bluuebunny/crossref_metadata_2025 huggingface dataset:
A Comparison of Histamine Secretion from Isolated Peritoneal Mast Cells of the Mouse and Rat
Authors: Barrett K.E. , Pearce F.L.
Published: August 2009

The effect of different histamine liberators on isolated peritoneal mast cells of the mouse and rat has been examined. Both cell types responded to the lectin, concanavalin A, and the release was in each case potentiated by phosphatidylserine. The rat cells released histamine on treatment with dextran but the mouse cells were essentially refractory to the polysaccharide. The mouse cells were significantly more responsive to the actions of the ionophore A23187 and adenosine 5′-triphosphate but much less reactive towards the polycations, compound 48/80 and peptide 401 (the MCD-peptide from bee venom). The reactivity of the mouse cells towards the latter two agents was enhanced in the absence of extracellular calcium. These results further emphasize the functional heterogeneity of mast cells from different sources.
Monoclonal Hybridoma Antibodies against Human IgE and Their Use in a Rapid and Sensitive Enzyme Immunoassay for the Semiquantitative Assessment of Total IgE Levels in Human Blood
Authors: Chandler H.M. , Coulter A.R. , Healey K. , Kornitschuk M. , MacGregor A. , Hurrell J.G.R.
Published: August 2009

A simple semiquantitative enzyme immunoassay (EIA) for the rapid estimation of IgE levels in specimens of human blood, plasma or serum is described. The test requires little labour input and does not require highly trained personnel or instrumentation. By using two monoclonal antibodies of different anti-IgE specificities it is possible, with a single incubation of 20 min at ambient temperature, to detect elevated IgE levels (≥ 33 IU/ml) within a total test time of 25 min, and low levels of IgE (≤ 10 IU/ml) within 35 min. For diagnosis of elevated/normal IgE levels only, a single incubation of 10 min. at ambient temperature may be used with a total test time of less than 20 min. The EIA system utilizes glass capillary tubes and urease-labelled antibodies, a system that has proven satisfactory in other applications.
Granular and Globular Acid α-Naphthyl-Acetate-Esterase Staining Pattern of Resting and Mitogen-Activated Murine Lymphocytes
Authors: Treves S. , Ali-Khan Z.
Published: August 2009

Available methods for the identification of T-lymphocytes in tissue sections and in single cell suspensions are often tedious and require a lot of manipulation. Here we confirm previous findings that nonspecific α-naphthyl-acetate esterase (ANAE) is a reliable method for the identification of resting thymus-derived lymphocytes but not for concanavalin-A-stimulated lymphocytes, as a small proportion of lipopolysaccharide-stimulated lymphoblasts exhibit ANAE activity as well. We have also shown that there is a relationship between the globular staining pattern and the degree of cell activation. Finally, we noted that prolonged incubation of lymphocytes at 37 °C reduces the number of ANAE-positive cells.
Adjuvancy of Cremophor El° in Rodents
Authors: Descotes J. , Tachon P. , Laschi-Locquerie A. , Evreux J.C.
Published: August 2009

Cremophor El° proved as potent as Freund’s complete adjuvant for eliciting skin reactivity to bovine serum albumin in guinea pigs. It enhanced delayed hypersensitivity to sheep erythrocytes in mice while hemagglutinin antibody titers were not affected. This adjuvant activity is likely to play a critical role in hypersensitivity reactions to Cremophor-containing drugs, e.g. alphadione.
Effects in Mice of Rat Bromelain-Treated RBC and Lipopolysaccharide on Autoantibody Production against Bromelain-Treated Isologous RBC
Authors: Cox K.O. , Ramos T. , Cox J. , Samcewicz B.
Published: August 2009

Autoantibody production against mouse bromelain-treated (brom) red blood cells (RBC) was significantly increased in mice injected with rat brom RBC. These autoantibodies were not adsorbed by rat brom RBC in serological assays and did not lyse rat brom RBC in plaque-forming cell (PFC) assays using mixtures of rat brom RBC and mouse brom RBC as targets. These data suggest that the increased response induced by rat brom RBC is not due to the presence of common, or similar, antigens on the two types of RBC. The spleens of mice injected with lipopolysaccharide (LPS), or with both LPS and rat brom RBC, had a markedly increased number of PFC lysing mouse brom RBC. About 20% of the PFC induced by LPS and rat brom RBC also lysed rat brom RBC. The autoimmune response was not increased in mice injected twice with rat brom RBC and the secondary response induced by two injections of LPS was lower than that induced by one injection of LPS. However, injection of LPS after an initial challenge with rat brom RBC induced an autoimmune response similar in size to that induced by LPS alone. The decreased secondary response against mouse brom RBC following a second injection of rat brom RBC was associated with decreased production of antibodies of various specificities as detected in a reverse PFC assay. These results do not support the hypothesis that the poor secondary responses against mouse brom RBC following a second injection of rat brom RBC are due to the exhaustive differentiation of autoimmune B cells as part of a fail-safe mechanism to prevent autoimmunity.
Genetic Dependence of IgE Antibody Production in Mice Infected with the Nematode Nippostrongylus brasiliensis
Authors: Pfeiffer P. , König W. , Bohn A.
Published: August 2009

We investigated the IgE-antibody response in 36 inbred mouse strains during infection with Nippostrongylus brasiliensis. With regard to the N. brasiliensis-specific IgE-antibody activity responder and nonresponder mice were obtained. Mice with the H-2-f haplotype (A.CA, B10.M, A.TFR 5) are high specific responded. It is suggested that one Ir gene for the N. brasiliensis-induced IgE-antibody response is localized within the K to J region of the mouse major histocompatibility complex. To obtain a medium IgE-antibody response it appears that a complementation between two Ir genes is necessary. An additional genetic control beyond the H-2 complex cannot be excluded. As to the total nonspecific serum IgE levels the parasitic infection leads to a 20- to 40-fold increase in high and low specific responder strains as well. Pretreatment of high responder mice (A.TFR 5, B10.M) with sera that were obtained from normal or complete Freund’s adjuvant treated low (B10.G) and nonresponder (A.TFR 1) mice led to significant inhibition or enhancement of the N. brasiliensis-specific IgE-antibody response.
Genetic Susceptibility of Rats to Some Forms of Delayed Hypersensitivity
Authors: de Brito F.B. , Hanahoe T.H.P.
Published: August 2009

The F1 generation, produced by mating Wistar rats from the Tuck colony which react to dextran (R rats) with Wistar rats from the NELP colony which do not react (NR rats) all responded to dextran with an anaphylactoid oedema reaction, and developed arthritis after adjuvant injections (AIA), but failed to show ovalbumin-induced delayed hypersensitivity reactions (OA) and experimental allergic encephalomyelitis (EAE). Back-crossing the F1 generation with NR rats produced animals, only half of which responded to dextran, though all the offspring reacted well to AIA, but poorly to OA and EAE. This confirms that resistance to AIA is not associated with resistance to dextran.
Adjuvant Effects of ι, κ, and λ Carrageenans on Antibody Production in BALB/c Mice
Authors: Mancino D. , Minucci M.
Published: August 2009

Carrageenan, in its three fractionated forms ι, κ, and λ, is able to stimulate in BALB/c mice the production of IgE, IgG1 and those antibodies producing passive hemagglutination to ovalbumin. The adjuvant effect of carrageenan is more evident in the secondary response, and the ι form is more effective than the κ and the λ forms. The data are compatible with the possibility that adjuvanticity of carrageenan can be, at least partly, correlated to its antimacrophage activity.
Distribution of 3H-Dextran in Two Colonies of Wistar Rats after Intravenous Injection
Authors: Hanahoe T.H.P. , Wright J.D.
Published: August 2009

The rate of elimination of 3H-dextran from the blood after intravenous administration was found to be considerably faster in rats which react to dextran with an anaphylactoid reaction than in rats which do not. Correspondingly, the rate of appearance of 3H-dextran in the anaphylactoid target organs such as the footpad was faster in reactive rats, approximately three times as much radioactivity being located in their footpads, 30 min after injection. This difference in distribution may reflect differences in the sensitivity of mast cells of the two rat colonies to dextran.
Allergic Reactivity and Helminthic Infection in Amerindians of the Amazon Basin
Authors: Lynch Neil R. , López Reina , Istúriz Guillermo , Tenias-Salazar Elsa
Published: August 2009

Various aspects of the allergic reactivity of Amerindians in the Venezuelan sector of the Amazon basin were examined. The frequency of positivity in immediate hypersensitivity skin tests with extracts of common local environmental allergens was found to be extremely low (6.7%). As sera from significantly higher proportions of the study group contained specific IgE antibody against the test allergens, and their histamine-induced skin responses were normal, these results support previous suggestions of an inhibited expression of allergic reactivity in such populations. Indeed, the intense helminthic infections detected, and the extremely high total serum IgE levels measured (geometric mean 13,088 IU/ml) indicate the possible occurrence of mast cell saturation by parasite-induced IgE. However, despite a similar lack of agreement between the in vivo and in vitro tests for allergic reactivity against Ascaris lumbricoides in these subjects (43.5 and 97.9% positive, respectively), their extremely high responsiveness to the helminth allergens presents a marked contrast to that against the other environmental materials. Factors other than helminthiasis (e.g. racial, cultural, nutritional) might, therefore, also modulate the expression of allergic reactivity in such populations.
Comparison of the Anti-Anaphylactic Effects of Milrinone, Sulmazole and Theophylline in the Rat
Authors: Post Mark J. , te Biesebeek Jan Dirk , Wemer Johan , van Rooij Hans H. , Porsius Arijan J.
Published: August 2009

Milrinone and sulmazole, two recently developed cardiotonic drugs, inhibit specific fractions of the phosphodiesterase (PDE) isozyme system, while theophylline inhibits the PDE complex aspecifically. Since these drugs were shown to have anti-allergic effects in isolated lungs, we wanted to study milrinone and sulmazole in comparison with theophylline in an in vivo model of anaphylaxis. For this purpose, actively sensitized rats were intravenously challenged with antigen, while tracheal pressure, oesophageal pressure, blood pressure and airflow were continuously monitored. Antigen challenge induced a transient bronchoconstriction and a persistent fall in blood pressure until death occurred. Anaphylactic shock was consistently followed by metabolic acidosis. Milrinone, 3 mg/kg, and theophylline, 40 mg/kg (but not 10 mg/kg), reduced anaphylactic bronchoconstriction. Sulmazole, 40 mg/kg, almost abolished bronchoconstriction. Although milrinone, sulmazole and theophylline, 40 mg/kg, caused a fall in blood pressure prior to antigen challenge, total fall in blood pressure (drug-induced and antigen-induced) was not affected by these drugs. Surprisingly, sulmazole reduced mortality significantly. This phenomenon is probably related to the less severe metabolic acidosis in the presence of sulmazole.
Induction of IgE Synthesis in Anti-IgM-Activated Nonatopic Human B Cells by Recombinant Interleukin-3
Authors: Matsumoto Tomoaki , Clark Steven , Rocklin Ross E.
Published: August 2009

The signals required to induce purified normal human B cell subpopulations into IgE production were studied. Pokeweed mitogen (PWM)-stimulated T cell supernatant induced IgE synthesis in low-density but not high-density Percoll-gradient-separated resting B cells. The PWM supernatant also enhanced (greater than 2-fold) IgE synthesis by anti-IgM (but not Staphylococcus A Cowan I)-activated high-density B cells (but not low-density B cells) and had affinity for lentil lectin. Subsequent studies were carried out using purified human recombinant interleukins (rIL). Human rIL-3 consistently (10/19 experiments) augmented IgE synthesis by anti-IgM-activated normal B cells. rIL-4 did not consistently (4/23 experiments) induce IgE synthesis by normal B cells or mixtures of T and B lymphocytes ± monocytes. Furthermore, IL-1, IL-2, IL-5, IL-6, granulocyte, macrophage-colony stimulating factor (GM-CSF), granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF) and interferon-γ also failed to induce IgE synthesis. Identification of IL-3 as the factor(s) in the PWM supernatant that was responsible for inducing IgE synthesis was inconclusive since its effect could not be reversed by the addition of anti-IL-3 antibody. Thus, our results suggest that at least two distinct soluble factors are involved in the induction of IgE synthesis by nonatopic B cells.
Human IgE Response: Virus-Activated IgE Secretors Are Interleukin-2-Dependent Cells
Authors: Chan Marcia A. , Dosch Hans-Michael
Published: August 2009

The activation of human IgE-secreting B lymphocytes was difficult until the use of low cell density limiting dilution cultures, which are more efficient and eliminate regulatory growth restraints, became possible. The transformation of IgE-producing cells by Epstein-Barr virus unexpectedly required the presence of small supplements of mature T lymphocytes early in culture. Interleukin 2 (IL2, 10 U/ml) was able to completely replace T-cell supplements when added during a brief (1–2 cell cycles) but critical period, 5–8 days after infection with the virus. The observation that IgE-committed B cells are principally able to express receptors for and respond to IL2 is of experimental utility and implies the possibility that this T-cell product could be part of the complex T-cell controls which govern IgE-committed B cells.
Fc-Epsilon-Receptor-Bearing Lymphocytes in Patients with Clonorchiasis
Authors: Watanabe Naohiro , Yanagihara Yukiyoshi , Joh Kosuke , Hamada Atsuo , Tomita Yusuke , Kobayashi Akio
Published: August 2009

Expression of Fc-ε-receptors (FcεR) on peripheral blood lymphocytes was examined in patients with clonorchiasis. Mean serum IgE levels in these patients was 2,518 IU/ml. FcεR was detected by flow cytometry analysis with monoclonal antibody. The frequency of FcεR+ lymphocytes and the density of FcεR on the cells in the patients were similar to those in normal subjects. Most of the FcεR+ lymphocytes were B cells. The number of FcεR+ cells decreased after incubation in serum-free medium at 37 °C for 3 h. The amount of regulatory molecules of IgE production, IgE binding factor, was not significantly different between the patients and normal subjects. The number of FcγR bearing T or B cells in the patients was also similar to that of normal subjects. These results indicate that the mechanisms of elevated serum IgE in patients with clonorchiasis might be different from other diseases with hyperglobulinemia E.
Natural Killer Susceptibility of Brain Tumor Cell Lines Inversely Correlates with the Degree of Cell Differentiation and Not with the Level of Human Histocompatibility Antigen Expression
Authors: Alonso Corona , Serrano Rafael , Solana Rafael , Peña José
Published: August 2009

The purpose of this paper is to investigate the natural killer (NK) sensitivity of 15 cell lines derived from human brain tumors expressing different levels of major histocompatibility complex (MHC) antigens. Although it has recently been reported that NK susceptibility varies inversely with target cell class-I human leukocyte antigen (HLA) expression, our results show no correlation between class-I HLA, class-II HLA or β2-microglobulin expression on target cells and NK sensitivity, whereas a significant inverse correlation between NK susceptibility and the degree of tumor cell differentiation has been found. Thus, poorly differentiated tumor cells are highly susceptible to NK lysis, whereas well-differentiated ones are NK resistant. These results suggest that sensitivity to lysis by NK cells of cell lines derived from human brain tumors is primarily determined by the stage of differentiation and not by the level of MHC antigen expression on target cell.
Characteristic Nucleolar-Reticular Immunofluorescence Staining Pattern on Touch Prints of Hamster Liver: A Marker for Scl-70 Antibody
Authors: Kühn G. , Jarząek-Chorzelska M. , Kolacinska-Strasz Z. , Blaszczyk M. , Chorzelski T.P. , Jablonska S.
Published: August 2009

Sera from 48 patients with systemic sclerosis (SSc) and from 62 patients with other connective tissue diseases were studied for nuclear staining patterns by indirect immunofluorescence (IIF) using touch prints of hamster liver, HEp-2 and cryostat sections of monkey esophagus as substrates. Immunodiffusion studies performed in all cases disclosed Scl-70 antibody in 36 SSc patients. The IIF pattern of Scl-70 antibody is characterized by diffuse granular and nucleolar staining on HEp-2, a combined nucleolar and diffuse reticular staining on imprints of hamster liver and large speckled (nucleolar) fluorescence on cryostat sections of monkey esophagus. Touch prints of hamster liver revealed a partially nucleolar fluorescence as a dominant sign of the combined staining. Hamster liver substrate which is easily available might replace the expensive HEp-2 cells for detecting Scl-70 antibody.
Gliadin Antibody Production by Small Intestinal Lymphocytes from Patients with Coeliac Disease
Authors: Ciclitira P.J. , Hooper L.B. , Ellis H.J. , Freedman A.R.
Published: August 2009

Small intestinal lymphocytes (SIL) were isolated from jejunal biopsies from 9 non-coeliac controls, 6 treated and 5 untreated coeliac patients. Six-day cell culture supernatants were assayed for total IgG, IgM and IgA and specific anti-casein and anti-gliadin antibodies by ELISA. SIL from the untreated patients secreted more total IgM and IgA than SIL from the treated patients and more total IgG, IgM and IgA than SIL from the controls. The untreated patients’ cells secreted more specific anti-gliadin IgA than those from the treated patients and more anti-gliadin IgG, IgM and IgA than the controls. SIL from the untreated group released more anti-casein IgM than those from either the treated or control groups. There were no differences in total immunoglobulin, or specific anti-casein or anti-gliadin antibody secretion by SIL from the treated and control groups. Comparison of immunoglobulin release from cells lysed prior to culture and that secreted both after 30-min and 6-day cultures showed that the majority of immunoglobulin had been synthesised in vitro and could not be accounted for by carry-over.
Lipopolysaccharide-Reactive B Cells against Bromelain-Treated Mouse Erythrocytes: Preferential Migration to and Survival in the Peritoneal Cavity
Authors: Kawaguchi Susumu
Published: August 2009

The ratio of lipopolysaccharide (LPS)-reactive B cells specific for bromelain-treated mouse erythrocytes (BrMRBC) to general LPS-reactive B cells is far higher in the peritoneal cavity of normal mice than in their spleen. To investigate the high concentration of anti-BrMRBC LPS-reactive B cells in the peritoneal cavity, spleen and peritoneal cells from (CBA/N × C3H/He)F1 female normal mice were injected intravenously and intraperitoneally into F1 male X-linked immunodeficient mice. Both groups of B cells in the intravenously transferred cell population were able equally to home in the spleen, but only anti-BrMRBC LPS-reactive B cells could be detected migrating into the peritoneal cavity. About half the anti-BrMRBC LPS-reactive B cells in the intraperitoneally transferred cell population could be recovered from the peritoneal cavity, but general LPS-reactive B cells were eliminated rather rapidly from the peritoneal cavity. Neither group of B cells could be detected migrating from the peritoneal cavity into the spleen. These findings suggest that the high concentration of anti-BrMRBC LPS-reactive B cells in the peritoneal cavity may be caused by their preferential ability to penetrate into the peritoneal cavity through circulation and survive there.
Age as a Factor Determining Susceptibility for Experimental Autoimmune Encephalomyelitis in the Rat
Authors: Källén Bengt , Nilsson Olle
Published: August 2009

The effect of age on the susceptibility to experimental autoimmune encephalitis was studied in different strains of rats and their hybrids. In the Brown-Norway (BN) strain no susceptibility was seen at any age. In the nearly resistant Fischer (F) strain, no change in clinical disease but an increase in histological scoring could be seen with increasing age. When hybrids between F and BN were studied, a clear-cut increase in clinical susceptibility was seen with age but no change in histological scores, while the F1 hybrid between BN and the susceptible Lewis strain showed no change in clinical susceptibility but an increase in histological scores with age. The Findings are discussed together with previous data on these strains in terms of different mechanisms of resistance in the F and BN strains.
Inhibition of Allergic and Nonallergic Leukotriene C4 Formation and Histamine Secretion by Azelastine: Implication for Its Mechanism of Action
Authors: Chand Naresh , Pillar John , Nolan Kevin , Diamantis William , Sofia Duane
Published: August 2009

Azelastine, an orally effective antiasthmatic agent, has been reported to inhibit antihistamine-resistant, leukotriene-mediated allergic bronchoconstriction in guinea pigs. This suggests that azelastine might act through inhibition of leukotriene (LT) C4/D4 synthesis. We have examined the effect of azelastine on allergic and nonallergic histamine secretion and LTC4 formation. Azelastine and the known 5-lipoxygenase inhibitors, nordihydroguaiaretic acid and AA-861, exerted concentration-dependent inhibition of allergic LTC4 formation in chopped lung tissue from actively sensitized guinea pigs and calcium ionophore A23187-stimulated LTC4 synthesis in mixed peritoneal cells from rats. Azelastine also produced concentration-dependent inhibition of allergic and nonallergic histamine secretion from rat peritoneal mast cells. The ability of azelastine to inhibit allergic and nonallergic histamine secretion and LTC4 generation may contribute to its mode of action and its therapeutic efficacy.
Accessory Cell Function of Human Alveolar Macrophages in Antigen-Induced T Lymphocyte Proliferation
Authors: Ohtsuka Morio , Yoshizawa Yasuyuki , Sato Tetsuo , Yano Heiichi , Mukai Ryozaburo , Hasegawa Shizuo , Moore Vernon L.
Published: August 2009

We compared the accessory cell function of human alveolar macrophages (AM) to that of human blood monocytes (Mo) obtained by bronchoalveolar lavage and venipuncture from normal volunteers. Graded numbers of either AM or Mo were added to autologous peripheral blood T lymphocytes that were stimulated with a purified protein derivative of tuberculin (PPD). Either AM or Mo were cocultured with allogeneic T lymphocytes in mixed lymphocyte reaction (MLR) experiments. Both AM and Mo supported the PPD-induced T lymphocyte proliferation and allogeneic MLR at low ratios of AM or Mo to T lymphocytes with similar efficiency. However, AM showed marked suppressive effects at higher ratios of AM to T lymphocytes (1:1). PPD-pulsed AM, but not AM killed by physical treatments (heat, freeze-thaw, sonication), induced T lymphocyte proliferation. An indirect immunofluorescent study demonstrated that most AM express HLA-DR antigens. Furthermore, AM synthesized DR antigens with molecular weights of 33,000 and 29,000–31,000 daltons. When AM were treated with both anti-DR monoclonal antibody and complement, PPD-induced T lymphocyte proliferation and MLR were diminished. These results suggest that human AM function as accessory cells in the antigen-induced T lymphocyte proliferation and DR antigens on AM play an important role in the accessory cell function.
Effect of Human Mast Cell Tryptase on Human Plasma Proenzymes
Authors: Harvima Ilkka T. , Harvima Rauno J. , Penttilä Ilkka M. , Eloranta Terho O. , Horsmanheimo Maija , Fräki Jorma E.
Published: August 2009

The effect of human skin mast cell tryptase on human plasma proenzymes (prothrombin, coagulation factor XII, complement C1s, protein C and plasminogen) was investigated. Tryptase had no effect on these proenzymes, when incubated with them at 37°C for up to 90 min, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by the ability to hydrolyze specific peptide p-nitroanilide substrates. After prolonged treatment with tryptase, proenzymes could be fully activated with their specific activators. The results indicate that tryptase neither activates these plasma proenzymes nor inactivates the corresponding active enzymes. As a positive control, the tryptase preparation was also incubated with human fibrinogen and rat thymus histones. Prolonged treatment with tryptase increased the thrombin-induced clotting time of fibrinogen. Tryptase also efficiently hydrolyzed histone H1 from rat thymus. Histones H3/H2B and H2A were hydrolyzed less efficiently than H1, and no hydrolysis of histone H4 by tryptase was detected under the experimental conditions.
Receptor-Operated, but Not Voltage-Operated, Calcium Channels Are Involved in Basophil Leucocyte Activation and Histamine Release
Authors: Tedeschi A. , Miadonna A. , Lorini M. , Arquati M. , Zanussi C.
Published: August 2009

Transmembrane calcium flux is a critical step in basophil and mast cell activation and subsequent histamine release. This calcium flux is likely to take place through specialized membrane ion channels. Two types of calcium channels have been described so far: the first type is voltage operated and the second type is receptor operated. Depolarization of cell membrane by K+-rich solutions is followed by voltage-operated channel opening in excitable cells, such as smooth muscle cells. We evaluated whether high K+ extracellular concentrations can trigger basophil activation and histamine release. We found that human basophil leucocytes, showing a normal response to activating signals, such as anti-IgE antiserum and formylmethionine peptide, release no histamine when exposed to K+-rich media, alone or in combination with the K+ carrier valinomycin. These results are consistent with there being receptor-operated, but not voltage-operated, calcium channels in the basophil leucocyte plasma membrane.
Fractionation and Analysis of Allergenicity of Allergens from Prosopis juliflora Pollen
Authors: Thakur Indu Shekhar
Published: August 2009

Prosopis juliflora pollen allergen extract was prepared, and its crude allergen extract was fractionated by Sephadex G-100 gel filtration. Six different fractions were obtained which was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Protein and carbohydrate content of each fraction were estimated. Fraction E (MW 20,000) showed a 25% carbohydrate concentration. The amino acid analysis indicated that this fraction was rich in glutamic acid and alanine. Antigenicity or allergenicity of fractionated allergens were checked by gel diffusion test, rocket immunoelectrophoresis, skin prick test, and radioallergosorbent test. All these tests indicated that fraction E consisted mainly of allergenic molecules (MW 20,000) of P. juliflora pollen.
Interactions of Hemopoietic Cytokines on Differentiation of HL-60 Cells
Authors: Tsuda Takashi , Switzer Jan , Bienenstock John , Denburg Judah A.
Published: August 2009

Nerve growth factor (NGF) is a neurotropic polypeptide which has broad biological activity other than support of growth and survival of sympathetic, sensory and central neurons. NGF promotes rat mast cell hyperplasia in vivo and human granulopoiesis in vitro, selectively augmenting basophil/mast cell differentiation in the presence of T cells or conditioned medium derived from a human T cell line (Mo-CM), a source of granulocyte-macrophage colony-stimulating factor (GM-CSF). NGF also synergizes with GM-CSF to promote human basophil/mast cell differentiation in both methylcellulose and suspension cultures of myeloid progenitors. In the current studies, we examined the interactions of NGF and several cytokines considered to be involved in human basophil/mast cell and eosinophil growth and differentiation, including interleukin (IL)-3, IL-4, IL-5, GM-CSF and granulocyte colony-stimulating factor (G-CSF). NGF synergistically enhanced IL-5 induced dose-dependent increases in histamine content and basophilic cell differentiation of myeloid leukemic HL-60 cells, but was only additive to similar effects of IL-3. In contrast, IL-4 and G-CSF did not promote basophilic differentiation of HL-60 cells in the presence or absence of NGF. Various combinations of GM-CSF, G-CSF, IL-3, IL-4 and IL-5 could not reproduce the synergy observed between NGF and either IL-5 or GM-CSF. NGF appears to represent a class of lineage-specific co-factors, in this case being involved in GM-CSF- or IL-5-induced basophilic lineage differentiation, thus contributing to tissue inflammation or repair.
Histamine Receptors on Leukocytes Are Expressed Differently in vitro and ex vivo
Authors: Leino L. , Lilius E.-M.
Published: August 2009

The effect of histamine and histamine antagonists on the respiratory burst activity of leukocytes was studied. The activity was measured as zymosan-induced luminol-dependent chemiluminescence (CL) of isolated leukocytes and 1:2,500 diluted whole blood (WB). Histamine caused a dose-dependent inhibition of CL. For separated leukocytes the ID50 was 8 × 10––5M and for WB it was 1.5 × 10––3M. Diphenhydramine, an H1-antagonist increased the inhibitory effect of histamine while H2-antagonist cimetidine blocked the inhibition of CL of separated leukocytes. Cimetidine was not capable of reversing the effect of histamine on WB-CL. These data suggest that the histamine receptors on leukocytes are associated with Fc and/or complement receptors and the expression and function of histamine receptors can be studied by measuring the respiratory burst activity. The results also provide evidence of differences in histamine action in vitro and ex vivo conditions in WB.
Increased Autoreactive T Cell Frequency in Tuberculous Patients
Authors: Del Gallo Flavia , Lombardi Giovanna , Piccolella Enza , Montani Saveria Gilardini , Del Porto Paola , Pugliese Orsola , Antonelli Guido , Colizzi Vittorio
Published: August 2009

The development of putative self-MHC-reactive T cells and their precursor frequency was estimated in peripheral blood lymphocyte cultures stimulated in vitro with PPD. The role of foreign antigen in the generation of self-MHC-reactive T cells in vivo was analyzed by comparing the frequency of autoreactive T cells in the peripheral blood of tuberculous patients with that observed in healthy individuals. It was found that PPD in vitro and Mycobacterium tuberculosis infection in vivo increased substantially the generation of autoreactive T cells. Autoreactive T cell clones were shown (1) to recognize self MHC class II products; (2) to release gamma interferon in the absence of exogenous antigen, and (3) to express autocytotoxic activity. All these findings suggest that self-MHC-reactive T cells may be involved in the inflammatory response to M. tuberculosis.
Immunoreactive Leukotrienes in Nettle Plants (Urtica urens)
Authors: Czarnetzki Beate M. , Thiele Thomas , Rosenbach Thomas
Published: August 2009

In order to clarify the mechanisms of urtication after contact with stinging plants, nettle (Urtica urens) hair and whole-plant extracts were examined for the presence of leukotriene (LT) B4 and LTC4 by reverse phase high-pressure liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) and for in vitro neutrophil chemotactic activity and histamine contents. Both hair and plant extracts contained high levels of LTB4 and LTC4 by RIA as well as histamine. The presence of LTB4 was supported by RP-HPLC elution profiles and by in vitro chemotaxis. Nettle hairs therefore resemble insect venoms and cutaneous mast cells with regard to their spectrum of mediators.
Serological Analysis of Experimental Autoimmune Thyroiditis in the Buffalo Strain Rat
Authors: Cohen S.B. , Weetman A.P.
Published: August 2009

Previous experiments have indicated that experimental autoimmune thyroiditis (EAT) induced in the Buffalo strain of rat by neonatal thymectomy closely resembles Hashimoto’s thyroiditis. In these experiments we have analysed the serology of this type of EAT. Anti-thyroglobulin (Tg) antibodies were the only thyroid autoantibodies detected by immunoblotting. The IgG subclass distribution, analysed by a monoclonal anti-rat IgG subclass ELIS A, altered during the course of disease. The proportion of IgG 1 subclass Tg antibodies rose and the proportion of IgG2b subclass antibodies fell as disease progressed and the relative functional affinities of the anti-Tg IgG subclasses increased as disease progressed. The sera from animals with EAT were not cytotoxic in the presence of complement to a cultured rat thyroid cell line. In conclusion, EAT does not result in the production of antibodies against heterologous autoantigens as in Hashimoto’s thyroiditis. Tg antibodies are produced which have similar properties to human Tg autoantibodies, including lack of cytotoxicity, and subclass restriction; these appear to have little pathogenic role in the disease.
Gm Allotype Genes and Gene Dosage Affecting Both IgG Subclass and IgE Levels in Atopic Patients
Authors: Oxelius Vivi-Anne
Published: August 2009

The imbalanced IgG subclass levels of 50 atopic patients with IgE > 600 kU/1 reflected the Gm expression of the patients. IgG1 was significantly increased but only in patients with the phenotypes Gm(ffnn) and Gm(a, a,XXX, XXX)·The typical IgG4 increase was found in the most frequent Gm(ffnn) and Gm(a, f, n,”) phenotypes. Significant increase of IgG2, IgG3 and IgG4 was found in patients homozygous for G2m(n) compared to those lacking this allotype. IgG2 decrease was found in those with the Gm(f, f,XXX,XXX) phenotype and IgG3 decrease in those with the Gm(a, a,XXX,XXX) phenotype. Also the IgE levels were influenced by the Gm allotypes with increased IgE levels in the Glm(ff) patients compared to the G1m(aa) patients.
Enhancement of Antigen-Induced Bronchoconstriction in the Guinea Pig after Intravascular Complement Activation with Cobra Venom Factor
Authors: Regal Jean F.
Published: August 2009

The purpose of the present studies was to begin to test the hypothesis that the complement system is important for antigen-induced bronchoconstriction in the guinea pig. The effect of the complement-depleting agent cobra venom factor (CVF) on antigen-induced bronchoconstriction in guinea pigs passively sensitized with IgG or IgE antibody to ovalbumin was determined. Intravenous injection of CVF significantly reduced total hemolytic complement activity (CH50), caused a transient decrease in dynamic lung compliance, an increase in pulmonary resistance, and a decrease in circulating white blood cells with a sustained decrease in platelets. Antigen-induced bronchoconstriction was not inhibited in either IgG- or IgE-sensitized guinea pigs depleted of complement. Thus, a role for the complement system as a contributing factor in antigen-induced bronchoconstriction was not supported. On the contrary, our studies revealed an enhanced antigen-induced bronchoconstriction in guinea pigs depleted of complement with CVF. Our studies were then directed towards characterizing the enhanced response to begin to determine the mechanism responsible. The enhanced antigen-induced bronchoconstriction occurred in both IgG- and IgE-sensitized guinea pigs and was most apparent at lower antigen doses. The enhanced response still occurred in animals treated for 5 min with a dose of CVF (10 U) which caused no demonstrable decrease in CH50, a 30% level of conversion of C3, and a normal response to C5a. These data suggested that the enhanced response was not related to the level of C5a responsiveness of the animal. In addition, CVF-treated animals were normally responsive to various purported mediators of antigen-induced bronchoconstriction: histamine, leukotriene C4, platelet activating factor, the thromboxane-mimetic U-46,619, and prostaglandin F2α. The response to prostaglandin D2 was slightly enhanced. Thus, CVF treatment did not result in a generalized airway hyperreactivity. Since antigen-induced bronchoconstriction was not inhibited in animals depleted of complement, these studies suggest that an intact complement system is not necessary for this antigen-induced event. However, the enhanced antigen-induced bronchoconstriction seen after CVF treatment suggests that complement depletion and/or activation in vivo may be an important modulator or determinant of the severity of a subsequent antigen-induced bronchoconstriction.
Mouse Autoantibodies Bind to a Phospholipase-C-Sensitive Structure on Red Blood Cells
Authors: Hardy S.J. , Cox K.O.
Published: August 2009

After culturing mouse peritoneal cells in vitro for 4 days, high numbers of cells can be detected that secrete autoantibodies against isologous red blood cells (RBC), modified with the proteolytic enzyme bromelain (Brom). Plaque-forming cell numbers against mouse Brom RBC were significantly reduced by pre-treating mouse Brom RBC prior to haemolytic assay with phospholipase C, an enzyme that hydrolyzes phospholipids, notably phosphatidylcholine. In contrast, further treatment of mouse Brom RBC with Brom, neuraminidase, β-chymotrypsin, trypsin, or papain had no effect on plaque-forming cell numbers. These results show that phosphatidylcholine is an integral part of the mouse RBC autoantigen exposed by Brom treatment.
Fibronectin Levels in Plasma after Platelet-Activating Factor Inhalation
Authors: Nabe Makoto , Miyagawa Hidefumi , Hopp Russell J. , Agrawal Devendra K. , Bewtra Againdra K. , Townley Robert G.
Published: August 2009

We evaluated fibronectin levels in plasma and blood leukocyte counts after platelet-activating factor-acether (PAF) inhalation to examine whether PAF may cause pulmonary vascular injury. PAF inhalation challenges were performed on 5 normal subjects and 5 atopic asthmatics. Fibronectin levels in plasma were significantly increased at 5 min (342.5 ± 36.9 μg/ml) after PAF challenge as compared with the baseline values of 277.1 ± 33.0 μg/ml (p < 0.024). Total white blood cell and neutrophil counts were significantly decreased 5 min after PAF inhalation (WBC from 5,360 ± 460 to 4,260 ± 380/mm3, p < 0.028; neutrophils from 3,113 ± 441 to 2,214 ± 344/mm3, p < 0.043). Our study shows that plasma fibronectin levels transiently increase and that peripheral total WBC counts decrease after PAF inhalation. Elevated fibronectin levels may provide evidence of pulmonary vascular injury due to neutrophils which might be primed and recruited to pulmonary circulation by PAF.
Isolation of cDNA Coding for the Major Mite Allergen Der p II by IgE Plaque Immunoassay
Authors: Chua K.Y. , Doyle C.R. , Simpson R.J. , Turner K.J. , Stewart G.A. , Thomas W.R.
Published: August 2009

A lambda gt11 library made with cDNA from the house dust mite Dermatophagoides pteronyssinus was screened with human allergic serum by IgE plaque radioimmunoassay. This resulted in the isolation of clones coding for the major allergen Der p II. The cDNA coded for a 129-residue protein of 14,131 daltons with no N-glycosylation sites. No sequence homology with other proteins was evident. The Der p II expressed in Escherichia coli reacted with IgE in 14 of 17 sera from mite-allergic patients giving clonal evidence for its designation as a major allergen. This, along with previous work, has resulted in the cloning of the two major mite allergens.
Characterization of Four Major Allergens of Hen Egg-White by IEF/SDS-PAGE Combined with Electrophoretic Transfer and IgE-Immunoautoradiography
Authors: Holen E. , Elsayed S.
Published: August 2009

The antigenic and allergenic epitope maps of the hen egg-white ovalbumin, ovomucoid, ovotransferrin and lysozyme were obtained using one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis and two-dimensional sodium dodecyl sulfate gel electrophoresis following isoelectric focusing. The isoelectric points of egg-white proteins were assigned by the isoelectric focusing technique. Two-dimensional electrophoresis following isoelectric focusing provided further information regarding epitope mapping. Furthermore, electrophoretic transfer of the proteins to nitrocellulose and subsequent immunoautoradiography, clearly demonstrated the allergenicity of these proteins. An important benefit of these methods was confirming that lysozyme bound strongly to IgE in all the human sera from egg-allergic individuals and that lysozyme, in addition to ovalbumin, ovomucoid and ovotransferrin, was one of the major allergens of hen egg-white.
A Contiguous Network of Dendritic Antigen-Presenting Cells within the Respiratory Epithelium
Authors: Holt Patrick G. , Schon-Hegrad Michael A. , Oliver Jane , Holt Barbara J. , McMenamin Paul G.
Published: August 2009

This study utilises a simple technique to section airway epithelium in a plane parallel to the basement membrane, thus providing a unique plan view of the intra-epithelial cell populations. Immunoperoxidase staining of these tissue sections for class II major histocompatibility complex la antigen reveals a virtually contiguous network of Ia-positive dendritic cells (DC) within the epithelium. These DC are shown to be capable of binding inhaled antigens in vivo in a form suitable for presentation to T cells. The strategic location of these cells and the fact that they account for virtually all staining in the airway epithelium during the steady state is convincing evidence that the DC network functions as the ‘first line of defence’ in surveillance for inhaled antigens and further suggests a major role for the intra-epithelial DC in allergic and infectious disease(s) in the respiratory tract.
Histological Study of Mast Cells in the Actively Sensitized Guinea Pig Lung and after Challenge: Effect of a Corticosteroid
Authors: Bachelet Claude-Marie , Bernaudin Jean-François , Fleury-Feith Jocelyne
Published: August 2009

Although mast cells are sometimes considered to play a minor role in hypersensitivity reactions, we used a histological method to show the modifications in the guinea pig lung mast cell population in the course of such reactions. We sensitized guinea pigs with ovalbumin and studied the effect of the challenge with and without corticosteroid treatment. We observed that the mast cell count was not modified after sensitization but was decreased after challenge. Twenty-four hours after challenge, the number of mast cells returned to the control value, indicating a renewal of the mast cell pool. A second challenge, 1 week after the first, did not provoke the same mast cell degranulation, suggesting a non-responsiveness to aerosol antigen. Betamethasone dipropionate treatment protected mast cells against challenge: in treated guinea pigs, mast cell degranulation was prevented, and we did not observe any change in mast cell count after challenge. The present study was useful to show an effect of corticosteroids on mast cell degranulation in immediate hypersensitivity reactions in vivo.
Glucocorticoid-Induced Appearance of the Macrophage Subtype RM 3/1 in Peripheral Blood of Man
Authors: Zwadlo-Klarwasser Gabriele , Bent Stefan , Haubeck Hans-Dieter , Sorg Clemens , Schmutzler Wolfgang
Published: August 2009

The effect of intravenous administration of the glucocorticoid prednylidene on the macrophage subtype RM 3/1 in the peripheral blood of man was studied. Injection of 60 mg steroid resulted in an increase in the proportion of RM 3/1 positive monocytes 12 h after application from the basic level to about 80%. After 24 h the number of RM 3/1 positive cells decreased but remained elevated over the basis rate for a period of at least 15 days. Similar results were obtained after administration of 30 or 6 mg prednylidene even if the peak value occurred with a delay of 12 h after 6 mg. A dose- and time-dependent induction of the RM 3/1 positive subtype could also be demonstrated in vitro by the addition of prednylidene or dexamethasone to cultured purified human monocytes/macrophages. Deoxycortone or indomethacin had no effects. These results suggest that glucocorticoids exert their influence on cells of the mononuclear phagocytic lineage by inducing a distinct monocyte/macrophage subpopulation which seems to be associated with anti-inflammatory functions.
Immune Response to Blomia kulagini and Dermatophagoides pteronyssinus in Sweden and Brazil
Authors: van Hage-Hamsten Marianne , Machado Lavinia , Barros Myrthes T. , Johansson S.G.O.
Published: August 2009

The immune responses to the non-pyroglyphid mite Blomia kulagini and the pyroglyphid mite Dermatophagoides pteronyssinus were compared in 440 Swedish farmers and 34 allergic subjects from Brazil using the radioallergosorbent test (RAST). Both B. kulagini and D. pteronyssinus were shown to be a common cause of sensitization, particularly in Brazil. The highest RAST scores were found against D. pteronyssinus in sera from both countries. Allergenic cross-reactivity between the two mites was studied by the RAST inhibition technique. Both B. kulagini and D. pteronyssinus possess their own unique allergens as well as allergens in common. Thus, besides mites belonging to the Dermatophagoides genus, other pyroglyphid and non-pyroglyphid mites in different countries should be taken into consideration before starting hyposensitization treatments.
Abolished Net Output of Lymphocytes from the Spleen after Immunization with Salmonella typhi H
Authors: Sandberg Göran
Published: August 2009

Cell release from the spleen was studied by comparing the content of mono- and polynuclear leucocytes in splenic afferent and efferent blood. A difference in the content of mononuclear cells was registered in normal animals, indicating a release of such cells from the spleen. Immunization with Salmonella typhi H antigen reduced this veno-arterial difference and abolished it totally after 11 days. Normal values were obtained again 28 days after immunization. The results indicate that the antigen causes an altered relation between input and output of mononuclear cells, mainly lymphocytes.
Western Blot Analysis of Water-Soluble Wheat Flour (Triticum vulgaris) Allergens
Authors: Pfeil T. , Schwabl U. , Ulmer W.T. , König W.
Published: August 2009

Water-soluble wheat flour allergens were analyzed using the western blot technique. The sera of 130 bakers (42 with wheat flour allergy, 88 without allergy) were analyzed for IgE, IgG, and IgG4 binding towards wheat flour allergens. Three major allergens with molecular weights of 47, 17, and 15 kD were identified (Tri v Bd 47, Tri v Bd 17, and Tri v Bd 15; IUIS allergen nomenclature) by their ability to bind IgE from approximately 50% of the allergic sera. The specificity of the IgG was mostly directed against three high molecular weight components (134, 122, and 98 kD). In contrast, IgE and IgG4 was found frequently against the major allergens. 40% of healthy donors versus 12% of the asthmatic bakers had no detectable IgG4 against wheat flour extract. The biological activity of the water-soluble wheat flour components has been shown by their ability to induce histamine release from basophil leukocytes obtained from patients with bakers’ asthma. The release of histamine from basophil leukocytes significantly correlated with the presence of IgE antibodies against wheat flour components of 47, 17 and 15 kD. A regulatory role of wheat flour specific IgG4 in allergic diseases could not be evaluated by western blot analysis.