bluuebunny/crossref_metadata_2025 huggingface dataset:
Primary and Secondary Effector Cells in the Pathogenesis of Bronchial Asthma
Authors: Holgate Stephen T. , Twentyman Orion P. , Rafferty Paul , Beasley Richard , Hutson Penny A. , Robinson Clive , Church Martin K.
Published: August 2009

The immediate and late asthmatic reactions provoked by inhaled allergens have provided useful models enabling the dissection of individual inflammatory cells and their mediators that may contribute to the pathogenesis of asthma. The immediate reaction is considered to be mast cell-mediated on the basis that about 50% of the response is inhibitable by potent and selective H1-receptor antagonists such as terfenadine and astemizole. Additional inhibition (∼30%) by the potent cyclooxygenase inhibitor flurbiprofen implies an important role for prostanoids in the immediate response, the most likely mast cell-derived product being prostaglandin (PG) D2. In man, PGD2 is selectively metabolised to 9α1 1β-PGF2, a unique prostaglandin which shares with PGD2 contractile properties on guinea-pig and human airways smooth muscle. The inability of piriprost, a potent leukotriene synthesis inhibitor, to influence the allergen-provoked immediate reaction raises the possibility that sulphidopeptide leukotrienes play a minor role in this response. The late asthmatic reaction is considered to resemble clinical asthma since it is accompanied by increased responsiveness of the airways to a wide range of stimuli. The late reaction in man is inhibited by nedocromil sodium (4 mg) but only marginally attenuated by salbutamol (200 μg) if both drugs are administered prior to allergen challenge. Since salbutamol, in the dose administered, is a potent mast cell-stabilising agent, these findings must question the obligatory role of mast cell mediator release in the pathogenesis of the late response. A model of the late reaction in conscious guinea-pigs demonstrates a selective influx of neutrophils followed by eosinophils 6–12 h after challenge in relation to the late reaction, suggesting a role for these secondary effector cells in mediating this response and possibly the acquisition of increased bronchial responsiveness. These human and animal studies demonstrate a complex interaction of primary and secondary effector cells in the pathogenesis of bronchoconstrictor responses to allergen and should help to clarify the contribution of ‘inflammation’ to clinical asthma.
Urinary Trypsin Inhibitors Afford Cardioprotective Effects through Activation of PI3K-Akt and ERK Signal Transduction and Inhibition of p38 MAPK and JNK
Authors: Kim Seok J. , Yoo Kyung Y. , Jeong Cheol W. , Kim Woong M. , Lee Hyung K. , Bae Hong B. , Kwak Sang H. , Li Mei , Lee JongUn
Published: August 2009

Background: We determined the effect of urinary trypsin inhibitors (UTI) in regional myocardial ischemia/reperfusion (I/R) injury and its underlying mechanisms involving the role of prosurvival kinases such as phosphatidylinositol-3-OH kinases (PI3K)-Akt and extracellular signal-regulated kinases (ERK 1/2) and apoptotic kinases such as p38 and JNK. Methods: The rats were anesthetized and subjected to an I/R insult consisting of 30-min left anterior descending coronary artery (LAD) occlusion followed by reperfusion. Infarct size was measured after 120 min of reperfusion. UTI was given alone or along with wortmannin (inhibitor of PI3K) or PD098059 (inhibitor of ERK1/2) before LAD occlusion. The phosphorylation of Akt, ERK1/2, p38 and JNK was determined by immunoblotting after 5 min of reperfusion. UTI was administered 10 min before LAD occlusion, and wortmannin and PD098059 were administered 20 min before LAD occlusion. Results: UTI significantly reduced the infarct size compared with the control. Wortmannin or PD098059 alone did not affect the infarct size, but they abolished the UTI-induced cardioprotective effect. UTI significantly reduced the phosphorylation of p38 and JNK, while it enhanced that of Akt and ERK1/2. Conclusions: UTI has a protective effect against regional myocardial I/R injury through activation of survival kinases PI3K-Akt and ERK1/2 and attenuation of p38 and JNK.
Modification of House Dust Mite Allergens by Monomethoxypolyethylene Glycol
Authors: Mosbech H. , Dreborg S. , Påhlman I. , Stahl Skov P. , Steringer I. , Weeke B.
Published: August 2009

In animal models, allergen modification by coupling to monomethoxypolyethylene glycol (mPEG) molecules can reduce allergenicity of the extract and makes the allergen capable of suppressing boosted IgE response. To investigate in a human system the degree of attenuation implied by a mPEG modification of a house dust mite (Dermatophagoides pteronyssinus) extract, 55 adults with asthma caused by house dust mites were tested by skin prick test (SPT) and histamine release assay (HR). RAST inhibition was performed on sera from 6 additional patients. Modified extract containing 0.42 mmol mPEG/g protein was used for the analyses. In order to get the same response of the two extracts when assessed by HR and SPT, a median increase in concentration of 10-fold of the mPEG-modified extract compared to the unmodified extract was needed. Interindividual variation was limited. Sixty-four to 72% needed a dose increase within ± half a decade from this value. In 42–49% of the patients, results from SPT and HR deviated less than half a decade. The relative potency of the modified extract as measured by RAST inhibition was reduced to 17–78% (mean 39%). Reduced allergenicity would by itself mean less side effects in immunotherapy. When planning such therapy it is important to know that mPEG modification reduces the allergenicity to a similar extent in a majority of patients.
Genetic Control of Delayed-Type Hypersensitivity to D-Penicillamine Antigen
Authors: Ishii Norihisa , Ichiyama Shinichi , Ishii Takayo , Aoki Ichiro , Okuda Kenji , Nakajima Hiroshi
Published: August 2009

For determining the mechanism of genetic control of D-penicillamine free-base-specific delayed-type hypersensitivity (DTH) in mice, footpad swelling response was employed. Studies with various congenic and recombinant inbred strains of mice revealed that D-penicillamine free-base-specific DTH was controlled by the I-A subregion. The footpad swelling response showed a highly antigen-specific pattern, and revealed that D-penicillamine free base was not cross-reactive with D-penicillamine disulfide or L-penicillamine, but with D-penicillamine HCl. Treatment of immune lymphoid cells with monoclonal antibodies plus complement revealed that the cells responsible for DTH transfer were Lyt-1+2––, L3T4+ and la–– T cells.
Carbohydrate-Specific Human Heterophile Antibodies in Normal Human Sera That React with Xenogeneic Cells
Authors: Laus R. , Ulrichs K. , Müller-Ruchholtz W.
Published: August 2009

Human heterophile antibodies (HHA) that are present in normal human sera (NHS) play an important role in hyperacute xenograft rejection. The aim of this study was to analyze the occurrence, mode of action and molecular specificity of HHA in NHS that are directed against xenogeneic lymphocytes (isolated from mouse, rat, guinea pig, rabbit, cattle and pig) and isolated rat pancreatic islets. All sera contained variable amounts of HHA that killed the target cells via the classical complement pathway. The cytotoxic activity of these HHA was specifically inhibited by certain carbohydrates (α-D-melibiose, β-lactose, β-gentiobiose, β-cellobiose, D-mannose, N-acetyl-β-D-mannosamine and α-D-rhamnose) and by rat IgM. By means of affinity chromatography with immobilized inhibitors we obtained an antibody preparation of mainly IgG type from NHS (up to 3.5 mg/10 ml serum) that reacted strongly with rat lymphocytes and isolated rat pancreatic islets. Though thus far residual xenospecifíc antibody activity has remained in the sera even after multiple affinity chromatography, these data suggest that specific elimination of HHA is feasible and that it may be thus possible to overcome a major obstacle to xenotransplantation.
Decreased T Helper Cell Function in Patients with Cystic Fibrosis
Authors: Knutsen Alan P. , Slavin Raymond G. , Roodman Stanford T. , Mueller Kathleen R. , Marino Nicholas L.
Published: August 2009

T cell immune function was further evaluated in cystic fibrosis (CF) patients. CF patients, regardless of severity of pulmonary disease or colonization with Pseudomonas aeruginosa, had lower percentages of T helper cells (p < 0.01) and decreased T helper function as demonstrated by diminished T help for control B cell pokeweed mitogen-stimulated IgG (p < 0.01) and IgM (p < 0.01) synthesis. Increased T suppressor function, measured by co-culture with control B and T cells, was noted in only 25% of CF patients. CF patients’ T cells exhibited decreased allogeneic T cell cytotoxicity compared to normal controls (p < 0.01). This study extends previous studies demonstrating decreased T cell functions in CF patients and indicates a specific decrease in T helper function.
A Nasal Allergy Model Developed in the Guinea Pig by Intranasal Application of 2,4-Toluene Diisocyanate
Authors: Tanaka Ken-ichi , Okamoto Yutaka , Nagaya Yoshiko , Nishimura Fumiko , Takeoka Akemi , Hanada Satoshi , Kohno Shigekatsu , Kawai Mitsuru
Published: August 2009

An experimental model of nasal allergy has been developed in guinea pigs by intranasal application of 2,4-toluene diisocyanate (TDI). A 10% TDI solution in ethyl acetate was painted onto the nasal vestibuli of the animals once a day for 5–10 days. During the course of repeated application of TDI, the number of animals which secreted rhinorrhea containing eosinophils increased. Morphological survey of the nasal mucosa showed infiltration of eosinophils and some other changes indicative of acute inflammation. Moreover, mast cells were found not only in the subepithelial connective tissue but also in the epithelial layer. Nasal mucus obtained from the mucosa has been found to be an effective test material for studies of nasal allergy. A striking decrease of specific granules was found in some mast cells contained in the mucus. In parallel with the symptomatology, biochemical and serological studies suggested the involvement of type I allergy in the experimental system; TDI-specific histamine release from the nasal mucosa and positive passive cutaneous anaphylaxis were found 3 weeks after the application of TDI.
Function of Exudate Neutrophils from Skin in Psoriasis
Authors: Coble Britt-Inger , Briheim Gunnar , Dahlgren Claes , Molin Lars
Published: August 2009

Human neutrophils harvested from skin chambers containing autologous serum in psoriatic patients were compared with peripheral blood neutrophils by examining migration, phagocytosis, and oxidative activation. Random migration was reduced in exudate cells, whereas the chemotactic response to yeast-activated serum was evident not only in blood neutrophils, but also in exudate cells. The metabolic activation of exudate neutrophils, when stimulated with formyl-methionyl-leucyl-phenylalanine and measured as chemiluminescence, was enhanced by 50–400% compared to blood neutrophils. The chemiluminescence response to phorbol myristate acetate was on the other hand reduced to 35%. In the phagocytic assay, using C3bi- and IgG-opsonized yeast particles, exudate neutrophils from non-psoriatic healthy controls showed enhanced uptake of C3bi-coated yeast compared to blood neutrophils. In psoriatics, the blood neutrophils showed enhanced C3bi-mediated phagocytosis compared to non-psoriatic control cells. No further increase in C3bi-mediated phagocytosis was then seen in exudate cells from these patients. IgG-mediated phagocytosis was in contrast to C3bi similar between blood and exudate neutrophils in psoriatics and non-psoriatic controls. These experiments show that during exudation priming of different receptor-mediated processes can occur. However, no significant difference was observed between different functional capacities in exudate neutrophils from psoriasis patients and non-psoriatic controls.
In vitro Immunosuppressive Properties of the Plant Alkaloid Tetrandrine
Authors: Seow W.K. , Ferrante A. , Goh D.B.H. , Chalmers A.H. , Si-ying Li , Thong Y.H.
Published: August 2009

Tetrandrine is an extract of the creeper Stephania tetrandra used in traditional Chinese medicine for the treatment of rheumatic diseases. It has recently been shown to retard and even reverse the lesions of silicosis in humans and rats. Data presented in this report indicate that tetrandrine has potent immunosuppressive properties. Mitogen-induced lymphoproliferative responses were markedly reduced even when tetrandrine was added after the initiation of cultures. In vitro antibody synthesis by B cells was also suppressed by tetrandrine, as was the natural killer cell-mediated lysis of K562 cells. It does not interfere with receptor-ligand binding, but does affect the inositol triphosphate second messenger system. These effects of tetrandrine were observed at nontoxic concentrations as shown by lymphocyte viability studies. These results indicate that tetrandrine possesses potent immunosuppressive properties, and may warrant further study in animal models of chronic inflammatory conditions, autoimmune diseases and transplant rejection.
Effect of Deuterium Oxide on Leukotriene C4 Generation in Immunologically Stimulated Human Leukocytes
Authors: Mita Haruhisa , Yui Yasuo , Yasueda Hiroshi , Shida Takao
Published: August 2009

Addition of deuterium oxide (D2O), 6–36%, resulted in a dose-dependent increase in allergen- or anti-IgE-induced leukotriene C4 (LTC4) generation from human basophils. In the presence of 36% D2O, the enhancement was 260 ± 135% for allergen stimulation and 480 ± 152% for anti-IgE stimulation as compared with the control incubated in normal buffer. The increasing effect of D2O on LTC4 generation from basophils was completely reversed by washing the cells before incubation with allergen. Vinblastine as well as colchicine, at a concentration of 100 μM, counteracted the effect of D2O. The enhanced release of histamine and LTC4 from basophils challenged with allergen was suppressed by Dimaprit, a histamine H2 receptor agonist, at a concentration required to inhibit the release by 50% of 5 × 10––5M for histamine and 10––5M for LTC4. These observations suggest that microtubules may be involved in LTC4 generation from immunologically stimulated basophils.
Effects of AHR-5333, a New Potential Antiallergy Compound, in in vivo Models of Immediate Hypersensitivity
Authors: Yanni John M. , Foxwell Marianne H.
Published: August 2009

AHR-5333 [1-[4-[3-[4-[bis(4-fluorophenyl)hydroxymethyl]-1-piperidinyl]propoxy]-3-methoxyphenyl]ethanone] possessed potent, long-acting activity in rat and guinea pig in vivo models of immediate hypersensitivity. AHR-5333 was more potent than azatadine (2×), ketotifen (∼3 ×), oxatomide (∼5 ×), albuterol (6 ×) and aminophylline (∼ 50 ×) in a passive, foot anaphylaxis model in rats and more potent than diphenhydramine (∼237 ×), oxatomide (∼5.6 ×) and theophylline (∼295 ×) in guinea pigs challenged with aerosolized antigen. A long duration of action was noted after oral dosing of guinea pigs (24 h PD50, 0.78 mg/kg). Administration by aerosol (1%) to sensitized, spontaneously breathing, conscious guinea pigs protected against antigen-induced anaphylactic collapse; this protection persisted through 8 h. When administered prior to antigen challenge, AHR-5333 (10 mg/kg, p.o.) effectively inhibited ascaris antigen-induced skin hypersensitivity reactions in both dogs and cynomolgus monkeys.
Isolation of a Thymocyte Growth Peptide from Human Thymus
Authors: Ernström Ulf , Karlsson Perry , Söder Olof
Published: August 2009

A thymocyte growth peptide (TGP), recruiting immature thymocytes into the S phase without affecting immunologically mature thymocytes or peripheral T cells, has previously been purified from bovine and rodent thymus. We now report the isolation of a corresponding human activity. Aqueous extracts of human thymus were found to stimulate the DNA synthesis of human thymocytes in vitro. Homogenates of suspended thymocytes displayed higher activity than extracts of intact thymus, whereas extracts of thymic stroma depleted of thymocytes contained much weaker activity. The results indicate the existence of a human TGP originating from the thymocytes and not from the stroma, implying an autocrine or paracrine growth regulation. Human TGP was purified from a formic acid extract of human thymus and found to have a molecular ratio between 1,000 and 2,000 and an isoelectric point of 4.5–4.9. The results demonstrate the existence of a human TGP similar but not identical to bovine TGP and it is proposed that this peptide acts as a progression factor for the intensely proliferating immature cortical thymocytes.
L-Ascorbic Acid Modulates 5-Lipoxygenase Activity in Human Polymorphonuclear Leukocytes
Authors: Schmidt K.H. , Steinhilber D.S. , Moser U. , Roth H.-J.
Published: August 2009

Leukotriene B4 (LTB4), a metabolite of the arachidonic acid pathway mediated by 5-lipoxygenase, is released by stimulated polymorphonuclear neutrophils (PMNL) and has been postulated to be an important mediator of the inflammatory response. Extracellular L-ascorbate at concentrations in the range of 0.5–5.0 mM effectively inhibits the biosynthesis of LTB4 and 5-hydroxyeicosatetraenoic acid (5-HETE) stimulated by the calcium ionophore A 23187. The ionophore-activated LTB4 production is reduced after incubation of PMNL with opsonized zymosan as a phagocytic stimulus. Extracellular L-ascorbate at concentrations above 0.1 mM reverses the zymosan-induced deactivation of 5-lipoxygenase, resulting in significantly higher LTB4 and 5-HETE yields. The inhibitory effect of zymosan preincubation on LTB4 production is independent of calcium or free arachidonic acid in the incubation medium. Interaction of L-ascorbate with the catabolism of LTB4 was excluded by measuring the trihydroxy metabolites which were unchanged. Furthermore, the formation of glutathione derivatives of leukotriene A4 can be excluded due to the lack of glutathione transferase activity in PMNL. In order to link the intracellular function of L-ascorbate with the serum level, the ascorbate uptake has been studied in more detail. The L-ascorbate transport into PMNL is stereospecific and can best be described by kinetics consisting of a saturation part, Km and Vmax being 39 μM and 0.28 nmol/l08 cells min, respectively, plus passive diffusion, the diffusion coefficient P being 1.75 μl/l08 cells min. Furthermore, the uptake is inhibited by the isomers D-ascorbic acid and D-erythorbic acid. Therefore, it can be concluded that low concentrations of L-ascorbate preserve 5-lipoxygenase from oxidative deactivation, resulting in a stimulation of LTB4 production and release which subsequently acts as an efficient amplifier of the chemotactic activity of PMNL. The data suggest further that L-ascorbate accumulation in PMNL may play a role in the down-regulation of inflammatory processes. This dual effect of L-ascorbate in PMNL provides for the first time a molecular mechanism for the efficacy of vitamin C as a modulator of the 5-lipoxygenase activity.
Superiority of Solenopsisinvicta Venom to Whole-Body Extract in RAST for Diagnosis of Imported Fire Ant Allergy
Authors: Butcher Brian T. , deShazo Richard D. , Ortiz Ana A. , Reed Margaret A.
Published: August 2009

RAST determinations using imported fire ant (IFA) whole-body extract (IFAWBE) and venom (IFAV) were performed with the sera of 29 sting-sensitive patients. Only 48% (14/29) of sera were positive (percent binding ≥ 6%) in RAST to IFAWBE compared with 79% (23/29) positive to IFAV, and there was significantly greater binding to IFAV than to IFAWBE (p≤0.0002). Thus, RAST with IFAV identified 31% more sting-sensitive individuals than RAST with IFAWBE, a finding similar to that noted in studies using IFAV and IFAWBE for skin testing. We conclude that IFAV is the more appropriate allergen for use in in vitro tests to detect IFA-specific IgE antibody in allergic patients.
Irritant Symptoms and Immunologic Responses to Multiple Chemicals: Importance of Clinical and Immunologic Correlations
Authors: Patterson Roy , Harris Kathleen E. , Stopford Woodhall , Van der Heiden Gloria , Grammer Leslie C. , Bunn William
Published: August 2009

An evaluation of workers in a plant was conducted because of multiple complaints of ocular, nasal, skin and chest symptoms. Antibody activity against 4 different chemicals was identified: an aliphatic diisocyanate, 4-vinylcyclohexene dioxide, trimellitic anhydride (TMA) and an unknown chemical present in a plasticizing ester known as n-octyl-n-decyl-trimellitate. The source of TM A which resulted in immunization in the plant is unknown. The presence or absence of antibodies did not correlate with the presence or absence of symptoms and it was concluded that no occupational allergic disease was present in these workers. Antibody studies alone do not make a diagnosis of occupational allergic disease and clinical correlation is required. Immunoassays may be useful in identifying exposures to immunizing chemicals in the workplace for potential clinical correlation or for exposure monitoring in the workplace.
Clinical and Serological Survey of Pulmonary Aspergillosis in Patients with Cystic Fibrosis
Authors: Schønheyder H. , Jensen T. , Høiby N. , Koch C.
Published: August 2009

A 5.5-year survey revealed 9 patients with allergic bronchopulmonary aspergillosis (ABPA) and 1 patient with a highly probable diagnosis among 200 Danish patients with cystic fibrosis (CF). The incidence of ABPA was 0.9 (0.4–1.7, 95% confidence limits) per 100 patients per year. All the patients with definite or probable ABPA had serum antibodies to the Aspergillus fumigatus catalase antigen, and in 5 patients the appearance of catalase antibodies during the incipient phase of ABPA was documented. 29 patients without ABPA also had catalase antibodies, but in lower levels than ABPA patients (p = 0.006). The 6-month prevalence rate of A. fumigatus in sputum was 80 and 72% in the 2 groups, respectively. The finding of catalase antibodies in some CF patients without ABPA may indicate the occurrence of a symptom-poor form of pulmonary aspergillosis.
Inhibition of Histamine Release from Rat Mast Cells by the Plant Alkaloid Tetrandrine
Authors: Teh B.S. , Seow W.K. , Chalmers A.H. , Playford S.P. , Ioannoni B. , Thong Y.H.
Published: August 2009

The plant alkaloid tetrandrine was shown to have significant inhibitory effects on receptor-ligand-mediated histamine release from rat mast cells at concentrations similar to or lower than that observed with theophylline and sodium cromoglycate. Inhibition of histamine release did not occur when non-specific stimulants such as aspririn, A23187 or adenosine triphosphate were used. Inhibition of ovalbumin-IgE and concanavalin A-mediated histamine release was reversible by washing the cells, showing that tetrandrine does not bind tightly to the cell membrane or cytoplasmic components. These results, taken together with previous reports of its anti-phagocytic, anti-oxidant and immunosuppressive properties, suggest that tetrandrine may be a broad spectrum non-steroidal drug of potential value in the treatment of allergic diseases.
Microculture Assay for Human Macrophage Maturation in vitro
Authors: Andreesen Reinhard , Mackensen Andreas , Osterholz Jürgen , Brugger Wolfram , Löhr Georg W.
Published: August 2009

The terminal differentiation of monocytes (mo) to macrophages (MO) is essential for the complex functioning of the mononuclear phagocyte system. Similar to in vivo MO maturation, blood mo transform to MO when cultured in vitro. This offers a model system for experimental studies of functional defects within this cell lineage. In order to develop a microassay for the evaluation of MO maturation in vitro we monitored the antigenic phenotype of MO during differentiation in vitro. The cells were grown either in suspension on hydrophobic Teflon foils, or as plastic adherent monolayers. Surface antigens were visualized on single cells with the immunoperoxidase slide technique or quantitated with the enzyme-linked immunosorbent assay (ELISA). As MO mature in vitro they increase in cell size, in the amount of beta-2-microglobulin, the myeloid antigen CD14 and HLA-DR antigen expression as measured with cell-ELISA, both in microwell cultures and on single cells. Upon terminal differentiation mo-MO express differentiation-specific antigens (transferrin receptor, surface transferrin and antigens of the MAX series) which are indicative of a successful MO maturation. Monocyte cultures in microplates and the subsequent evaluation of differentiation antigen expression could be used as an experimental system to study the modulation of MO differentiation in vitro and may serve as an in vitro parameter of MO function in vivo.
Modulation of Pulmonary Inflammation after Endotoxin Inhalation with a Platelet-Activating Factor Antagonist (48740 RP)
Authors: Rylander Ragnar , Beijer Lena , Lantz Clark , Burrell Robert , Sedivy Pierre
Published: August 2009

An acute pulmonary response was induced in guinea pigs and hamsters by inhalation of bacterial endotoxin in the form of a purified lipopolysacchande (LPS). Pretreatment with the platelet-activating factor (PAF) antagonist, 48740 RP, inhibited damage to endothelial cells, decreased vascular permeability and the number of neutrophils in the airways 24 h after exposure to LPS. The increase in the number of platelets in the airways caused by endotoxin was not affected. The results suggest that PAF modulates early inflammation after endotoxin inhalation.
Concerning the Assays for Autonomic Nerve Receptors
Authors: Hayashi Kouichi , Sugiyama Satoru , Taki Fumio , Shindow Joe , Takagi Kenzo , Satake Tatsuo , Ozawa Takayuki
Published: August 2009

Alterations in autonomic nerve receptors play an important role in various pathological conditions, including bronchial asthma. Nevertheless, receptor assay conditions such as incubation temperature and incubation time are not consistent among various investigators. This study was designed to clarify the effects of incubation temperature and time on alterations in the number of autonomic nerve receptors. Guinea pig lung membranes were divided into five groups which were incubated under different incubation temperatures and over different incubation times. After incubation, the following experiments were performed. Beta-, alpha-1-adrenergic, and muscarinic acetylcholine receptor assays were performed by direct binding technique using L-3H-dihydroalprenolol, 3H-bunazosin, and L-3H-quinuclidinyl benzilate, respectively. Elevation of incubation temperature and prolongation of incubation time caused a significant decrease in the number of beta-adrenergic receptors and an increase in the number of alpha-1-adrenergic receptors. The number of muscarinic acetylcholine receptors did not change significantly in spite of changes in incubation temperature and time. Adenylate cyclase activity was measured by following the synthesis of cyclic adenosine monophosphate from nonradioactive adenosine triphosphate. Isoproterenol-stimulated adenylate cyclase activity decreased significantly in correspondence with the elevation of incubation temperature and prolongation of the incubation time. Contents of free fatty acids in lung membranes were measured by high-performance liquid chromatography. Free fatty acid contents increased significantly in accordance with elevation of incubation temperature and prolongation of incubation time which reflected on the degradation of membrane phospholipids. Changes in membrane phospholipids are known to alter the number of receptors. Therefore, it is concluded that to minimize artifacts associated with the degradation of membrane phospholipids, receptor assays should be performed under low incubation temperature and for a short incubation time.
Measuring Human IgG4 by a Two-Site Immunoradiometric Assay with Monoclonal Antibodies
Authors: Bui R.H.D. , Molinaro G.A. , Eby W.C. , Jauhar R. , Mace J.W.
Published: August 2009

By using 2 monoclonal antibodies, we developed a solid-phase 2-site immunoradiometric assay for measuring human IgG4. The measuring range (0.05–20 μg/ml) covered more than 2 orders of magnitude. The sensitivity level should make this assay especially useful when IgG4 concentrations are too low to be detected by conventional methods.
Effect of Chemotactic Agents on Rat and Guinea Pig Eosinophil Cytotoxicity in vitro
Authors: Musgrove N.R.J. , Cook R.M.
Published: August 2009

Purified guinea pig and rat peritoneal eosinophils were examined for their ability to mediate an antibody-dependent cell cytotoxic reaction against 51Cr-labelled chick red blood cells in the presence of N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP), leukotriene B4, eosinophil chemotactic factor of anaphylaxis, or histamine. Guinea pig eosinophils were induced either by repeated intraperitoneal injections of polymyxin B sulphate or by a single intraperitoneal injection of saline. Using subagglutinating concentrations of antibody, the antibody-dependent cell cytotoxic response was significantly enhanced with FMLP, leukotriene B4, and eosinophil chemotactic factor of anaphylaxis. Histamine was without effect. Rat eosinophils were induced either by three intravenous injections of Sephadex G-200 followed by saline intraperitoneally, by a single intraperitoneal injection of saline, or by infection with the parasite Mesocestoides corti. None of the materials examined was able to enhance the cytotoxic activity of the rat eosinophil preparations over the range tested, in the presence of subagglutinating concentrations of antibody. The antibody-dependent cell cytotoxic response of eosinophils from infected animals was significantly reduced compared with those from Sephadex/saline-treated rats. Incubation of eosinophils obtained from infected animals with FMLP in the presence of agglutinating concentrations of antibody restored eosinophil cytotoxicity. None of the other materials was active. It is concluded that the ability of rat and guinea pig eosinophils to be activated in vitro is partly species related and partly due to the method of cell induction. Differences between the two species may be important when establishing animal models of eosinophil function.
Isolation of a Chemotactic Protein for Neutrophils from Human Ovarian Follicular Fluid
Authors: Seow W.K. , Thong Y.H. , Waters M.J. , Walters M. , Cummins J.M.
Published: August 2009

Human follicular fluid was found to contain a factor chemotactic for human neutrophils. This chemotactic activity was shown to be heat labile and trypsin sensitive. Gel permeation chromatography experiments indicate that the chemotactic protein has an approximate molecular weight of 100 kd. Isoelectric focussing suggests an isoelectric point for this protein of around 5.8. The biologic and clinical significance of this apparently unique chemotactic protein remains to be determined.
Effect of Irradiation and Depletion of C3-Complement Component on the Course of Treponema pallidum Infection in a Resistant Guinea Pig Strain
Authors: Wicher Konrad , Wicher Victoria , Jakubowski Adam , Bartholomew William , Nakeeb Shaheen
Published: August 2009

The role of complement and ionizing radiation in the natural resistance to Treponema pallidum infection of Albany guinea pigs was explored. Depletion of C3 by cobra venom factor for a period of 14 days affected neither the host’s susceptibility to infection nor the humoral response. Total body irradiation with 420 or 800 R was fatal within 20–30 days and there was no multiplication of treponemes in the infected host. Animals showing lethal signs were euthanized and tissues removed for examination. Exposure to a nonlethal dose of 300 R increased the susceptibility to infection (46% symptomatic lesions) and facilitated multiplication of treponemes at the site of inoculation and in the lymphoid organs, but the humoral response was not different from that of non-irradiated controls. The results seem to suggest a defect in antigen recognition by the immunocompetent cells in the resistant Albany guinea pigs.
Molecular Interactions between Human IgG, IgM Rheumatoid Factor and Streptococcal IgG Fc Receptors
Authors: Schröder A.K. , Gharavi A.E. , Christensen P.
Published: August 2009

Group A streptococci type M15 were previously shown to bind both human IgG via the Fc component and a purified monoclonal IgM kappa rheumatoid factor (IgM RF). Using 125I-labelled IgG and 125I-la-belled IgM RF, the present study gave association constants of 2.2 × 107 and 2.9 × 108M––1respectively. The binding of 125I-IgG to the streptococci was inhibited by unlabelled IgG, IgG Fc and fragment D of staphylococcal protein A but not by the IgM RFor F(ab′)2 of anti-idiotype antibodies to RF (anti-Id RF). Inversely, unlabelled IgM RF and anti-Id RF inhibited the binding of 125I-IgM RF markedly and unlabelled human IgG and IgG Fc only slightly or moderately, respectively. Thus, group A streptococci type M15 showed different binding sites for IgG Fc and the antibody combining sites of a human monoclonal RF. The findings were still more complex on a background of previous reports showing that streptococcal IgG Fc receptors and RFs bind to the same amino acids on the Fc molecule. This complex pattern may play a role in the pathogenesis of rheumatoid arthritis.
Modulation of Systemic and Mucosal Immune Responses to Inhaled Ragweed Antigen in Experimentally Induced Infection with Respiratory Syncytial Virus Implication in Virally Induced Allergy
Authors: Leibovitz Eugene , Freihorst Joachim , Piedra Pedro A. , Ogra Pearay L.
Published: August 2009

Groups of BALB/c mice were either sham-infected or infected intranasally with respiratory syncytial virus (RSV). On the third day following intranasal inoculation, all groups of mice were exposed by inhalation to ragweed antigen for 5 consecutive days and rechallenged with ragweed on day 31. Development of antibody activity to ragweed antigen was examined in serum and bronchial washings at regular intervals employing an ELISA assay for IgG and IgA antibody activity and passive cutaneous anaphylaxis for IgE-specific responses. The serum IgG and IgE antibody response to ragweed following primary exposure developed at significantly higher levels in mice previously infected with RSV, compared to sham-infected controls. In addition, an earlier rise in serum IgE response to ragweed occurred in the RSV-infected animals. IgG and IgA anti-ragweed antibody activity in bronchial washings was also observed with significantly higher levels in the RSV-infected animals when compared to the controls, although a similar increase in antigen-specific IgE activity in bronchial washings was found in both groups of animals. These findings support the possibility that mucosally restricted virus infections of the respiratory tract may enhance the development of sensitization and the magnitude of antibody responses to other inhaled allergens found concomitantly in the respiratory tract during acute infection.
Immunoblot Analysis of IgE-Binding Antigens in Spices
Authors: van Toorenenbergen A.W. , Huijskes-Heins M.I.E. , Leijnse B. , Dieges P.H.
Published: August 2009

The IgE response to coriander and other spices was studied by immunoblotting, after separation of the spice extracts by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A major IgE-binding component from coriander had an isoelectric point of pH5. After incubation of SDS-PAGE-separated spice extracts with serum from a patient with an occupational allergy to spices, a closely related pattern of IgE binding to coriander, dill and anise extract was observed. These results suggest that the botanically related spices, coriander, anise and dill, contain common IgE-binding structures.
Effect of Influenza Virus Infection on Allergic Sensitization to Inhaled Antigen in Mice
Authors: Holt Patrick G. , Vines Julia , Bilyk Natalie
Published: August 2009

Repeated exposure of healthy mice to an aerosol of ovalbumen (OA) leads to the development of IgE-isotype-specific immunological tolerance. However, when initial OA exposure occurs during the acute phase of influenza infection, tolerance does not occur, and the mice instead develop high titres of OA-specific IgE in response to subsequent challenge with the allergen.
Bronchoconstriction Induced by Platelet-Activating Factor in the Guinea Pig and Its Inhibition by CV-3988, a PAF Antagonist: Serial Changes in Findings of Lung Histology and Bronchoalveolar Lavage Cell Population
Authors: Takizawa Hajime , Ishii Akira , Suzuki Shuji , Shiga Junji , Miyamoto Terumasa
Published: August 2009

Platelet-activating factor (PAF), 1-o-alkyl-2-acetyl-sn-glyceryl-3-phosphoryl choline, induced acute bronchoconstriction in the guinea pig when intravenously administered in a very small dosage (30 ng/ kg). Serial changes in findings of lung histology and bronchoalveolar lavage (BAL) cell population were studied. Severe bronchial smooth muscle contraction and hypersecretion were followed by eosinophil infiltration. Increase in the number of eosinophils was also elucidated by BAL. Prominent damage to the ciliated bronchial epithelial cells was suggested from the lung histology, which was also supported by the fact that a number of ciliated cells was recovered by BAL. Late-onset bronchial constriction after PAF infusion was not observed. CV-3988, one of the specific PAF antagonists, completely inhibited the PAF-induced bronchoconstriction when administered before the PAF infusion. We concluded that PAF is a potent in vivo agent which brings about hypersecretion, eosinophil accumulation, bronchial epithelial damage as well as bronchoconstriction in the guinea pig, and that its action is completely blocked by CV-3988.
Production and Characterization of a Murine Monoclonal Antibody to Aspergillus fumigatus Antigen Having IgG- and IgE-Binding Activity
Authors: Kurup Viswanath P.
Published: August 2009

By employing hybridoma technology, a monoclonal antibody against Aspergillus fumigatus was produced. This antibody, isotyped as IgM k, reacted with 12 of 16 antigens extracted from 9 different A. fumigatus strains. This antibody also reacted with all 3 Aspergillus flavus antigens studied, but not with Aspergillus niger, Aspergillus terreus, Penicillium notatum or Candida albicans antigens. Western blot analysis indicated that this antibody reacted with two concanavalin A (Con-A) binding bands of the A. fumigatus antigen extract. The specific binding antigens were isolated using monoclonal antibody affinity chromatography. When used in immunoassay this fraction demonstrated strong IgG and IgE antibody-binding activities against patient sera. The antibody levels against the purified fraction were significantly higher in patients’ sera than in normal controls. The purified fraction demonstrated comparable reactivity with the crude A. fumigatus extract against patient sera, but the former has the added advantage of being pure and standardizable for dependable and reproducible results.
Relationship between Leukotriene C4 and an Uteroglobin-Like Protein in Nasal and Tracheobronchial Mucosa of Children
Authors: Volovitz Benjamin , Nathanson Ian , DeCastro Giovanna , Kikukawa Tadahiro , Mukherjee Anil B. , Brodsky Linda , Fox Molly J. , Ogra Pearay L.
Published: August 2009

Secretions from the nose and nasopharynx (NPS) are more readily accessible than tracheobronchial secretions (TBS) for clinical investigations, but it is unclear whether changes in mediator release in the nasopharynx reflect similar changes in the tracheobronchial tree. In order to clarify this question, NPS and TBS were taken from 20 children with tracheotomy and tested for the presence of leukotriene C4 (LTC4) and uteroglobin-like protein (UTG-LP). LTC4 and UTG-LP were measured both when the children were healthy and when they had clinical evidence of an acute respiratory tract illness. The mean concentration of LTC4 in NPS and TBS increased during illness although the mean concentration in NPS was significantly higher than in TBS during respiratory illness. In healthy children UTG-LP was detected only in TBS. During acute respiratory illness the concentration of UTG-LP in TBS decreased but remained significantly higher than in NPS. Data presented in this study indicate that changes in the LTC4 concentration in NPS appear to reflect changes in LTC4 concentration in TBS although the levels of LTC4 in NPS were significantly higher than in TBS. An inverse correlation between the concentrations of LTC4 and UTG-LP in NPS and TBS was demonstrated.
Discrepancy between Effects of in vivo and in vitro Administration of Gammaglobulin on Phagocytic Killing of Streptococcus pneumoniae in an Antibody-Deficient Serum
Authors: Braconier J.H. , Prellner K. , Sjöholm A.G.
Published: August 2009

Phagocytic killing of Streptococcus pneumoniae serotypes 6A, 14, 18C, 19F and 23F was investigated in the dysgammaglobulinemic serum of a patient with recurrent pneumococcal infections. Previous studies with this serum had established combined IgG2, IgG4 and IgA deficiency, deficiency with regard to specific antipolysaccharide antibodies and essentially normal complement functions. Phagocytic killing of all serotypes was reduced in the patient’s serum. Addition of immunoglobulin in vitro enhanced both classical and alternative complement pathway mediated opsonization. In constitution experiments neither purified C1q nor CRP influenced phagocytic killing. Surprisingly, intramuscular administration of a fairly small dose of gammaglobulin to the patient was associated with a rapid increase in the serum opsonic activity for serotype 23 F. The increased opsonization occurred before specific anticapsular antibodies were detectable in serum. The findings suggest that the possible effects of gammaglobulin treatment may not exclusively be related to the acquisition of serotype-specific antibodies.
Sensitivity of Passive Cutaneous Anaphylaxis in Rats
Authors: Watanabe Naohiro , Kobayashi Akio
Published: August 2009

The sensitivity of passive cutaneous anaphylactic (PCA) reactions was examined in rats infected with the nematode parasite, Nippostrongylus brasiliensis (Nb). PCA reactions with anti-DNP IgE antibody were remarkably suppressed by Nb infection already 10 days after infection and persisted for at least 28 weeks. 10 days after infection, no anti-Nb IgE antibody was present in the sera of infected rats, whereas 18 or 28 weeks after infection, circulating anti-Nb IgE antibody was present. It was concluded that two different mechanisms might explain the suppression. 10 days after infection, nonspecific IgE induced by potentiated IgE production inhibited passive sensitization with anti-DNP IgE antibody by occupying the IgE receptors on mast cells, whereas 18 or 28 weeks after infection, the mast cell IgE receptors were occupied by IgE antibody specific for the Nb antigen. The reactivity through non-immunological processes (skin sensitivity to compound 48/80) was not modified by Nb infection at any time after infection.
Effect of a Leukotriene D4 (LTD4) Antagonist on LTD4 and Ascaris Antigen-Induced Airway Responses in Rhesus Monkeys
Authors: Patterson Roy , Harris Kathleen E. , Krell Robert D.
Published: August 2009

An acute airway response to aerosolized leukotriene D4 (LTD4) qualitatively simulates an IgE-mediated ascaris antigen-induced airway response. The LTD4 airway response is completely inhibited by the LTD4 antagonist, ICI 198615, in normal monkeys. This LTD4 antagonist was then evaluated to determine whether it could inhibit the IgE-mediated ascaris antigen response using the threshold antigen dose-response system or the single antigen dose-response system. In 6 ascaris airway-reactive monkeys, the LTD4 antagonist demonstrated partial inhibition in 2 animals by either the threshold dose system in 1 animal or the single dose system in another animal. In the remaining animals there was no inhibition of the antigen-induced response by the LTD4 antagonist.
Stimulation of Human Neutrophil Degranulation by Mefloquine
Authors: Bates Edna J. , Ferrante A.
Published: August 2009

The effect of the antimalarial drug mefloquine on human neutrophil degranulation, chemiluminescence, superoxide production and viability was examined in vitro. Mefloquine was found to significantly stimulate the release of lysozyme, β-glucuronidase and myeloperoxide at a concentration of 10 μg/ml (2.5 × 10––5M) without loss of cell viability. At 40 μg/ml mefloquine (1 × 10––4M) cell viability was significantly decreased. Mefloquine at 10 μg/ml also significantly increased the release of lysozyme and β-glucuronidase but not myeloperoxidase when neutrophils were stimulated with opsonized zymosan. At a lower zymosan concentration myeloperoxidase release was also increased. Enzyme activity was not directly stimulated by mefloquine. Mefloquine at 10 μg/ml significantly increased luminol-dependent chemiluminescence but significantly inhibited lucigenin-dependent chemiluminescence when neutrophils were stimulated with opsonized zymosan. Under these conditions superoxide release, measured by cytochrome c reduction, was inhibited to a lesser degree. These results are discussed with reference to our previous report that mefloquine inhibits the neutrophil iodination reaction [Immunology 58: 125–130, 1986] and the use of mefloquine as an anti-inflammatory drug.
Human Basophilic Cell Differentiation Promoted by 2.5S Nerve Growth Factor
Authors: Matsuda Hiroshi , Switzer Jan , Coughlin Michael D. , Bienenstock John , Denburg Judah A.
Published: August 2009

In liquid cultures of human cord blood mononuclear cells, the activities of the 2.5S nerve growth factor (NGF) inducing basophil and eosinophil differentiation were investigated. Various concentrations of immunopurified 2.5S NGF derived from murine submaxillary glands were added to cultures with or without conditioned medium from a human T cell line (Mo-CM), which has previously been shown to produce activities stimulating granulocyte-macrophage colonies. Addition of NGF led to significant increases in differentiation of basophilic cells accompanied by histamine synthesis at 2 weeks in vitro; eosinophil differentiation was not increased in these cultures. In addition, NGF could be shown to amplify basophil differentiation induced by Mo-CM, and the activity of NGF inducing basophil differentiation was dependent on the presence of T lymphocytes. These results indicate that NGF stimulates T-lymphocyte-dependent basophilic cell differentiation from human cord blood progenitors and may in this way support differentiation of basophils or mast cells in vivo at sites of allergic tissue inflammation.
Reactivity of Mouse Antibodies against Bromelain-Treated Mouse Erythrocytes with Various Mouse Cells before and after Protease Treatment
Authors: Kawaguchi S.
Published: August 2009

A mouse monoclonal antibody against bromelain-treated mouse erythrocytes (BrMRBC) was conjugated with fluorescein isothiocyanate. Various cells from the blood and lymphoid tissues of mice were stained before and after protease treatment with the fluorescent antibody. Without protease treatment, only the platelets were specifically, though dully, fluorescent. Protease treatment made all the erythroctes, the majority of platelets, thymus and bone marrow cells and a small part of the spleen cells brightly fluorescent. The reactivity of the antibody with cells depended on the temperature, being stronger at 0 than at 37°C. The present findings demonstrate that various mouse cells besides erythrocytes bear the epitopes for anti-BrMRBC antibodies in exposed or hidden form.
Seasonal Patterns of Circulating Basophils
Authors: Chavance Michel , Herbeth Bernard , Kauffmann Francine
Published: August 2009

The seasonal patterns of circulating basophils were investigated in a sample of 33,000 French subjects, aged 20–60 years. Circannual variations (minimum in October, maximum in July) which might be related to pollens were observed. Periods with cold weather (temperature below 0°C) correlated to a temporary rise in basophils.