beings, we examined its expression in keratinocytes both in culture and in situ, and studied the relationship between NF-kappaB activation and the inhibition of keratinocyte proliferation induced by known modulators of keratinocyte growth. METHODS: The expression of subunits of the NF-kappaB family was examined in human skin, primary cultured keratinocytes and an immortalized keratinocyte line by immunohistochemistry and reverse transcriptase-polymerase chain reaction analysis. NF-kB activation was examined in keratinocytes treated with various modulating agents by electrophoretic mobility shift assay (for DNA-binding activity) and by immunocytochemistry (nuclear translocation). The proliferative capacity of treated keratinocytes was also examined by 3H-thymidine incorporation, cell cycle analysis, and expression of Ki-67, a nuclear marker for cell proliferation. The involvement of NF-kappaB was assessed using sodium salicylate, which inhibits NF-kappaB activation. RESULTS: The NF-kappaB subunits, p50, p65, RelB, and c-Rel (but not p52), were detected in keratinocytes and in normal epidermis at mRNA and protein levels. The four subunits were expressed in a cytoplasmic (rather than a nuclear) pattern in both basal and suprabasal keratinocytes. Phorbol myristate acetate (PMA), tumour necrosis factor alpha, and interferon gamma each activated NF-kappaB and inhibited keratinocyte proliferation. Lipopolysaccharide and dexamethasone did not activate NF-kappaB and had the least effect on proliferation. Finally, a high concentration of calcium (Ca2+) and retinoic acid each failed to activate NF-kappaB, but were potent inhibitors of keratinocyte proliferation, respectively. PMA-induced cell cycle arrest of keratinocytes was blocked by pretreatment with sodium salicylate. CONCLUSIONS: NF-kappaB is constitutively expressed in a resting state in both human cultured keratinocytes and the epidermis. Activation of NF-kappaB is required for PMA-induced keratinocyte growth arrest.


Subject(s)
Epidermis/metabolism , Keratinocytes/metabolism , NF-kappa B/metabolism , Cell Cycle/drug effects , Cell Division/drug effects , Cells, Cultured , Gene Expression , Humans , Immunoenzyme Techniques , Infant, Newborn , Keratinocytes/cytology , Male , NF-kappa B/genetics , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sodium Salicylate/pharmacology , Tetradecanoylphorbol Acetate/antagonists & inhibitors , Tetradecanoylphorbol Acetate/pharmacology
7.
J Immunol ; 167(12): 6893-900, 2001 Dec 15.
Article in English | MEDLINE | ID: mdl-11739507

ABSTRACT

Despite their critical function as APCs for primary immune responses, dendritic cells (DC) and Langerhans cells (LC) have been rarely used as targets of gene-based manipulation because well-defined regulatory elements controlling LC/DC-specific expression have not been identified. Previously, we identified dectin-2, a C-type lectin receptor expressed selectively by LC-like XS cell lines and by LC within mouse epidermis. Because these characteristics raised the possibility that dectin-2 promoter may direct LC/DC-specific gene expression, we isolated a 3.2-kb nucleotide fragment from the 5'-flanking region of the dectin-2 gene (Dec2FR) and characterized its regulatory elements and the transcriptional activity using a luciferase (Luc) reporter system. The Dec2FR contains a putative TATA box and cis-acting elements, such as the IFN-stimulated response element, that drive gene expression specifically in XS cells. Dec2FR comprises repressor, enhancer, and promoter regions, and the latter two regions coregulate XS cell-specific gene expression. In transgenic mice bearing a Dec2FR-regulated Luc gene, the skin was the predominant site of Luc activity and LC were the exclusive source of such activity within epidermis. By contrast, other APCs (DC, macrophages, and B cells) and T cells expressed Luc activity close to background levels. We conclude that epidermal LC are targeted selectively for high-level constitutive gene expression by Dec2FR in vitro and in vivo. Our findings lay the foundation for use of the dectin-2 promoter in LC-targeted gene expression systems that may enhance vaccination efficacy and regulate immune responses.


Subject(s)
Epidermal Cells , Gene Targeting/methods , Langerhans Cells/immunology , Lectins/genetics , 5' Flanking Region , Animals , Base Sequence , Cell Line , Cells, Cultured , DNA-Binding Proteins/metabolism , Lectins, C-Type , Leukocytes/metabolism , Luciferases/genetics , Luciferases/metabolism , Mice , Mice, Inbred BALB C , Mice, Transgenic , Molecular Sequence Data , Promoter Regions, Genetic , Regulatory Sequences, Nucleic Acid , Tissue Distribution , Transcription, Genetic
8.
Gene ; 272(1-2): 51-60, 2001 Jul 11.
Article in English | MEDLINE | ID: mdl-11470510

ABSTRACT

Previously we identified the novel type II lectin receptor, dectin-1, that is expressed preferentially by murine antigen presenting dendritic cells (DC) and is involved in co-stimulation of T cells by DC. To identify the human homologue (DECTIN-1), we employed degenerative PCR amplification of mRNA isolated from DC and subsequent cDNA cloning. DECTIN-1 is a type II lectin receptor with high homology to type II lectin receptors expressed by natural killer (NK) cells. It contains an immunoreceptor tyrosine-based activation motif within the cytoplasmic domain. Human DECTIN-1 mRNA is expressed predominantly by peripheral blood leukocytes and preferentially by DC. The mRNA likely encodes a 33 kDa glycoprotein. In human epidermis, the protein is expressed selectively by Langerhans cells, which are an epidermal subset of DC. A truncated form of DECTIN-1 RNA (termed T beta) encodes for a polypeptide lacking almost the entire neck domain, which is required for accessibility of the carbohydrate recognition domain to ligands. Genome analysis showed the deleted amino acid sequence in T beta to be encoded by an exon, indicating that T beta RNA is produced by alternative splicing. DECTIN-1 gene maps to chromosome 12, between p13.2 and p12.3, close to the NK gene complex (12p13.1 to p13.2) which contains genes for NK lectin receptors. Our results indicate that human DECTIN-1 shares many features with mouse dectin-1, including the generation of neck domain-lacking isoforms, which may down-regulate the co-stimulatory function of dectin-1.


Subject(s)
Dendritic Cells/metabolism , Membrane Proteins/genetics , Nerve Tissue Proteins/genetics , Alternative Splicing , Amino Acid Sequence , Animals , Blotting, Northern , COS Cells , Cell Line , Chromosome Mapping , Chromosomes, Human, Pair 12/genetics , Cloning, Molecular , Conserved Sequence , DNA, Complementary/chemistry , DNA, Complementary/genetics , Gene Expression , HL-60 Cells , Humans , In Situ Hybridization, Fluorescence , Jurkat Cells , Langerhans Cells/metabolism , Lectins, C-Type , Membrane Proteins/metabolism , Mice , Molecular Sequence Data , Nerve Tissue Proteins/metabolism , Phylogeny , Protein Isoforms/genetics , Protein Isoforms/metabolism , Psoriasis/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Tissue Distribution , Tumor Cells, Cultured
9.
J Biol Chem ; 276(11): 8125-34, 2001 Mar 16.
Article in English | MEDLINE | ID: mdl-11114299

ABSTRACT

We isolated a novel molecule (DC-HIL) expressed abundantly by the XS52 dendritic cell (DC) line and epidermal Langerhans cells, but minimally by other cell lines. DC-HIL is a type I transmembrane protein that contains a heparin-binding motif and an integrin-recognition motif, RGD, in its extracellular domain (ECD). A soluble fusion protein (DC-HIL-Fc) of the ECD and an immunoglobulin Fc bound to the surface of an endothelial cell line (SVEC). This binding induced adhesion of SVEC to its immobilized form. Sulfated polysaccharides (e.g. heparin and fucoidan) inhibited binding of soluble DC-HIL-Fc and adhesion of SVEC. By contrast, an integrin inhibitor (RGDS tetramer) had no effect on binding to SVEC, but prevented adhesion of SVEC. This differential RGD requirement was confirmed by the finding that DC-HIL-Fc mutant lacking the RGD motif can bind to SVEC but is unable to induce adhesion of SVEC. Furthermore, DC-HIL appears to recognize directly these sulfated polysaccharides. These results suggest that DC-HIL binds to SVEC by recognizing heparan sulfate proteoglycans on endothelial cells, thereby inducing adhesion of SVEC in an RGD-dependent manner. We propose that DC-HIL serves as a DC-associated, heparan sulfate proteoglycan-dependent integrin ligand, which may be involved in transendothelial migration of DC.


Subject(s)
Dendritic Cells/chemistry , Heparan Sulfate Proteoglycans/metabolism , Membrane Glycoproteins/genetics , Membrane Proteins/genetics , Oligopeptides/physiology , Receptors, Immunologic/genetics , Amino Acid Sequence , Animals , Cell Adhesion , Cell Line , Cloning, Molecular , Dendritic Cells/physiology , Endothelium, Vascular/cytology , Eye Proteins , Membrane Glycoproteins/chemistry , Membrane Glycoproteins/physiology , Membrane Proteins/chemistry , Membrane Proteins/physiology , Mice , Mice, Inbred BALB C , Molecular Sequence Data , RNA, Messenger/analysis , Receptors, Immunologic/chemistry , Receptors, Immunologic/physiology
10.
Gene Ther ; 8(22): 1729-37, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11892841

ABSTRACT

Langerhans cells (LC), which are a skin-specific member of the dendritic cell (DC) family of antigen presenting cells, play critical roles in the initiation of cellular immune responses in the skin. We developed a LC-targeted gene therapy format in this study, aimed at the establishment of in situ protocols for genetic manipulation of LC function. Dectin-2 is a unique C-type lectin that is expressed selectively by DC, including epidermal LC. A 3.2 kb 5' flanking fragment isolated from the mouse dectin-2 gene, termed the dectin-2 promoter (pDec2), exhibited significant transcriptional activities in epidermal-derived DC lines of the XS series, but not in any of the tested non-DC lines. When pDec2-driven luciferase gene (pDec2-Luc) or enhanced green fluorescence protein gene (pDec2-EGFP) was delivered to mouse skin using the gene gun, expression of the corresponding gene product was observed in the epidermal compartment almost exclusively by the IA+ population (ie LC). LC in the gene gun-treated sites showed features of mature DC and they migrated to the draining lymph node, suggesting that LC-targeted gene expression may lead to the development of immune responses. In fact, EGFP-specific cellular immune responses became detectable after gene gun-mediated delivery of pDec2-EGFP plasmid. These results introduce a new concept that LC function can be genetically manipulated in situ by the combination of gene gun-mediated DNA delivery and a DC-specific promoter.


Subject(s)
Gene Expression Regulation , Genetic Therapy/methods , Langerhans Cells/metabolism , Lectins/genetics , Promoter Regions, Genetic , Animals , Biolistics , Cell Movement , Female , Green Fluorescent Proteins , Interferon-gamma/biosynthesis , Langerhans Cells/immunology , Lectins, C-Type , Luciferases/genetics , Luminescent Proteins/genetics , Mice , Mice, Inbred BALB C , Transcription, Genetic
11.
Photochem Photobiol ; 74(6): 805-10, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11783936

ABSTRACT

To determine whether ultraviolet B (UVB) irradiation leads to activation of HIV in human skin, we conducted prospective and controlled studies in two academic medical centers in Texas from July 1995 to April 1999. HIV-positive patients with UV-treatable skin diseases were enrolled at each center, 18 subjects at one and 16 at the other. In one center, specimens from lesional and nonlesional skin biopsies were taken before and after sham- or UVB-irradiation administered in vivo or in vitro. In the other center, UVB phototherapy was administered three times weekly and specimens from skin biopsies were taken before and after 2 weeks (six treatments). Cutaneous HIV load was assessed using reverse transcriptase-polymerase chain reaction and reverse transcriptase-polymerase chain reaction in situ hybridization. UVB irradiation led to a 6-10-fold increase in the number of HIV in skin. To ascertain a role for nuclear factor kappa B (NFkappaB) in UVB-inducible HIV activation, two types of blockers, NFkappaB oligonucleotide decoy and sodium salicylate, were tested; each inhibited UVB-inducible HIV activation in skin partially. We conclude that UVB irradiation leads to increased numbers of HIV in human skin via processes that include release of cytoplasmic NFkappaB.


Subject(s)
HIV/radiation effects , NF-kappa B/antagonists & inhibitors , Skin/radiation effects , Skin/virology , Ultraviolet Rays/adverse effects , HIV/genetics , HIV/isolation & purification , HIV Infections/therapy , HIV Infections/virology , Humans , Phototherapy/adverse effects , Prospective Studies , Skin/drug effects , Sodium Salicylate/pharmacology
12.
J Biol Chem ; 275(16): 11957-63, 2000 Apr 21.
Article in English | MEDLINE | ID: mdl-10766825

ABSTRACT

Using a subtractive cDNA cloning strategy, we isolated previously five novel genes that were expressed abundantly by the murine dendritic cell (DC) line XS52, but not by the J774 macrophage line. One of these genes encoded a unique, DC-associated C-type lectin, termed "dectin-1." Here we report the characterization of a second novel gene that was also expressed in a DC-specific manner. Clone 1B12 encoded a type II membrane-integrated polypeptide of 209 amino acids containing a single carbohydrate recognition domain motif in the COOH terminus. The expression pattern of this molecule, termed "dectin-2," was almost indistinguishable from that for dectin-1; that is, both were expressed abundantly at mRNA and protein levels by the XS52 DC line, but not by non-DC lines, and both were detected in spleen and thymus, as well as in skin resident DC (i.e. Langerhans cells). Interestingly, reverse transcriptase-polymerase chain reaction and immunoblotting revealed multiple bands of dectin-2 transcripts and proteins suggesting molecular heterogeneity. In fact, we isolated additional cDNA clones encoding two distinct, truncated dectin-2 isoforms. Genomic analyses indicated that a full-length dectin-2 (alpha isoform) is encoded by 6 exons, whereas truncated isoforms (beta and gamma) are produced by alternative splicing. We propose that dectin-2 and its isoforms, together with dectin-1, represent a unique subfamily of DC-associated C-type lectins.


Subject(s)
Alternative Splicing , Dendrites/chemistry , Lectins/genetics , Amino Acid Sequence , Animals , Base Sequence , Cell Line , Cloning, Molecular , Lectins, C-Type , Mice , Mice, Inbred BALB C , Molecular Sequence Data , RNA, Messenger/metabolism
13.
J Biol Chem ; 275(26): 20157-67, 2000 Jun 30.
Article in English | MEDLINE | ID: mdl-10779524

ABSTRACT

Dendritic cells (DC) are special subsets of antigen presenting cells characterized by their potent capacity to activate immunologically naive T cells. By subtracting the mRNAs expressed by the mouse epidermus-derived DC line XS52 with the mRNAs expressed by the J774 macrophage line, we identified five novel genes that were expressed selectively by this DC line. One of these genes encoded a type II membrane-integrated polypeptide of 244 amino acids containing a putative carbohydrate recognition domain motif at the COOH-terminal end. This molecule, termed "dectin-1," was expressed abundantly at both mRNA and protein levels by the XS52 DC line, but not by non-DC lines (including the J774 macrophage line). Dectin-1 mRNA was detected predominantly in spleen and thymus (by Northern blotting) and in skin-resident DC, i.e. Langerhans cells (by reverse transcription-polymerase chain reaction). Affinity-purified antibody against dectin-1 identified a 43-kDa glycoprotein in membrane fractions isolated from the XS52 DC line and from the dectin-1 cDNA-transfected COS-1 cells. His-tagged recombinant proteins containing the extracellular domains of dectin-1 showed marked and specific binding to the surface of T cells and promoted their proliferation in the presence of anti-CD3 monoclonal antibody at suboptimal concentrations. These in vitro results suggest that dectin-1 on DC may bind to as yet undefined ligand(s) on T cells, thereby delivering T cell co-stimulatory signals. Not only do these results document the efficacy of subtractive cDNA cloning for the identification of unique genes expressed by DC, they also provide a framework for studying the physiological function of dectin-1.


Subject(s)
Dendritic Cells/metabolism , Membrane Proteins/chemistry , Membrane Proteins/genetics , Nerve Tissue Proteins/chemistry , Nerve Tissue Proteins/genetics , Amino Acid Sequence , Animals , Blotting, Northern , COS Cells , Cell Culture Techniques/methods , Cell Line , Cell Membrane/metabolism , Cloning, Molecular , DNA, Complementary/metabolism , Epithelial Cells/metabolism , Female , Flow Cytometry , Immunoblotting , Lectins/metabolism , Lectins, C-Type , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Phenotype , Rabbits , Recombinant Fusion Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , T-Lymphocytes/metabolism , Tissue Distribution
14.
Jpn J Thorac Cardiovasc Surg ; 48(11): 730-2, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11144094

ABSTRACT

Cryoablation in the left ventricular outflow tract and aortic valve replacement were conducted in a 70-year-old man with frequent premature ventricular contraction and nonsustained ventricular tachycardia complicated by Sellers grade 4 aortic regurgitation. The earliest endocardial activation site during premature ventricular contraction was confirmed by a preoperative electrophysiological study of the left sinus of Valsalva. Cryoablation preceded valvulectomy followed by aortic valve replacement. Clinical symptoms of premature ventricular contraction disappeared immediately after surgery.


Subject(s)
Aortic Valve Insufficiency/surgery , Cryosurgery , Sinus of Valsalva , Ventricular Outflow Obstruction/surgery , Ventricular Premature Complexes/surgery , Aortic Valve/surgery , Heart Valve Prosthesis Implantation , Humans
15.
Int Angiol ; 19(3): 242-9, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11201593

ABSTRACT

BACKGROUND: We have adopted an all-autogenous-vein-graft policy in carotid reconstruction for Takayasu arteritis, namely an ascendo-right carotid and right subclavian (axillary) arteries bypass using a pantaloon vein graft for patients all of whose arch branches are occluded, and an extra-anatomical bypass from the right subclavian artery for patients whose brachiocephalic artery is the only arch branch that remains patent. This report is to elaborate on these operations and to assess the long-term outcome. METHODS: Six patients were operated on according to this policy; (5 women, 1 man, age range: 14 to 59 years (mean: 30). The indications for surgery were severe cerebral ischaemia that significantly interfered with their daily lives. The pantaloon vein graft bypass was performed in four patients, and an extra-anatomical bypass in two. The specific management protocol to prevent the "postbypass hyperperfusion syndrome" and cerebral oedema included a shunt procedure to the internal carotid artery using one limb of the pantaloon vein graft, induced hypotension just before the completion of the carotid reconstruction and the administration of a glycerine-fructose solution. RESULTS: Cerebral ischaemic symptoms disappeared in all patients. All but one, who died of a ruptured thoraco-abdominal aneurysm on the 35th postoperative month, are living a normal life with a patent graft. No suture line complications have as yet been encountered (follow-up: 10 to 205 months, mean: 126 months). CONCLUSIONS: Carotid vein bypass for Takayasu arteritis, particularly, the pantaloon vein graft bypass is recommended for those of whom all aortic arch branches are occluded, resulting in severe brain ischaemia. Perioperative blood pressure control is important for prevention of the hyperperfusion syndrome.


Subject(s)
Brain Ischemia/surgery , Cerebral Revascularization/methods , Takayasu Arteritis/surgery , Veins/transplantation , Adolescent , Adult , Angiography , Axillary Artery/diagnostic imaging , Axillary Artery/surgery , Brachiocephalic Trunk/diagnostic imaging , Brachiocephalic Trunk/surgery , Brain Ischemia/diagnostic imaging , Female , Follow-Up Studies , Humans , Male , Middle Aged , Postoperative Complications/diagnostic imaging , Postoperative Complications/etiology , Subclavian Artery/diagnostic imaging , Subclavian Artery/surgery , Takayasu Arteritis/diagnostic imaging , Transplantation, Autologous
16.
J Immunol ; 158(11): 5393-9, 1997 Jun 01.
Article in English | MEDLINE | ID: mdl-9164960

ABSTRACT

UV light is a potent stimulus for keratinocytes to release several cytokines. Recently, UV light was shown to inhibit keratinocyte release of IL-7, a growth factor for dendritic epidermal T cells. Since to date IL-7 is the only keratinocyte-derived cytokine down-regulated by UV light, we addressed the molecular mechanisms involved. IFN-gamma treatment of the murine keratinocyte cell line Pam 212 resulted in an up-regulation of IL-7 mRNA, while IL-7 transcripts were suppressed in cells exposed to UV before IFN-gamma. Because IFN-gamma induces IL-7 via activation of an IFN-stimulated response element (ISRE) located in the 5' upstream region of the IL-7 gene, bandshift assays were performed using the ISRE sequence from the IL-7 gene. Nuclear extracts from untreated cells revealed two bands, a slower migrating band identified by supershift analysis as IFN regulatory factor-2 (IRF-2), a transcriptional repressor, and a more rapidly migrating band identified as IRF-1, a transcriptional activator. IFN-gamma significantly induced IRF-1 binding, whereas UV treatment plus IFN-gamma decreased IRF-1 binding, suggesting that UV light suppresses IFN-gamma-induced expression of IL-7 by interfering with IRF-1. Chloramphenicol transferase assay confirmed functional relevance, showing that the minimal promoter sequence for the ISRE explicitly responded to IFN-gamma, which was suppressed by UV irradiation. Northern blot analysis using an IRF-1 cDNA probe revealed that UV light reduced IFN-gamma-induced IRF-1 mRNA. This study demonstrates that UV light can inhibit cytokine activities by interference with transcriptional activators. This newly described ability of UV light may contribute to its immunosuppressive properties.


Subject(s)
DNA-Binding Proteins/immunology , Gene Expression Regulation/drug effects , Interferon-gamma/pharmacology , Interleukin-7/immunology , Keratinocytes/immunology , Phosphoproteins/immunology , Ultraviolet Rays , Animals , Cells, Cultured , DNA-Binding Proteins/genetics , Interferon Regulatory Factor-1 , Interleukin-7/biosynthesis , Interleukin-7/genetics , Keratinocytes/radiation effects , Mice , Phosphoproteins/genetics
19.
J Immunol ; 157(8): 3312-6, 1996 Oct 15.
Article in English | MEDLINE | ID: mdl-8871626

ABSTRACT

Ultraviolet B irradiation of skin leads to immunologic tolerance, rather than immunity against newly introduced Ag, by altering the function of Langerhans cells, skin-specific members of the dendritic cell (DC) family. Using the murine epidermal-derived DC line, XS52, which retains important features of resident Langerhans cells, we have tested the hypothesis that UV radiation delivers a signal leading to apoptosis. XS52 cells, when exposed to modest fluences (25-100 J/m2) of radiation, underwent apoptosis during a subsequent 6-h incubation with LPS or upon 6-h coculture with the keyhole limpet hemocyanin-specific Th1 clone HDK-1 in the presence of Ag. Specifically, XS52 cells treated in this way exhibited diminished cell viability, DNA laddering, and condensed staining of DNA. By contrast, none of these changes was induced by radiation alone, LPS alone, or coculture with T cells and Ag. Likewise, neither UV radiation plus T cells nor radiation plus Ag were sufficient to induce apoptosis, indicating that both T cells and Ag are required to induce apoptosis in the UV-sensitized cells. XS52 cells remained fully susceptible to T cell-mediated apoptosis even 16 h after irradiation, indicating the persistence of the sensitized state. These observations establish a model in which UV radiation induces a first event in which DC become sensitive to a second, apoptotic signal that is delivered by Ag-specific interaction with T cells or by LPS. We suggest that DC undergoing apoptosis deliver unusual activation signals to T cells during Ag presentation, signals that lead to cellular unresponsiveness rather than to effective immunity.


Subject(s)
Apoptosis/radiation effects , Langerhans Cells/immunology , Langerhans Cells/radiation effects , Animals , Antigen Presentation , Cell Line , Immune Tolerance/radiation effects , Langerhans Cells/cytology , Lipopolysaccharides/pharmacology , Lymphocyte Activation , Mice , Signal Transduction , Skin/immunology , Skin/radiation effects , T-Lymphocytes/immunology , Ultraviolet Rays
20.
J Immunol ; 157(4): 1529-37, 1996 Aug 15.
Article in English | MEDLINE | ID: mdl-8759735

ABSTRACT

Murine epidermis contains two leukocyte populations: Langerhans cells (LC), which are APC of dendritic cell (DC) lineage, and dendritic epidermal T cells (DETC), which are members of the tissue-type gamma delta T cell family. Despite close physical approximation in vivo, the extent to which LC and DETC affect each other's function has remained unknown. We addressed this question using the long term DC line XS52 and the gamma delta T cell line 7-17, both of which were established from mouse epidermis, and both of which retain important features of the resident populations from which they were derived. XS52 DC proliferated maximally when cocultured with gamma-irradiated 7-17 DETC. They also proliferated in response to culture supernatants collected from anti-CD3- or Con A-activated 7-17 DETC, but not from nonstimulated DETC. In both systems, DETC-induced XS52 DC growth was inhibited partially (up to 70%) by Abs against granulocyte/macrophage CSF (GM-CSF) or CD115 (CSF-1 receptor) and nearly completely (up to 90%) by both together. Among 28 tested cytokines, only GM-CSF, CSF-1, IL-4, and IL-13 promoted XS52 DC growth significantly. Anti-IL-4 failed to inhibit DETC-induced XS52 cell growth, and IL-4 was not detectable in DETC supernatants. Thus, we conclude that GM-CSF and CSF-1 (and perhaps IL-13) account for the DC growth-promoting activity secreted by DETC. These results suggest that during coculture, XS52 DC activate 7-17 DETC to secrete both GM-CSF and CSF-1. In fact, when cultured with XS52 DC, 7-17 DETC also elevated their expression of the gamma c receptor and acquired proliferative responsiveness to their own growth factor IL-15. We propose that LC and DETC in situ may interact with each other in a similar manner, thereby regulating their residence and function.


Subject(s)
Cell Communication/physiology , Cytokines/physiology , Dendritic Cells/cytology , Epidermis/immunology , Langerhans Cells/cytology , T-Lymphocyte Subsets/cytology , Animals , Cell Division/drug effects , Cell Line , Coculture Techniques , Concanavalin A/pharmacology , Culture Media, Conditioned , Dendritic Cells/metabolism , Dendritic Cells/radiation effects , Epidermal Cells , Granulocyte-Macrophage Colony-Stimulating Factor/antagonists & inhibitors , Granulocyte-Macrophage Colony-Stimulating Factor/physiology , Growth Substances/metabolism , Interleukin-13/physiology , Langerhans Cells/drug effects , Lymphocyte Activation , Macrophage Colony-Stimulating Factor/physiology , Mice , Muromonab-CD3/pharmacology , Receptor, Macrophage Colony-Stimulating Factor/immunology , Receptor, Macrophage Colony-Stimulating Factor/physiology , Receptors, Antigen, T-Cell, gamma-delta/analysis , Signal Transduction , T-Lymphocyte Subsets/metabolism , T-Lymphocyte Subsets/radiation effects
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