bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro Thymidine Uptake and Incorporation into Thymic and Bursal Lymphocytes from Young Hypothyroidic Chickens
Authors: Scott Thomas R. , Glick Bruce
Published: August 2009

Experiments were conducted to evaluate the ability of thymic and bursal lymphocytes from hypothyroidic chicks to take up 3H-thymidine. To produce hypothyroidism, chicks were fed a diet containing thiouracil (0.1 %) from day of hatch to 3 weeks of age. At 1, 2 and 3 weeks of age, body weights were significantly lower for hypothyroid chicks, and at 2 and 3 weeks of age, the relative weights of the thymus and bursa were significantly less than those of controls. Furthermore, chicks treated with thiouracil had significantly lower serum concentrations of 3,5,3′-triiodothyronine (T3) and thyroxine (T4). Thymic cells from hypothyroid chicks incorporated less 3H-thymidine at 1 and 3 weeks of age as assessed by liquid scintillation counting and autoradiography. Liquid scintillation counts for labeled bursal cells from hypothyroid chicks were not lower at 1 week of age, but the counts were lower at 3 weeks of age. Autoradiographic studies with bursal cells revealed that 3H-thymidine incorporation was less, but not significantly less, in medium-size lymphocytes from thiouracil-treated chicks at both 1 and 3 weeks of age. Thymidine incorporation into large bursal cells was affected more by thiouracil treatment at 1 than 3 weeks of age.
The Influence of Aluminum on Resistance to Listeria monocytogenes in Swiss-Webster Mice
Authors: Yoshida Steven , Gershwin Eric , Keen Carl L. , Donald James M. , Golub Man S.
Published: August 2009

Female, adult Swiss-Webster mice were fed, during gestation and subsequent lactation, diets containing either 25 (control), 500 or 1,000 μg Al/g diet in the form of Al lactate, followed by challenge with Listeria monocytogenes. Mice in the groups fed 1,000 and 500 μg Al/g had significantly lowered resistance to bacterial infection when compared to control animals (p < 0.025). The animals given 1,000 μg Al/g also had higher concentrations of liver Al (p < 0.05) than controls. Pups derived from these dams showed no differences in mortality rates or Al tissue levels. In contrast, nonpregnant animals fed 1,000 μg Al/g for 6 weeks had a slight decrease in mortality rate when compared to control animals. Dose-related changes in tissue Al levels were not observed in these adult, virgin mice. Finally, adult nonpregnant mice were injected subcutaneously with PBS only, or with 1 5 or 10 mg of Al (in the form of Al lactate) per kilogram body weight followed by bacterial challenge. Animals which received 5 mg Al/kg had lower mortality rates to L. monocytogenes when compared to other treatment groups (p < 0.025). This data suggests that Al has the potential to influence host resistance to bacterial infection depending on the physiological state of the host; it provides additional evidence for the role of Al as an environmental toxin.
Synthetic Allergenic Epitopes from the Amino-Terminal Regions of the Major Allergens of Hazel and Birch Pollen
Authors: Elsayed S. , Holen E. , Dybendal T.
Published: August 2009

Five immunogenic peptides from the major allergens of tree pollen extracts (residues 23–38) were synthesized by Merrifield solid-phase peptide synthesis. The primary structures of these peptides were deduced from the known N-terminal amino acid sequence of the major allergen of hazel, HIa, and birch, Bet v I. In a segment of 16 amino acids (23–38), 2 residues were the difference between birch and hazel. The selection of the peptides was on the basis of optimal hydropathicity. The segment 29–34 of these peptides was predicted to prefer a helical rather than β-sheet conformational state. The insertion of amino acid residues during the synthesis was confirmed by the analysis of amino acid composition prior to coupling the preceding residue. The linearities of the synthetic chains were assigned by the primary structure analysis of the entire chain by automatic Edman degradation. The synthetic peptides were purified by gel filtration chromatography and reversed-phase high-performance liquid chromatography, resulting in satisfactory homogeneity of the preparations. The immunogenicity of the peptides was tested in rabbits using conjugated peptide-bovine serum albumin through a carbodiimide spacer arm. The results indicated that all constituents of the conjugate could elicit an immune response. The peptides gave precipitation lines in crossed immunoelectrophoresis when using rabbit antibodies against the conjugated preparation. All the synthetic peptides and analogues from region 23–38 could inhibit, in dose-response patterns, the binding of specific IgE to the intact molecules. Similar results were shown for the shorter segment 25–34, suggesting that the C-terminal tetrapeptide did not contribute to the activity. The allergenicity of these peptides and their analogues is currently examined by Prausnitz-Küstner inhibition, histamine release and provocation tests.
Establishment of a Chicken × Chicken Hybridoma Secreting Specific Antibody
Authors: Nishinaka Shigeyuki , Matsuda Haruo , Murata Masayoshi
Published: August 2009

Chicken B lymphoblastoid cell mutants lacking thymidine kinase (TK) were selected from the parental cell line treated with ethyl methanesulfonate. To obtain chicken hybridomas secreting immunoglobulin against the defined antigen, a TK–– mutant was fused with inactivated Newcastle disease virus (NDV)-immunized chicken spleen cells using polyethyleneglycol. Finally, one hybridoma secreting IgM specific for NDV was established.
Enhancing Effect of Suspended Particulate Matter on the IgE Antibody Production in Mice
Authors: Takafuji Shigeru , Suzuki Shuji , Koizumi Kazuhiro , Tadokoro Kenji , Ohashi Haruo , Muranaka Masaharu , Miyamoto Terumasa
Published: August 2009

Suspended particulate matter (SPM), suspended in the polluted environmental atmosphere, are perpetually inhaled into the human body and are considered to have profound effects on human health. This study investigated the enhancing effect of SPM on the IgE antibody production in mice. The IgE antibody responses in mice immunized with intranasal administration of ovalbumin (OA) plus SPM at 3-week intervals were higher than responses in the animals immunized with OA alone. When the dose of OA administered as an antigen was 0.25 μg, the time course and magnitude of enhancement by SPM was comparable to those by killed Bordetella pertussis, a common adjuvant. SPM had an enhancing effect on IgE antibody production even in a small dose such as 0.25 μg administered at 3-week intervals. The possibility cannot be excluded that the natural exposure of humans to SPM in the environmental atmosphere may explain the high prevalence rate of allergic rhinitis caused by pollens in polluted districts in Japan.
Multispecificity of Monoclonal Rheumatoid Factor
Authors: Milgrom F. , Swierczynska Z.
Published: August 2009

The study involved a monoclonal rheumatoid factor referred to as RF-AN, obtained by Steinitz and his associates from IgG-reactive human lymphocytes ‘immortalized’ by infection with Epstein-Barr virus. RF-AN combined with red blood cells (RBC) sensitized by either human or rabbit IgG antibodies. The monoclonal character of RF-AN strongly suggested that the same molecule of this factor combined with IgG of both species. Additional evidence for this contention was obtained from absorption and mixed agglutination experiments. RBC sensitized by either human or rabbit antibodies would remove serological activity of RF-AN for both human and rabbit IgG. RF-AN produced exclusively mixed agglutinates when reacted with a mixture of human RBC sensitized by human antibodies and chicken RBC sensitized by rabbit antibodies. Furthermore, it was shown that inhibition with aggregated human Fraction II gave strictly ‘specific’ results in that it abolished the reaction of RF-AN with RBC sensitized by human antibodies, but not by rabbit antibodies. This result was interpreted as indicating that RF-AN has a multispecific character, i.e., has separate combining sites for human and rabbit IgG. Interestingly, specific inhibition could not be achieved with aggregated rabbit Fraction II and this preparation affected reactivity of RF-AN with RBC sensitized by human antibodies as well as by rabbit antibodies.
Production of Murine Monoclonal Antibodies against Francisella tularensis Antigens and Characterization of Antibody-Reactive Epitopes
Authors: Lilliehöök Bo , Sandström Gunnar
Published: August 2009

Eight monoclonal antibodies were produced by immunizing mice with a high molecular-weight antigen from Francisella tularensis. The antigen consists of carbohydrates and proteins and was found to contain several different epitopes. By treating the antigen with periodate or proteinase K, it was found that the majority of antibody-reactive epitopes was mainly dependent on the carbohydrate part of the antigen. On intact F. tularensis bacteria the epitopes seem to be exposed at the bacterial surface. Both main biovars of F. tularensis, tularensis (type A) and palaearctica (type B), exhibited the epitopes.
Cloning of cDNA Coding for an Allergen of Cocksfoot Grass (Dactylis glomerata) Pollen
Authors: Walsh David J. , Matthews Jayne A. , Denmeade Russell , Walker Matthew R.
Published: August 2009

Messenger RNA isolated from Cocksfoot grass (Dactylis glomerata) anthers has been used to generate a cDNA library in λt11. Three cDNA clones (7.8, 8.1, and 8.3) were demonstrated to be recognized by human IgE antibodies in atopic serum and by rabbit polyclonal antiserum raised to a crude aqueous extract of Cocksfoot pollen. The size of the cDNA inserts was determined as approximately 700 bp, and restriction mapping demonstrated them to be identical sequences. Lysogens obtained in Escherichia coli Y1089 allowed expression of a 140 kD β-galactosidase fusion protein containing 24 kD of cloned allergen protein. Fusion proteins were recognized by IgE antibodies in 75% (6/8) of atopic sera tested, but were not detected by non-atopic sera. On the basis of size and frequency of recognition in the atopic population, the cloned protein may represent a major allergen. Monoclonal antibodies specific for the major allergen of Cocksfoot pollen were not reactive with the fusion proteins. Reactivity of human IgE antibodies with the fusion protein could be blocked by crude Cocksfoot pollen extract, but not by the major allergen DG3 purified from the extract by affinity chromatography. Human and rabbit antibodies affinity purified against fusion protein 7.8 did not allow identification of the native protein component in crude extract encoded for by the cDNA clones.
Purification of Par j I, a Major Allergen from Parietaria judaica Pollen
Authors: Cocchiara Roberta , Locorotondo Giovanni , Parlato Alessandra , Guarnotta Gabriella , Ronchi Severino , Albeggiani Giuseppe , Amoroso Saverio , Falagiani Paolo , Geraci Domenico
Published: August 2009

A major allergen, the Par j I, was purified to homogeneity from Parietaria judaica pollen by means of ultrafiltration dialysis, preparative polyacrylamide gel chromatography and affinity chromatography through a column of Sepharose-monoclonal antibody specific for Par j I. The homogeneity of the Par j I was assessed by one single arc of immunoprecipitation both in crossed immunoelectrophoresis (CIE) and crossed radioimmunoelectrophoresis, by one single band of radiostaining after a sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transfer to nitrocellulose and by one single peak after a size exclusion chromatography on high-performance liquid chromatography (HPLC). The homogeneity was further supported by crossed Laurell immunoelectrophoretic analysis, in that only one arc of precipitation was magnified in CIE after addition of the purified allergen. The purified Par j I allergen was capable of interacting in vitro with 70% of the human IgE specific for a crude P. judaica extract, as determined by radioallergosorbent test inhibition. The purified Par j I was capable of inducing positive reactions in vivo in skin prick tests, and of inducing release of histamine from blood containing basophils as determined by a histamine release assay. The amino acid analysis of the Par j I showed 118 amino acid residues per monomer analyzed and, among other residues, three raethionine residues were detected. The molecular weight of the Par j I estimated by HPLC and amino acid composition was 26 kilodaltons.
The Tachykinins Neurokinin A and Physalaemin Stimulate Murine Thymocyte Proliferation
Authors: Söder Olof , Hellström Per M.
Published: August 2009

The tachykinins constitute a family of neuropeptides that are released from sensory neurons, mediating a variety of responses termed neurogenic inflammation. The present study investigates the possibility that tachykinins are also involved in immune-regulatory mechanisms. The mammalian tachykinins neurokinin A (NKA), neurokinin B, neuropeptide K and substance P, as well as the nonmammalian tachykinins physalaemin (PHY) and eledoisin, were analysed in 10-pM to 1.0-μM concentrations for regulatory influences in several lymphocyte proliferation assays. NKA, and to a lesser extent PHY, but none of the other tachykinins tested, displayed a stimulatory action in murine thymocyte cultures, utilised as an interleukin-1 (IL-1) bioassay. The effect was apparent only at a concentration of 0.1 μM or higher. No further stimulatory effect of the tachykinins could be observed in thymocyte cultures already suboptimally stimulated to proliferation by addition of IL-1. The tachykinins had no effect in direct and co-mitogenic T and B lymphocyte proliferation assays with rat spleen cells, in a thymocyte growth peptide assay with mouse thymic lymphoblasts or in an interleukin-2 (IL-2) bioassay with IL-2-dependent rat splenoblasts. Our findings indicate that NKA and PHY can act as immune regulators. The results are relevant for the understanding of the interaction between the nervous and the immune system, and are of particular interest in view of the inflammatory actions of both tachykinins and IL-1.
In vitro Proliferation of Murine Spleen Cells
Authors: Zhou P. , Quackenbush L.J. , Zaleski M.B.
Published: August 2009

Proliferative responses of normal (not immunized intentionally) spleen cells from inbred mouse strains to human recombinant interleukin 2 (L-2) were studied. It was shown that inbred strains vary in their responses to IL-2. The differences were observed for various doses of IL-2 and at various times after stimulation. Experimental data suggested that the strain differences in the magnitude of proliferative responses may result from an interplay of several interrelated factors such as differences in the frequency of the IL-2-responsive cells, differences in inducibility of the expression of the gene for the α-chain of the IL-2R, and genetic control of the responses. The latter was found to be exerted by two independent genes with the dominant alleles determining good responses to IL-2. The magnitude of the responses to IL-2 correlated with the magnitude of responses to concanavalin A, but seemed to have no effect on the interferon production or on the primary response to sheep red blood cells. The described phenomenon may have a practical significance in studies aimed at improving tumor immunotherapy with lymphokine-activated killer cells.
Effect of Tetrandrine on Immunological Responses and Cardiac Transplant Rejection in Mice
Authors: Si-ying Li , Teh B.S. , Seow W.K. , Ling Li-hua , Thong Y.H.
Published: August 2009

Mice treated with the plant alkaloid tetrandrine have a reduced capacity to mount delayed-type hypersensitivity (DTH) responses. This suppressive effect on DTH was more pronounced when treatment was begun 2 days prior to immunization with sheep eryhtrocytes. Similar dosage regimens significantly delayed the rejection of cardiac transplants. By contrast, there was significant enhancement of antibody production. These immunomodulatory properties of tetrandrine may account for some of its potential in the treatment of chronic inflammatory diseases, where immunological mechanisms are known to play a major role in pathogenesis.
Stability Studies of Calvatia cyathiformis Basidiospore Allergens
Authors: Horner Elliott , Ibanez M.D. , Lehrer S.B.
Published: August 2009

Calvatia cyathiformis allergens in unfractionated extract (crude), and in extract sequentially fractionated by gel filtration (GF) and hydrophobic interaction chromatography (HIC) were tested for stability. C. cyathiformis allergen sources (crude, GF, HIC) were sampled immediately (0 h), or incubated at 4, 24 or 37 °C and then sampled at 8, 24 or 96 h. Polyacrylamide gel-isoelectric focusing immunoprints revealed 3 allergen(s) groups, or bands (Bds) with respective pi of 3.6–4.6, 6.6 and 9.3. Only Bds 3.6–4.6 were stable at 37 °C. At 24°C, Bds3.6–4.6 persisted to 96 h, Bd6.6 persisted 24 h, and Bd9.3 waned in 8 h. At 4°C all 3 allergens in HIC were stable for 8 and 24 h; Bd9.3 was reduced at 96 h. All allergen activity was labile to low pH conditions except for Bds3.6–4.6. Proteinase K degraded Bd9.3 more rapidly than Bd6.6. Immunoprint patterns corresponded to the stained gels and were consistent among different sources. Bd9.3 is very labile, but reactive with 63% of sera tested. Since 10–15% of C. cyathiformis reactors bind IgE only to Bd9.3, this is notable variability, and significant for diagnosis and treatment.
Novel Abnormality in Subpopulations of Circulating Lymphocytes
Authors: Lehmann Paul F. , Sawyer Tom , Donabedian Haig
Published: August 2009

A patient, who had developed anergy after disseminated histoplasmosis and who had gone on to develop a cardiomyopathy, demonstrated an unusually large proportion of T lymphocytes bearing the γ and δ chains of the T cell resceptor (TCR-γ/δ). These γδ+ cells composed over half of the circulating lymphocytes and were clearly divisible into two subgroups based on the level of expression of the γ and δ chains and their ability to bind one of the monoclonal antibodies reactive with the TCR-δ chain. In addition, the patient showed an abnormally high proportion of circulating lymphocytes having the CD2––, 3+, 4––, 8–– phenotype which has been only recently described. The pattern of lymphocyte subpopulations changed over the 9 years that the patient was followed since his original infection and appears to characterize a novel abnormality.
A Positive Feedback Loop for Staphylococcal Enterotoxin-A-Stimulated IFN-Gamma Production Requires Macrophage Immune-Associated Antigen Upregulation
Authors: Russell Jeffry K. , Pontzer Carol H. , Johnson Howard M.
Published: August 2009

The C57B1/6-derived T cell line, L12-R4, produced murine interferon-gamma (IFNγ) in response to mitogenic stimulation by phorbol myristate acetate (PMA) or concanavalin A (Con A), but not by staphylococcal enterotoxin A (SEA). Low levels of IFNγ were produced by SEA stimulation of L12-R4 cells cocultured with C57B1/6 bone marrow macrophages (BMM). Significantly increased yields of IFNγ resulted from 48-hour pretreatment of the BMM with recombinant IFNγ (100 U/ml) prior to coculture. Polyclonal anti-IFNγ and anti-IFNα/β were used to characterize the interferon as IFNγ. Paraformaldehyde (0.1%) treatment of IFNγ-pretreated BMM did not affect IFNγ production, suggesting that processing of SEA was not required. IFNγ treatment of BMM resulted in significantly increased expression of immune-associated (la) antigen as determined by flow cytometric analysis, suggesting that the accessory cell role of BMM involved la antigen. Polyclonal anti-Ia antibody selectively inhibited the production of IFNγ by SEA-stimulated whole spleen cell cultures, consistent with the necessity of la antigen for BMM help in SEA induction of IFNγ. These findings suggest that la antigen is necessary for BMM accessory function in SEA induction of IFNγ. More interestingly, the results implicate class II molecules in a positive feedback loop for IFNγ production by SEA.
Assessment of the Functional Activity of Antigen-Bearing Dendritic Cells Isolated from the Lymph Nodes of Contact-Sensitized Mice
Authors: Jones David A. , Morris Alan G. , Kimber Ian
Published: August 2009

The immunostimulatory properties of dendritic cells (DC) which rapidly accumulate within the draining lymph nodes of mice topically exposed to contact allergens have been assessed as a function of their ability to initiate proliferative responses by sensitized lymph node cells (LNC) in vitro. A proportion of such DC bear antigen which is in a highly immunogenic form. Thus, at low stimulator :responder ratios, DC-enriched populations prepared from the draining nodes of mice exposed to the skin-sensitizing ñuorochrome fluorescein isothiocyanate (FITC) induced significant proliferative responses by FITC-primed responder lymphocytes. Under the same experimental conditions, unfractionated or DC-enriched LNC isolated from naive mice and coated with at least equivalent amounts of FITC in vitro were without effect. The influence of DC populations on lymphocyte proliferation was largely, but not wholly, antigen-specific in nature. Dendritic cell fractions isolated 18 h following topical exposure to high concentrations of FITC or oxazolone induced a marked proliferative response by lymphocytes sensitized to the homologous chemical and a significantly less vigorous response by lymphocytes primed to unrelated haptens. In contrast DC isolated from the nodes of mice sensitized with 2,4-dinitrochlorobenzene (DNCB) caused proliferation only of DNCB-primed lymphocytes. The relative potential of DC populations prepared from FITC- and oxazolone-primed mice to induce antigen-specific and non-specific stimulation was dependent upon both the concentration of chemical used for sensitization and the period of exposure.
Effect of the Late Complement Components C5b-9 and of Platelet-Derived Growth Factor on the Prostaglandin Release of Human Synovial Fibroblast-Like Cells
Authors: von Kempis Johannes , Torbohm Ingo , Schönermark Matthias , Jahn Bernhard , Seitz Michael , Hänsch Gertrud M.
Published: August 2009

We examined the prostaglandin E (PGE) synthesis of cultured adherent synovial fibroblast-like cells (SFC) from patients with osteoarthritis (OA) in the noninflammatory state as well as with rheumatoid arthritis (RA). In cells from RA patients the spontaneous PGE release was generally higher compared to that of OA patients, but decreased fast with time in culture. After cell passage, similar PGE baseline levels were seen in cells of the two patient groups. The cells could then be stimulated by the terminal complement components C5b-9 or C5b-8. PGE synthesis was also stimulated by the platelet-derived growth factor (PDGF), interleukin-1 (IL-1), or lipopolysaccharide (LPS). The amount of PGE synthesis after incubation with PDGF, LPS and IL-1 was comparable to that released after C5b-9. Thus, like other inflammatory mediators C5b-9 and PDGF trigger the increased PGE production by SFC and thus may participate in the development of synovial inflammation and contribute to the pathogenesis of RA.
Increased Delayed-Type Hypersensitivity Reaction in Rats Neuromanipulated with Capsaicin
Authors: Nilsson G. , Ahlstedt S.
Published: August 2009

We studied the neuroimmunomodulatory effect of capsaicin on the development of delayed-type hypersensitivity (DTH) reaction and antibody formation after reduction of neuropeptides in the peripheral sensoric nervous system. Rats were sensitized with picryl chloride on their shaved abdomen or by subcutaneous injections with picrylsulfonic acid (PSA). Manipulation of the content of neuropeptides in the sensory nerve system with capsaicin was performed 1 week before or after sensitization. An increase of the DTH reaction assessed as increased ear thickness after challenge was seen especially when the rats were given capsaicin after sensitization. The formation of specific antibodies was not affected, although the level of total IgG was decreased in PSA-sensitized rats treated with capsaicin after sensitization. The results demonstrated that the DTH reaction in rat was slightly but significantly affected by the abolishment of neuropeptides in sensoric nerve endings with capsaicin. Even if the direct effect of capsaicin on the immune system still has to be elucidated, these results indicate that neuropeptides in sensory nerves are involved in the pathogenesis of DTH.
Investigation of Mast Cell Differentiation in vivo by Use of Monoclonal Antibodies
Authors: Odarjuk Jutta , Rossow N. , Karawajew L.
Published: August 2009

In the present study mast cell differentiation/maturation was studied in vivo after depletion of mature mast cells from the peritoneal cavity by injection of distilled water. The reconstituting cell population was characterized by use of different staining methods. Additionally, the monoclonal antibody (MAb) IWF F2, which recognizes a membrane antigen of rat mast cells, was used to follow up mast cell differentiation/maturation in the course of the experiment. The antigen expression was studied both by immunofluorescence of antigen-bearing cells and by MAb inhibition of compound 48/80-stimulated histamine release from mast cells. In the course of the experiment the amount of antigen-positive cells increased continuously from less than 5% to control level (1st and 22nd days, respectively). The expression of the membrane antigen detectable by the MAb precedes the appearance of cytochemically identifiable mast cells for several days. The mast cells mature morphologically and functionally as indicated by increasing size, histamine content and MAb inhibition of compound 48/80-stimulated histamine release. The results obtained suggest the MAb IWF F2 to be a useful methodical tool for additional characterization of mast cell differentiation/maturation processes.
In vitro Generated Mast Cells Investigated by Monoclonal Antibodies
Authors: Rossow N. , Odarjuk J. , Karawajew L.
Published: August 2009

In the present study we have investigated the development of mast cells under in vitro conditions after depletion of mature mast cells of male Wistar rats by intraperitoneal application of distilled water. The studies were carried out both by monoclonal antibodies (MAb) against rat peritoneal mast cells and by cytochemical techniques. During cultivation the amount of MAb-positive cells increased from less than 5% to 64% after 3 weeks, which was accompanied by increased histamine levels and cell size. After 2 weeks the first Alcian blue stained mast cells were detectable, while safranin-positive granules appeared only after 3 weeks. These findings suggest that there are mast cell precursors in the peritoneum which may differentiate into mature mast cells. The antigen on the cell surface recognized by the MAb seems to be expressed during mast cell maturation.
Immunoblot Analysis of Allergens in Crude Mosquito Extracts
Authors: Wu Chii-Huei , Lan Joung-Liang
Published: August 2009

Whole body extracts of Aedes albopictus (AAL), Aedes aegypti (AAE) and Culex quequectates (CQU) were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. At least 28, 37 and 26 polypeptides, molecular weights (MWs) ranging from 16 to 120 kilodaltons (kD), respectively, were visible following Commassie brilliant blue staining. Sera of two mosquito-sensitive patients were examined for IgE binding to mosquito proteins by immunoblotting. Sera from both patients revealed IgE binding to 52- and 56-kD proteins of AAL. In addition, serum from patient B also bound 48 kD of AAL and did not bind any components of AAE and CQU. Serum from patient A also bound 32 and 58 kD of AAE, and bound only 58 kD of CQU. Sera from 2 control subjects, nonatopic and mosquito-negative atopic, did not bind specific human IgE to any proteins, as demonstrated by negative autoradiographs. Immunoblot analysis showed unique individual IgE-binding patterns and suggested that both genus- and species-specific mosquito allergens exist.
Inherited Deficiencies of the Late-Acting Complement Components Other than C9 Found among Healthy Blood Donors
Authors: Inai Shinya , Akagaki Yohji , Moriyama Takeshi , Fukumori Yasuo , Yoshimura Keiji , Ohnoki Shiro , Yamaguchi Hideo
Published: August 2009

Among sera from 145,640 healthy blood donors in Osaka, 16 were found to have abnormalities in late-acting complement components other than C9. It was found that of these 16 sera, 2 were deficient in C5, 4 in C6, 6 in C7 and 4 in the C8 α-γ-subunit. The incidence of deficiency of each component among the Osaka blood donors was calculated as follows: C5 deficiency, 0.0014%; C6 deficiency, 0.0027%; C7 deficiency, 0.0041 %; C8 α-γ-subunit deficiency, 0.0027%. We confirmed that 13 donors were healthy and 12 had no past history related to a complement component deficiency. From these results, not only C9 deficiency but also deficiencies of the other late-acting complement components were found among the healthy blood donors, but no early-acting component deficiencies were noted.
Adenosine Potentiates Neurally- and Histamine-Induced Contraction of Canine Airway Smooth Muscle
Authors: Sakai Noritaka , Tamaoki Jun , Kobayashi Kenji , Katayama Michio , Takizawa Takao
Published: August 2009

We studied the effect of adenosine on airway reactivity of isolated canine bronchial smooth muscle under isometric conditions in vitro. Administration of adenosine and its analogs increased the contractile responses of bronchial segments to electrical field stimulation in a dose-dependent fashion, where the rank order potency was N-ethylcarboxamideadenosine > adenosine > N-cyclohexyladenosine, but had no effect on those to exogenous acetylcholine. This potentiation was more pronounced at relatively low than at high stimulus frequencies, the maximal increase from the baseline responses being 56.3 ± 9.6% at 1 Hz (mean ± SE, p < 0.01). Adenosine also increased the histamine-induced contraction causing a leftward shift of the histamine dose-response curves, an effect that was abolished in the presence of atropine. These results suggest that adenosine potentiates airway responsiveness to vagal stimulation and to histamine through the activation of prejunctional A2 receptor, probably involving the accelerated release of acetylcholine from the cholinergic nerve terminals.
Effect of Azelastine on Leukotriene Synthesis in Murine Peritoneal Cells and on Thromboxane Synthesis in Human Platelets
Authors: Nishihira J. , Hayakawa T. , Suzuki K. , Kato K. , Ishibashi T.
Published: August 2009

Azelastine, a newly synthesized antiallergic agent, strikingly inhibited the production of leukotriene B4 and C4 (LTB4 and LTC4) in murine peritoneal cells which had been stimulated by calcium ionophore A23187. The 50% inhibitory concentrations (IC50) of the agent were approximately 1.0 × 10––5M. In addition, azelastine significantly inhibited also 5-lipoxygenase activity in peritoneal cells with an IC50 of 1.0 × 10––5M, but not on LTC4 synthetase, LTA4 hydrolase or phospholipase A2 activity. Furthermore, azelastine showed little effect on either 12-lipoxygenase activity or thromboxane synthesis in human platelets. These results suggest that at least the drug’s antiallergic effects can be attributed to its inhibiting action of 5-lipoxygenase in regard to arachidonate metabolism.
Breast Milk Antibodies to Foods in Relation to Maternal Diet, Maternal Atopy and the Development of Atopic Disease in the Baby
Authors: Fälth-Magnusson Karin
Published: August 2009

Breast milk samples were collected from 152 women during the first week after delivery. The levels of IgG and IgA antibodies to beta-lactoglobulin, ovalbumin and gliadin were assessed with an enzyme-linked immunosorbent assay (ELISA). The breast milk antibody levels did not differ significantly between mothers on a strictly cow’s milk and egg-free diet, and mothers taking these foods. Moreover, the colostral food antibody levels did not differ significantly between atopic and non-atopic mothers. Neither was there any correlation between the colostral antibody levels and the development of atopic disease in the baby. I conclude that maternal antigen avoidance during late pregnancy does not affect the food antibody levels in colostrum. High levels of food antibodies in a colostrum sample seem not to offer protection against food allergy in the child.
Variability of Allergenicity in Eight Strains of the Green Algal Genus Chlorella
Authors: Tiberg Ebba , Einarsson Roland
Published: August 2009

Biochemical and immunological properties of allergen extracts from eight strains within four species of the unicellular green alga Chlorella were compared. The total protein and carbohydrate contents ranged from 13 to 29 and 18 to 47% of the dry weight, respectively. It was shown, both with RAST inhibition and immunoblotting that Chlorella contain components which specifically bind to patient IgE antibodies. The relative allergen potency as assayed by RAST inhibition varied fivefold. Between seven and twenty IgE-binding components were identified in the various strains with molecular weights from 13 to 80 kD. Thus, the use of thoroughly characterized Chlorella strains is mandatory for preparation of relevant allergen extracts.
Effect of Intravenously Administered Lipoxygenase Metabolites on Rat Tracheal Mucous Gel Layer Thickness
Authors: Yanni John M. , Foxwell Marianne H. , Whitman Laura L. , Smith William L. , Nolan Joseph C.
Published: August 2009

The effect of intravenous injections of 5-, 12- and 15-hydroxyeicosatetraenoic acids (HETE), leukotrienes D4 and E4 (LTD4, LTE4) on tracheal mucous gel layer (TMGL) thickness was assessed in rats. When administered in doses ranging from 0.03 pg to 33 ng per rat, the lipoxygenase metabolites produced significant increases in TMGL thickness. The order of potency of the metabolites was 15-HETE > 12-HETE ≥ 5-HETE > LTD4 ≥ LTE4. Imidazole (31.6 mg/kg), intravenously, significantly decreased this response. These findings suggest that the mono-HETEs, especially 15-HETE, may be important modulators of airway mucus in the rat.
Granulomatous Response in Selective IgE-Deficient SJA/9 Mice Infected with Schistosoma japonicum
Authors: Owhashi Makoto , Nawa Yukifumi , Watanabe Naohiro
Published: August 2009

Granulomatous response was examined in seletive IgE-deficient SJA/9 mice infected with Schistosoma japonicum. The average size of granulomas formed around newly deposited eggs in SJA/9 mice was significantly smaller than that in control C57BL/6 or SJL/J mice, although the numbers of adult worms recovered from each strain were comparable among them. Thus, IgE antibody specific to allergenic components of S. japonicum eggs seems to act as an amplifier for the formation of granulomatous lesions.
Immunosuppression and in vivo T Cell Function in Granulomatous Inflammation
Authors: Fujioka Akira , Pincelli Carlo , Hashimoto Akihiko , Fukuyama Kimie , Epstein William L.
Published: August 2009

Subcutaneous grafts of fresh or freeze-dried hepatic schistosome egg granulomas in inbred mice caused increased numbers of total cells, T cell subsets, and elevated DNA synthesis in regional lymph nodes. This in vivo T cell activation was not apparent by in vitro tests. Spleen cells were unaffected both in vivo and in vitro. Immunosuppressive therapy with ciclosporin or monoclonal antibodies to T cell subsets profoundly depressed the T cell numbers and function, even when primed by a prior graft. Subsequent grafting partly reversed these effects, and T cells became activated. The findings indicate that in vitro assessment of activation and suppression of T cell function does not necessarily correspond to the events occurring in vivo.
Demonstration of Human, Rye Grass Pollen Extract-Specific, Helper and Suppressor Stimulating Activity of Modified Allergens by Effects on in vitro Hapten-Specific Antibody Production
Authors: Wheeler A.W. , Johansson S.G.O.
Published: August 2009

An in vivo system is described in which penicilloyl antibody was produced from peripheral leucocytes of a grass pollen-sensitive patient who had received penicillin therapy, by challenge of the cells with penicilloyl-grass pollen extract conjugate. Incubation of these leucocytes with a number of modified preparations of grass pollen extract with various T-cell-stimulating properties was shown to affect penicilloyl antibody production. Both chymotryptically fragmented rye grass pollen extract and a conjugate of f met-leu-phe and rye grass pollen extract enhanced penicilloyl-specific antibody similarly to the enhancement induced by unmodified extract, though at high concentration some suppression was seen. A conjugate of polysarcosine and rye grass pollen extract, previously shown to cause antibody suppression in mice, was similarly suppressive for penicilloyl-specific antibody. The system therefore shows potential for the evaluation of the effects of modified allergen treatment on antibody levels via T-cell mechanisms.
Passive Air-Pouch Anaphylaxis in Rats
Authors: Konno-Amano Satoshi , Hirao Tetsuji , Kohno Hiroyuki , Mori Hiroshi , Sato Yoshihisa
Published: August 2009

The effects of some antiallergic agents on passive air-pouch anaphylaxis (PAPA) in the dorsal skin of rats were investigated by measuring plasma exudation and histamine content in the pouch fluid. Antiallergic agents, disodium cromoglycate (DSCG), tranilast and ketotifen, dose dependently inhibited both plasma exudation and histamine release, except that ketotifen showed a dose-unrelated inhibition of histamine release. An antihistamine, pyrilamine, suppressed plasma exudation without affecting histamine release. A cyclooxygenase inhibitor, indomethacin, and a 5-lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), showed no significant effect. A slow-reacting substance of anaphylaxis antagonist, FPL55712, exerted no effect at 0.3 and 3.0 μg/ml, but at 50 μg/ml it suppressed not only plasma exudation but also histamine release. Dexamethasone suppressed plasma exudation dose dependently without inhibiting histamine release significantly. A β-stimulant, isoproterenol, and prostaglandin E2 also exerted significant inhibitory effects on plasma exudation but not on histamine release at 1 mg/ml. Theophylline inhibited both plasma exudation and histamine release. These observations indicate that PAPA is useful for studying antiallergic agents.
Production of Eosinophil-Activating Factor (EAF) by Peripheral Blood Mononuclear Cells from Asthma Patients
Authors: Thorne Kareen J.I. , Richardson B.A. , Butterworth A.E. , Hay I. , Jackson M. , Higenbottam T.W.
Published: August 2009

Mononuclear cells from blood samples from asthmatic patients were tested for their ability to produce two eosinophil activating factors, EAF and TNF. High levels of EAF were produced by some but not by all patients. The influence of external factors on EAF production was evaluated. Patients receiving prednisolone (10–30 mg/day) tended to produce only low levels of EAF. Prednisolone has been shown to inhibit production of EAF in vitro by isolated mononuclear cells, at concentrations of 0.1–1 μg steroid/ml. Incubation of mononuclear cells with certain allergens enhances EAF production in sensitive individuals. Little or no production of TNF was observed in the patients examined.
Differential Release of Histamine and Prostaglandin D2 in Rat Peritoneal Mast Cells Activated with Peptides
Authors: Levi-Schaffer Francesca , Shalit Meir
Published: August 2009

Rat peritoneal mast cells co-cultured with mouse 3T3 fibroblasts (MC/3T3) are fully responsive to immunologic stimuli. To assess their nonimmunologic activation MC/3T3 were challenged with various peptides. Optimal concentrations of substance P (10––4M) and bradykinin (5 × 10––5M) induced histamine release of 58.219.3 and 66.8 ± 6.6%, respectively, while neurotensin (10––4M) released only 16.6 ± 3.7% histamine. Freshly isolated mast cells (F-MC) challenged with the same concentrations of peptides released lower percentages of histamine (substance P 45.6 ± 5.1%, bradykinin 32.5 ± 5.3%, neurotensin 11.3 ± 6.0%). In both MC/3T3 and F-MC, only minute amounts of prostaglandin D2 (PGD2) were produced. In contrast, activation with anti-IgE antibodies and compound 48/80 caused both histamine release and PGD2 generation. Compound 48/80-stimulated MC/3T3 and F-MC released 80.2 ± 3.4 and 51.8 ± 6.2% histamine, respectively, and produced 15.4 ± 2.8 ng/106 mast cells and 3.9 ± 1.4 ng/106 mast cells PGD2, respectively. These findings indicate that peptides and bradykinin induce selective release of histamine with no PGD2 production in both F-MC and MC/3T3. Moreover, MC/3T3 preserve their functional characteristics of connective tissue mast cells since they are fully responsive to these peptides as F-MC.
Direct Activation of Human Polymorphonuclear Leukocytes by Pseudomonas aeruginosa
Authors: Mai G.T. , Seow W.K. , McCormack J.G. , Thong Y.H.
Published: August 2009

Brief exposure of polymorphonuclear leukocytes (PMNs) to unopsonized Pseudomonas aeruginosa resulted in significant enhancement of PMN adherence and 3H-deoxyglucose uptake, two important indicators of PMN activation. Dose-response and time-course experiments show that maximal stimulation of PMNs occurred at a PMN: bacteria ratio of 10:1 and exposure time of 10 min. The PMN-stimulatory capacity of P. aeruginosa was completely or partially abolished by treatment with formalin, heat, ultrasonication and ultraviolet (UV) irradiation, all of which affect the integrity of ligands on the bacteria surface. The putative glycoprotein ligand(s) contains galactose rather than glucose as shown by the ability of galactose in the medium to abrogate PMN-stimulatory activity. This PMN-stimulatory activity was not mediated by soluble factor(s) released from either P. aeruginosa or PMNs as shown by experiments with appropriate supernatants. Finally, PMN-stimulating activity was also observed with P. maltophilia and P. pseudomallei, but not by the other Pseudomonas spp. such as P. cepacia and P.Buorescens. These results suggest that P. aeruginosa and other Pseudomonas spp. may have little capacity to circumvent PMN host defences. This may account in part for their limited pathogenic role as opportunistic micro-organisms, for the severe inflammatory lesions seen in Pseudomonas infections when they do occur, and is consistent with the fact that infection with these organisms is commonly associated with impaired quantity or function of PMNs.
Relation between Splenic Antibody Formation and Serum Antibody Levels to Sheep Erythrocytes as Studied at the Level of Individual Mice
Authors: Hol Cees , Hofhuis Frans M.A. , van Dijk Hans
Published: August 2009

The relation between splenic antibody formation and serum antibody levels after intraperitoneal immunization of mice with sheep erythrocytes was studied in individual animals. A strong correlation between plaque-forming cell numbers in the spleen and haemolysin and haemagglutinin titres in the serum was observed. These results are at variance with recent data on the development of splenic auto-haemolysin-forming cells and corresponding serum levels after injection of mice with Escherichia coli endotoxin. The background behind the discrepancy between the antigen-driven immune response and the more polyclonal stimulation of B cells with endotoxin is being discussed.
Studies on the Osmophilic Fungus Wallemia sebi as an Allergen Evaluated by Skin Prick Test and Radioallergosorbent Test
Authors: Sakamoto Tatsuo , Urisu Atsuo , Yamada Masanori , Matsuda Yoshio , Tanaka Kenji , Torii Shimpei
Published: August 2009

Recently, Wallemia sebi, a species of osmophilic fungi, has been abundantly detected in housedust using low water activity media. In this study, allergenic activity of W. sebi was assessed by skin prick testand radioallergosorbent test (RAST) in 74 asthmatic patients ranging from 6 to 32 years of age. Aspergillus fumigatus and house dust were used for comparison. In skin prick test, W. sebi extract, A. fumigatus extract andhouse dust extract elicited positive reactions in 4 (5.4%), 4 (5.4%) and 51 (68.9%) patients, respectively. RASTshowed positive results in 14 subjects (18.9%) for W. sebi extract, in 8 (10.8%) for A. fumigatus extract and in 59 (79.7%) for house dust extract. These results indicated that some asthmatic individuals showed immediate-typehypersensitivity to W. sebi, and which means this fungal species may be of importance to atopic diseases as acausative agent.
Phorbol Myristate Acetate Stimulates Formation of Diphosphoinositide in Rat Mast Cell Granules
Authors: Kurosawa Motohiro , Okayama Yoshimichi , Kobayashi Setsuo
Published: August 2009

Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. Diphosphoinositide (DPI) synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [γ32P]ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HCl and were separated by thin-layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of phorbol myristate acetate (PMA) to the granules caused an increase of 32P incorporation from [γ32P]ATP in the DPI fraction, which can be catalyzed by PI kinase. This effect of PMA in the DPI synthesis was dose dependent and maximal effects were observed at 10 ng/ml.
Induction of Polyclonal and Monoclonal Antibodies with a Single Allergen Separated from Birch Pollen Extract by SDS-PAGE and Nitrocellulose Blots
Authors: Wheeler A.W. , Deards M. , Fox B. , Hickman B.E. , Spackman V.M.
Published: August 2009

A Gold staining allergenic band, Bet v 1, was excised from a nitrocellulose blot following SDS-PAGE of birch pollen extract. This was used to raise a polyclonal rabbit antiserum and monoclonal mouse antibodies specific only for the Bet v 1. The method offers practical advantages for the production of antibodies to individual allergens in complex extract mixtures, without laborious purification methods.
Comparison of Histamine and 5-Hydroxytryptamine Content and Secretion in Rat Mast Cells Isolated from Different Anatomical Locations
Authors: Purcell W.M. , Cohen D.L. , Hanahoe T.H.P.
Published: August 2009

Rat mast cells, isolated from different anatomical locations, were found to exhibit heterogeneity with respect to histamine and 5-hydroxytryptamine (5-HT) content. Unlike peritoneal mast cells, those derived by enzymatic digestion of rat lung and mesentery tissues contained amounts of 5-HT in excess of histamine. Both compound 48/80 and A23187 were more effective liberators of histamine than 5-HT in each cell type, despite their differences in relative amine content. However, in response to a mixture of specific antigen and phosphatidylserine, the pattern of secretion of both amines from each mast cell type was observed to be essentially collinear. These results are discussed in terms of amine release mechanisms and post-secretory events.
Contribution of Post-Secretory Mechanisms to the Observed Pattern of Histamine and 5-Hydroxytryptamine Secretion from Peritoneal Rat Mast Cells in Response to Compound 48/80
Authors: Purcell W.M. , Cohen D.L. , Hanahoe T.H.P.
Published: August 2009

Compound 48/80 induces a greater percentage release of histamine than 5-HT from purified peritoneal rat mast cells such that the log-concentration effect curves diverge. Mast cells were shown to remove histamine and 5-HT applied exogenously, although uptake of 5-HT was in preference to that of histamine. At concentrations of clomipramine and fluoxetine that were shown to inhibit 5-HT uptake, the net percentage release of 5-HT increased in a corresponding manner. Likewise, pargyline and nialamide, which inhibit monoamine oxidase, potentiated the release of 5-HT, although they were without effect upon 5-HT uptake. Observed differences in the pattern of histamine and 5-HT secretion are proposed to be the result of post-secretory events involving amine reuptake and metabolism rather than a reflection of differential release mechanisms.