bluuebunny/crossref_metadata_2025 huggingface dataset:
Bleomycin-Induced Pulmonary Fibrosis in Genetically Mast Cell-Deficient WBB6F1-W/Wv Mice and Mechanism of the Suppressive Effect of Tranilast, an Antiallergic Drug Inhibiting Mediator Release from Mast Cells, on Fibrosis
Authors: Mori Hiroshi , Kawada Kenji , Zhang Peng , Uesugi Yuki , Sakamoto Osami , Koda Akihide
Published: August 2009

It has been well known that the number of mast cells increases during the development of fibrosis in various tissues including the lung. However, the role of mast cells in fibrosis still remains obscure. In the present paper, we evidenced that pulmonary fibrosis could be induced in genetically mast cell-deficient WBB6F1 W/Wv mice as well as WBB6F1-+/+ mice having mast cells normally by the treatment with bleomycin (BLM, 5 mg/kg, i.v., 10 days), and there was not much difference in the histological changes of lungs between the two strains. An increase in the hydroxyproline content of the lung of WBB6F1-W/Wv mice was rather higher than that of WBB6F1-+/+ mice. Previously, we reported that tranilast, an antiallergic drug inhibiting chemical mediator release from mast cells, suppressed the development of BLM-induced pulmonary fibrosis in ICR mice, suggesting the possibility that mast cells play certain roles in fibrosis. However, it was evidenced in the present report that tranilast suppressed BLM-induced fibrosis in WBB6F1-W/Wv mice. Tranilast neither suppressed the cytotoxic activity of BLM against KB cells and L-929 cells in vitro, nor inhibited the antitumor activity of BLM against Sarcoma-180 transplanted subcutaneously into ICR mice. Tranilast may act through suppressing BLM-induced activation of lymphoid cells including macrophage and neutrophil. These results indicate an inconsequential role of mast cells in the development of fibrosis. Increases in the number of mast cells and in histamine content of the lung, which were widely reported in the lungs of BLM-treated mice, may be the result of fibrosis.
Anaphylaxis Is Enhanced by Increased Absorption of Antigen in Postinflammatory State
Authors: Levine Seymour , Saltzman Arthur
Published: August 2009

Hemagglutinating antibodies and cell-mediated immunity are increased when antigens are injected intraperitoneally (i.p.) into rats during the healing phase of a chemical peritonitis. In the present work, the anaphylactogenic effects of either sensitization or challenge were increased when any of three different antigens were injected i.p. in the postinflammatory state. The postinflammatory state made it possible to sensitize rats for anaphylaxis without any adjuvants at all. Lymph nodes draining the peritoneal cavity had evidence of enhanced absorption of inoculum in the postinflammatory state.
Bradykinin Induces Eosinophil Accumulation in the Rat Pleural Cavity
Authors: Pasquale Claudia P. , Martins Marco A. , Bozza Patricia T. , Silva Patricia M.R. , Faria Neto Hugo C. Castro , Pires Ana L.A. , Cordeiro Renato S.B.
Published: August 2009

Intrathoracic injections of bradykinin (1–100 μg/cavity) induced a dose-dependent increase in the number of eosinophils recovered from the rat pleural cavity 24 h later. Eosinophilia by bradykinin was preceded by a marked pleural neutrophil influx within 6 h and was absent only 72 h following stimulation. Bradykinin (10––9––10––5M) failed to induce in vitro eosinophil chemotaxis, indicating that its in vivo effect must be mediated by an intermediate messenger. BW 755C (25 mg/kg) and the more selective lipoxygenase inhibitor BW A4C (20 μg/cavity) suppressed the pleural eosinophilia induced by bradykinin (50 μg/cavity), whereas the platelet-activating factor (PAF)-acether antagonist BN 52021 was inactive. We conclude that bradykinin is able to attract eosinophil in vivo by a mechanism independent of PAF-acether and sensitive to the blockage of the lipoxygenase pathway.
Faecally Derived Hydrolytic Enzymes from Dermatophagoides pteronyssinus: Physicochemical Characterisation of Potential Allergens
Authors: Stewart Geoffrey A. , Lake Fiona R. , Thompson Philip J.
Published: August 2009

The previous findings that the group I and III mite allergens, and amylase present in mite faeces are hydrolytic enzymes has prompted a study to determine whether this material contains other enzymes which could be allergenic. Thus, spent growth medium devoid of whole Dermatophagoides pteronyssinus mites was shown to contain glucoamylase, lipase and lysozyme in addition to the cysteine protease, serine protease and amylase activities associated with the above allergens, respectively. All of these enzymes are probably associated with mite digestive processes. They were rapidly solubilised, heterogeneous with regard to charge (pI in the range 4–8) and demonstrated maximum biochemical activity in the neutral pH range. Three serine proteases were detected and comprised a chymotrypsin-like, a trypsin-like and an unclassified enzyme with pI of 4.1 and 5.3, 8.5 and 7.1, respectively. Only one cysteine protease was observed, which paralleled immunochemically identified Der p I in a variety of assays. It was shown to cleave at lysyl residues and could be inhibited by the specific cysteine protease inhibitor, E-64. The remaining serine proteases, glucoamylase, lipase and lysozyme represent potential allergens.
Beta-2-Adrenoceptors of Polymorphonuclear Leukocytes in Children with Atopic Dermatitis
Authors: Pohl A. , Otto J. , Urbanek R.
Published: August 2009

[125I]-cyanopindolol (ICYP) was used in a binding assay to compare the number and affinity of β2-adrenoceptor binding sites on circulating polymorphonuclear leukocytes (PMN) of 21 children with atopic dermatitis (AD) and 23 age-matched controls. We found no correlation between the β2-adrenoceptor density and the age of the children in either collective. Furthermore, the number and affinity of β2-adrenoceptor binding sites on PMN was neither influenced by sex nor severity of disease, serum IgE level or percentage of eosinophils in the differential blood count. There was a tendency towards a lower number of β2-adrenoceptors in AD subjects (1,429 ± 587 receptors/PMN) compared to controls (1,753 ± 596 receptors/PMN), but this difference was statistically not significant.
Time Course of Cellular Infiltration in the Nasal Mucosa during the Immediate Allergic Reaction
Authors: Lozewicz S. , Gomez E. , Chalstrey S. , Gatland D. , Davies R.J.
Published: August 2009

In 23 patients with allergic rhinitis, biopsies of the nasal mucous membrane were taken at one of the following times after challenge of one nostril with allergen: 0 (baseline) (n = 7), ½ h (n = 6), 1 h (n = 5), and 2 h (n = 5). In the nostril stimulated by allergen there was a transient early phase influx of eosinophils while the numbers of stainable mast cells decreased, probably due to their degranulation. In the contralateral unstimulated nostril, there was no change in numbers of eosinophils but the numbers of stainable mast cells decreased. These results support the proposed role in allergic rhinitis of the mast cell and eosinophil, and suggest that the eosinophil may be a rapidly mobilized effector cell.
Effect of Histamine on Tumor Necrosis Factor Production by Human Monocytes
Authors: Hotermans G. , Bury T. , Radermecker M.F.
Published: August 2009

This study was aimed at evaluating the effect of histamine on tumor necrosis factor (TNFα) secretion by purified human blood monocytes. TNFα was measured by radioimmunoassay. Histamine caused a dose-dependent inhibition of lipopolysaccharide-induced TNFα production from human blood monocytes, averaging maximally 50% at 10––5M. Preincubation of mononuclear cells with an H2 antagonist (cimetidine), but not with an H1 antagonist (promethazine) prevented this inhibitory effect of histamine. In conclusion, histamine causes, in vitro, a depression of TNFα secretion by human monocytes through activation of H2 receptors.
Increased Expression of FcεR2/CD23 on Peripheral Blood B Lymphocytes and Serum IgE Levels in Kawasaki Disease
Authors: Furukawa Susumu , Matsubara Tomoyo , Motohashi Toshikazu , Sasai Keiko , Nakachi Seigi , Umezawa Yoshimi , Yabuta Keijiro
Published: August 2009

We analyzed the expression of FcεR2/CD23 on the peripheral blood B lymphocytes of 10 patients with Kawasaki disease (KD) using a fluorescence-activated cell sorter. The absolute count of CD23-positive B lymphocytes and the ratio of CD23-positive B lymphocytes to B lymphocytes were high during the acute stage of KD in comparison to those during the convalescent stage and in control subjects. In addition, both the increased number of CD23-positive B lymphocytes and the increased serum IgE levels appeared during the latter part of the acute stage. These results suggest that the increased number of CD23-positive B lymphocytes during the latter part of the acute stage in KD reflect the activation of B lymphocytes and have an important role in IgE-mediated immunity.
Ability of Polymorphonuclear Leukocytes to Generate Active Oxygen Species in Children with Bronchial Asthma
Authors: Kato Masahiko , Nakano Minoru , Morikawa Akihiro , Kimura Hirokazu , Shigeta Makoto , Kuroume Takayoshi
Published: August 2009

Airway inflammation with polymorphonuclear leukocytes (PMN) may play an important role in bronchial hyperresponsiveness (BHR). PMN generate superoxide anion (O2––) and other oxygen radicals that can damage lung tissue. We investigated the ability of peripheral PMN of children with bronchial asthma and control subjects to generate O2–– and other active oxygen species using a 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-α]pyrazin-3-one, a highly sensitive and specific chemiluminescence (CL) probe for O2––, and luminol-dependent CL. The ability of PMN of subjects with asthma to generate O––2 and other active oxygen species was significantly greater than that of PMN of control subjects when stimulated with opsonized zymosan (OZ), phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine. Furthermore, in the same asthmatic children, the generation of O2–– and other active oxygen species was significantly higher with attacks than without attacks when PMN were stimulated with OZ. We also demonstrated that O2–– generation correlated with the degree of BHR to inhaled histamine. These results suggest that PMN of asthmatic children, especially those with attacks, generate more active oxygen species than that of control subjects and that airway inflammation caused by O2–– may be closely related to BHR in subjects with bronchial asthma.
Distribution of Substance P-Immunoreactive and Calcitonin Gene-Related Peptide-Immunoreactive Nerves in Normal Human Lungs
Authors: Komatsu Takeshi , Yamamoto Masashi , Shimokata Kaoru , Nagura Hiroshi
Published: August 2009

The distribution of substance P (SP)-immunoreactive (IR) and calcitonin gene-related peptide (CGRP)-IR nerves in normal human lungs was immunohistochemically investigated. SP-IR and CGRP-IR nerves were found in lamina propria and adjacent to blood vessels, and also in some of the ganglia and nerve bundles in the submucosal area. Comparison of serial consecutive sections revealed that SP-IR and CGRP-IR nerves represented similar distribution patterns. CGRP-IR nerves were scarce in bronchial smooth muscles, whereas SP-IR nerves were always present therein. Neither SP-IR nor CGRP-IR nerves were found, however, among submucosal glands or in alveolar septa. These localization patterns were closely related to the functions of SP and CGRP in normal lungs.
A New Method of the Measurement of Nasal Secretion in Guinea Pigs
Authors: Namimatsu Akio , Yamaura Satomi , Go Koichiro , Tanimoto Hideji , Okuda° Minoru
Published: August 2009

A simple quantitative method to measure nasal secretion in guinea pigs is described. Nasal secretion was measured with a piece of cotton thread dyed with fluorescein at one end which was inserted into an anterior naris and kept there for 60 s. The stretch of color of a thread dyed with fluorescein was proportional to fluid volume and to increase in weight of a thread due to absorbed nasal secretion induced by nasal provocation. In addition, the stretch of color due to nasal secretion was associated with the score of rhinorrhea. Thus, it is considered that the amount of nasal secretion can be reflected to the length of the stretch of color. Each secretion on the ipsilateral and the contralateral sides induced by nasal provocation could be separately measured by this method. The amount of nasal secretion induced by allergen in passively sensitized guinea pigs could be reduced by pretreatment with ketotifen or flutropium. These results suggest that our method may serve as a quantitative test for nasal secretion in guinea pigs, which would be useful in the study of hypersecretory response in the allergic model or in evaluating the effect of antiallergic drugs on nasal allergy.
Characterization of Methylated Bovine Serum Albumin-Induced Allergic Inflammation in Rats
Authors: Hasegawa Takashi , Hirasawa Noriyasu , Watanabe Masako , Arakawa Masaaki , Tsurufuji Susumu , Ohuchi Kazuo
Published: August 2009

An air pouch type allergic inflammation in rats was induced using an insoluble cationic protein, methylated bovine serum albumin (MeBSA), as an antigen. Changes in vascular permeability, local tissue edema, histamine contents in the pouch fluid, and number of infiltrated leukocytes and chemotactic activity in the pouch fluid were analyzed during an 8-hour period after injecting the antigen solution into the air pouch of the immunized and nonimmunized rats. Vascular permeability during the first 30-min interval in the immunized rats was higher than that in the nonimmunized rats, reflecting a higher histamine level in the pouch fluid. However, both the increase in vascular permeability and histamine level in the immunized rats in this period were much lower than those induced by a soluble, noncationic antigen, azobenzenearsonate-conjugated acetyl bovine serum albumin. In the MeBSA-induced allergic inflammation model, a second peak of vascular permeability was induced at 2 h, and local tissue edema formation became apparent at 2 h, reaching a plateau at 4 h. A prominent increase in leukocyte infiltration, especially neutrophils, into the pouch fluid was induced at 4 h in accordance with an increase in chemotactic activity in the pouch fluid. These observations indicate that the acute phase of MeBSA-induced allergic inflammation is characterized by a weak anaphylactic response and a prominent neutrophil infiltration.
Ongoing IgE Synthesis by Atopic B Cells Is Enhanced by Interleukin-3 and Suppressed Directly by Interferon-Gamma in vitro
Authors: Matsumoto Tomoaki
Published: August 2009

Human recombinant interleukin-3 (rIL-3; 10 U/ml) consistently augmented spontaneous IgE synthesis by isolated atopic B cells in vitro, whereas rIL-4 (1–1,000 U/ml) failed to induce IgE synthesis by these cells. Recombinant interferon-γ (rIFN-γ) suppressed ongoing IgE production by atopic B cells in a dose-dependent manner. IFN-γ also inhibited IgE synthesis by a human myeloma cell line (U-266), demonstrating the direct effect of IFN-γ on the terminal differentiation of IgE-secreting plasma cells. IL-3 and IFN-γ from different sources displayed the same effects on IgE synthesis. Neutralizing antibodies toward IL-3 or IFN-γ abolished their activities toward IgE synthesis, supporting the specificity of the effect of these cytokines. The quantity of endogenous IFN-γ produced by stimulated T cells was significantly decreased in atopic patients compared to nonatopic controls, which might be responsible for the propensity of a high blood IgE level in atopic patients.
Induction of FcεRII/CD23 on Human T Cells by Excretory and Secretory Antigen of Dirofilaria immitis
Authors: Yamaoka Kunio A. , Tsukidate Setsuko , Higaki Megumu , Miyasaka Nobuyuki , Fujita Koichiro
Published: August 2009

We investigated the capacity of excretory and secretory antigen (ES) derived from living filarial worms in the induction of CD23 expression on human peripheral blood T cells by using flow cytometry. ES (10 μg/ml) significantly induced the expression of CD23 on human T cells. Moreover, increased CD23 expression was completely abolished by preincubation with specific antibody to ES. The results suggest that ES might play a certain role in IgE antibody production by induction of CD23 expression on T cells.
Aspirin-Sensitive Asthma: Significance of the Cyclooxygenase-Inhibiting and Protein-Binding Properties of Analgesic Drugs
Authors: Williams W.R. , Pawlowicz Anna , Davies B.H.
Published: August 2009

The in vitro release of endogenous and exogenous PgF2α from plasma and serum proteins by aspirin and other analgesic drugs has been studied by RIA and equilibrium-dialysis techniques, respectively. Before aspirin addition, the mean plasma level of PgF2α mesured by RIA was significantly lower in aspirin-sensitive asthma (ASA) patients (11.3 ± 6.5 pg/ml; n = 8) than in aspirin-tolerant asthma (ATA) patients (25.0 ± 11.4 pg/ml; n = 21). After aspirin addition (50 μg/ml) the mean PgF2α level detected in plasma by RIA was higher in ASA patients (97.6 ± 5.5 pg/ml) than in ATA patients (66.9 ± 4.5). The binding of [3H]PgF2α to serum protein was significantly inhibited by NSAIDs but not by paracetamol (0.2–1.0 mM). These results implicate PgF2α and the protein-binding property of analgesic drugs in the pathogenesis of aspirin-sensitive asthma.
Identification of Reactive Synthetic Gliadin Peptides Specific for Coeliac Disease
Authors: Devery J.M. , Bender V. , Penttila I. , Skerritt J.H.
Published: August 2009

Gluten intolerance (coeliac disease) is characterised by the development of a small intestinal lesion following exposure to the gliadin fraction after consumption of wheat and related cereals. Cellular immune mechanisms are thought to be responsible for gliadin toxicity, but the toxic sequence/s within gliadin have not been clearly established. A panel of synthetic gliadin peptides was tested using peripheral blood mononuclear cells from coeliac patients and two assays for cell-mediated immunity. Using the indirect leucocyte migration inhibition factor and the macrophage procoagulant activity assays, gliadin peptides which were located in the aminoterminal or the proline-rich domain of the alpha/beta gliadin molecule were coeliac-active. Peptides predicted by T cell algorithms or on the basis of homology to adenovirus Ad12 Elb protein and which were located in the proline-poor gliadin domains were inactive. Protein sequence studies which indicate significant homology in the proline-poor gliadin domains with a number of non-coeliac-toxic seed proteins also supported the hypothesis that the proline-rich domains may be more important in the pathogenesis of coeliac disease.
Detection of Antidrug IgG Antibodies in Patients with Adverse Drug Reactions to Amodiaquine
Authors: Clarke J.B. , Neftel K. , Kitteringham N.R. , Park B.K.
Published: August 2009

Antidrug IgG antibodies have been detected in some patients receiving amodiaquine (AQ). Antidrug antibodies were detected in 6/7 patients who experienced serious well-defined adverse drug reactions during malaria prophylaxis and in 7/22 patients who received comparable doses of the drug (at least 400 mg weekly ×6) but did not present with clinical adverse drug reactions. In contrast antidrug antibodies were not detected in 7 patients who received the drug for treatment (1.0–1.2 g total over 3 days). The specificity of the IgG response was defined by hapten inhibition experiments (IC50 value for AQ ranged between 0.050 and 0.282 μM) which suggest that the antibody recognised the drug linked to cysteine residues in protein via the 4-hydroxyanilino side chain. The data show that AQ is immunogenic in man and are consistent with the hypothesis that idiosyncratic adverse reactions to the drug have an immunological aetiology.
Human Upper Airway Epithelial Cell-Derived Granulocyte-Macrophage Colony-Stimulating Factor Induces Histamine-Containing Cell Differentiation of Human Progenitor Cells
Authors: Ohtoshi Takayuki , Tsuda Takashi , Vancheri Carlo , Abrams John S. , Gauldie Jack , Dolovich Jerry , Denburg Judah A. , Jordana Manel
Published: August 2009

Nasal polyps and allergic rhinitis are upper airway inflammatory conditions characterized by increased numbers of eosinophils and metachromatic cells in the epithelial layer of the nasal mucosa. The objective of the current studies was to investigate the potential contribution of epithelial cells to the accumulation of inflammatory cells in the tissue. We have established pure cultures of human upper airway epithelial cells from normal and inflamed nasal polyps and allergic rhinitis tissue and examined the ability of conditioned medium from these cells (EpCM) to induce differentiation of human hemopoietic progenitors in vitro. We show that, under appropriate culture conditions, EpCMs, particularly those from cells derived from inflamed tissues, induce histamine-containing cell differentiation of cells of the human HL-60 myeloid leukemia cell line. These EpCMs also induce the emergence of both eosinophil/basophil and granulocyte/macrophage colonies in methylcellulose cultures of human peripheral blood mononuclear cells. We also show that CMs from epithelial cells derived from inflamed tissues contain greater amounts of granulocyte-macrophage colony-stimulating factor (GM-CSF) compared to CMs from normal epithelial cells. Finally, we show that the histamine-containing cell differentiation of HL-60 cells as well as the colony growth induced by EpCM can be fully inhibited by preincubating this CM with a monoclonal neutralizing antibody to human GM-CSF. These studies: (a) illustrate the ability of human upper airway epithelial cells to secrete GM-CSF in vitro; (b) demonstrate differences between normal and inflamed tissue-derived epithelial cells, and (c) suggest that epithelial cells may, via the release of cytokines such as GM-CSF, contribute to the large tissue accumulation to inflammatory cells which characterizes nasal polyps, allergic rhinitis and, by extension, asthma.
Release of Mediators from Human Gastric Mucosa and Blood in Adverse Reactions to Benzoate
Authors: Schaubschläger W.W. , Becker W.-M. , Schade U. , Zabel P. , Schlaak M.
Published: August 2009

A study was carried out on 29 patients to investigate the amount of histamine liberation and release of platelet-activating factor and 6-keto-prostaglandin F1α from gastric mucosa and whole blood or mononuclear cells by sodium benzoate. The patients suffered from asthma (10), atopic dermatitis (7) and chronic urticaria (4). 8 patients with unrelated, non-immunologic diseases served as controls. In the oral provocation test (OPT) 3 patients experienced a recurrence of their original disease, whilst 1 asthmatic patient reacted with abdominal disorder. The release of histamine and prostaglandin from mucosa was significantly increased by sodium benzoate in comparison to the spontaneous release observed in patients. The mucosa of the control persons did not react to sodium benzoate. Furthermore, there was a significant difference in prostaglandin release between patients with positive OPT and the control persons. No difference could be found between patients with negative OPT and those with positive OPT. Additionally, in the mediator release from whole blood or mononuclear cells there was no obvious difference apparent. These results suggest a possible involvement of prostacyclin and histamine in adverse reactions to benzoate. Due to the sensitivity of the method, a mediator release from mucosa can already be demonstrated in a preclinical state of the pseudoallergic reaction in the absence of clinical symptoms.
Nasal Mucosal Hypersensitivity in Guinea Pigs Intermittingly Exposed to Cold
Authors: Namimatsu Akio , Go Koichiro , Hata Taeko
Published: August 2009

To develop an animal model for experimental nasal hypersensitivity and hyperreactivity, guinea pigs were subjected to intermittent exposure to cold temperature (intermittent cold stress, SART stress) for 5 consecutive days. In SART-stressed guinea pigs, nasal mucosal hypersensitivity to histamine evoking sneeze response and nasal hypersecretion in response to methacholine were observed. The hypersensitivity remained for further 7 days after being released from SART stress. On the other hand, such nasal mucosal hypersensitivity was not caused by a continuous cold stress alone, suggesting that intermittent exposure to cold may be of importance for the appearance of nasal mucosal hypersensitivity. In passively sensitized SART-stressed guinea pigs, the quantity of nasal secretion induced by an allergen was significantly increased compared with that of a group of normal animals. The expression of muscarinic acetylcholine receptor (m-ACh·R) became higher in SART-stressed guinea pigs. Thus, hypersensitivity and hyperreactivity in this system were found to be associated with an increase in density of m-ACh·R. SART-stressed guinea pigs will serve as an animal model for hypersensitivity in nasal mucosa, which would be useful for the study of nasal allergy.
Experimental Lung Injury Induced by Trimellitic Anhydride Inhalation on Guinea Pigs
Authors: Tao Yoshiaki , Sugiura Tsutomu , Nakamura Hiroshi , Kido Masamüsu , Tanaka Isamu , Kuroiwa Akio
Published: August 2009

Trimellitic anhydride (TMA) causes lung injury by inhalation exposure in humans. In order to investigate more precisely the mechanism of lung injury by TMA, an experimental animal model of TMA-induced lung injury was established. Guinea pigs were intramuscularly injected with trimellitylated bovine serum albumin (TM BSA) with complete Freund’s adjuvant (CFA). Appreciable amounts of antibodies against TM epitopes were detected in the sera of these animals. Guinea pigs that were passively sensitized with anti-TM antisera as well as the actively immunized animals developed hemorrhagic pneumonitis after TMA inhalation. It is well recognized that hyperimmune antisera of guinea pigs contain two subclasses of IgG antibodies, IgG1 and IgG2, which are known as homocytotropic and heterocytotropic antibodies, respectively. To determine the role of these antibodies in the airway injury with alveolar hemorrhages, they were isolated by gel filtration and by ion exchange column chromatography, and the guinea pigs that were sensitized with each of these antibodies were exposed to TMA inhalation. The extent of lung injury in these animals was quantitatively determined by the measurements of lung extravasation of Evans blue dye injected intravenously after TMA inhalation. Significant increases in the extravasation of dye were observed in both animal groups sensitized with IgG1 and IgG2 antibodies. In addition, results obtained with heat-treated antisera and IgG1 antibody did not significantly differ from those obtained with untreated samples. These results suggest that the lung injury resulting from TMA inhalation exposure can be induced with humoral antibodies, not only IgG1 but also IgG2, and that at least two types of allergic reactions are involved in the pathogenesis of lung injury.
Alpha-2-Macroglobulin-Kallikrein Complex: A Temperature-Sensitive Mediator in Contact-System-Induced Inflammation with a Potential Role in Late and Delayed Hypersensitivity Responses
Authors: Lasser E.C. , Lyon Sandra G. , Negrete Sylvia
Published: August 2009

Maximal complexing of α2-macroglobulin (α2M) and kallikrein (KK) occurs at a temperature of 22–24 rather than at 37 °C. The protease expressivity of the complex is also maximal at 22–24 °C. α2M-KK complex, sustained permeability changes in guinea pig skin. These findings suggest that the complex, rather than free KK, could play a role in the kinin release reported in some late-phase reactions, some instances of delayed-type hypersensitivity and some cold-induced reactions.
Characterization of a Monoclonal Antibody (RJ5) against the Immunodominant 41-kD Antigen of Candidaalbicans
Authors: Shen Horng-Der , Choo Kung-Bung , Yu Kwok-Woon , Ling Win-Lin , Chang Fu-Chung , Han Shou-Hwa
Published: August 2009

A 41-kD component of Candida albicans was identified to be the major antigen radioimmunoprecipitated by antibodies with increased titers in the sera of patients with invasive candidiasis. A mouse monoclonal antibody (RJ5) was generated which, by immunoblotting, showed positive reactivity to the immunoprecipitated 41-kD component. By two-dimensional gel electrophoresis and immunoblotting, MoAb RJ5 was shown to react with different isoforms of the 41-kD component with pI values from 6.1 to 6.9. Furthermore, MoAb RJ5 showed positive reactivity to cytoplasmic antigens of C. albicans by frozen section and immuno-peroxidase staining. By SDS-polyacrylamide gel electrophoresis and immunoblotting, MoAb RJ5 showed no cross-reactivity to antigens of Candida tropicalis and Candida parapsilosis. The epitope of the 41-kD molecule recognized by MoAb RJ5 was susceptible to treatment of proteinase K at concentrations of ≥ 5 μg/ml, and was relatively resistant to periodate oxidation with concentration of NaIO4 up to 20 mM. This MoAb may be useful in the purification and characterization of the immunodominant 41-kD antigen of C. albicans, and as a probe in the detection of Candida antigens in the sera of patients with invasive candidiasis.
Immunocytochemical Determination of the Role of Alveolar Macrophages in Endotoxin Processing in vitro and in vivo
Authors: Keller, III George E. , Dey Richard D. , Burrell Robert
Published: August 2009

Endotoxin (lipopolysaccharide or LPS) inhalation has been implicated in increased pulmonary edema, most likely due to activation of an inflammatory response. The purpose of this study was to determine the cell types in the lung responsible for binding inhaled lipid A from Enterobacter agglomerans LPS. Five-hour exposures of aerosolized lipid A resulted in measurable pulmonary edema in hamsters, as determined by the accumulation of lung water. Immunocytochemistry was used to localize the inhaled lipid A in the cell types in the lung. Alveolar macrophages had decreased levels of lipid A as compared to unexposed controls, suggesting a possible metabolism by the macrophages. In vitro exposure of macrophages to lipid A resulted in a time-dependent clearance of lipid A which was inversely related to its concentration. Alveolar macrophages thus appear to be responsible for the removal of inhaled lipid A in this model and may initiate the physiological events which bring about pulmonary edema.
Successful Induction of Allergic Contact Dermatitis to Mercury and Chromium in Mice
Authors: Vreeburg Karel J.J. , Groot Klaasde , van Hoogstraten Ingrid M.W. , von Blomberg Mary E. , Scheper Rik J.
Published: August 2009

Availability of reproducible mouse models for allergic contact dermatitis (ACD) to the metal allergens nickel, mercury and chromium, would be of great value for pathogenetic and preventive studies. We explored epicutaneous sensitization to nickel, mercury and chromium in mice in which oral grooming of the sensitization site was prevented by a plaster cast around the abdomen and lower thorax. This procedure was based on earlier findings that oral ingestion of allergen could prevent contact sensitization. The present results show that BALB/c mice can be readily sensitized to mercury and chromium using this epicutaneous casting method, without the further use of adjuvants. With nickel, however, neither this method, nor conventional methods involving the use of Freund’s complete adjuvant (FCA) were effective.
Soluble CD23 Increases IL-3 Induction of Histamine Synthesis by Human Bone Marrow Cells
Authors: Arock M. , Michel L. , Dalloul A.H. , Guillosson J.J. , Debré P. , Mossalayi M.D.
Published: August 2009

Interleukin 3 (IL-3) and soluble CD23 (sCD23/IgERII) have similar capacities to induce basóphil development from human bone marrow (BM) precursors. IL-3 also has the capacity to induce de novo histamine synthesis from human BM cells. In the present study, we examined the effect of sCD23 on histamine synthesis by human BM cells and its relationship with IL-3. Our data showed that recombinant 25 KDa sCD23, although on its own failed to induce hitamine release, significantly enhanced IL-3-stimulated histamine synthesis by BM cells. These data provide further support for the cytokine-like role of sCD23 on hematopoietic cells.
IgM in the Airways of Asthma Patients
Authors: Hol B.E.A. , van de Graaf E.A. , Out T.A. , Hische E.A.H. , Jansen K.M.
Published: August 2009

In order to obtain information on the role of IgM in the pathogenesis of asthma, we assayed IgM in the broncho-alveolar lavage fluid (BALF) from 14 non-smoking patients with asthma and 9 non-smoking healthy persons, using an ELISA. The concentrations of IgM in the epithelial lining fluid (ELF) were calculated on the basis of urea in serum and BALF. The concentrations of IgM in ELF after correction for serum IgM, cQIgM, from the patients as a group were higher than those from controls (Mann-Whitney U test, p < 0.05). The IgM in BALF and serum, analysed by sucrose density gradient centrifugation was pentameric. The transudation of IgM from blood into ELF was compared with that of α2 macroglobulin (A2M), a protein with a similar molecular mass. In 6 patients the cQIgM/cQA2M value was above the upper value of the range for controls suggesting abnormal local production of IgM. The presence of the secretory component (SC) linked to IgM was determined by ELISA. The percentage of SC-IgM in BALF was increased in 5 out of 7 patients tested. The increased local IgM concentrations and its increased local production may suggest a role for local IgM-mediated reactions in the inflammatory events associated with asthma.
Allergenic and Antigenic Cross-Reactivities of Group IX Grass Pollen Allergens
Authors: Zhang Lei , Kisil Fred T. , Sehon Alec H. , Mohapatra Shyam S.
Published: August 2009

The allergenic and antigenic cross-reactivities between a major recombinant Poa pratensis(Poa p) IX allergen, rKBG8.3, and its corresponding proteins of different grass pollens were examined. Immunoblotting of the proteins of thirteen different grass pollens using anti-rKBG8.3 antibodies indicated that Poa p IX-like proteins are present in ten other grass pollens, albeit in variable amounts and polymorphic forms. These proteins ranged in size from 20 to 88 kDa in different grass pollens. The percent relative binding determined for each grass pollen extract using allergic human sera showed a significant correlation (r = 0.891) with that of anti-rKBG8.3 antiserum. Moreover, there was a strong association (r = 0.901) between the Kentucky bluegrass extract and rKBG8.3 with respect to their inhibition of the binding of human IgE antibodies to allergens in grass pollen extracts. Taken together, these results suggest that the allergenic and antigenic epitopes of the Poa p IX-related proteins in some but not all grass pollens are similar in structure and specificities. It is concluded that the group IX allergens constitute a major family of homologous proteins in several grass pollens.
Role of Interleukin-5 in Local Accumulation of Eosinophils in Mouse Allergic Peritonitis
Authors: Kaneko Masayuki , Hitoshi Yasumichi , Takatsu Kiyoshi , Matsumoto Shuu
Published: August 2009

The injection of ragweed pollen extracts into the peritoneal cavity of actively immunized mice caused an accumulation of eosinophils in the cavity. We investigated the role of interleukin-5 (IL-5), one of the T-cell-derived factors, in this in vivo local accumulation. Ciclosporin, an immunosuppressant that mainly suppresses T cell functions, dose-dependently inhibited the in vivo local accumulation. Injections of the neutralizing antibody to IL-5 into the cavity decreased the number of the accumulating eosinophils. When the peritoneal cells of the immunized mice were cultured in the presence of the antigen in vitro, IL-5 activity was detected in the culture supernatant. The results indicate that IL-5 plays an important role in the in vivo local accumulation of eosinophils after an antigen challenge.
In vitro Formation of Complement Activation Products by Lipopolysaccharide Chemotypes of Salmonellaminnesota
Authors: Gardiner James S. , Keil Lynn B. , DeBari Vincent A.
Published: August 2009

We have applied immunoassays for complement activation products C4d, fragment Bb and the protein S-C5b-9 neoantigen (S-MAC) to assess activation of classical, alternative and terminal pathways, respectively, by lipopolysaccharides (LPS) from the smooth strain (SS) of Salmonella minnesota and the shallow rough (core) mutants R60, R345, R5 and R7. Incubations of sera (n = 6) with LPS generated small and insignificant quantities of Bb and S-MAC in the case of Rb, Re and Rd chemotypes and slightly greater quantities with Ra. SS-LPS brought about significant (p = 0.01) increases in the formation of both Bb and S-MAC. No significant changes were observed in the concentration of C4d. Polymyxin B enhanced Bb and S-MAC production by SS-LPS, optimally at the lowest concentration of polymyxin B studied, 10 ng/ml. These data confirm and extend observations about complement activation by LPS and suggest that immunoassay may be useful in studying mechanisms of complement activation.
Activation of Human Vascular Endothelial Cells by IFN-γ: Acquisition of HLA Class II Expression, TSST-1-Binding Activity and Accessory Activity in T Cell Activation by the Toxin
Authors: Araake Minako , Uchiyama Takehiko , Imanishi Ken’ichi , Yan Xiao-Jie
Published: August 2009

Human umbilical vascular endothelial cells (HUVEC) stimulated with recombinant human IFN-γ were investigated for expression of HLA class II molecules, toxic shock syndrome toxin-1 (TSST-l)-binding activity and accessory cell (AC) activity in TSST-1-induced T cell activation. HUVEC stimulated with recombinant human IFN-γ ranging from 10 to 1,000 units/ml for 72 h express HLA class II molecules. Intensity of the expression was dependent on the concentration of IFN-γ. HUVEC stimulated with 1,000 units/ml of IFN-γ for 72 h exhibited 125I-TSST-l-binding that was blocked by the unlabeled toxin and monoclonal antibodies (mAb) to DR/DP. The activity was not removed by paraformaldehyde fixation. The IFN-γ-stimulated HUVEC exhibited AC activity in TSST-1-induced IL-2 production by T cells from human peripheral blood mononuclear cells. The activity was blocked by mAb to DR. The above two activities were also observed in L cells transfected with DR2 genes but not in the unstimulated HUVEC and control L cells. In view of the fact that TSST-1 binds directly to HLA class II molecules and activates human T cells in association with HLA class II molecules on AC, it is likely that the acquisition of TSST-1-binding activity and AC activity in the toxin-induced T cell activation is mediated by the expression of HLA class II molecules. Vascular endothelial cells may play an important role in the development of pathological changes in TSS patients.
Tumor Necrosis Factor Alters Cytoskeletal Organization and Barrier Function of Endothelial Cells
Authors: Camussi G. , Turello E. , Bussolino F. , Baglioni C.
Published: August 2009

Treatment of human umbilical cord vein endothelial cells with tumor necrosis factor results in marked changes in cell shape and cytoskeletal organization. After 4 h of treatment, these cells loose reciprocal contacts with the formation of intercellular gaps. This retraction reaches a maximum after 6 h when most stress fibers staining for F-actin disappear and vinculin becomes diffused in the cytoplasm. Such changes spontaneously reverse after 24 h in the presence of tumor necrosis factor or after 2 h of incubation in fresh medium. After treatment with tumor necrosis factor, endothelial monolayers become permeable to albumin because of gaps that form between cells. Normal human serum, plasma α1-proteinase inhibitor and an anti-inflammatory peptide that decrease synthesis of platelet-activating factor inhibit the changes induced by tumor necrosis factor. Furthermore, receptor antagonists of platelet-activating factor have the same effect. These findings suggest that platelet-activating factor is a secondary mediator responsible for the changes in cell shape and cytoskeletal organization, and for the leakiness of endothelial monolayers.
Serum Levels of Interleukin 4, Interleukin 6 and Interferon-Gamma following in vivo Isotype-Specific Activation of IgE Synthesis in Humans
Authors: Lebedin Yuri S. , Raudla Ludmila A. , Chuchalin Alexandre G.
Published: August 2009

It is now generally accepted that interleukin 4 (IL4), interleukin 6 (IL6) and interferon-γ (IFNγ) play main roles in the regulation of human IgE synthesis. This concept is based mainly on in vitro data. To obtain corresponding in vivo data, we determined IL4, IL6 and IFNγ by immunoassays in sera collected from 4 atopic patients following a clinical trial of selective IgE apheresis (plasmaimmunoadsorption). This treatment removes several milligrams of IgE from patient’s blood and is suggested to induce strong and isotype-specific activation of the IgE system. Serum IgE levels restored rapidly within 3–5 days after IgE apheresis. However, very low and constant levels of IL4 (from < 50 to 130 pg/ml) and IL6 (from < 300 to 920 pg/ml) were detected in the sera of the treated patients. Serum IFNγ was absent before treatment (concentrations less than 0.5 U/ml) and increased to low but detectable levels (0.90 and 8.05 U/ml) on the day following the last IgE apheresis in 2 of 4 patients. In our opinion, the data presented argue against in vivo participation of IL4 and IL6 in the activation of the human IgE system, at least in atopic patients under constant allergen exposure.
Macrophage Reverse Cholesterol Transport in Mice Expressing ApoA-I Milano
Authors: Alexander Eric T. , Weibel Ginny L. , Joshi Michelle R. , Vedhachalam Charulatha , de la Llera-Moya Margarita , Rothblat George H. , Phillips Michael C. , Rader Daniel J.
Published: August 2009

Objective—

To compare the abilities of human wild-type apoA-I (WT apoA-I) and human apoA-I
Milano
(apoA-I
M
) to promote macrophage reverse cholesterol transport (RCT) in apoA-I–null mice infected with adeno-associated virus (AAV) expressing either WT apoA-I or apoA-I
M
.

Methods and Results—

WT apoA-I– or apoA-I
M
–expressing mice were intraperitoneally injected with [H
3
]cholesterol-labeled J774 mouse macrophages. After 48 hours, no significant difference was detected in the amount of cholesterol removed from the macrophages and deposited in the feces via the RCT pathway between the WT apoA-I and apoA-I
M
groups. Analysis of the individual components of the RCT pathway demonstrated that the apoA-I
M
–expressing mice promoted ATP-binding cassette transporter A1 (ABCA1)-mediated cholesterol efflux as efficiently as WT apoA-I but that apoA-I
M
had a reduced ability to promote cholesterol esterification via lecithin cholesterol-acyltransferase (LCAT). This resulted in reduced cholesteryl ester (CE) and increased free cholesterol (FC) levels in the plasma of mice expressing apoA-I
M
compared to WT apoA-I. These differences did not affect the rate of delivery of labeled cholesterol to the liver via SR-BI–mediated selective uptake or its subsequent excretion in the feces.

Conclusion—

Within the limits of the in vivo assay, WT apoA-I and apoA-I
M
are equally efficient at promoting macrophage RCT, suggesting that if apoA-I
M
is more atheroprotective than WT apoA-I it is not attributable to an enhancement of macrophage RCT.
Laminar Shear Stress Regulates Endothelial Kinin B1 Receptor Expression and Function
Authors: Duchene Johan , Cayla Cécile , Vessillier Sandrine , Scotland Ramona , Yamashiro Kazuo , Lecomte Florence , Syed Irfan , Vo Phuong , Marrelli Alessandra , Pitzalis Costantino , Cipollone Francesco , Schanstra Joost , Bascands Jean-Loup , Hobbs Adrian J. , Perretti Mauro , Ahluwalia Amrita
Published: August 2009

Objective—

The proinflammatory phenotype induced by low laminar shear stress (LSS) is implicated in atherogenesis. The kinin B1 receptor (B1R), known to be induced by inflammatory stimuli, exerts many proinflammatory effects including vasodilatation and leukocyte recruitment. We investigated whether low LSS is a stimulus for endothelial B1R expression and function.

Methods and Results—

Human and mouse atherosclerotic plaques expressed high level of B1R mRNA and protein. In addition, B1R expression was upregulated in the aortic arch (low LSS region) of ApoE
−/−
mice fed a high-fat diet compared to vascular regions of high LSS and animals fed normal chow. Of interest, a greater expression of B1R was noticed in endothelial cells from regions of low LSS in aortic arch of ApoE
−/−
mice. B1R was also upregulated in human umbilical vein endothelial cells (HUVECs) exposed to low LSS (0 to 2 dyn/cm
2
) compared to physiological LSS (6 to 10 dyn/cm
2
): an effect similarly evident in murine vascular tissue perfused ex vivo. Functionally, B1R activation increased prostaglandin and CXCL5 expression in cells exposed to low, but not physiological, LSS. IL-1β and ox-LDL induced B1R expression and function in HUVECs, a response substantially enhanced under low LSS conditions and inhibited by blockade of NFκB activation.

Conclusion—

Herein, we show that LSS is a major determinant of functional B1R expression in endothelium. Furthermore, whereas physiological high LSS is a powerful repressor of this inflammatory receptor, low LSS at sites of atheroma is associated with substantial upregulation, identifying this receptor as a potential therapeutic target.
Foxp3 Regulates Megakaryopoiesis and Platelet Function
Authors: Bernard Jamie J. , Seweryniak Kathryn E. , Koniski Anne D. , Spinelli Sherry L. , Blumberg Neil , Francis Charles W. , Taubman Mark B. , Palis James , Phipps Richard P.
Published: August 2009

Objective—

Platelets are crucial for hemostasis and are vital regulators of inflammation. Foxp3 is a key transcription factor for T regulatory cell development. Humans with IPEX (immune dysregulation, polyendocrinopathy, enteropathy, x-linked) and the
scurfy
(

Foxp3
sf

) mouse have mutations in the
Foxp3
gene that lead to a host of pathologies including autoimmunity and skin diseases. Scurfy mice and some humans with IPEX are also thrombocytopenic. The purpose of this study was to determine whether the absence of functional Foxp3 leads to defects in megakaryocytes and platelets.

Methods and Results—

We discovered that human and mouse megakaryocytes express Foxp3 mRNA and protein. Using shRNA and

Foxp3
sf

mice, we demonstrated that Foxp3-deficient mouse and human megakaryocyte progenitors exhibited proliferation defects. Striking platelet abnormalities were observed in both an IPEX patient and

Foxp3
sf

mice. Impaired platelet spreading and release of TGF-β and CD40 ligand (CD40L), and abnormal levels of plasma CD40L were observed in a case of IPEX syndrome.

Foxp3
sf

mice were thrombocytopenic and had increased platelet volume and altered serum levels of CD40L, TXB
2
, and TGF-β.

Conclusion—

These findings provide compelling new evidence that Foxp3 is needed for proper megakaryopoiesis and plays a role in regulating platelet function including spreading and release.