bluuebunny/crossref_metadata_2025 huggingface dataset:
Subepithelial Hydrostatic Pressure May Regulate Plasma Exudation across the Mucosa
Authors: Persson Carl G.A. , Erjefält Ingrid , Gustafsson Birgitta , Luts Anders
Published: August 2009

This study demonstrated in guinea pig tracheal tubes in vitro that small increases in serosal hydrostatic pressure caused significant mucosal crossing of serosal macromolecules. Reversibility and repeatabilityof this passage agree with inflammatory stimulus-induced appearance of exuded plasma in airway lumen invivo. Bradykinin, histamine, and terbutaline, which induce and inhibit, respectively, plasma exudation in vivo, were without effect on the present in vitro permeability. Carbachol, similar to histamine, contracted the trachea, and did not increase, but rather decreased the pressure-induced luminal entry of serosal macromolecules.It is proposed that a plasma-exudation-induced hydrostatic pressure load transiently separates epithelial cells, providing a direction-selective and non-injurious intercellular pathway for passage of bulk plasma exudateinto the airway lumen. This mechanism would allow potent plasma protein systems to operate on mucosal surfaces at sites of insults without compromising the mucosa as a barrier to luminal solutes.
IgE-Mediated Allergic Reactions to Potatoes
Authors: Wahl R. , Lau Susanne , Maasch H.J. , Wahn U.
Published: August 2009

We investigated sera of 12 patients with IgE-mediated hypersensitivity reactions to potato, using different in vitro methods. Radioallergosorbent test classes 2–4 (7.6–46.5% binding) were measured with potato allergen disks. Immunoblot detected allergen bands with an isoelectric point range of approximately 4.5–5.2 and mainly in a molecular weight range between 16.00 and 30.00 kD. But also at molecular weights of 43.00 and 65.00 kD allergen bands (IgE specificities) could be detected. Histamine release assay performed with isolated fractions of the potato extract showed a great individual variation and positive results of fractions of molecular weights between 10.00 and 80.00 kD. From our data we conclude that raw potato is a ‘multiallergenic’ vegetable.
Obtention of Monoclonal Antibodies against Human IgG4 using Two Different Immunization Strategies: Development of a Biotin-Based ELISA for IgG4 Quantitation
Authors: Nogales Lucia Jimeno , Carreira José , Lombardero Manuel
Published: August 2009

Several monoclonal antibodies (MAbs) against human IgG4 have been produced in two fusion experiments. In fusion I, Balb/c mice were conventionally immunized and most of the antibodies obtained recognized IgG class-specific epitope(s). Therefore, a second fusion experiment was performed using IgG4-im-munized mice that had been made tolerant to epitopes shared by the four IgG subclasses, and most of the hybridomas derived from this second fusion secreted antibodies specific for IgG4. Five MAbs which were found to be specific for both IgG4 isoallotypes (4a and 4b) were selected from the two fusion experiments. These antibodies define at least two non-overlapping epitopes located on the Fab and Fc regions of IgG4 molecule, respectively. Two MAbs were used to develop a two-site ELISA for IgG4 quantitation, in which one MAb (anti-Fab) was immobilized in the solid phase and the other one (anti-Fc) was biotin-labeled. The assay had a detection limit below 1 ng/ml and a practical working range between 4 and 200 ng/ml. The use of these MAbs should improve our understanding of the role of IgG4 in different clinical conditions.
Gut Hypersensitivity Reactions in Guinea Pigs following Vaccination or Infection with Trichostrongylus colubriformis
Authors: Ward Colin W. , Wagland Barry M.
Published: August 2009

A quantitative approach to measuring gut hypersensitivity responses to parasite antigens is described. Guinea pigs exposed to infection with Trichostrongylus colubriformis, were sensitised to excretory-secretory products and homogenates of third-stage larvae (L3), fourth-stage larvae (L4) and adult worms. Εhe nature of these responses and their modification by pharmacological agents were typical of immediate-type hypersensitivity reactions. Gut sensitivity was greatest in the anterior half of the small intestine, persisted for at least 12 months and was more pronounced against L3 antigens than against L4 or adult preparations. Guinea pigs injected with homogenates of L3, L4 or adult worms were sensitised to these homogenates, but not to T. colubriformis excretory-secretory products.
Characteristics of MTT as an Indicator of Viability and Respiratory Burst Activity of Human Neutrophils
Authors: Pruett Stephen B. , Loftis Audrey Y.
Published: August 2009

In the present study, we have examined the suitability of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) for assessing the viability and respiratory burst activity of human neutrophils. MTT is a good indicator of the relative viability of neutrophils which have been in culture up to 24 h, if sufficient serum (10% fetal calf serum) is present in the medium and the cells being compared have been in culture for a similar period of time. However, it is not suitable for exact assessment of percent viable cells in a cell population with low viability. The effectiveness of MTT as an indicator of respiratory burst activity is demonstrated by the sharp increase in MTT reduction induced by respiratory burst stimuli and by the ability of superoxide dismutase to inhibit 75% of MTT reduction by stimulated neutrophils. Unlike nitroblue tetrazolium (which is reduced primarily intracellularly) and cytochrome c (which is reduced extracellularly), MTT is apparently reduced both intracellulary and extracellulary by activated neutrophils.
Elevation of Neutrophil Chemotactic Activity in the Human Serum and the Decreased Number of Neutrophils after Platelet-Activating Factor Inhalation
Authors: Miyagawa Hidefumi , Nabe Makoto , Hopp Russell J. , Agrawal Devendra K. , Bewtra Againdra K. , Townley Robert G.
Published: August 2009

We previously reported a significant decrease in neutrophils at 5 min and an increased number of neutrophils at 15 min after PAF-acether inhalation. To investigate the mechanism, we measured the neutrophil chemotactic activity of the sera. Neutrophil chemotactic activity (NCA) of the serum at 5 min was greater than at baseline (p < 0.01) or at 15 min (p < 0.05). Furthermore, we investigated the correlation between the change in NCA and decreased number of neutrophils. A significant correlation was found between these two at 5 min (r = 0.751 p < 0.05). These data suggest that an elevation of NCA may play an important role in the decrease of circulating neutrophils after PAF-acether inhalation.
Histamine Release Induced by Radiographic Contrast Media
Authors: Amon Ulrich , Ennis Madeleine , Lorenz Wilfried , Schnabel Michael , Schneider Claus
Published: August 2009

Intravascular application of radiographic contrast media (RCM) can cause adverse allergic/pseudoallergic reactions in certain individuals. In view of the increased risk for the reactions associated with cardiovascular diseases, we have investigated histamine release from isolated mast cells derived from the peritoneal cavity and the lung of normotensive and spontaneously hypertensive rats. Six commonly used RCM were tested in their clinical formulations: Angiographin® (amidotrizoate), Ray vist® (ioglycate), Telebrix® (ioxithalamate), Hexabrix® (ioxaglate), Solutrast® (iopamidol), and Ultravist® (iopromide). The three RCM with low osmolality (Hexabrix®, Solutrast® and Ultravist®) released little histamine from all cell populations tested. Mast cells derived from spontaneously hypertensive rats released significantly more histamine following challenge with high osmolar RCM than those derived from normotensive rats. Higher concentrations were required to elicit release from the peritoneal cells than from the pulmonary cells. These results indicate a role for underlying cardiovascular diseases in mediator release and should be followed up by investigations on basophils or mast cells obtained from control subjects and patients with cardiovascular disease.
Serum Antibodies Reactive with Saccharomyces cerevisiae in Inflammatory Bowel Disease: Is IgA Antibody a Marker for Crohn’s Disease?
Authors: Barnes R.M.R. , Allan Susan , Taylor-Robinson C.H. , Finn R. , Johnson P.M.
Published: August 2009

Sera from patients with Crohn’s disease (CD) and ulcerative colitis (UC) have been evaluated for antibodies reactive with Saccharomyces cerevisiae (anti-Sacc antibodies) using an enzyme-linked immunoassay (ELISA). IgG anti-Sacc antibodies were detected in 63% (25/40) of CD patients, compared with 15% (4/27) of UC patients (p <0.001) and 8% (5/60) healthy adult controls (p <0.001). Furthermore, the prevalence of detectable IgG anti-Sacc antibodies in adult patients with coeliac disease, dermatitis herpetiformis, irritable bowel syndrome or atopic eczema was not significantly different to controls. In comparison, the prevalence of detectable IgG anti-Escherichia coli antibodies was not significantly different between CD (75%) or UC (79%) patients. More particularly, elevated levels of serum IgA anti-Sacc antibodies were detected in 17/40 CD patients, but in none of the 27 UC patients. These data confirm that serum antibodies reactive with S. cerevisiae are strongly associated with CD and further show that serum IgA anti-Sacc antibodies may be specific for this disorder.
Nedocromil Sodium Prevents the Release of 15-Hydroxyeicosatetraenoic Acid from Human Bronchial Epithelial Cells Exposed to Toluene Diisocyanate in vitro
Authors: Mattoli Sabrina , Mezzetti Maurizio , Fasoli Angelo , Patalano Francesco , Allegra Luigi
Published: August 2009

The in vivo exposure to an asthmogenic stimulus, toluene diisocyanate (TDI), causes airway epithelial damage associated with inflammation and increased bronchial hyperresponsiveness. The latter mechanisms might partly be mediated by the release of eicosanoids from bronchial epithelial cells. We have previously demonstrated that the in vitro exposure of bronchial epithelial cells to TDI results in the release of immunoreactive 15-hydroxyeicosatetraenoic acid (15-HETE), a product of activation of the 15-lipoxygenase pathway with inflammatory properties. In the present study we show that TDI-induced release of 15-HETE from epithelial cells can be prevented by nedocromil sodium, an anti-asthmatic drug with anti-inflammatory properties. This mode of action of the compound may explain its clinical effectiveness in asthma.
Preparation of Allergen Extracts from the Green Alga Chlorella
Authors: Tiberg Ebba , Rolfsen Wenche , Einarsson Roland
Published: August 2009

Allergen extracts were prepared from different raw materials of three strains of the unicellular green alga Chlorella (C. vulgaris, C. homosphaera and C. saccharophila). Growth variation, batch variation and composition of partially purified extracts were studied by analyzing total protein and carbohydrate content, protein and IgE-binding patterns and allergenic potency. Chlorella allergens were produced in the same proportions in early and late exponential growth phases but appeared partly degraded in the stationary phase. Uniform growth and extraction from Chlorella cells in the late exponential phase were demonstrated. Partially purified extracts from cells in late exponential phase contained 50% protein of the dry weight compared to 20% in crude extracts. The carbohydrate content was 20–30% of the dry weight of both crude and purified extracts. The allergenic potency of the purified extracts was 150–340% of the crude extracts, as assayed by RAST inhibition. The purified C. vulgaris extract showed a reduced IgE binding compared to crude extracts, while C. homosphaera and C. saccharophila extracts exhibited consistent IgE binding and are therefore suitable for in vivo and in vitro studies of allergenic properties of Chlorella.
Anti-Inflammatory Effects of Onions: Inhibition of Chemotaxis of Human Polymorphonuclear Leukocytes by Thiosulfinates and Cepaenes
Authors: Dorsch W. , Schneider E. , Bayer T. , Breu W. , Wagner H.
Published: August 2009

Seven different synthetic thiosulfinates, and cepaene- and/or thiosulfinate-rich onion extracts were found to inhibit in vitro the chemotaxis of human granulocytes induced by formyl-methionine-leucinephenylalanine in a dose-dependent manner and at a concentration range of 0.1–100 μM. Diphenylthiosulfinate showed the highest activity and was found to be more active than prednisolone. The anti-inflammatory properties of onion extracts are related, at least in part, to the inhibition of inflammatory cell influx by thiosulfinates and cepaenes.
Relationship between LTC4 Generation of Hypodense Eosinophils and Bronchial Hyperreactivity in Asthmatic Children
Authors: Schauer U. , Daume U. , Müller R. , Riedel F. , Gemsa D. , Rieger C.H.L.
Published: August 2009

n 19 asthmatic children aged 6–16 years, the degree of bronchial hyperreactivity was determined in relationship to the concentration of inhaled histamine which caused a fall of the specific conductance (sGaw) to 60% of the baseline value PC60sGaw.· At the time of lung function testing, a sample of heparinized blood was obtained from each patient. Eosinophils were purified and separated into a normodense and hypodense fraction by Percoll gradient centrifugation. After in vitro stimulation by ionophore A 23187, the leukotriene C4 (LTC4) content was determined in the culture supernatants. Hypodense eosinophils of the 13 children with a histamine threshold lower than 1 mg/ml generated significantly (p <0.01) larger amounts of LTC4 (0.8–36.3 ng/106 cells) when compared to 6 children with a histamine threshold higher than 1 mg/ml (0.7–12.1 ng/106 cells) and 12 healthy controls (0.4–8.2 ng/106 cells). Preincubation of eosinophils with platelet activating factor (PAF) induced an enhanced LTC4 production, not only in hypodense cells from both asthmatic groups but also in normodense cells from patients with severe hyperresponsiveness. These results are consistent with other results which suggest an important role of eosinophils, their activation by PAF and enhanced release of spasmogenic LTC4 in the pathogenesis of asthma.
Modulation of the Immune Response to Allergens: Phospholipase A Degradation Products Suppress IgG and IgE Response in Mice
Authors: Pesce Amadeo J. , Freisheim James M. , Litwin Allen , Michael Gabriel
Published: August 2009

Degradation products of phospholipase A2 (PLA2), the major allergen of honeybee venom, were prepared by peptic digestion of the enzyme and fractionated by molecular exclusion chromatography. The largest of the isolated products, P-l, was found to be a centrally nicked PLA2 molecule with a loss of several amino acids. It was examined for its antigenic, immunogenic and immunosuppressive properties. This peptic product was unable to block antigen-antibody reactions between PLA2 and anti-PLA2 antibodies and was incapable of inducing passive cutaneous anaphylaxis in rat skin sensitized with mouse anti-PLA2 IgE antibodies. BDF1 female mice injected intravenously with P-1 exhibited suppressed IgG and IgE anti-PLA2 responses to a subsequent PLA2 immunization. Intravenous administration of P-1 to mice with ongoing immune response to PLA2 resulted in decline of antigen-specific IgE titers. Transfer of splenic T cells from mice injected with P-1 into naive recipients caused profound suppression of the response to subsequent immunization with PLA2 indicating presence of PLA2-specific T suppressor cells. These results indicate that a degradation product of PLA2 prepared by peptic digestion lost antigenic and immunogenic properties of the intact enzyme but retained immunosuppressive properties, thus offering a novel approach to allergen immunotherapy.
CD1-Positive Epidermal Langerhans Cells in Regressed Tuberculoid and Lepromatous Leprosy Lesions
Authors: Narayanan R.B. , Girdhar A. , Girdhar B.K.
Published: August 2009

A comparison was made of the numbers of epidermal Langerhans cells in active and regressed lesions of tuberculoid and lepromatous leprosy using the OKT6 and OKIa monoclonal antibodies. A reduction in the numbers of CD1+ epidermal Langerhans cells was noticed in the regressed lesions of both the tuberculoid and lepromatous leprosy lesions unlike the active lesions. The majority of infiltrates in both types of regressed lesions were HLA-DR+ and CD1––.
Interleukin-4 Gene Expression in High and Low IgE Responder Mice
Authors: Mori Akio , Yamamoto Kazuhiko , Suko Matsunobu , Watanabe Naohiro , Ito Mamoru , Miyamoto Terumasa , Okudaira Hirokazu
Published: August 2009

Interleukin-4 (IL-4) gene expression in murine spleen cells was examined by stimulation with con-canavalin A (Con A). Spleen cells from a DBA/2 strain of mice, a high IgE responder, expressed IL-4 mRNA within 3 h after incubation with Con A. Maximal IL-4 mRNA expression was observed 6–9 h after stimulation. The amount of IL-4 mRNA induced by Con A was greatest in spleen cells obtained from high IgE responder strains of mice. A trace amount of mRNA was induced in spleen cells from low IgE responder (SJL) mice. The amount of mRNA induced in spleen cells from an intermediate IgE responder (C57BL/6) was smaller than that from high responders, but significantly greater than that from low responders. Spleen cells from IgE nonresponders (SJA/9) developed only a negligible amount of IL-4 mRNA after stimulation with Con A. Time course and optimal concentration of Con A for the expression of IL-4 mRNA were essentially the saηie in high (DBA/2) and low (SJL) responder strains of mice. The decreased expression of IL-4 mRNA in low IgE responder spleen cells upon stimulation was not due to the decrease in Thy-1-positive cells or L3T4-positive cells in the spleen. The results obtained in the present study may indicate that high and low IgE responder traits are determined depending on their levels of IL-4 mRNA expression.
Remarkable Increases of Salivary IgE Levels in Allergic Syndromes
Authors: Negretti F. , Casetta P.
Published: August 2009

Serum and salivary IgE levels were tested by indirect ELISA in patients with allergic contact dermatitis and allergic asthma or rhinitis. The remarkable increases of salivary IgE (from 928.4% mean augmentation in allergic asthma or rhinitis to 2686.2% in allergic contact dermatitis) suggest the idea of using saliva in IgE titration.
Determination of Cow Milk Formula Allergenicity in the Rat Model by in vitro Mast Cell Triggering and in vivo IgE Induction
Authors: Fritsché R. , Bonzon M.
Published: August 2009

The allergenicity of cow milk formulas was assessed in vitro by their ability to trigger prelabelled 3H-serotonin release from normal rat mast cells sensitized passively with rat reaginic anti-cow milk protein antisera. It was found that enzymatically hydrolyzed milk formulas had 1,000 (BEBAHA) to over 100,000 (AL-FARE) times lower mast cell-triggering capacities than a standard milk formula (NAN). In vivo, these hydrolyzed formulas induced 100 (BEBAHA) to 10,000 (ALFARE) times less IgE antibodies than the standard milk formula.
Expression of Intercellular Adhesion Molecule-1 in Murine Allergic Contact Dermatitis
Authors: Goebeler Matthias , Gutwald Johannes , Roth Johannes , Meinardus-Hager Georg , Sorg Clemens
Published: August 2009

Intercellular adhesion molecule-1 (ICAM-1) plays an important role in the interaction of immunocompetent cells in inflammation. It contributes to lymphocyte recruitment, antigen presentation and T-cell activation. Recently, the murine homologue of ICAM-1, originally designated as MALA-2, has been identified. The monoclonal antibody YN1/1.7 for murine ICAM-1 enables the analysis of ICAM-1 expression in murine allergic contact dermatitis, a prototype of cutaneous T-cell-mediated inflammatory response. At 0, 1, 8, 24, 48, 72 h, and 7 and 14 days after challenge with 2,4-dinitrofluoro-1-benzene (DNFB) cryostat sections of ear skin were immunostained for ICAM-1, I-A and mononuclear cells (L3T4, Lyt-2, BM8). In normal skin ICAM-1 labeling was restricted to endothelial cells and dermal dendritic cells/macrophages; keratinocytes (KCs) did not express ICAM-1. The dermal cellular infiltrate increased progressively from 8 to 72 h after DNFB challenge. The majority of infiltrating cells were BM8+ macrophages (75%) and L3T4+ (10%) or Lyt-2 + T cells (10%); maximally 30% of those stained positive for ICAM-1. At 24 h, focal ICAM-1 expression on KCs developed, reached a maximum at 72 h and faded therafter. Migration of T cells into the epidermal layer started at 48 h at sites which had already expressed ICAM-1. Our data provide evidence that expression of ICAM-1 by epidermal cells precedes infiltration of the epidermis by T lymphocytes as shown before in human cutaneous disorders. Thus, a mouse model may be useful to investigate the role of ICAM-1 in inflammation further.
Effect of Locally Administered Heparins on Delayed-Type Hypersensitivity Reactions
Authors: Kakakios A.M. , Ryan J. , Geczy C.L.
Published: August 2009

Inflammatory reactions involved in delayed-type hypersensitivity (DTH) are associated with extravascular coagulation and fibrin deposition. Heparin and other anticoagulants administered systemically inhibit DTH reactions but the direct effect of intradermally injected heparin on the developmentof DTH skin responses has not been reported. The effects of heparin on the DTH reaction elicited by ovalbumin (OVA) in guinea pigs 1–3 weeks after sensitization were examined. Unfractionated heparin, low affinity heparin (LAH; non-anticoagulant) and high affinity heparin (HAH; anticoagulant) were injected together with suboptimal amounts of OVA. Heparin and LAH enhanced skin induration, LAH (0.5 μg) by an average of 50% above that due to OVA alone at 24 h (p < 0.01). In contrast, HAH (0.5 μg) significantly reduced skin induration at 24 h. Heparin and LAH also significantly increased cellular infiltration with LAH having the greater effect. At 4 h the infiltrate consisted mainly of neutrophils whereas at 24 h mononuclear cells predominated. Fibrin deposition, assessed both by immunohistology and quantitation of radioactive fibrin extracted from skin test sites, was increased by 30% when OVA was tested in the presence of LAH. Mast cell heparin released locally at sites of DTH has the potential to modulate these reactions in either a pro- or anti-inflammatory manner. This study is the first to demonstrate differences in the capacities of LAH and HAH to modulate cell-mediated inflammation.
Role of Mucosal Mast Cells in Intestinal Graft-versus-Host Reaction in the Mouse
Authors: Newlands G.F.J. , Mowat A.McI. , Felstein Michelle V. , Miller H.R.P.
Published: August 2009

Hyperplasia of mucosal mast cells (MMC) is found in many enteropathies which are caused by T lymphocytes, but their exact role is unknown. In this study we have investigated whether MMC play a part in the immunologically mediated enteropathy which occurs in mice with graft-versus-host reaction (GvHR). There were simultaneous increases in the numbers of jejunal MMC and in the concentrations of mouse intestinal mast cell proteinase both in serum and in the intestine in two separate models of GvHR. Although these changes developed in parallel with the evolving GvHR, there was no correlation between the degree of MMC activation and the severity of the intestinal pathology. In addition, mast cell deficient W/Wv mice developed systemic and intestinal GvHR as severe as their normal congenic littermates, despite a markedly deficient MMC response. We conclude that the role of MMC in enteropathy may be to regulate or repair T lymphocyte mediated immunopathology.
How Similar Are in vitro Differentiated, Cord Blood Derived Eosinophils to Peripheral Blood Eosinophils?
Authors: Bach Michael K. , Brashler John R. , Sanders Martin E. , Furitsu Takuma , Ishizaka Teruko
Published: August 2009

In this study we attempted to compare cord blood derived, in vitro differentiated eosinophils to peripheral blood eosinophils with respect to their capacity to respond to various activators and, therefore, their potential ability to contribute to an inflammatory response. The cells were compared with respect to their density, content of eosinophil peroxidase, and eosinophil-derived neurotoxin, and with respect to their responses to various activators. The in vitro cultured, cord blood derived eosinophils were distinctly lighter than the freshly isolated peripheral blood cells. This difference in cell density was reflected in a slightly reduced content of both eosinophil peroxidase (1.17 ± 0.29 compared to 2.03 ± 0.22 arbitrary units/2,000 cells) and eosinophil-derived neurotoxin (18.7 ± 4.0 vs. 26.4 ± 4.5 ng/l04 cells). We compared the cells with respect to two different activation end points; the production of activated oxygen metabolites (superoxide anion) and the secretion of cationic proteins from their granules (eosinophil peroxidase and eosinophil-derived neurotoxin). In general, these responses were either the same in the two cell populations, or they were only slightly lower in the cord blood derived cells. There were, however, a few notable exceptions. Thus the secretory responses of the cultured cells to C5a and C3a anaphylatoxins and O2–– production with the chemotactic peptide, formyl-methionyl-leucyl-phenylalanine, and with aggregated IgG were consistently greater than those of the normodense eosinophils. The possible implications of these differences on the state of maturation of the in vitro differentiated eosinophils are briefly discussed.
Production of Superoxide Anion and Hydrogen Peroxide by Human Neutrophilic Granulocytes during Chemotactic Migration towards f-Met-Leu-Phe, C5a, Leukotriene B4, Monocyte-Derived Chemotaxin/IL-8 and Platelet-Activating Factor
Authors: Kownatzki Eckhard , Uhrich Sibylle
Published: August 2009

During the chemotactic migration of human neutrophilic granulocytes towards the chemotactic factors f-Met-Leu-Phe, C5a, leukotriene B4 (LTB4), monocyte-derived chemotaxin (MOC/IL-8) and platelet-activating factor (PAF) in Boyden chambers, the production of superoxide anion and hydrogen peroxide was measured by superoxide dismutase-inhibitable cytochrome C reduction and oxidation of p-OH-phenylacetic acid, respectively. With the exception of 10––6M PAF, none of the factors at optimal chemotactic concentrations induced the production of O2 or H2O2 in amounts significantly different from neutrophilic granulocytes migrating at random. At 20–50 times the optimal chemotactic concentration some O2 and H2O2 production was observed with f-Met-Leu-Phe, C5a and LTB4, but not with MOC/IL-8. Superoxide dismutase, catalase or a combination of the two added to both compartments of the Boyden chambers did not affect the random or chemotactic migration towards any of the chemotactic factors. The results suggest that chemotactic migration and the production of reactive oxygen metabolites by human neutrophilic granulocytes are unrelated events.
Differential Effect of Hypertonicity on FMLP-, Anti-IgE- and Ca2+ Ionophore A23187-Induced Histamine Release from Human Basophil Leukocytes
Authors: Tedeschi A. , Arquati M. , Lorini M. , Miadonna A.
Published: August 2009

Effects of different extracellular Na+ and K+ concentrations (respectively, 135, 155, 220, 260 mM NaCl, and 2.7, 20, 50, 100 mMKCl) on IgE-dependent and IgE-independent histamine release from human basophils were examined. High extracellular Na+ and K+ concentrations were shown to reduce N-formyl-methionyl-leucyl-phenylalanine- (FMLP), but not anti-IgE- or Ca2+ ionophore A23187-induced histamine release. A high extracellular Ca2+ (7.2 mM CaCl2) concentration increased basophil response to anti-IgE and FMLP. The enhancement of FMLP- but not of anti-IgE-induced histamine release was antagonized by high extracellular Na+ and K+ concentrations. When leukocytes were suspended in isotonic choline chloride solutions (choline is a nonpermeant monovalent cation), an enhancement of anti-IgE- and FMLP-induced histamine release was observed. This suggests that monovalent cations, namely Na+ ions, at physiological concentrations, downregulate histamine release from human basophils. At high choline chloride concentrations, FMLP-, but not anti-IgE-induced histamine release was inhibited. Thus, the reduction of FMLP-evoked histamine secretion from human basophils seems to be due to hypertonicity and not to the type of monovalent cation, either permeant or nonpermeant, contained in extracellular milieu. The different effects of a hypertonic solution on anti-IgE and FMLP-induced histamine release are probably related to the different cell activation pathways triggered by the two stimuli.
Immunology of Celiac Disease: Tissue and Species Specificity of Endomysial and Reticulin Antibodies
Authors: Valeski Edward , Kumar Vijay , Beutner Ernst H. , Lerner Aaron , Chorzelski Tadeusz P.
Published: August 2009

Patients with celiac disease and dermatitis herpetiformis have circulating antibodies to reticulin and endomysium, the ‘extracellular matrix’ components as defined by their detection on rodent and primate tissues, respectively. Because both types of antibodies occur in both forms of gluten-sensitive enteropathy, studies were conducted to determine if the two types of antibodies can be distinguished by species and organ specificity. The results of these studies indicate that distinct endomysium-specific and reticulin-specific antibodies can be found and that these differ in their species specificity; i.e., endomysium antigen occurs in primate and not in rodent tissues while the reticulin antigen occurs in rodent and not in primate tissues. However, the activity of both endomysium and reticulin antibodies demonstrates similar tissue distribution, in that both react to antigens associated with gastrointestinal smooth muscles, peritubular and periglomerular areas of the kidney, and sinusoidal and periportal areas of the liver. Also, both antigens seem to be present in sheep and goat tissue. These studies indicate that the antigens reactive with endomysial antibodies are distinct from those reactive with reticulin antibodies.
Density Distribution and Density Conversion of Neutrophils in Allergic Subjects
Authors: Miyagawa Hidefumi , Okada Chiharu , Sugiyama Haruhito , Hopp Russell J. , Agrawal Devendra K. , Nabe Makoto , Gordy Cathy , Bewtra Againdra K. , Townley Robert G.
Published: August 2009

We investigated the density distribution of neutrophils in peripheral blood of allergic subjects. We divided neutrophils into four groups: fraction 1 (density > 1.085), fraction 2 (1.081 < density ≤ 1.085), fraction 3 (1.077 < density ≤ 1.081) and fraction 4 (density ≤ 1.077). The percentage of neutrophils in fraction 2 in allergic rhinitis (AR) subjects or asthmatics was lower than that in normals (p < 0.01). The percentage of neutrophils in fraction 3 and fraction 4 from AR or asthmatics was greater than that in normals (fraction 3, p < 0.01; fraction 4, p < 0.05). In neutrophils from AR subjects (fraction 3), chemotaxis to N-formyl-methionyl-leucyl-phenylalanine or platelet-activating factor (PAF) was enhanced compared to fraction 2. PAF (10––7M) changed the density of neutrophils (p < 0.01), which were inhibited by WEB 2086 (p < 0.05). Furthermore, granulocyte-macrophage colony-stimulating factor changed the density of neutrophils (p < 0.01). These findings suggest that biological agents may activate neutrophils and convert their density resulting in neutrophils with lower density in allergic subjects.
Correlation between Lymphocyte Proliferative Responses and Dendritic Cell Migration in Regional Lymph Nodes following Skin Painting with Contact-Sensitizing Agents
Authors: Kimber I. , Kinnaird A. , Peters S.W. , Mitchell J.A.
Published: August 2009

We have investigated whether there exists a correlation between the induction of draining lymph node cell (LNC) proliferation in contact allergy and the accumulation of dendritic cells (DC) within such lymph nodes. CBA/Ca mice, which compared with mice of BALB/c strain, mount a more vigorous lymphocyte proliferative response following sensitization, also exhibited a more marked accumulation of DC in draining lymph nodes 24 h following skin painting. Moreover, studies with the skin-sensitizing fluorochromes fluorescein isothiocyanate (FITC) and rhodamine B isothiocyanate (RITC) revealed that DC-enriched fractions of draining LNC prepared from CBA/Ca mice contained a higher percentage of antigen-bearing cells than did those from BALB/c mice. A relationship between DC migration into lymph nodes and the magnitude of the induced LNC proliferative response was also indicated by experiments performed in BALB/c mice with a variety of contact allergens. It was observed that there was a direct correlation between the vigour of the proliferative response measured 3 days following exposure and the frequency of DC in draining nodes at 24 h. Collectively these data suggest that following skin sensitization the migration of DC into the draining lymph nodes influences quantitatively the primary immune response and the development of contact allergy.
T Cell Cytotoxicity in Cystic Fibrosis: Relationship to Pulmonary Status
Authors: Knutsen Alan P. , Mueller Kathleen R.
Published: August 2009

T cell cytotoxicity (CTL) to an allogeneic lymphocyte target was evaluated in patients with cystic fibrosis (CF) before and during pulmonary exacerbations (group 1) compared to another group of CF patients who had stable pulmonary disease activity during their two study periods (group 2). CTL activity was significantly decreased in group 1 subjects studied prior to their pulmonary flares and in group 2 CF patients compared to normal controls at effector:target ratios of 12.5:1 and 6.25:1 (p < 0.05 and p < 0.05, respectively). Furthermore, in group 1, CTL lysis was significantly decreased during pulmonary flares compared to before flares at the 25:1 and 12.5:1, effector:target, (p < 0.05 and p < 0.05, respectively). T suppressor cell activity as measured by effect on in vitro control B cell IgM synthesis was significantly increased during pulmonary flares (p < 0.05). Diminished CTL may be partially responsible for persistent colonization of Pseudomonas aeruginosa in CF.
Reactivity of Lymphoid Cells Isolated from the Tracheobroncheal Lymph Nodes of Two Guinea-Pig Strains with High or Low Susceptibility to Respiratory Anaphylaxis
Authors: Westphal Petersen J. , Hasløv K. , Lundberg L. , Weis Bentzon M. , Gehring T.
Published: August 2009

The mechanisms underlying the different sensitivity to respiratory anaphylaxis were studied in two inbred strains of guinea pigs with high (IMM/S) or low (IMM/R) sensitivity to ovalbumin (OA) aerosol-induced respiratory anaphylaxis. Guinea pigs were immunized with OA in Freund’s complete adjuvant and lymphocyte stimulation tests were done with cells from the tracheobroncheal lymph nodes (TBL) draining the airways, from peripheral blood lymphocytes and from the axillary and inguinal lymph nodes. The TBL of the IMM/R strain had a signficantly lower response than those of the IMM/S strain. This selectively low response, which was not antigen specific, could be increased by cyclophosphamide pretreatment. The high TBL response in the IMM/S strain was decreased by OA inhalations to the level of the IMM/R strain. OA inhalations decreased the IgE response in the IMM/S but not the IMM/R strain. Different interpretations of these findings are discussed, with emphasis on the possibility that an inherent suppressor cell population in the TBL are involved.
Characterization and Quantification of Cellular Infiltrates in Nasal Mucosa of Patients with Grass Pollen Allergy, Non-Allergic Patients with Nasal Polyps and Controls
Authors: Fokkens Wytske J. , Holm Adriaan F. , Rijntjes Evert , Mulder Paul G.H. , Vroom Thea M.
Published: August 2009

Little is known about cellular infiltrates in nasal mucosa and the differences between these infiltrates in allergic and non-allergic patients. A reproducible and objective method making use of monoclonal antibodies for the quantification and characterization of cellular infiltrates in biopsy specimens of nasal mucosa is described. This method was used to study quantitative differences in cellular infiltrates in the epithelium and lamina propria of the nasal mucosa of patients with isolated grass pollen allergy, non-allergic patients with nasal polyps, and controls. A surprisingly wide variation was found in all groups. In all groups the T lymphocytes were much more numerous than the B lymphocytes. The number of CD8+ cells exceeded the number of CD4+ cells in the epithelium but in the lamina propria the numbers were approximately equal. Significant differences between the three groups were found with respect to the number of CD1+, IgE+, neutrophils and cytoplasmic IgG4+ cells. No significant differences were found in the numbers of CD4+, CD8+, CD14+, CD22+, HLA-DR+, IgGl-3+ cells or eosinophils. The use of biopsy in combination with monoclonal antibodies is an easy and well-tolerated method to study immunological reactions in the nasal mucosa. The results of this study indicate a possible role for a T-cell-mediated response in allergic rhinitis.
Skin Reactivity to Substance P, Not to Neurokinin A, Is Increased in Allergic Asthmatics
Authors: Iwamoto Itsuo , Kimura Akira , Tanaka Makoto , Tomioka Hisao , Yoshida Sho
Published: August 2009

The tachykinins substance P (SP) and neurokinin A are believed to be major mediators of neurogenic inflammation. To determine whether the skin reactivity to tachykinins is increased in asthmatics, we examined the erythemas and wheals induced by intradermal injections of SP, the C-terminal peptide SP6–11, the N-terminal peptide SP1–9 and neurokinin A (10––7––10––5M) in 10 allergic asthmatics and 9 normal subjects. SP and SP1–9 induced both erythemas and wheals in a concentration-dependent manner in allergic asthmatics and in normal subjects, whereas SP6–11 and neurokinin A induced only wheals in both groups. SP induced greater erythemas and wheals in allergic asthmatics than in normal subjects. However, the wheals induced by neurokinin A were not significantly different between the two groups. SP1–9 also induced greater erythemas and wheals in allergic asthmatics than in normal subjects, whereas the wheals induced by SP6–11 were not significantly different between the two groups. Therefore, the increased skin reactivity to SP was dependent on the N-terminal peptide but not on the C-terminal peptide. We conclude that the skin reactivity to SP but not to neurokinin A is increased in allergic asthmatics.
Neutral Endopeptidase Modulates Substance P-Induced Activation of Human Neutrophils
Authors: Iwamoto Itsuo , Kimura Akira , Yamazaki Hiroomi , Nakagawa Noriaki , Tomioka Hisao , Yoshida Sho
Published: August 2009

Neutral endopeptidase (NEP; EC 3.4.24.11) is well recognized as a regulatory peptidase for substance P (SP)-induced responses in various tissues. To determine whether NEP regulates SP-induced activation of human neutrophils, we examined the effect of the NEP inhibitor phosphoramidon on SP-induced superoxide generation and chemotaxis in human blood neutrophils. SP (10––6––10––4M) induced superoxide generation and chemotaxis in the neutrophils dose dependently. The NEP inhibitor enhanced the SP-induced responses. Thus, phosphoramidon (10––6M) shifted the dose-response curves of SP-induced superoxide generation and chemotaxis of the neutrophils to the left by 0.5–0.6 log. Phosphoramidon prevented the hydrolysis of SP by the neutrophils, the NEP activity of the neutrophils being assessed as 125 ± 13 pmol of SP/min/106 cells. The N-terminal peptide SP1–9 (up to 3 × 10––4M), which was a major degrading product by NEP of the neutrophils, did not activate the neutrophils. We conclude that NEP modulates SP-induced activation of human neutrophils.
Effects of Ozone Exposure on Experimental Asthma in Guinea Pigs Sensitized with Ovalbumin through the Airway
Authors: Sumitomo Midori , Nishikawa Masanori , Fukuda Tsutomu , Kaneko Tamotsu , Ikeda Hirotada , Suzuki Shunsuke , Okubo Takao
Published: August 2009

As ozone (O3) is known to cause airway inflammation and hyperresponsiveness, we examined the effects of 03 exposure (1, 3, or 5 ppm, 2 h) on sensitization and provocation in guinea pigs sensitized with ovalbumin (OA) through the airway. In groups exposed to O3 before sensitization, 5 ppm increased the production of IgG1 antibodies and decreased the OA sensitization threshold from 0.01 to 0.002%. In those exposed before provocation, 1, 3, or 5 ppm of O3 decreased the OA provocation threshold from 0.5 to 0.02%, and this enhancement appeared to depend on airway hyperresponsiveness. We conclude that O3 exposure may play an important role in causing asthmatic attacks rather than enhancing allergic sensitization.
Effects of a Single-Dose Pretreatment with Captopril on the Immediate Response to Nasal Challenge with Allergen
Authors: Proud David , Naclerio Robert M. , Meyers Deborah A. , Kagey-Sobotka Anne , Lichtenstein Lawrence M. , Valentine Martin D.
Published: August 2009

We performed a double-blind, placebo-controlled, crossover study using 12 subjects to determine the effects of a single 50-mg dose of captopril on the response to nasal challenge with increasing doses of allergen. Levels of kinins, histamine and N-α-p-tosyl-L-arginine methyl ester (TAME)-esterase activity were measured in nasal lavages, and symptom scores and the number of sneezes were recorded. Captopril had no significant effects on histamine, TAME-esterase, sneezing or symptom scores. Peak postchallenge kinin levels, however, were significantly elevated (p < 0.05) compared to placebo, while an increase in the magnitude of the dose-response curve was of marginal significance (p = 0.058). Thus, captopril causes increases in the kinin levels in nasal secretions during the allergic response. If increased kinin levels persist or worsen with chronic therapy, it is possible that they could exacerbate allergic symptoms during repeated seasonal exposure.
Ontogeny of Mast Cells in the Ventral Prostate of the Rat
Authors: Hammel I. , Roizman P. , Massas R. , Abramovici A.
Published: August 2009

The ontogenic evolution of mast cells in the rat ventral prostate was studied using the Grimelius silver impregnation method. The mast cell density was highest during the pubertal period and later, it declined significantly with age. Most mast cells were identified in the fibrovascular stroma in close proximity to nerve fibers and blood vessels. The total number of mast cells seems to be constant when correlated with prostatic weight.
Basic Mechanisms of Cellular Priming and Release of Inflammatory Mediators
Authors: Brom J. , Brom C. , König W.
Published: August 2009

Simultaneous stimulation of human neutrophils (PMN) with the receptor-mediated activator formyl-met-leu-phe (FMLP) and the G protein activator sodium fluoride (NaF) resulted in the synergistic generation of leukotrienes. Activation of human platelets with thrombin and NaF showed an additive formation of 12-HETE. This enhancement in lipid mediator generation correlated with a time-dependent synergism in G protein activation after sequential stimulation of intact cells with FMLP and NaF or thrombin and NaF, respectively. In addition, polymerization of actin, an early event in cell activation, was enhanced after incubation with cytokines and FMLP.
IgG Anti-IgE Autoantibodies in Immunoregulation
Authors: Stadler Beda M. , Gang Qiu , Vogel Monique , Jarolim Erika , Miescher Sylvia , Aebischer Ivan , de Weck Alain L.
Published: August 2009

Human sera contain anti-IgE autoantibodies with diverse biological functions in vitro. Opposite functions can be shown for triggering of IgE-mediated histamine release from human basophils in terms of anaphylactogenic or nonanaphylactogenic autoantibodies. Furthermore, autoantibodies are either capable of removing IgE from the surface of CD23-positive cells of binding more IgE to such cells. A similar dichotomy seems also to exist for the effect of autoantibodies on human IgE mRNA synthesis as well as IL-4-induced proliferation of human mononuclear cells. Thus, anti-IgE autoantibodies may represent a key factor in the manifestation and the development of allergic disease.
Isolation and Characterization of a 60-kDa IgE-Binding Component Derived from Sera of Atopic Patients (Atopic Dermatitis)
Authors: Bujanowski-Weber J. , Knöller I. , König W.
Published: August 2009

The low-affinity receptor for IgE (FcεRII, CD23) and the related soluble IgE-binding factors (IgE-BF; sCD23) play an important role in IgE regulation. Sera of patients suffering from atopic dermatitis (AD) were reported to contain an IgE-binding component with a molecular weight of 60 kD. The aim of our studies was the isolation and characterization of the 60 kD component. Sera of patients with AD were fractionated by ammonium sulfate precipitation (10–90%). The fractions were analyzed with regard to their IgE and their IgE-BF contents. Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and subsequent autoradiography with 125I-labeled human IgE (PS) was performed to detect IgE-binding activity. The major amount of IgE as well as IgE-BF was obtained within the 30–50% ammonium sulfate precipitation. In addition, IgE-binding activity was precipitated at 60% saturation. Separation by gel filtration under physiological conditions indicated IgE-BF with molecular weight of > 100, 60, 25 and 15 kD. Rechromatography of the > 100-kD fraction led to IgE-binding activity with a molecular weight of 60 kD which is not present within normal sera. The data demonstrate that the 60-kD component is partially bound to serum IgE. One may suggest that the complex is involved in the induction and persistence of allergic disorders.
Role of CD8 T Cells in Rat IgE Responses
Authors: Kemeny D.M. , Diaz-Sanchez D.
Published: August 2009

The role of the cytokines interleukin-4 and interferon-γ in the regulation of IgE responses in the mouse and man have focused on the role of CD4 T cells. In the rat, antigen-specific CD8 T cells, generated following inhalation of antigen, have been shown to be capable of suppressing IgE responses. Repeated intraperi-toneal injections of 1 ng ricin and 1 μg antigen established a long-lived IgE response in both low- and high-IgE responder rat strains (Wistar and Brown Norway). The duration of the IgE antibody response was 204 and 248 days, respectively. Total IgE levels rose from 30 ± 20 to 39,000 ± 7,500 ng/ml in the Wistar rat and from 120 ± 100 to 47,000 ± 8,000 ng/ml in the Brown Norway rat. An even greater (104-fold) increase was seen in antigen-specific IgE antibody levels. Ricin alone had no effect and concomitant or prior stimulation with antigen was required. The proportion of CD4+ and CD8+ cells present in the spleen at the peak of the IgE response was markedly increased compared with animals given ricin or antigen alone. Furthermore, CD8 T cells were approximately 100 times more sensitive to ricin than CD4 T cells. These data suggest that enhancement of IgE responses in ricin-treated animals results from the selective deletion of T cells which suppress IgE and are of the CD8 phenotype.
Control of Eosinophilia
Authors: Sanderson Colin J.
Published: August 2009

Experiments in vitro have suggested that IL5 is a late-acting factor in eosionophil production, and that other factors such as IL3, G-CSF and GM-CSF are required for the production of committed eosinophil progenitors. Furthermore, work in vitro indicates that in addition to IL5, both IL3 and GM-CSF are capable of stimulating eosinophil differentiation. Thus, there would appear to be both considerable redundancy in cytokine actions in eosinophilia as well as a complex network of cytokine activities to induce eosinophilia. Experiments in vivo, however, suggest a less complicated control mechanism, dominated by IL5.