bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of Secretory IgA, Secretory Component, and Eosinophils in Mucosal Inflammation
Authors: Motegi Youichi , Kita Hirohito , Kato Masahiko , Morikawa Akihiro
Published: April 2003

Eosinophils and their products are important in the pathophysiology of allergic inflammation in mucosal tissues. Secretory component (SC) bound to IgA mediates transepithelial transport of IgA. As another biological activity of SC, we have reported that secretory IgA (sIgA) and SC preferentially activate human eosinophils. When eosinophils were stimulated with immobilized sIgA, degranulation and superoxide production were greater than when stimulated with serum IgA. In contrast, neutrophils responded similarly to sIgA and serum IgA. Superoxide production by eosinophils stimulated with cytokines was enhanced synergistically by immobilized SC, while SC showed no effect on neutrophil activation. Eosinophil superoxide production stimulated with sIgA was abolished by anti-CD18 mAb, suggesting that β2 integrins might be crucial for this reaction. There are several reports that SC and sIgA may play important roles in regulating eosinophil functions in vivo in diseases associated with mucosal eosinophilia and in various allergic diseases. It is speculated that eosinophils in the mucosa are activated by SC or sIgA, and that subsequent degranulation and superoxide production are induced.
Soybean Glycinin G1 Acidic Chain Shares IgE Epitopes with Peanut Allergen Ara h 3
Authors: Beardslee Tom A. , Zeece Michael G. , Sarath Gautam , Markwell John P.
Published: April 2003

Background: The identification of IgE epitopes for proteins is the first step in understanding the interaction of allergens with the immune system. Proteins from the legume family have shown in vitro cross-reactivity in IgE-binding assays, but this cross-reactivity is rarely clinically significant. Resolution of this discrepancy requires IgE epitope mapping of legume family protein allergens. Methods: We constructed six fusion proteins representing overlapping regions of soybean glycinin G1 acidic chain. These fusion proteins were used in immunoblotting and a novel sandwich ELISA with pooled sera from soy-allergic individuals to reveal a common IgE-binding region. This region was the focus for IgE epitope mapping using overlapping synthetic peptides. Results: Data from the fusion protein experiments revealed an IgE-binding region consisting of residues F192–I265. Analysis of the overlapping synthetic peptides to this region indicated that IgE epitopes to glycinin G1 acidic chain consist of residues G217–V235 and G253–I265. The epitopes identified for glycinin G1 acidic chain are homologous to IgE epitopes previously identified for the peanut allergen Ara h 3 [1]. However, residues identified by alanine scanning in the peanut epitopes as being important for IgE binding are different in the natural soybean epitopes. Conclusions: The IgE epitopes identified for glycinin G1 acidic chain apparently represent an allergenic region of several legume family seed storage proteins. Our findings indicate that the identification of IgE epitopes and structural analysis of legume family proteins will provide valuable information to the study of food allergies.
Effect of Aerosolized Docosahexaenoic Acid in a Mouse Model of Atopic Asthma
Authors: Yokoyama Akihito , Hamazaki Tomohito , Ohshita Akira , Kohno Nobuoki , Sakai Kimiko , Zhao Guang-Dong , Katayama Hitoshi , Hiwada Kunio
Published: April 2003

Background: Docosahexaenoic acid (DHA) found in fish oil is known to depress inflammation-related mediators. We investigated a novel delivery method of tridocosahexaenoyl-glycerol (DHA-TG). Methods: BALB/c mice (6–8 weeks old) were primed intraperitoneally with ovalbumin (OVA) and Al(OH)3 on days 0 and 7, and with aerosolized OVA on day 7. Primed mice were challenged by repeated exposure to aerosolized OVA on days 15–17. Just before each exposure to aerosolized OVA, the mice were also exposed to an aerosol of emulsified DHA-TG or soybean oil, or saline (days 7 and 15–17). Bronchial hyperresponsiveness (BHR) to methacholine was measured, and bronchoalveolar lavage fluid was obtained 24 h after the last challenge (day 18). Lungs were histologically examined. Results: Bronchoalveolar lavage fluid of saline-treated mice showed an increased cellularity with predominant eosinophils. Exposure to DHA-TG significantly reduced the total cell number and the eosinophil percentage in lavage fluid, whereas soybean oil did not. Conclusion: DHA but not soybean oil exposure reduced BHR and cell infiltration to bronchovascular bundles. This type of DHA administration could be studied in clinical trials.
Cross-Reactivity of TNP Immune Effector T Cells That Mediate Contact Hypersensitivity and Inflammatory Bowel Disease in the Mouse
Authors: Ptak Włodzimierz , Szczepanik Marian , Bryniarski Krzysztof , Stachura Jerzy , Gryglewski Andrzej , Szewczyk Katarzyna , Ptak Maria
Published: April 2003

Background: Experiments were aimed to test the cross-reactivity of immune Th1 cells that mediate contact hypersensitivity (CHS) or inflammatory bowel disease (IBD) to TNP in the mouse. Methods: CBA/J mice were immunized either epicutaneously or intrarectally with TNP and after appropriate time intervals were challenged with antigen in a crossed manner. The CHS reaction was measured by the ear swelling test. IBD was quantified by increase of colon weight and myeloperoxidase level. Both reactions were confirmed histologically. In passive-transfer experiments, mesenteric lymph node cells of animals sensitized intrarectally and peripheral lymph node and spleen cells of mice immunized epicutaneously were used. In some experiments, before being immunized mice were made either unresponsive to the TNP hapten by induction of suppressor T cells, or resistant to suppression after induction of upregulatory T cells. Results: Irrespective of the mode of sensitization upon appropriate challenge with antigen all mice developed a good CHS reaction as well as significant IBD. This cross-reactivity could be passively transferred by immune cells. In mice in which antigen-specific down- or upregulatory cells were induced before sensitization both CHS and IBD to TNP were modulated accordingly. Conclusion: TNP hapten deposited on skin or on mucosal surfaces induces effector cells that recognize antigen independent of its tissue localization, and produce a local inflammatory reaction. TNP-specific up- and downregulatory cells, shown before to regulate the CHS reaction, similarly modulate the generation and development of hapten-induced IBD.
Identification of a Novel Cat Allergen – Cystatin
Authors: Ichikawa Kunio , Vailes Lisa D. , Pomés Anna , Chapman Martin D.
Published: April 2003

Background: Cat allergen is an important cause of sensitization among children with asthma in Japan. Although there is good evidence that cats produce other allergens, only one major allergen, Fel d 1, has been studied in detail. Aims: To identify and define the molecular structure of the other potential cat allergens. Methods: A cat skin cDNA library was screened using IgE antibodies to cat dander and selected clones were sequenced and expressed. Results: One cDNA clone contained an open reading frame encoding a 98-amino acid residue protein. Sequence homology searches revealed a high degree of identity with bovine and human cystatin A, 79 and 75%, respectively. This cat cystatin clone contained the conserved cysteine protease motif and two of three lipocalin motifs. By plaque immunoassay, 60–90% of cat allergic sera had IgE Ab to cat cystatin. This cysteine protease inhibitor motif was partially conserved in dog allergens, Can f 1 and Can f 2, which are lipocalins. Recombinant cystatin was produced in Escherichia coli cells and purified as an 11-kD protein, corresponding to the predicted MW of cystatin. The structure of cat cystatin was modeled on human cystatin B using the SWISS-MODEL. Conclusion: A newly identified allergen, cystatin, has been cloned from cat skin and is a member of the cysteine protease inhibitor family.
The Roles of Mitogen-Activated Protein Kinase Pathways for Human Cultured Mast Cells
Authors: Nagai Hiroichi , Kimata Masahiro , Tanaka Hiroyuki , Inagaki Naoki
Published: April 2003

Background: Human cultured mast cells release histamine, sulfidoleukotrienes and prostaglandin D2, and produce some cytokines after an allergic reaction. As concerns the mitogen-activated protein kinase family, extracellular signal-regulated kinases, c-Jun NH2-terminal kinases and p38 mitogen-activated protein kinase are known. The present study has been conducted to investigate the role of mitogen-activated protein kinase in the mediator release from human cultured mast cells. Material and Methods: To investigate the role of these kinase pathways, we examined the participation of this kinase activation for mediator release by making use of U0126, a signal transduction, extracellular signal-regulated kinase pathway inhibitor, and SB203580, a p38 mitogen-activated protein kinase pathway inhibitor. Results: Histamine release is not related to mitogen-activated protein kinase pathways. Sulfidoleukotrienes and prostaglandin D2 release are mediated by signal transduction, extracellular signal-regulated kinase pathway. Granulocyte macrophage-colony-stimulating factor production may be induced by both signal transduction, extracellular signal-regulated kinase and c-Jun NH2-terminal kinase pathways. Conclusion: Mitogen-activated protein kinase pathways play an important role in mediator release from human mast cells.
Effect of Polymorphism of the β2-Adrenergic Receptor on Response to Regular Use of Albuterol in Asthma
Authors: Israel Elliot , Drazen Jeffrey M. , Liggett Stephen B. , Boushey Homer A. , Cherniack Reuben M. , Chinchilli Vernon M. , Cooper David M. , Fahy John V. , Fish James E. , Ford Jean G. , Kraft Monica , Kunselman Susan , Lazarus Stephen C. , Lemanske, Jr. Robert F. , Martin Richard J. , McLean Diane E. , Peters Stephen P. , Silverman Edwin K. , Sorkness Christine A. , Szefler Stanley J. , Weiss Scott T. , Yandava Chandri N. ,   for the National Heart, Lung, and Blood Institute’s Asthma Clinical Research Network
Published: April 2003

Background: Regular use of inhaled β-adrenergic agonists may have adverse effects in some asthma patients. Polymorphisms of the β2-adrenergic receptor (β2-AR) can affect its regulation; however, results of smaller studies of the effects of such polymorphisms on response to β-agonist therapy have been inconsistent. Methods: We examined the possible effects of polymorphisms at codons 16 (β2-AR-16) and 27 (β2-AR-27) on response to albuterol by genotyping 190 asthmatics who had participated in a trial of regular versus as-needed albuterol use. Results: During the 16-week treatment period, patients homozygous for arginine (Arg/Arg) at β2-AR-16 who used albuterol regularly had a small decline in morning peak expiratory flow (AM PEF). This effect was magnified during a 4-week run-out period, when all patients returned to as-needed albuterol only. By the end of the study, Arg/Arg subjects who had used albuterol regularly had an AM PEF 30.5 ± 12.1 liters/min lower (p = 0.012) than Arg/Arg patients who had used albuterol as needed only. Subjects homozygous for glycine at β2-AR-16 showed no such decline. Evening PEF also declined in the Arg/Arg regular but not in as-need albuterol users. No significant differences between regular and as-needed treatment were associated with polymorphisms at β2-AR-27. Conclusions: Polymorphisms of the β2-AR may influence airway responses to regular inhaled β-agonist treatment.
Invited Lecture: Activation of the Epithelial Mesenchymal Trophic Unit in the Pathogenesis of Asthma
Authors: Holgate Stephen T. , Lackie Peter M. , Howarth Peter H. , Roche William R. , Puddicombe Sarah M. , Richter Audrey , Wilson Susan J. , Holloway John W. , Davies Donna E.
Published: April 2003

Background: A recent NIH Workshop and an ERS Task Force concluded that more work was needed to understand mechanisms of severe and chronic asthma. This report describes a series of studies that identify aberrant epithelial mesenchymal signalling in the airways as an important event in maintaining inflammation and driving remodelling in response to environmental injury. Methods: Immunohistochemistry, genotyping and functional studies conducted on cultured asthmatic cells and mucosal biopsies were used to identify biochemical pathways involved in epithelial injury and repair in asthma and their relationship to disease severity. Results: Our findings suggest that the asthmatic state results from an interaction between a susceptible epithelium and Th-2-mediated inflammation to alter the communication between the epithelium and the underlying mesenchyme – the epithelial mesenchymal trophic unit – leading to disease persistence, airway remodelling and refractoriness to corticosteroid treatment. Conclusions: Asthma is more than an inflammatory disorder, but requires engagement of important signalling pathways involved in epithelial repair and tissue remodelling. These pathways involving EGFRs and TGF-βRs provide targets against which to develop novel therapies for chronic asthma.
Mosquito Allergy: Recombinant Mosquito Salivary Antigens for New Diagnostic Tests
Authors: Simons F. Estelle R. , Peng Zhikang
Published: April 2003

In patients with mosquito allergy, lack of a readily available, sensitive, specific, safe test is the major obstacle to accurate diagnosis. Three recombinant mosquito salivary antigens, rAed a 1 (68 kD), rAed a 2 (37 kD) and rAed a 3 (30 kD), from Aedes aegypti have been cloned, expressed, purified and characterized. All three recombinant antigens are shared by Aedes vexans and other mosquito species, and all have been found to have biologic activity in humans. In recent studies, 43% of 28 mosquito bite test-positive subjects had a positive skin test to rAed a 1, 11% to rAed a 2 and 32% to rAed a 3. The sizes of the skin test reactions to the recombinant antigens correlated with the sizes of the A. aegypti bite test reactions. None of 15 A. aegypti bite test-negative subjects had a positive skin test to any of the recombinant antigens. Recombinant mosquito salivary antigens will facilitate the diagnosis of mosquito allergy.
Dendritic Cells as Regulators of the Immune Response to Inhaled Allergen: Recent Findings in Animal Models of Asthma
Authors: Lambrecht Bart N. , Hoogsteden Henk C. , Pauwels Romain A.
Published: April 2003

Antigen-presenting dendritic cells are essential for the recognition and presentation of allergens to the cells of the immune system. Airway dendritic cells capture allergen in the mucosa and present it to naive T cells after migration into the draining lymph nodes. In this review article, we discuss the most recent findings from animal models of asthma, which highlight an essential role for these cells in the induction and maintenance of eosinophilic airway inflammation. This increasing knowledge might lead to the identification of new targets for the prevention and therapy of asthma.
(S)-Albuterol Increases the Production of Histamine and IL-4 in Mast Cells
Authors: Cho Seong H. , Hartleroad Jeffrey Y. , Oh Chad K.
Published: April 2003

Background: Racemic albuterol is an equimolar mixture of (R)-albuterol and (S)-albuterol. Previous studies indicated that (S)-albuterol may exert proinflammatory effects. We investigated the effect of (S)-albuterol in the production of mast cell mediators such as histamine and interleukin (IL)-4. Methods: Murine mast cells were either unstimulated or stimulated by IgE-receptor crosslinking. Both groups of mast cells were pretreated with either (R)- or (S)-albuterol. Histamine release and IL-4 secretion were measured by ELISA. Expression of L-histidine decarboxylase (L-HDC) and IL-4 was analyzed by semiquantitative reverse transcription-polymerase chain re- action. Results: In the overnight IgE-stimulated group, secreted histamine and total histamine were approximately 19.9 and 18.3% greater in cells co-treated with (S)-albuterol than untreated cells, respectively (n = 4, p < 0.002, p < 0.02), whereas there was no significant difference between the cells treated with (R)-albuterol and untreated cells. When the IgE-stimulated cells were treated with (S)-albuterol for 6 and 24 h, histamine release was approximately 18.3 and 24% greater, respectively (n = 4, p < 0.01). L-HDC is an essential enzyme for synthesizing histamine and its message was significantly induced in mast cells treated with (S)-albuterol. Both IL-4 message and protein were also significantly increased after treatment with (S)-albuterol. In the overnight IgE-stimulated group, IL-4 secretion was increased by approximately 58.8% upon exposure to (S)-albuterol (n = 5, p < 0.01). (R)-albuterol had no effect on mast cell mediator release. Conclusion: (S)-albuterol may have adverse effects in asthma control by activating mast cells to produce inflammatory mediators such as histamine and IL-4.
Lymphocyte Diversity in a 9-Year-Old Boy with Idiopathic CD4+ T Cell Lymphocytopenia
Authors: Frühwirth Martin , Clodi K. , Heitger Andreas , Neu Nikolaus
Published: April 2003

Since CD4+ lymphocytopenia can be caused by disturbed thymic T-cell maturation, we investigated the T-cell subsets of a 9-year-old boy fulfilling the diagnostic criteria for CD4+ lymphocytopenia in a follow-up period of 4 years. We found (I) reduced CD45RA expression, (II) enhanced CD45RO expression and (III) a significant increase in γδTCR-bearing T cells. An accelerated apoptosis (11%) was observed in the CD45RO+, but not CD45RA+ subset. These findings provide evidence that a disturbed thymic T-cell maturation process might play a role in the pathogenesis of CD4+ lymphocytopenia.
Anaphylaxis due to Hypersensitivity to Anisakis simplex
Authors: Domínguez-Ortega J. , Alonso-Llamazares A. , Rodríguez L. , Chamorro M. , Robledo T. , Bartolomé J.M. , Martínez-Cócera C.
Published: April 2003

Anaphylactic reactions are among the most frequent causes for requests for emergency medical care. It is possible that no clear precipitating factor can be found delaying the onset of adequate treatment and increasing the risk of new episodes of anaphylaxis in the future. Hypersensitivity to Anisakis simplex, an intestinal parasite found in fish, is an unusual cause of anaphylaxis but it should always be borne in mind in countries where a great deal of fish is consumed, especially if it is eaten raw or undercooked. 3 cases of anaphylaxis due to hypersensitivity to A. simplex are described.
Are Mast Cells MASTers in HIV-1 Infection?
Authors: Marone Gianni , de Paulis Amato , Florio Giovanni , Petraroli Angelica , Rossi Francesca W. , Triggiani Massimo
Published: April 2003

HIV-1 gp120 interacts with IgE VH3+ on the surface of human basophils and mast cells (FcΕRI+ cells), acting as a viral immunoglobulin superantigen. gp120 from different clades induces mediator release from FcΕRI+ cells. gp120 also induces IL-4 and IL-13 synthesis in human basophils. The chemokine receptors CCR3 and CXCR4, which are coreceptors of HIV-1 infection, are expressed by human FcΕRI+ cells. HIV-1 Tat protein is a potent chemoattractant for basophils and lung mast cells, interacting with CCR3. Incubation of basophils with Tat protein upregulates the surface expression of the CCR3 receptor. There is evidence that human FcΕRI+ cells could be infected in vitro by M-tropic HIV-1 strains.
Characterization of α5-Integrin-Dependent Mast Cell Adhesion following FcεRI Aggregation
Authors: Dastych Jaroslaw , Wyczółkowska Janina , Metcalfe Dean D.
Published: April 2003

Background: Integrin receptors are engaged in the upregulation of mast cell adhesion to extracellular matrix components upon stimulation with cytokines and antigen. Fibronectin receptor containing the α5-integrin subunit is critical for mast cell interaction with the extracellular matrix protein fibronectin (FN). Methods: The murine MCP5/L mast cell line was employed to investigate the process of FcΕRI-mediated mast cell adhesion to FN. RT-PCR and cytofluorimetric analysis were used to assess the expression of α5 integrin in MCP5/L mast cells. Radiolabelled mast cells were sensitized with monoclonal IgE and used in adhesion assays. Anti-α5-integrin antibody (Ab), monovalent hapten and metabolic inhibitors were used to characterize antigen-mediated mast cell adhesion to FN. Results: Addition of antigen to IgE-sensitized cells resulted in transient upregulation of mast cell adhesion to FN with a maximum adhesion following 30 min of incubation. Mast cell adhesion was inhibited with anti-α5-integrin monoclonal antibodies blocking FN receptor or with excess monovalent hapten preventing antigen-mediated IgE cross-linking. The presence of the protein kinase C (PKC) inhibitor staurosporine also inhibited mast cell adhesion in a dose-dependent fashion. The process of FcΕRI-mediated upregulation of mast cell adhesion to FN was not associated with an increase in surface expression of mast cell FN receptors. Conclusion: The major FN receptor on MCP5/L mast cell surface, an integrin containing the α5 subunit mediates a transient change in mast cell adhesiveness following IgE cross-linking. FcΕRI-derived signals engage PKC and upregulate mast cell adhesion in a process which might involve changes in integrin avidity rather than integrin expression.
Underdiagnosis and Undertreatment of Asthma: A 9-Year Study of Italian Conscripts
Authors: Ciprandi Giorgio , Vizzaccaro Andrea , Cirillo Ignazio , Tosca Mariangela , Passalacqua Giovanni , Canonica Giorgio Walter
Published: April 2003

Background: During the last 20 years, the epidemiology of asthma has been investigated in various populations, thus providing interesting data including the accuracy of diagnosis and the adequacy of treatment. Methods: We studied for 9 years the epidemiological features of asthma in a homogeneous population of 18-year-old males referred to La Spezia Military Navy Hospital for a call-up visit (conscripts). Results: During the period of 1990–1998, 91,700 men were screened at their call-up visit; out of them 5,371 (7%) had asthma, but in 7.4% the disease had not been previously diagnosed. During the observation period there was a significant decrease of the level III severity of asthma, but about one quarter of the asthmatic subjects received no treatment at all. On the other hand, a significant increase in the prescription of inhaled steroids was observed. Conclusion: Despite educational efforts, a not negligible underdiagnosis and inadequate treatment of asthma in young adults still persist.
Chemoattractant Receptors Expressed on Type 2 T Cells and Their Role in Disease
Authors: Cosmi Lorenzo , Annunziato Francesco , Maggi Enrico , Romagnani Sergio , Manetti Roberto
Published: April 2003

The existence of two functionally distinguished populations among T cells has been established in both mice and humans. Type 1 T helper (Th1) cells are involved in the defense against intracellular bacteria and many viruses, while type 2 Th cells (Th2) are the major actors in the response against parasites and play a central role in allergic inflammation. More recently, several data have suggested that some chemokine receptors are tightly regulated on T cells, and in accordance with this selective expression, Th1 and Th2 cells can be differentially recruited by specific chemokines to the inflammatory sites. Among Th2-associated chemokine receptors, CCR3, CCR4 and CCR8 have been described to play a central role in allergic inflammation. However, CCR3 is mainly expressed on basophils, eosinophils and mast cells, but it is poorly expressed by Th2 cells, and CCR4 is also expressed by Th subsets different from Th2 cells. So far, the chemoattractant receptors which among T cells appear to be selectively expressed by Th2 cells or their subsets are CCR8 and CRTH2. The ligand for CRTH2 is not a chemokine, but is prostaglandin (PG)D2, which is able to attract basophils, eosinophils, Th2 cells and type 2 cytotoxic (Tc2) CD8+ T lymphocytes. The selective expression of CRTH2 on Th2 and Tc2 cells may be useful to develop new therapeutic strategies against allergic diseases and against other immune disorders. Additional studies, however, are required to understand its effective importance in the induction and maintenance of Th2- or Tc2-mediated response and inflammation.
Theophylline Inhibits TNF-α-Induced CD4 Expression on Human Eosinophils and CD4+ Eosinophil Migration
Authors: Tsukadaira Akihiro , Okubo Yoshio , Horie Shiro , Koyama Sekiya
Published: April 2003

Background: Increasing evidence regarding asthma suggests that CD4+ cells are preferentially recruited to sites of bronchial inflammation. Interleukin (IL)-16 has been reported as playing an important role in the accumulation of CD4+ cells. We have shown that the CD4 molecule is expressed on normal human eosinophils by tumor necrosis factor (TNF)-α stimulation. Methods: We evaluated the effects of theophylline, KF19514 [a selective phosphodiesterase (PDE) IV inhibitor] and dexamethasone on CD4 expression on eosinophils and eosinophil migration in response to IL-16, a natural soluble ligand of the CD4 molecule. Results: The maximum eosinophil migration was observed when eosinophils were cultured with TNF-α at 10 ng/ml for 18 h and the concentration of IL-16 was 10 pg/ml. CD4+ eosinophil migration in response to IL-16 was mostly, if not fully, chemokinetic and this migration was significantly inhibited by Fab of anti-CD4 monoclonal antibody. Theophylline (10–4–10–3M), KF19514 (10–7–10–6M) and dexamethasone (10–8– 10–6M) significantly inhibited CD4 expression on eosinophils induced by TNF-α. Theophylline (10–3M) and KF19514 (10–6M) inhibited CD4+ eosinophil migratory responses induced by IL-16, but 10–6M dexamethasone did not. Theophylline and KF19514 augmented the intracellular adenosine-3′,5′-cyclic monophosphate (cAMP) concentration in eosinophils, suggesting modulation by cAMP of CD4 expression and eosinophil migration. Conclusions: These data suggest that TNF-α-induced CD4+ eosinophils may contribute to eosinophil migratory responses induced by IL-16. Theophylline and selective PDE IV inhibitor may prevent airway inflammation by downregulating CD4 expression on eosinophils and inhibiting eosinophil migration through CD4 and IL-16 interaction.
Oligoclonal Analysis of the Atopic T Cell Response to the Group 1 Allergen of Cynodon dactylon (Bermuda Grass) Pollen: Pre- and Post-Allergen-Specific Immunotherapy
Authors: Eusebius Nirupama P. , Papalia Lina , Suphioglu Cenk , McLellan Susan C. , Varney Michael , Rolland Jennifer M. , O’Hehir Robyn E.
Published: April 2003

Background: Bermuda grass pollen (BGP) is an increasingly important seasonal aeroallergen in Australia and other subtropical and temperate regions. BGP shares minimal allergenic cross-reactivity with pollens of rye grass or other Pooideae grasses often used for desensitization regimens in grass pollen allergy. Current allergen immunotherapy is seldom used in asthmatic patients due to IgE-mediated side effects. Since clinically effective immunotherapy is linked with altered allergen-specific T cell response, characterisation of human T cell reactivity to Cyn d 1, the major B cell allergen of BGP, should permit the design of effective and safe immunotherapy for BGP allergy. Methods: Short-term BGP-specific CD4+ T cell lines were established from peripheral blood of 14 BGP-sensitive patients before and after conventional 50% BGP and 50% 7-grass mix subcutaneous specific allergen immunotherapy (SIT). T cell diversity of antigen specificity and function was assessed by proliferation and cytokine production to BGP, Cyn d 1 and Cyn d 1 peptides. Results: Three highly immunogenic regions of Cyn d 1 were identified in 13/14 patients pre-SIT: Cyn d 1 (109–128), (181–209) and (217–241). The SIT regimen was clinically efficacious. Following SIT, decreased proliferation to BGP, Cyn d 1 and Cyn d 1 peptides was observed with a marked decrease in the IL-5:IFN-γ ratio. Conclusions: Cyn d 1 is a major T cell allergen of BGP. Decreased Cyn d 1-specific IL-5 dominant T cell responses were observed in association with clinically effective treatment with the 50% BGP and 50% 7-grass mix. Identified dominant T cell regions of Cyn d 1 should facilitate safer vaccine development for BGP-induced asthma in addition to rhinitis.
Altered Expression of Extracellular Matrix in Human-Cytomegalovirus-Infected Cells and a Human Artery Organ Culture Model to Study Its Biological Relevance
Authors: Schaarschmidt Peter , Reinhardt Barbara , Michel Detlef , Vaida Bianca , Mayr Klaus , Lüske Anke , Baur Regine , Gschwend Jürgen , Kleinschmidt Klaus , Kountidis Margaritis , Wenderoth Ulrich , Voisard Rainer , Mertens Thomas
Published: April 2003

The influence of human cytomegalovirus (HCMV) on the transcription of 11 selected, representative extracellular matrix genes was investigated in cell culture. Northern blot hybridization indicated the downregulation of all mRNAs investigated. Based on our results and the known repression of other extracellular matrix transcripts and the β-actin transcription during HCMV infection, we suggest that one molecular mechanism contributing to the cytopathic effect may be the transcriptional downregulation of genes encoding proteins involved in cell structure and intercellular connection. To further study the biological relevance of this and other pathogenetic mechanisms, we established a human renal artery organ culture system and characterized this new infection model for HCMV. Our model is a new suitable system for the investigation of molecular as well as functional consequences of HCMV infection in a more physiological microenvironment.
Diagnostic Value of Nucleic-Acid-Sequence- Based Amplification for the Detection of Cytomegalovirus Infection in Renal and Liver Transplant Recipients
Authors: Goossens Valère J. , Blok Marinus J. , Christiaans Maarten H.L. , van Hooff Johannes P. , Sillekens Peter , Höckerstedt K. , Lautenschlager Irmeli , Middeldorp Jaap M. , Bruggeman Cathrien A.
Published: April 2003

To evaluate the diagnostic value of nucleic-acid-sequence-based amplification (NASBA) for the detection of cytomegalovirus (CMV) infection in transplant recipients, we compared immediate early 1 (IE1) and late pp67 mRNA detection by NASBA with the antigenemia assay, PCR and viral culture in 72 renal transplant (RTx) recipients and with antigenemia and serology in 25 liver transplant (LTx) recipients. Antigenemia, viral culture and pp67 NASBA were almost equivalent for the detection of CMV in RTx recipients. In LTx recipients, antigenemia detected more positive samples and more positive recipients compared to pp67 NASBA. In RTx recipients, PCR detected more positive samples and positive recipients compared to pp67 NASBA, antigenemia and viral culture. Also the first day of detection was slightly earlier for PCR. However, IE1 NASBA was the most sensitive test and detected 96% of all positive samples and positive transplant recipients. In addition, IE1 NASBA preceded PCR and all other positive results. This makes IE1 NASBA a very attractive screening test for the early detection of CMV infection.
New Advances in the Diagnosis of Congenital Cytomegalovirus Infection
Authors: Lazzarotto T. , Varani S. , Gabrielli L. , Spezzacatena P. , Landini M.P.
Published: April 2003

With the advances in anticytomegalovirus (anti-CMV) serology, the new recombinant IgM tests seem likely to become the screening tests for pregnant women whose prepregnancy serological status for CMV is unknown. When a woman is found to be IgM-positive, further diagnostic evaluation focused on determining whether this is due to a primary infection should be carried out. Maternal primary infections that were difficult to determine until a few years ago unless documented by seroconversion can now be readily diagnosed from the presence of low-avidity anti-CMV antibody which persists for approximately 20 weeks after primary infection. In primarily infected mothers prenatal diagnosis can be performed between 21 and 23 weeks of gestation, and the amniotic fluid (AF) represents the pathological material of choice to determine intrauterine virus transmission. In AF, the virus can be detected by culture and/or PCR. Both procedures differentiate uninfected from infected fetuses, but cannot predict fetal outcome. The determination of the viral load in AF carried out by quantitative PCR is more promising and could represent an important starting point for preemptive fetal therapy.
Proinflammatory Potential of Cytomegalovirus Infection
Authors: Cinatl Jr. J. , Vogel J.-U. , Kotchetkov R. , Scholz M. , Doerr H.W.
Published: April 2003

We observed the effects of antiviral therapy on CMV and/or oxidative-stress-induced stimulation of proinflammatory molecules including interleukin-8 (IL-8), melanoma growth stimulatory activity-α (GRO-α) and intercellular adhesion molecule 1 (ICAM-1) using human foreskin fibroblasts. Ganciclovir, foscarnet or cidofovir completely suppressed virus replication, as demonstrated by CMV late (L) antigen production. These drugs did not influence CMV immediate-early (IE) antigen expression and had no effects on CMV-induced cellular changes in IL-8, GRO-α and ICAM-1 levels. Phosphorothioate oligonucleotide (ISIS 2922) suppressed both CMV IE and L antigen by 99%. ISIS 2922 completely suppressed CMV-induced upregulation of both chemokines and ICAM-1. Induction of oxidative stress by H2O2 upregulated IL-8 expression. Oxidative stress and CMV infection showed synergistic effects on IL-8 expression. ISIS 2922 only partially inhibited the upregulation of IL-8 in infected cells treated with H2O2, whereas cotreatment with ISIS 2922 and antioxidants inhibited the upregulation almost completely. The results showed that inhibition of CMV IE expression alone or in combination with antioxidants is promising for the treatment of CMV disease.
DNA Vaccination for HIV-1 and SIV
Authors: Barouch Dan H. , Letvin Norman L.
Published: April 2003

Control of the worldwide AIDS epidemic will only be achieved with a safe and effective prophylactic HIV-1 vaccine. DNA vaccination has recently emerged as a promising vaccine modality that can elicit both humoral and cellular immune responses. HIV-1- and SIV-specific immune responses have been elicited by DNA vaccines in both mice and nonhuman primates. However, these immune responses have not been capable of protecting nonhuman primates against pathogenic AIDS virus challenges. A number of approaches are therefore being investigated to augment DNA vaccine-elicited immune responses.
Leukocyte Adhesion after Oxidant Challenge in the Hamster Cheek Pouch Microcirculation
Authors: Bouskela E. , Cyrino F.Z.G.A. , Lerond L.
Published: April 2003

Oral administration of S-5682 (Daflon 500 mg, 90% diosmin, 10% hesperidin) inhibits oxidant-induced increase in macromolecular permeability in the postcapillary venules of the hamster cheek pouch microcirculation. In this study, the effect of S-5682 on leukocyte-endothelium interaction was evaluated using the same experimental model. Hamsters kept on a standard diet were divided into 5 groups (n = 6) and treated orally, twice a day, with placebo (10% lactose solution), S-5682, 5, 20 or 80 mg/kg/day (suspended in 10% lactose solution) or α-tocopherol, 1 mg/kg/day, for 10 days prior to the oxidant challenge with tert-butylhydroperoxide (TBOOH). Topical application of TBOOH (10–4 M for 5 min) to hamsters given acridine orange prior to TBOOH resulted in increases in the number of rolling and sticking (no movement for at least 30 s) leukocytes in postcapillary venules. No changes in the number of rolling leukocytes could be observed in the treated groups compared with the placebo group (p > 0.05). On the contrary, leukocyte adhesion was inhibited in groups treated with S-5682 (5, 20 and 80 mg/kg/day) or α-tocopherol: placebo 105 ± 3/6 mm2 (mean ± SEM); S-5682, 5 mg/kg/day 68 ± 3/6 mm2 (p < 0.01), 20 mg/kg/day 55 ± 3/6 mm2 (p < 0.001) and 80 mg/ kg/day 39 ± 2/6 mm2 (p < 0.001) and α-tocopherol 36 ± 1/6 mm2 (p < 0.001). The inhibition of oxidant-induced leukocyte adhesion by S-5682 was similar to that seen for ischemia-reperfusion and the higher dose of S-5682 was as effective as α-tocopherol in inhibiting it.
Venous Ulcer Reappraisal: Insights from an International Task Force
Authors: Clement D.L.
Published: April 2003

An international task force made up of a panel of 16 experts was mandated to review and objectively evaluate all aspects of chronic venous disease of the leg. All available publications on chronic venous disease of the leg from 1983 to 1997 were identified through computerized search. Three different screenings were then performed in order to select only relevant papers providing moderate to strong scientific evidence. Final conclusions and further therapeutic recommendations were made based on these publications. Compression, medications, local therapies, sclerotherapy and surgery are the existing therapeutic options for which the following recommendations can be made. Compression: Properly applied bandages both fixed and stretched have been shown to be effective. Compression stockings may be used. Compression needs to be in excess of 35 mm Hg. Medications: Although preliminary results have shown a beneficial effect of several vasodilators and oral micronized purified flavonoid fraction, the evidence for the efficacy of medications on venous ulcer healing is still limited and further studies are required before recommendations can be made. There is no evidence to routinely administer antibiotics. Local therapies have an as yet unproven adjunctive role. Sclerotherapy is unlikely to be effective unless there is superficial venous insufficiency, in which it may have an as yet unproven role. Surgery: Patients with active venous ulcer and sapheno-femoral or sapheno-popliteal junction incompetence benefit from surgical treatment. In the absence of reflux or following deep vein thrombosis, there is no evidence to support surgical treatment.
Expression and Function of Recombinant Endothelial Nitric Oxide Synthase in Human Endothelial Cells
Authors: Zanetti Michela , Katusic Zvonimir S. , O’Brien Timothy
Published: April 2003

Endothelial dysfunction is frequently involved in the pathogenesis of vascular disease. While nitric oxide (NO) inhibits smooth muscle cell proliferation, its effect on endothelial cell proliferation is unclear. The aim of this study was to determine if adenoviral-mediated gene transfer of endothelial NO synthase (eNOS) to human umbilical vein endothelial cells (HUVECs) would result in increased generation of NO and affect endothelial cell proliferation. HUVECs were transduced with adenoviral vectors encoding eNOS (AdeNOS) or β-galactosidase (Adβgal) or exposed to diluent (control). AdeNOS-transduced cells showed increased eNOS expression as detected by Western blot analysis, and increased concentrations of cGMP (control 0.7 ± 0.1; Adβgal 0.9 ± 0.2; AdeNOS 3.1 ± 0.5 pmol/mg protein; p < 0.001) and nitrite (control 11.8 ± 1.2; Adβgal 13.3 ± 1.7; AdeNOS 21.1 ± 2.2 nmol/mg protein/hour; p < 0.01). DNA synthesis as assessed by [3H]thymidine incorporation and cell counts were significantly reduced (by ∼30%) in AdeNOS-transduced HUVECs. Expression of mitogen-activated protein kinase was also decreased in AdeNOS-transduced cells. This study shows that adenoviral-mediated gene transfer of eNOS to HUVECs inhibits endothelial cell proliferation.
Transition of Apoptotic Resistant Vascular Smooth Muscle Cells to Troptotic Sensitive State Is Correlated with Downregulation of c-FLIP
Authors: Imanishi Toshio , Hano Takuzo , Nishio Ichiro , Liles W. Conrad , Schwartz Stephen M. , Han David K.M.
Published: April 2003

Fas and its ligand, FasL, are a receptor-ligand pair identified as promoting cell death in several tissues. Vascular smooth muscle cells (VSMCs) are resistant to FasL or anti-Fas antibody (Ab) signal, and a number of in vitro studies show that VSMC death can only be induced by anti-Fas Ab or FasL in the presence of protein inhibitor or additional inflammatory mediators. It remains to be clarified whether known, constitutively expressed cytoprotective molecules are reduced by protein inhibitor, thereby accounting for sensitization to cell death by Fas/FasL signaling. We found that Fas mRNA and protein exist in several primary VSMCs, as previously reported. We also demonstrated (1) that critical death-signaling molecules, such as FADD, caspase-1/ICE, and caspase-3/YAMA, are present in these VSMCs, (2) that human VSMCs contain high concentrations of c-FLIP (3) and that following treatment with the protein inhibitor, CHX, cell extracts showed a decrease in c-FLIP protein that was dose- and time-dependent on the degree of apoptosis and inversely correlated with both caspase-8 and -3 activity. In contrast, there was neither a change nor an even modest upregulation of Bcl-2 family, even after 12 h of treatment with CHX. Taken together, these results may provide a novel insight into atherogenesis and suggest that c-FLIP may contribute to an apoptosis-resistant state of VSMC, and that a downregulation of c-FLIP may render VSMCs susceptible to apoptosis.
Early Remodeling of Saphenous Vein Grafts: Proliferation, Migration and Apoptosis of Adventitial and Medial Cells Occur Simultaneously with Changes in Graft Diameter and Blood Flow
Authors: Kalra M. , Miller V.M.
Published: April 2003

Background: This study was designed to determine how migration and proliferation of adventitial and medial cells correlate temporally with hemodynamic characteristics including changes in diameter and blood flow in vein grafts. Methods: Male mongrel dogs underwent end-to-side reversed saphenous vein bypass grafting across a ligated femoral artery. Hemodynamic parameters were assessed by duplex ultrasound. Proliferating cells were labeled 24–48 h after grafting with 5-bromo-2′-deoxyuridine (BrdU). Grafts were removed on postoperative days 2, 5, 7 and 14. Immunohistochemistry was performed on graft sections with antibodies to nuclear antigen Ki-67 (MIB-1), BrdU and smooth muscle cell markers. Apoptosis was identified by modified TUNEL staining. Proliferating/apoptotic cells were quantified digitally. Results: Mean luminal cross-sectional area, blood flow and velocity increased from day 2 to 7. Cell proliferation was evident in adventitia and media on day 2, maximum on day 5 and decreased significantly by day 14. Apoptosis was maximum on day 5. Early proliferating cells (BrdU labeled) localized in the adventitia and media on day 2 were more prevalent in the neointima on days 5–14 suggesting inward migration. On day 2, proliferating cells stained positive only for vimentin. By days 5–14, proliferating cells stained for α-smooth-muscle actin, a phenotype characteristic of myofibroblasts. Conclusion: These results indicate that cell proliferation and apoptosis occur simultaneously within the adventitia and media of the vein during the first week following grafting, when changes in diameter and blood flow are greatest. In addition, proliferating adventitial cells subsequently migrate inwards to contribute to the formation of neointima.
Deep Venous Thrombosis: Leukocyte Rheology at Baseline and after in vitro Activation
Authors: LoPresti R. , Ferrara F. , Canino B. , Montana M. , Caimi G.
Published: April 2003

We evaluated leukocyte rheology, expressed as leukocyte filtration, polymorphonuclear (PMN) membrane fluidity and cytosolic Ca2+ concentration in subjects with acute deep venous leg thrombosis (DVT). In 14 subjects with leg DVT we examined the leukocyte filtration [unfractionated, mononuclear cells (MN), PMNs], PMN membrane fluidity and PMN cytosolic Ca2+ concentration. Subsequently, we evaluated the same PMN variables after in vitro chemotactic activation with 4-phorbol-12-myristate-13-acetate. At baseline, we observed a significant difference in the filtration of unfractionated and MNs and in PMN cytosolic Ca2+ concentration. After PMN activation, a significant variation, greater in DVT subjects, was present in PMN filtration at 5 and 15 min. In normals, no variation was present in PMN membrane fluidity or cytosolic Ca2+ concentration after activation, while in subjects with DVT we found a significant variation in both PMN parameters. These results underline that there is a systemic leukocyte functional alteration in DVT.
A Chronopharmacodynamic Study on Standard Heparin, a Low Molecular Weight Heparin (Nadroparin) and Danaproid: Establishing and Comparing the Daily Variations of These Drugs in Rats
Authors: Abrial David , Blanc Alain , Réhailia Mohamed , Mismetti Patrick , Bouchut Colette , Laporte-Simitsidis Silvy , Decousus Hervé , Buisson Bernard
Published: April 2003

The effects of dosing time on the anticoagulant activity of unfractionated heparin, low molecular weight heparin (nadroparin) and danaproid were investigated. The chronopharmacological comparisons of the drugs were done on the anti-Xa, anti-IIa activities and activated partial thromboplastin time assays. Several dosing times were considered and an analysis based on a population approach was adopted. Under unfractionated heparin, the pharmacological activities did not exhibit significant daily variations. In contrast, significant daily profiles were observed in all the biological assays performed with low molecular weight heparin. Anti-Xa and anti-IIa activities showed some fluctuations over a 24-hour period with a peak at noon. As for the variations of the activated partial thromboplastin time, two peaks were noted early in the morning and at the beginning of nightfall. As for danaproid, only a daytime maximum of anti-Xa activity could be found.
Absence of Transplacental Passage of the Low Molecular Weight Heparin Enoxaparin
Authors: Dimitrakakis Constantine , Papageorgiou Panayiotis , Papageorgiou Ioannis , Antzaklis Aris , Sakarelou Nafsika , Michalas Stylianos
Published: April 2003

Background: Venous thromboembolism (VTE) during pregnancy and puerperium remains a major cause of maternal morbidity and mortality. The use of low molecular weight heparin (LMWH) constitutes a promising alternative for the prevention of VTE instead of unfractionated heparin as it can be administered subcutaneously once daily and without coagulation measurement. Unfortunately, the safety of LMWHs administration for the mother and fetus has not been well established. Study Design: In order to examine the safety of enoxaparin to the fetus, 24 women were recruited and 40 mg of enoxaparin was administered in 14 of them. All 24 women were going to have an early termination of pregnancy due to major fetal malformations. Maternal blood samples were drawn before and after the injection of enoxaparin, while fetal blood samples were taken only after the drug administration. Anti-IIa and anti-Xa activities were measured. Results: A statistically significant increase of anti-Xa activity in the mothers studied was pointed out, while there was no detection of anti-IIa and anti-Xa activities in the fetuses. Conclusions: Since no anti-IIa and anti-Xa activities were detected in the fetuses’ blood samples, it is concluded that enoxaparin does not cross the placenta and therefore appears safe for the fetus.
Severe Thrombotic Complications Associated with Activated Protein C Resistance Acquired by Orthotopic Liver Transplantation
Authors: Gillis Shmuel , Lebenthal Avi , Pogrebijsky Galina , Levy Yizhar , Eldor Amiram , Eid Ahmed
Published: April 2003

We report a patient who developed recurrent hepatic artery thrombosis and deep venous thrombosis following orthotopic liver transplantation. Investigations revealed the presence of activated protein C (APC) resistance due to a mutation in the factor V gene in the transplanted liver. The patient’s own peripheral blood cells did not carry the mutation. Although part of factor V is located in the platelets and may be endogenously synthesized by megakaryocytes, this case demonstrates the major clinical importance of hepatic-derived factor V. It may be reasonable to screen liver donors with a history of a thrombotic event for APC, and to consider anticoagulation in the recipients of livers positive for this defect.