bluuebunny/crossref_metadata_2025 huggingface dataset:
Purification and N-Terminal Amino Acid Sequence of Two Birch Pollen Isoallergens (Bet v I and Bet v II)
Authors: Elsayed S. , Vik H.
Published: August 2009

The major allergen of birch pollen BV45 (Bet v I) was previously isolated by molecular weight exclusion chromatography and eluted in the molecular weight region of 15–29 KD. Further purification of this fraction on an SP-Trisacryl M cation exchange matrix allowed 6 peaks of which the 4th (BV4A4) and 6th (BV4A6) included two dominant IgE-binding birch pollen isoallergens designated Bet v I and Bet v II. Final purification, using the ‘Applied Biosystems’ Peptide Micro Separation System, revealed two sharp peaks with a high degree of homogeneity. This was ascertained by automatic N-terminal amino acid (AA) sequence analyses which showed high average repetitive yields of the phenyl-thiohydantoin (PTH) AAs of the isoallergens sequenced. N-terminal AA analyses of the two fractions allowed 51 cleavages with correct identifications of PTH AAs for 3 replicates. The sequence data of the two isoallergens showed large homologies with the hazel pollen allergen, Cor a I, the birch pollen allergen, Ag 23, and the translated cDNA sequence derived from cloning birch pollen allergen genes. The sequence homologies support that Betula verrucosa allergens were derived from a gene family expressing several isologous allergens, 2 of which with 13 variable residues in a segment of 51 AAs. The antigenicity of the two fractions, Bet v I and Bet v II, was demonstrated by fused rocket immunoelectrophoresis (FRIE) and by crossed immunoelectrophoresis (CIE) giving single symmetrical antigenic precipitation. The specific IgE-binding capacity of the two isoallergens was supported by: (a) immunoautoradiography of the plates provided from FRIE; (b) specific IgE-binding inhibition in radioallergosorbent test (RAST) for fraction BV4A, and (c) similarly high RAST inhibition for the individual isoallergens. The in vivo allergenic activity was clearly shown by skin prick and nasal provocation tests. In conclusion, two isoallergens from birch pollen with proximate pi values (5.2 and 5.5) were isolated for the first time and the N-terminal sequence characterized.
Immunization of Rabbits with Mosquito Bites: Immunoblot Analysis of IgG Antimosquito Antibodies in Rabbit and Man
Authors: Brummer-Korvenkontio Henrikki , Lappalainen Pekka , Reunala Timo , Palosuo Timo
Published: August 2009

Rabbits immunized with 50–60 Aedes communis mosquito bites developed IgG antibodies recognizing a 21.5-kD mosquito antigen detected by the immunoblot technique. This protein seems to be a major immunogen in the saliva of A. communis mosquitoes. Studies on 60 human sera revealed that IgG antibodies recognizing the 21.5-kD antigen occured in about 50% of young children and adults but were not found in the sera of 20 unexposed infants. The present immunoblot method allows further characterization of antigens and antibodies specific to mosquito saliva and contributes to understanding their role in mosquito bite reactions.
Comparison of the Anaphylactoid Response Induced in Rats by Castanospermine and Dextran
Authors: Doherty N.S. , Beaver T.H.
Published: August 2009

The plant alkaloid castanospermine (10 mg/kg or higher, i.p.; 400 mg/kg, p.o.) induced in some rat strains an anaphylactoid reaction similar to that induced by dextran, i.e. erythema and edema of the snout, ears and paws, for several hours after administration. 86% of the rats from a responsive strain responded to castanospermine while 79% responded to dextran. Rats which responded to castanospermine showed marked, but transient, tachyphylaxis to a second dose of castanospermine or dextran. Rats maintained on a complex-carbohydrate-free diet also responded to castanospermine, excluding the possibility that the effect was due to absorption of dextran-like, dietary, complex carbohydrates. These data raise the possibility that some apparent food allergies in man could be due to the presence in the diet of plant alkaloids with properties similar to those of castanospermine.
Allergen-Induced Biphasic Bronchoconstriction in Rats
Authors: Watanabe Akihiko , Hayashi Haruki
Published: August 2009

The development of an allergic bronchoconstriction model in rats is described. In actively sensitized Donryu strain rats, there was a remarkable biphasic increase in airway resistance within 10 min after antigen challenge on day 9 to day 21. The increase in airway resistance, correlated with the IgE titer and the dose of antigen, was inhibited by disodium cromoglycate (DSCG) or by aminophylline. This bronchoconstriction was remarkably blocked by methysergide (25 and 100 μg/kg) while pyrilamine inhibited it partially at the same dose. Serotonin (> 30 μg/kg) but not histamine (< 1,000 μg/kg) induced a bronchoconstriction. FPL-55712 (1, 10 mg/kg) inhibited it significantly. The content of thromboxane B2 (TxB2) in plasma increased during the bronchoconstriction while the content of peptide-leukotrienes (p-LTs) in plasma did not increase significantly. OKY-046 inhibited not only allergic bronchoconstriction but also the increase in TxB2 levels in plasma. The late phase of the bronchoconstriction was more susceptible to OKY-046. In conclusion, this model seems to be useful for the screening of antiasthma drugs because of a relationship with the dose of antigen, IgE titer and the susceptibility to an antiallergic drug or a bronchodilator. It is demonstrated that the major part of this allergic bronchoconstriction depends on serotonin, and it is also suggested that thromboxane A2 may play an important role in the late phase of the bronchoconstriction.
T Lymphocyte Stimulation of Human Small Intestinal Glycoprotein Biosynthesis: Effects of Anti-CD3 Antibody on Normal and Coeliac Mucosa
Authors: Crabtree J.E. , Heatley R.V. , Trejdosiewicz L.K. , Losowsky M.S.
Published: August 2009

The effects of mucosal T-cell activation on human small intestinal glycoprotein biosynthesis have been examined during short-term culture in vitro. The incorporation of 3H-glucosamine into tissue and secreted glycoproteins was determined. Activation of mucosal T lymphocytes of normal duodenal biopsies with the monoclonal anti-CD3 antibody significantly increased both the total glucosamine incorporation into glycoproteins (p < 0.01) and the secretion of in vitro radiolabelled glycoproteins (p < 0.001). This effect was inhibited by ciclosporin. Secretion of glycoproteins was also stimulated by culture with pokeweed mitogen. In patients with coeliac disease, culture of small intestinal biopsies with anti-CD3 antibody significantly increased (p < 0.05) glycoprotein biosynthesis in treated patients, but had no stimulatory effect in untreated patients with villous atrophy. These results show that activation of mucosal T lymphocytes induces quantitative changes in intestinal glycoprotein synthesis and secretion and T lymphocytes therefore have an important role in non-specific intestinal defences. The results are consistent with the suggestion that increased glycoprotein synthesis and secretion in untreated coeliac mucosa result from T-cell activation.
IgE-Binding Proteins from Pine (Pinus radiata D. Don) Pollen: Evidence for Cross-Reactivity with Ryegrass (Lolium perenne)
Authors: Cornford C.A. , Fountain D.W. , Burr R.G.
Published: August 2009

A complex mixture of pine (Pinus radiata D. Don) pollen proteins are rapidly released into aqueous solutions. IgE-binding proteins have been identified in these extracts using combined SDS-PAGE immunoblotting techniques. These IgE-binding proteins were detected using atopic patient and commerical pooled human sera known to be high in ryegrass-specific IgE. Enzyme-immunoassay inhibition studies revealed that leached P. radiata pollen proteins could partially inhibit serum IgE binding to ryegrass RAST discs thus providing preliminary evidence for allergen cross-reactivity between these two unrelated species.
Oxygen Radical Generation in Human Platelets: Dependence on 12-Lipoxygenase Activity and on the Glutathione Cycle
Authors: Jahn B. , Hänsch G.M.
Published: August 2009

Stimulation of the arachidonic acid metabolism in human platelets also induces chemiluminescence, which indicates the formation of highly reactive oxygen species. To analyze the pathway by which oxygen radicals are generated, we tested the dependence of O2–– generation on the eicosanoid metabolism. We found that inhibition of lipoxygenase, but not of cyclooxygenase, prevented the O2–– production. Moreover, depletion of cellular glutathione peroxidase resulted in reduced synthesis of the lipoxygenase product 12-hydroxyeico-satetraenoic acid, as well as in reduced O2–– thus establishing a link between O2–– generation, the lipoxygenase pathway and the glutathione cycle.
Neutrophil Derived Interference in the Fluorometric Determination of Histamine
Authors: Siegel Paul D. , Lewis Daniel M. , Olenchock Stephen A.
Published: August 2009

Zymogen-stimulated neutrophils release many pre- and nonpreformed factors including proteolytic enzymes, oxygen free radicals and arachidonic acid metabolites. Measurement of the histamine concentration of a supernatant fluid commonly employs an organic extraction procedure followed by condensation to o-phthalaldehyde. Diamine oxidase digestion studies and high-performance liquid chromatographic analysis suggest that a factor(s) derived from zymogen-stimulated neutrophils can produce a positive interference in the fluorometric determination of histamine.
Experimental Cedar Pollinosis in Rhesus Monkeys
Authors: Chiba Nobuyuki , Tamura Tatsuya , Koizumi Kinya , Tanigawa Manabu , Shiba Motoharu
Published: August 2009

In order to develop an experimental model for human cedar pollinosis, 4 rhesus monkeys (Macaca mulatta) were subcutaneously immunized with crude cedar pollen antigen, which contained 1.0 μg or 10 μg protein with aluminium hydroxide gel, six times every month. All 4 animals showed immediate-type cutaneous reactivity. Serum IgE antibody responses against the pollen antigen, as quantitated by the Prausnitz-Küstner reaction, were seen in 3 of 4 monkeys. Radioallergosorbent test scores increased in 2 monkeys and remained at higher levels throughout the study. Histamine release from leukocytes was positive in all animals. In conjunctival provocation tests, they showed clinical signs such as eye-scratching after the antigen was dropped into their eyes. This reaction is similar to that observed in humans and suggests this animal model might be useful in studying human cedar pollinosis.
Mast Cells in the Ovine Lower Respiratory Tract: Heterogeneity, Morphology and Density
Authors: Chen W. , Alley M.R. , Manktelow B.W. , Davey P.
Published: August 2009

The lower respiratory tract (LRT) of 6 adult sheep was fixed in either isotonic formalin-acetic acid or neutral buffered formalin in order to study the heterogeneity, morphology and density of mast cells (MCs). Two subtypes of MCs were found, one histochemically similar to connective tissue MCs (formalin resistant) and the other similar to mucosal MCs as found in the intestine of the rat (formalin sensitive). Although both subtypes were present at all levels of the tract, formalin-sensitive MCs were significantly more abundant (p < 0.01) at all levels, and their density increased distally from the trachea to the peripheral lung. The formalin-sensitive MCs were predominantly located in the alveolar septa and in the superficial lamina propria of airways and less frequently within the airway epithelium. The MCs in the ovine LRT were found to be morphologically heterogeneous at both the light-microscopic and electron-microscopic levels. These findings indicate that ovine respiratory tract MCs have similarity to human lung MCs, and therefore potential for use as a model for the study of human allergic disease of the respiratory system.
Anti-Cephalexin Monoclonal Antibodies and Their Cross-Reactivities to Cephems and Penams
Authors: Nagakura Naoki , Shimizu Tadayori , Masuzawa Toshiyuki , Yanagihara Yasutake
Published: August 2009

Three cell lines producing monoclonal antibodies (MAbs), Cep1-2, 2-2 and 6, against cephalexin were established and the immunoglobulin class of the MAbs was IgM. The cross-reactions of the MAbs with penams and cephems were examined by enzyme-linked immunosorbent assay (ELISA). The cross-reactivities of Cep1-2 and 2–2 were scarcely influenced by the structures of acyl side chains of cephems and penams. The cross-reactivities of Cep1-2 were affected by the presence or absence of dihydrothiazolidine ring of cephem nucleus in hapten-protein conjugates which were prepared by alkaline method, MBS method and activated ester method but the cross-reactivities of Cep2-2 were not. The findings suggest that Cep1-2 recognize the degradate product(s) of cephem nucleus and Cep2-2 recognize a new antigenic determinant (NAD), which is formed by the conjugation of beta-lactam and carrier protein. On the other hand, the cross-reactivities of Cep6 were influenced by the structure of amino acyl side chain. It seems that Cep6 recognize specifically the acyl side chain at the C-7 of cephem. In ELISA inhibition test, three MAbs showed different inhibition pattern. The reaction of Cep1-2 with cephalexin-HSA was inhibited by cephalexin lysate. Cep2-2 and Cep6 were weakly inhibited by the binding to cephalexin-HSA by cephalexin lysate. Furthermore, the reactions of all MAbs were remarkably inhibited by penicillamine. The above results indicate that the MAbs can recognize at least three epitopes of the degradate product(s) of cephem nucleus, NAD and acyl side chain in cephalexin-protein conjugate.
Induction of Eczematous Skin Reaction in Experimentally Induced Hyperplastic Skin of Balb/C Mice by Monoclonal Anti-DNP IgE Antibody: Possible Implications for Skin Lesion Formation in Atopic Dermatitis
Authors: Katayama Ichiro , Tanei Ryoji , Yokozeki Hiroo , Nishioka Kiyoshi , Dohi Yoshitane
Published: August 2009

Biphasic skin reaction with peak response at 1 and 24 h with prominent mast cell degranulation was induced by intravenous application of a monoclonal anti-DNP IgE antibody and subsequent skin test. This reaction was hapten-specifíc and mast-cell-dependent because no reaction was observed when oxasolone was used as an elicitation antigen or skin test was elicited in genetically mast-cell-deficient mice (W/Wvv). A partial spongiotic reaction and mononuclear cell infiltration into the epidermis were observed in mice with hyperplastic epidermis induced by topical retinoic acid. Cotransfer of DNFB-sensitized lymph node cells with anti-DNP IgE antibodies failed to enhance the skin test reaction in unsensitized mice. These results suggest that, to some degree, IgE antibody may play some role in the development of eczematous skin lesions in the rodent system without the involvement of cellular hypersensitivity.
Expression of IgH Promoter/Enhancer Ly-1 Transgene in Hematopoietic Chimeric Mice Generated by Embryonic Stem Cell Line
Authors: Sumida Takayuki , Maeda Toshiro , Koike Takao , Tomioka Hisao , Yoshida Sho , Taniguchi Masaru , Rajewsky Klaus
Published: August 2009

We have produced hematopoietic chimeric mice from an embryonic stem (ES) cell line earring Ly-1 cDNA under the control of IgH promoter and enhancer. Various amounts of serum IgM (5–86% of total IgM) in chimeric mice were of ES origin and 30–60% of IgM-positive B cells from the chimeric mice analyzed were reconstituted from ES cells. Using these chimeric mice, the expression of the Ly-1 transgene on lymphoid tissues was examined by polymerase chain reaction assay with primers specific for the transgene, and by cell sorter analysis. Transcription of the Ly-1 transgene was detected in spleen cells, thymocytes and lymph node cells; however, the expression of the Ly-1 molecule was observed only on lipopolysaccharide (LPS)-stimulated splenic IgM-positive B cells but not on resting splenic B cells. There was no significantly increased expression of Ly-1 on splenic T cells and thymocytes. Thus, our findings demonstrate that conventional splenic B cells could express the Ly-1 transgene on their surface in vivo after LPS stimulation. Also discussed is the ES-derived chimeric hematopoietic system.
Mast Cells, Tissue Histamine and Eosinophils in Early- and Late-Phase Skin Reactions: Effects of a Single Dose of Prednisolone
Authors: Hammarlund Ann , Pipkorn Ulf , Enerbäck Lennart
Published: August 2009

Skin prick tests with allergen and histamine were performed on the volar aspect of the forearms in a double-blind, cross-over study with 40 mg of prednisolone and placebo in 16 pollen-allergic subjects. Skin biopsies were taken before any treatment and 15 min (group 1; n = 8) and 6 h (group 2; n =8) after local challenge with allergen, corresponding to the timing of an early- and late-phase reaction. The specimens were used for the histological evaluation of mast cell and eosinophil density as well as for the determination of the histamine and protein content. The size of the induced weal and flare area as well as of any late-phase reaction was determined using digitized planimetry. The single dose of prednisolone, given 2 h prior to challenge, did not affect the size of the weal and flare response. Only 4 of the individuals developed a visible late-phase response. Eosinophils were virtually absent before allergen exposure, but were already present 15 min after allergen challenge, largely associated with the blood vessels, and were numerous at 6 h. There was, however, no relationship between eosinophil density and the presence or extent of any visual late phase. The mast cells/basophils showed a tendency to increase at the 6-hour determination. The infiltration of eosinophils was blocked by the glucocorticoid. This treatment also induced a difference in the mast cell density at the 6-hour determinations, associated with a similar difference in the histamine content of the biopsy specimens. From the present study, we can conclude that the eosinophil infiltration is an early event and can be blocked by a single dose of prednisolone only 2 h after the administration of the glucocorticoid.
Modulation of Rat Peritoneal Mast Cell and Human Basophil Histamine Release by Estrogens
Authors: Cocchiara Roberta , Albeggiani Giuseppe , Di Trapani Giovanna , Azzolina Antonina , Lampiasi Nadia , Rizzo Federico , Geraci Domenico
Published: August 2009

This study was undertaken to investigate the effect of estrogens on the histamine release mediated by IgE in rat peritoneal mast cells (PMC) and in sensitized human basophils. The estrogens were found to enhance the histamine release of either rat PMC and sensitized human basophils upon stimulation with anti-IgE. The enhancement was estrogens dose-dependent reaching the maximum value of 23% for rat PMC and 41% for sensitized human basophils stimulated with anti-IgE upon preincubation with 10––8M estrogens. Moreover, when purified PMC were used, the enhancing effect was still detected, suggesting a direct interaction between estrogens and mast cells. The enhancing effect took place quite rapidly reaching plateau levels in about 60 min. Basophils preincubated at 4 instead of 37 °C did not give any appreciable enhancement, suggesting that it was temperature-dependent and that the effect observed was not due to cytotoxicity. Incubation of PMC or human basophils with estrogens alone, without challenge with anti-IgE, did not give any detectable histamine release. The enhancement of histamine release by estrogens is probably mediated by IgE molecules present on the cell membrane, since this effect was not observed on challenge with substance P or compound 48/80, two segretagogues known to induce histamine release not via IgE.
Increased Expression of High-Affinity Low-Density Lipoprotein Receptors on Human T-Blasts
Authors: Huber Lukas A. , Böck Günther , Jürgens Günther , Traill Karine N. , Schönitzer Dieter , Wick Georg
Published: August 2009

Like all cells, lymphocytes need cholesterol for proper function, a requirement met by a finely tuned homeostasis between intracellular synthesis and uptake from the environment via low-density lipoproteins (LDL). We used flow cytometry to analyze the receptor activity of resting cells and T blasts incubated/activated in serum-free culture medium, or in medium supplemented with 25–5,000 μg/ml LDL. Dioctadecyl-indocarbocyanine has proved to be a useful fluorescent probe for investigating the LDL receptor activity of lymphocytes. The results show the receptor activity of day-3 resting T cells to be reduced more than 50% by 50 μg LDL/ml, whereas 100-fold higher concentrations are necessary to achieve the same level of reduction in day-3 PHA blasts. The LDL receptor activities of individual blood donors’ resting T cells, in vitro cholesterol-deprived resting T cells, and activated T blasts, were compared using two analytical techniques: spectrofluorometric analysis of detergent-solubilized cell suspensions and flow cytometric analysis of single living cells. Receptor affinity was determined by Scatchard analysis of spectrofluorometric binding curves, and by Line-weaver-Burke plots of flow cytometric data. Both methods yielded essentially identical dissociation constants (Kd) for cholesterol-deprived resting T cells and mitogen-activated T blasts, which fell in the expected range for the high-affinity LDL receptor (4.1–8.9 nM). In addition, spectrofluorometric analysis, but not flow cytometry, permitted quantification of LDL uptake. The low receptor activity of freshly isolated T cells, however, could only be reliably measured by flow cytometry; this highly sensitive technique not only afforded specific binding curves, but also determination of Kd of the same high affinity (7.5 nM) as that of T cells with upregulated LDL receptor activity.
Carl Prausnitz Memorial Lecture
Authors: Sehon Alec H.
Published: August 2009

Some of the studies employing nonantigenic, nonallergenic and tolerogenic derivatives of antigens and allergens, synthesized by coupling onto them an optimal number (n) of molecules of monomethoxy-polyethylene glycol (mPEG), are briefly reviewed. Administration of antigen (mPEG)n conjugates into mice resulted in specific immunosuppression, which was mediated by antigen-specific suppressor T (Ts) cells and by suppressor factor(s) (TsF) produced by these cells. These Ts cells were cloned and shown to be Thy-1.2+, CD3+, CD4––, CD5––, CD8+, and expressed the αβ heterodimer of conventional T cell receptors (TCR). The TsF had the functional activity of Ts cells and possessed at least one epitope related to the α-chain of TCR. The results of clinical trials of mPEG conjugates of common allergens are briefly referred to; it is suggested that the time is opportune for the evaluation of the therapeutic efficacy of mPEG conjugates of pure allergens which can be synthesized on an industrial scale by recombinant DNA technology. Finally, possible applications of tolerogenic mPEG conjugates of xenogeneic monoclonal antibodies, immunotoxins and immunogenic recombinant lymphokines to the development of immunotherapeutic strategies in oncology, transplantation, autoimmunity and acquired immunodeficiency syndrome are discussed.
In vitro Development and Functions of Human Mast Cells
Authors: Ishizaka T. , Furitsu T. , Inagaki N.
Published: August 2009

A long-term co-culture of mononuclear cells of human umbilical cord blood with mouse embryo-derived 3T3 fibroblasts resulted in the development of human mast cells. These mast cells are morphologically and functionally mature cells, containing 1.4–2.8 μg histamine per 106 cells and bear approximately 105 FcεRI per cell. The mast cells sensitized with human IgE released histamine upon challenge with anti-IgE. Electron-microscopic analysis of the cells showed that these cells were mature human mast cells, and clearly different from basophilic granulocytes. Most of the mast cells contained some granules with regular crystalline arrays and both tryptase and chymase, resembling human skin mast cells. When mononuclear cells of cord blood were seeded in a millicell insert which was placed on 3T3 fibroblasts monolayer, the number of mast cells developed in the millicell inserts was comparable to those developed in the co-culture of the same cord blood cells with 3T3 fibroblasts. Recent observations that mast cells developed in the presence of concentrated culture supernatants of 3T3 fibroblasts without fibroblasts feeder layers, confirmed that soluble factors released from 3T3 fibroblasts are essential and sufficient for the differentiation of human mast cell progenitors in vitro. Analysis of functional characteristics of cultured mast cells revealed that they respond to anti-IgE, Ca2+ ionophore A23187 and substance P for histamine release, but failed to respond to compound 48/80 and FMLP. Upon anti-IgE challenge, sensitized mast cells generated approximately 80 ng PGD2 per 106 cells, and approximately 50 ng of LTC4 per 106 cells but no detectable generation of LTB4. Characterization of growth factors for human mast cells, followed by gene cloning of the growth factor(s) will provide us with a powerful tool for in vitro development of mature human mast cells, which will pave the way to further detailed studies on human mast cells in many aspects.
Increased Numbers of Th2-Like CD4+ T Cells in Target Organs and in the Allergen-Specific Repertoire of Allergic Patients
Authors: Romagnani S. , Maggi E. , Parronchi P. , Macchia D. , Piccinni M.P. , Ricci M.
Published: August 2009

Phytohemagglutinin (PHA)-induced human T cell clones (TCC) derived from conjunctival flogistic tissues of 3 patients with vernal conjunctivitis produced unusually high amounts of interleukin-4 (IL-4) and no, or limited amounts of, gamma-interferon (IFN-γ). Allergen (Dermatophagoides pteronyssinus or Lolium perenne group I)-specific TCC derived from peripheral blood of two atopic donors produced significantly higher amounts of IL-4 and significantly lower amounts of IFN-γ than TCC specific for bacterial antigens (tetanus toxoid and PPD) contemporarily established from the same donors. These data provide evidence for a compartimentalization of Th2-like helper T cells in target organs and in the allergen-specific T cell repertoire of allergic patients. Non-B, non-Tbone marrow cells could produce IL-4, but not IL-2 or IFN-γ, in response to cross-linkage of Fcε type I receptors. These cells may further contribute to the maintenance and amplification of allergic inflammation.
Histamine-Releasing Factors and Inhibitory Factors
Authors: Grant Andrew , Alam Rafeul , Lett-Brown Michael A.
Published: August 2009

Peripheral-blood mononuclear cells (MNC) synthesize several histamine-releasing factors (HRF) spontaneously and when stimulated. Some of the characterized cytokines have histamine-releasing activity, especially connective tissue activating peptide III, neutrophil-activating peptide 2 and interleukin-3 (IL-3). At least two species of HRF remain to be characterized. MNC also secrete a histamine release-inhibitory factor (HRIF), which is a highly specific inhibitor, because it antagonizes only HRF. IL-8 resembles the low-molecular-weight species of HRIF in terms of size and specificity.
IgE-Dependent Histamine-Releasing Factors
Authors: MacDonald Susan M. , Langdon Jacqueline M. , Greenlee Brian M. , Kagey-Sobotka Anne , Lichtenstein Lawrence M.
Published: August 2009

A cytokine, termed histamine-releasing factor (HRF) and produced by many cell types, has become the focus of research by many investigators due to its potential importance as a stimulus in chronic inflammation. We are producing and characterizing an HRF which causes IgE-mediated histamine release from human basophils. Following extensive purification procedures, the molecule will be sequenced and synthesized. A functional heterogeneity of IgE molecules was revealed by these studies. We are currently producing IgE antibody in vitro and testing the hypothesis that differential glycosylation is the basis for the heterogeneity. Knowledge of the structures and interactions of these molecules should advance our understanding of allergic and more chronic diseases.
Platelet-Activating Factor Modulates Interleukin-6 Production by Mouse Fibroblasts
Authors: Braquet Pierre , Pignol Bernadette , Maisonnet Thierry , Mencia-Huerta Jean Michel
Published: August 2009

We compared the effects of platelet-activating factor (PAF), interleukin-1β (IL-1β) and polyriboinositic-polyribocytidylic acid (poly-I:C) on IL-6 production by confluent L929 mouse fibroblasts. At concentrations above 1 μM, PAF dose-dependently enhanced IL-6 production; at 5 μM PAF this increase (72.7 ± 19.9 U/ml) was higher than that evoked by 100 U/ml IL-1β (5.7 ± 0.4 U/ml) or 50 μg/ml poly-I:C (39.3 ± 6.7 U/ml). The IL-6 production induced by 5 μM PAF was not inhibited by addition of the specific PAF antagonist BN 52021 (10 μM) to the incubation medium. These results demonstrate that, as this is the case for IL-1, PAF also modulates IL-6 production.
Platelet-Derived Growth Factor Can Activate Purified Primate, Phorbol Myristate Acetate-Primed Eosinophils
Authors: Bach Michael K. , Brashler John R. , Stout Barbara K. , Johnson Herbert G. , Sanders Martin E.
Published: August 2009

We are interested in the physiologic mechanisms of eosinophil activation because of the presumed participation of activated eosinophils in the inflammatory sequelae of asthma. Suspecting that other formed elements of the blood may contribute to such an activation, we examined the capacity of platelet-derived growth factor (PDGF), a product of activated platelets, to activate eosinophils. We found that highly purified monkey and human eosinophils, but not guinea pig eosinophils, were activated by PDGF (superoxide anion production) in a dose-dependent fashion. Moreover, this activation was further dependent on a prior ‘priming’ of the cells by a brief exposure to subthreshold concentrations of phorbol ester. The response was specific for the BB homodimer of PDGF suggesting it is receptor-dependent.
Interleukin-5 mRNA in Mucosal Bronchial Biopsies from Asthmatic Subjects
Authors: Hamid Q. , Azzawi May , Ying Sun , Moqbel R. , Wardlaw A.J. , Corrigan C.J. , Bradley B. , Durham S.R. , Collins J.V. , Jeffery P.K. , Quint Diana J. , Kay A.B.
Published: August 2009

Using in situ hybridization, we have investigated the expression of interleukin-5 (IL-5) mRNA in bronchial biopsies from asthmatics (n = 10) and controls (n = 9). The number of IL-5-mRNA-positive cells were compared with the number of CD25+ and EG2+ cells and total eosinophil counts. Specific hybridization signals for IL-5 mRNA were demonstrated in 6 out of the 10 asthmatic subjects but in none of the controls. The 6 IL-5-mRNA-positive asthmatics tended to have more severe disease and showed a significant increase in the degree of infiltration of the bronchial mucosa by activated T lymphocytes and eosinophils.
lnterleukin-4 Induced IgE and IgG4 Secretion by B Cells from Atopic Dermatitis Patients
Authors: Spiegelberg Hans L. , O’Connor Richard D. , Falkoff Reuben J.M. , Beck Luanda
Published: August 2009

Peripheral blood mononuclear cells from 8 normals and 8 patients with atopic dermatitis (AD) were cultured with recombinant interleukin-4 (IL-4) and the IgE and IgG subclass levels in the culture supernatants measured by radioimmunoassays. IL-4 induced IgE and IgG4 secretion by B cells from both normals and AD patients whereas it has no consistent effect on IgG1, IgG2 and IgG3 secretion. The IL-4 dose response was similar for IgE and IgG4 secretion by cells from both normals and AD patients. On the average, the patients’ cells secreted more IgE and less IgG4 than the cells from normals, but because of a large variation, the differences were not significant. However, the ratio of IgG4:IgE secretion was significantly greater for normals than AD patients (mean ± SEM 7.1 ± 1.6:1 vs. 1.5 ± 0.4:1; p < 0.01). The data demonstrate that IL-4 induces IgE and IgG4 secretion by B cells from both normals and AD patients and suggest that the IL-4 induced switch from IgM to IgG4 or IgE secretion may proceed preferentially to IgE in AD patients as compared to normals.
The Molecular Biology of Eosinophil Granule Proteins
Authors: Hamann Kimm J. , Barker Robert L. , Ten Rosa M. , Gleich Gerald J.
Published: August 2009

Here, we briefly review the molecular biology of the human eosinophil granule proteins, major basic protein (MBP), eosinophil peroxidase (EPO), eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). The nucleotide sequence of MBP cDNA indicates that MBP is translated as a 25.2-kilodalton preproprotein; the mpb gene consists of 6 exons and 5 introns spanning 3.3 kilobases (kb). The ∼2.1-kb nucleotide sequence of EPO cDNA corresponds to a prosequence, light chain and heavy chain in that order; similarities to other peroxidases suggest the existence of a multigene family. EDN and ECP cDNAs and genes are remarkably similar throughout, suggesting a relatively recent divergence. Promoter regions of the 4 genes show interesting differences and similarities which may be related to differential gene regulation.
Potential Implication of Endothelial Cells in Bronchial Asthma
Authors: Lassalle Ph. , Delneste Y. , Gosset Ph. , Tonnel A.B. , Capron A.
Published: August 2009

Inflammatory cells like eosinophils, neutrophils or mononuclear phagocytes have long been recognized as essential components in the pathophysiology of asthma. After recruitment in situ and subsequent activation, they are considered as responsible for epithelial and submucosal bronchial alterations. However, to access to the inflammatory site, these cells have to cross the endothelial wall, suggesting so a potential implication of endothelial cells (EC) in bronchial asthma. To test this hypothesis, we studied in a first step the modulation of vascular adhesions like intercellular adhesion molecule-1 (ICAM-1) on EC: supernatants of alveolar macrophages (AM) recovered by bronchoalveolar lavage in patients exhibiting a late asthmatic reaction, induced an enhanced expression of ICAM-1 on EC preparations; increase of ICAM-1 was clearly correlated to amounts of tumor necrosis factor-α (TNFα) present in AM supernatants, as shown by inhibition experiments with anti-TNFα antiserum. The second way to explore the possible role of EC in asthma was the detection of autoantibodies to EC in various allergic disorders: antibodies against a 120-kD EC antigen in patients with allergic granulomatosis and angiitis, antibodies towards a 55-kD component, common to human EC and platelets in patients with severe asthma, namely characterized by their corticosteroid dependence or by aspirin-induced intolerance. So our data suggest that bronchial asthma might result from either EC activation, through the induction of surface adhesion molecules or from an autoimmune process involving EC antigens.
T Cells and Asthma
Authors: Walker Christoph , Virchow, Jr. Johann-Christian , Iff Thomas , Bruijnzeel Pieter L.B. , Blaser Kurt
Published: August 2009

Abnormalities of peripheral-blood lymphocyte subsets and activation markers were detected in patients with both allergic and nonallergic asthma. Most allergic asthmatics were characterized by increased numbers of IL-2R+ helper T cells and CD23+B cells. In contrast, nonallergic asthmatics showed increased numbers of IL-2R+ and HLA-DR+ helper and cytotoxic T cells, and a clear redistribution from naive (CD45RA+) to memory (CD45RO+) cells. The number of IL-2R+ T cells correlated with the number of CD23+ B cells in allergic asthma. These changes in the distribution and activation state of T cells suggest an active role for T cells in the pathogenesis of both allergic and nonallergic asthma.
Basidiomycete Allergy: Identification and Characterization of an Important Allergen from Calvatia cyathiformis
Authors: Homer Elliott , Lopez M. , Salvaggio John E. , Lehrer Samuel B.
Published: August 2009

Basidiomycetes were not considered as major aeroallergen sources until spore traps revealed the prevalence of basidiospores, which were in some cases associated with epidemic outbreaks of asthma. More recently, we established that approximately one third of subjects with respiratory allergic disease were skin prick test positive to basidiospore extracts. Bronchial challenge with these extracts induced reversible immediate and late phase bronchospasm in sensitive subjects. Screening individual RAST-positive sera for reactivity to immunoprinted Calvatia cyathiformis spore extract indicated that a basic component with pI 9.3 reacted with 63% of test sera. Further analysis by RAST and immunoprint (IP) inhibition showed that this component, Calc Bd9.3, appeared to cross-react with spore extracts from 3 of 5 other species tested. To isolate this component for additional analysis, a protocol was used with two sequential stages of preparative isoelectric focusing (IEF) followed by hydrophobic interaction chromatography. This yielded purified Cal c Bd9.3 that retained IgE-binding activity by IP, was a single band by IEF and SDS-PAGE (16 kD) analysis when stained with Coomassie blue. Double diffusion in gel with rabbit antiserum to Cal c BD9.3 demonstrated a single precipitin band in crude extract that was identical to that obtained with purified Cal c Bd9.3. By IP this antiserum recognized pure Cal c Bd9.3 and the 9.3 band in crude extracts from C. cyathiformis and 3 other species, corroborating the cross-reactivity. However, other bands were also recognized. These studies indicate that basidiospores are major fungal aeroallergens, many of which contain a common basic allergen to which most sensitized subjects react.
Analysis of ‘Clean’ Air Samples by Scanning and Transmission Electron Microscopy and X-Ray Probe Micro-Analysis
Authors: Chavarria F. , Heap P. , Carswell F.
Published: August 2009

Air samples taken from the clean air over Bristol were examined using scanning, transmission and X-ray probe micro-analysis. A diversity of material was identified demonstrating the importance of man-made pollutants which have the potential to produce lung damage in addition to pollen moulds.
Ultrastructural Localization of the Allergen Der p I in the Gut of the House Dust Mite Dermatophagoides pteronyssinus
Authors: Thomas B. , Heap P. , Carswell F.
Published: August 2009

The sites and concentrations of Der p I within the house dust mite were determined. Highest concentrations were found in the epithelium of the posterior end of the mite stomach, implying that this is the site of Der p I synthesis and secretion.
Tribolium confusum (Confused Flour Beetle, Rice Flour Beetle) – An Occupational Allergen in Bakers: Demonstration of IgE Antibodies
Authors: Schultze-Werninghaus G. , Zachgo W. , Rotermund H. , Wiewrodt R. , Merget R. , Wahl R. , Burow G. , zur Strassen R.
Published: August 2009

Specific IgE to proteins from Tribolium confusum (TC), a flour beetle, was detected in 9/125 sera of subjects exposed to rye and wheat flour. TC RAST was not inhibited by Dermatophagoides pteronyssinus, rye or wheat flour. Immunoblot experiments showed specific binding to three proteins from adult TC or pupae, not present in rye or wheat flour. These findings suggest that TC might act as an occupational allergen in a proportion of bakers.
The Immune Response to Anti-Idiotype Antibodies Bearing an Internal Image Epitope of Tetanus Toxin/Toxoid
Authors: Poskitt D.C. , Turnbull S. , Macdonald L. , Jean-François M.J.B.. , Yasmeen D.
Published: August 2009

Mouse monoclonal (Ab1) anti-tetanus toxin/toxoid antibodies were used to raise Ab2β (tetanus toxin/toxoid internal image bearing) anti-idiotype antibodies in rabbits. Those rabbit serum antibodies (Ab2β) that did not bind to mouse serum proteins on an affinity column gave rise to an Ab3 anti-tetanus toxin/toxoid antibody response in mice. Rabbit serum antibodies that did bind to the affinity column, when eluted and used to inoculate mice also gave rise to an Ab3 anti-tetanus toxin/toxoid antibody response. It is suggested that one population of rabbit Ab2β anti-idiotype antibodies (unbound fraction) bears a partial or complete internal image of a tetanus epitope (Ab2β1) while others (bound fraction) bear a complete or partial mirror image of a mouse immunoglobulin epitope as well (Ab2β2).
Antiasthmatic Effects of Picrorhiza kurroa: Androsin Prevents Allergen- and PAF-induced Bronchial Obstruction in Guinea Pigs
Authors: Dorsch W. , Stuppner H. , Wagner H. , Gropp M. , Demoulin S. , Ring J.
Published: August 2009

In the Ayurvedic medicine, Picrorhiza kurroa Royle ex Benth. is used for the treatment of liver and lung diseases. Using different chemical and pharmacological methods, we could identify the phenol glycoside androsin as active compound preventing allergen and platelet-activating factor induced bronchial obstruction in guinea pigs in vivo (10 mg/kg p.o.; 1 h prior to the inhalation challenge). Histamine release from human polymorphonuclear leukocytes in vitro was inhibited by other compounds yet to be identified.
Influence of Mepacrine on the Reaction of Adoptive Cutaneous Anaphylaxis
Authors: Bashmakov Yu.K. , Sidorenko T.V. , Dugovskaya L.A.
Published: August 2009

In a test of adoptive cutaneous anaphylaxis, the influence of the phospholipase A2 inhibitor mepacrine, on the intensity of the local anaphylactic reaction was investigated in the skin of recipients following intracutaneous injection of syngenic immune splenocytes. Injection of the mepacrine solution with preincubated sensibilized splenocytes inhibits the cutaneous anaphylactic reaction after a single intravenous administration of allergen to recipients. The inoculation of immune splenocytes, preincubated in mepacrine but without the phospholipase A2 inhibitor, to the skin of syngeneic recipients is accompanied by less suppression of the local skin anaphylactic reaction than with a common injection of mepacrine with immune splenocytes.
Selective Impairment of T Lymphocyte Activation following Contact Sensitization with Oxazolone
Authors: Kimber Ian , Foster John R. , Baker David , Turk John L.
Published: August 2009

We have previously reported that topical exposure of mice to oxazolone results in the appearance of regulatory mechanisms which markedly depress lymph node cell (LNC) proliferative responses to subsequent challenge with the same chemical. In the present study, we have sought to identify the cellular targets for such immunoregulation. Autoradiographic analyses revealed that although pre-exposure to oxazolone caused a substantial reduction of paracortical hyperplasia following challenge, the frequency of proliferating cells in lymphoid follicles was slightly increased. That B lymphocyte responses are unaffected by oxazolone-induced immunoregulation was confirmed by investigation of anti-hapten antibody formation by draining LNC. Challenge with oxazolone resulted in an accelerated antibody response in mice previously exposed to the same chemical. These data reveal that the active immunoregulation induced following sensitization with oxazolone is selective for T lymphocytes. Evidence is presented that CD4+ and CD8+ T lymphocytes possess equivalent sensitivity to these mechanisms.
Effects of Cetirizine on Human Eosinophil and Neutrophil Activation in vitro
Authors: Walsh G.M. , Moqbel R. , Hartnell A. , Kay A.B.
Published: August 2009

The ability of cetirizine, a novel antihistamine agent, to inhibit the in vivo activation of human eosinophils, neutrophils and monocytes has been investigated using C3b- and IgG-dependent rosette formation, cytotoxicity against opsonised parasitic larvae and adherence to plasma-coated glass (PCG). The drug inhibited platelet-activating factor (PAF)-induced enhancement of eosinophil and neutrophil IgG (Fc) and complement (C3b) rosettes with an IC50 of 2 × 10––5M. There was also comparable inhibition of PAF-dependent enhancement of eosinophil cytotoxicity (for complement-coated schistosomula of Schistosoma mansoni). Cetirizine inhibited PAF-induced eosinophil, but not neutrophil, hyperadherence to PCG. These data support the view that cetirizine may exert some of its anti-allergic effects by inhibiting the activation of human granulocytes and that it may also selectively inhibit PAF-induced eosinophil hyperadherence.
Effect of Neutral Endopeptidase Inhibition on Substance-P-Induced Increase in Short-Circuit Current of Canine Cultured Tracheal Epithelium
Authors: Tamaoki Jun , Sakai Noritaka , Isono Kazuo , Kanemura Toshinori , Chiyotani Atsushi , Yamauchi Fumiko , Takizawa Takao , Konno Kimio
Published: August 2009

We studied the effect of substance P (SP) on the electric properties of cultured canine tracheal epithelium and its possible modulation by neutral endopeptidase (NEP) by Ussing’s short-circuited technique in vitro. Addition of SP (5 × 10––6M) to the mucosal side increased short-circuit current (SCC) from 5.1 ± 0.9 to 10.3 ± 2.2 μA/cm2 (mean ± SE; p < 0.01), which was accompanied by increases in transepithelial potential difference and conductance. The effect of the mucosal SP on SCC was dose-dependent, with the maximal increase from the baseline value being 5.8 ± 1.0 μA/cm2 observed at 5 × 10––5M. The NEP inhibitor phosphoramidon (10––5M)did not affect these responses. On the other hand, SCC was not altered by the addition of SP to the submucosal side. However, it was increased dose-dependently in the presence of phosphoramidon (10––5M) but not in the presence of captopril, bestatin or leupeptin. This stimulatory effect of submucosal SP was abolished by furosemide, diphenylamine-2-carboxylate and Cl-free medium, but not by amiloride. These results suggest that SP may selectively stimulate Cl secretion across the airway epithelium and that this effect may be modulated by submucosal NEP.
Characterization of Eosinophils in a Continuous Ambulatory Peritoneal Dialysis Patient with Eosinophilic Peritonitis
Authors: Sligh Julie M. , Gottlieb Steven J. , Domoto Douglass T. , Slavin Raymond G.
Published: August 2009

A continuous ambulatory peritoneal dialysis patient developed eosinophilic peritonitis and was followed for 7 months. After 1 month, the peritonitis resolved, with a concomitant drop in percentage of hypodense eosinophils (Eos) recovered from peritoneal dialysate (PD) as well as a drop in fluid major basic protein levels. Blood eosinophil differential percentages were low, but the percentage of hypodense Eos in the blood tended to be relatively increased. Stool samples showed no evidence of parasitic infection, and epicutaneous skin tests were negative. Leukotriene C4 levels remained relatively constant as did white blood cell counts. Flow cytometric analysis of lymphocytes and granulocytes from PD and blood revealed high levels of CD23-positive lymphocytes.
Cypress (Cupressus sempervirens) Pollen Allergens: Identification by Protein Blotting and Improved Detection of Specific IgE Antibodies
Authors: Ford S.A. , Baldo B.A. , Panzani R. , Bass D.
Published: August 2009

On the basis of results of an investigation of the effects of different treatments employed, a dialysed and reduced extract of Cupressus sempervirens was separated electrophoretically on sodium dodecylsulphate-polyacrylamide gels before being transferred and then fixed with glutaraldehyde to nitrocellulose membrane. Probing with sera from 91 subjects allergic to C. sempervirens pollen followed by detection of bound IgE antibodies with [125I]-labelled anti-human IgE revealed 17 IgE-binding proteins in the molecular weight range 14–96 kilodaltons (kDa). One component, of molecular weight ∼42 kDa, reacted with IgE antibodies in the sera of 81.3% of the allergic subjects and, for each of the subjects, this component bound the greatest quantity of IgE. Almost 50% of the sera recognized only the ∼42 kDa component, reinforcing the conclusion that this component is the major allergen of C. sempervirens pollen. A comparative study employing C. sempervirens pollen allergen discs prepared commercially or in the laboratory showed that values of the uptakes of [125I]-anti-IgE indicating the presence of pollen-reactive IgE antibodies obtained with the latter discs were consistently higher (means 4.5 vs. 0.88), and that false-negative results were obtained when many sera were used with the commercial discs. The results of this study provide an essential basis for the production of standardized, safe and effective C. sempervirens pollen extract applicable to diagnosis and therapy of cypress pollen allergy.