bluuebunny/crossref_metadata_2025 huggingface dataset:
Humoral Immune Response to Cow’s Milk in Children with Cow’s Milk Allergy
Authors: Firer M.A. , Hoskings C.S. , Hill D.J.
Published: August 2009

In 47 infants and children aged 4–66 months with clinically proven cow’s milk allergy and in a group of age-matched controls, serum IgG, IgA and IgM cow’s milk-specific antibodies were determined with ELISA assays while IgE cow’s milk-specific antibodies were measured with Pharmacia RAST. The patients were divided into three separate groups according to the time of clinical response to a standardized cow’s milk challenge protocol. Immediate reactions ( < 45 min after challenge), which were mainly accompanied by urticarial skin eruptions, were associated with elevated IgE milk-specific antibody levels, indicating the involvement of an immediate hypersensitivity mechanism. Alternatively, intermediate reactions (1–20 h after challenge), which were mainly accompanied by vomiting and diarrhea, were not IgE-mediated. In the late reactions ( > 20 h after challenge) both eczematous and gastrointestinal reactions were seen. Patients with eczema-tous eruptions also showed elevated IgE milk-specific antibody levels. IgG milk-specific antibody levels were similar in each of the patient groups but all groups were significantly lower than in the controls. Levels of IgA and IgM milk-specific antibodies were similar in patients and controls. The results indicate that different im-munopathogenic mechanisms are operative in these subgroups of patients with cow’s milk allergy.
Activation of the Alternative Pathway of Human Complement by Sulfhydryl Compounds of Analytic and Therapeutic Use
Authors: von Zabern I. , Nolte R.
Published: August 2009

Thiol-containing drugs (dimercaprol, dimercaptopropanesulfonate, captopril, penicillamine, N-acetylcysteine) and the standard reducing agent β-mercaptoethanol, activate the alternative pathway of complement as shown by in vitro experiments. Depending on the substance tested, at concentrations of 0.5–5 mM, cleavage products of C3 and factor B were demonstrable in serum by immunoelectrophoresis. The regulatory protein factor I proved to be very sensitive to thiols; this observation offers an explanation for the alternative pathway activating effect of these substances. At concentrations of thiols that initiate the alternative pathway, the classical pathway was not or only to a minor extent activated; however, the activity of C2, C5 and one or several of the components C6–9 was directly affected. Alkylation of the thiol group of the compounds tested, abrogated their effects on the complement system.
Eosinophil Chemotactic Lymphokine Produced by Spleen Cells of Schistosoma japonicum-lnfected Mice
Authors: Owhashi M. , Nawa Y.
Published: August 2009

Two distinctive types of eosinophil chemotactic lymphokines (ECF-L) directed against eosinophils obtained from bone marrow (BM-Eo) or from peritoneal cavity (PEC-Eo) were isolated from the media conditioned by spleen cells from mice infected with Schistosoma japonicum by a combination of anion-exchange chromatography on DE52, affinity chromatography on Procion Red agarose, and high-performance liquid chromatography. The molecular weight of ECF-L directed against BM-Eo was 10,000 and that against PEC-Eo was 46,000 by SDS-PAGE analysis. After isolation, selectivity of the two distinct ECF-L was examined using BM-Eo, PEC-Eo and circulating Eo. ECF-L directed against BM-Eo could attract only BM-Eo, whereas that directed against PEC-Eo could attract not only PEC-Eo but also circulating Eo. Synthetic ECF-A was chemotactic against both BM- and circulating-Eo but not against PEC-Eo. When BM-Eo were cultured with ECF-L selective to PEC-Eo, they became reactive to ECF-L selective to PEC-Eo, whereas ECF-L selective to BM-Eo did not have such effect. These results suggest that ECF-L selective to PEC-Eo is not simply a chemo-attractant but also is important in eosinophil differentiation.
Variability of Antigenicity/Allergenicity in Different Strains of Alternaria alternata
Authors: Steringer Ingrid , Aukrust Lars , Einarsson Roland
Published: August 2009

Biochemical and immunological properties were studied in crude mould extracts from 10 different strains of Alternaria alternata. The mould strains were grown on completely synthetic medium and harvested under identical conditions. The biochemical and immunological analyses (protein content, carbohydrate content, IEF, RAST-inhibition, CIE/CRIE and the content of a major allergen) showed significant variations between the individual strains of A. alternata. 32 antigens were identified in the CIE pattern of A. alternata and 19 different allergens were identified in CRIE analysis (1 major, 7 intermediate and 6 minor allergens). There was a significant variation in the content of the major A. alternata allergen (Ag-8, partially identical to Alt-I) between the different individual strains. Furthermore, a considerable variation in the content of Ag-8 was observed for a specific strain cultivated at different occasions under identical cultivation conditions during a time period of 3 years.
Detection of IgG Subclass-Specific Anti-IgE Antibodies in Normal and Atopic Individuals
Authors: Wilson Philip B. , Fairfield Janet E. , Beech Nick
Published: August 2009

Atopic dermatitis and atopic asthma patients were shown to have elevated IgG anti-IgE levels compared with those of controls. When the pattern of IgG subclass anti-IgE activity was studied, further differences between the three groups became apparent. Adults with atopic dermatitis had elevated IgG1 and IgG4 anti-IgE compared with those of controls but no corresponding increase in the IgG2 or IgG3 subclasses. For Sri Lankan children with atopic asthma and a high incidence of Nematoda infection, elevated anti-IgE was restricted to the IgG1 subclass. While IgG1 anti-IgE proved to be the predominant subclass for both atopic groups, IgG2 anti-IgE was the major subclass among controls.
Neuropeptide Regulation of Human Thymocyte, Guinea Pig T Lymphocyte and Rat B Lymphocyte Mitogenesis
Authors: Söder Olof , Hellström Per M.
Published: August 2009

The effect of 15 defined neuropeptides on the mitogenic activation of lymphocytes from human thymus, guinea pig lymph nodes and rat spleen was investigated. Lymphocytes were incubated in the absence or presence of polyclonal T and B cell activators together with increasing doses of the neuropeptides, and harvested at 48 h of culture after pulse-labeling with 3H-thymidine to assess the DNA synthesis. A dose-related stimulatory effect on the spontaneous 3H-thymidine incorporation of human thymocytes was obtained with methionine-enkephalin (met-enk), motilin and neurotensin. Vasoactive intestinal polypeptide (VIP) and peptide HI (PHI) were inhibitory. A similar responsiveness was observed in cultures of phytohemagglutinin P (PHA)-activated human thymocytes. The low level of basal DNA synthesis of guinea pig lymph node cells was stimulated by VIP and inhibited by neuropeptide Y (NPY) and PHI. PHA-activated lymph node T lymphocytes were stimulated by neurotensin, bombesin and motilin, whereas NPY inhibited the thymidine uptake. The low rate of spontaneous DNA synthesis of rat spleen cells was increased in the presence of VIP. Met-enk stimulated both basal and dextran sulfate-activated splenic B cell proliferation, whereas PHI was inhibitory in both cases. The following peptides were found to be inactive in all the above assays: substance P, cholecystokinin-octapeptide, somatostatin, galanin, oxytocin, pentagastrin and gastrin-releasing peptide 1–27 and 14–27. Although the responses were generally of low magnitude and observed at high peptide concentrations, the present study contributes to the understanding of possible mechanisms involved in interactions between the nervous and the immune system.
Mitogenic Effect of Beryllium Sulfate on Mouse B Lymphocytes but Not T Lymphocytes in vitro
Authors: Newmann Lee S. , Campbell Priscilla A.
Published: August 2009

Beryllium metal and its salts can produce disease in man and in animal models. Beryllium disease is thought to involve cell-mediated immunity and an antigen-dependent response by beryllium-specific T cells. Beryllium salts have been shown to stimulate lymphocyte proliferation and release of lymphokines, and to induce granuloma formation and delayed-type hypersensitivity reactions in mice, guinea pigs and man. The studies described here were designed to test the hypothesis that a second lymphocyte population, B cells, may be responding nonspecifically to beryllium. Different populations of BDF1 mouse lymphocytes were cultured in the presence of varying concentrations of beryllium sulfate (BeSO4), and the increase in 125-iodouracildeoxyriboside uptake after 72 h in culture was determined. The data show that BeSO4 is weakly mitogenic for normal mouse spleen cells. Furthermore, BeSO4 is mitogenic for normal and nude mouse spleen B cells and not for spleen T cells or thymocytes in vitro. These findings suggest that BeSO4 can stimulate B cells nonspecifically, and support the hypothesis that polyclonal activation of B cells by beryllium may occur.
Effect of a Fish-Oil-Supplemented Diet on Inflammation and Immunological Processes in Rats
Authors: Yoshino Shin , Ellis Earl F.
Published: August 2009

The effect of a diet enriched in fish-oil-derived fatty acids including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DCHA) in inflammation and immunological processes in rats has been investigated. Rats on a normal chow diet were given 500 mg/kg/day EPA and 333 mg/kg/day DCHA by gavage over a period of 50 days. Control groups received water, oleic acid or safflower oil. Acute and chronic phases of inflammation induced by antigenic challenge with bovine serum albumin were examined in the rat air-pouch model. In rats receiving fish-oil-derived fatty acids, there was a reduced production of the arachidonic acid metabolites, prostaglandin E2 and leukotriene B4, in the exudate in the chronic phase of inflammation, but not in the acute phase. This was associated with an increased infiltration of leukocytes, especially monocytes and macrophages, in the chronic inflammatory phase. The fish-oil-supplemented diet showed no effect on the volume of inflammatory exudate, the amount of protein in the exudate and connective tissue proliferation. Carrageenin-induced paw edema in the animals was not influenced by the diet. There was no effect of the dietary fish oil on production of antibodies to bovine serum albumin in the rats. However, the diet appeared to suppress a delayed-type skin reaction in the animals. These studies suggest that fish-oil-derived fatty acids may modulate chronic inflammation and a cellular-mediated immunological reaction by reducing the synthesis of arachidonic acid metabolites.
Cellular Composition of Peripheral Lymph and Skin of Sheep Defined by Monoclonal Antibodies
Authors: Hein W.R. , McClure S.J. , Miyasaka M.
Published: August 2009

The surface phenotype of cells in peripheral lymph collected from afferent lymphatics leading to the popliteal lymph node of sheep was determined using a panel of monoclonal antibodies (mAbs). The majority of lymphocytes (83.5%) expressed the sheep pan-T cell antigen and only 13.3% bore surface immunoglobulin molecules. All peripheral T cell subsets occurring in sheep were detected; 50.2% of lymphocytes were positive for mAb SBU-T4 (T helper), 7.3% were positive for mAb ST-8 (T cytotoxic), and 8.4 and 43.0% expressed T subset markers recognized by mAbs 197 and T-80, respectively. Major histocompatibility complex (MHC) class I antigens were detected on 71.1% of lymphocytes and MHC class II antigens on 21.8%. The macrophage/veiled cells found in peripheral lymph did not express lymphocyte subset markers but were positive for MHC class I and II antigens, the sheep homologue of T6 antigen, leukocyte common antigen and mAb 175 (myeloid/erythroid). Macrophage-like cells occurring in the epidermis of skin taken from the lower hindleg gave positive staining reactions to the same mAbs which stained the macrophage/veiled cells in peripheral lymph. These results illustrate differences between the migration of lymphocyte subsets through nonlymphoid as compared to lymphoid tissues and point to a possible developmental or migratory relationship between the macrophage-like cells in skin and those in afferent popliteal lymph.
Differential Effects of Histamine Receptor Antagonists on Human Natural Killer Cell Activity
Authors: Hellstrand Kristoffer , Hermodsson Svante
Published: August 2009

In this study we investigated the modulation of natural killer (NK) cell activity by various histamine receptor antagonists in vitro. The histamine H2-receptor antagonists cimetidine, ranitidine and tiotidine suppressed NK cell cytotoxicity (NKCC) at a high concentration (10––3M). Cimetidine enhanced NKCC of Ficoll-Hypaque-separated lymphocytes and of lymphocytes enriched for NKCC by Percoll density gradient centrifugation. The enhancing effect of cimetidine was dose-dependent at final concentrations of 10––4––10––7M and did not require the presence of adherent cells/monocytes. Ranitidine did not affect NKCC over a wide range of concentrations. Tiotidine strongly enhanced NKCC of low-density, large granular lymphocyte-enriched mononuclear cells (MNC) in the presence of adherent cells/monocytes, but was ineffective in nonadherent effector cells. All H2-receptor antagonists clearly antagonized histamine-induced NKCC enhancement in monocyte-containing effector cells. Clemastin, a specific H1-receptor antagonist, effectively suppressed NKCC. This effect was mimicked by a clemastin isomer with very low affinity for H1 receptors. We conclude that (1) cimetidine enhances NKCC in vitro by a mechanism of action that is not specifically related to antagonism of H2-receptors, (2) tiotidine displays mixed agonist/antagonist properties for MNC H2-receptors and (3) NK-suppressive properties of clemastin are unrelated to H1-receptor antagonism.
Depression of Lymph Node Cell Proliferation Induced by Oxazolone
Authors: Kimber Ian , Pierce Belinda B. , Mitchell Julie A. , Kinnaird Avril
Published: August 2009

The influence of topical exposure to two sensitizing chemicals on draining lymph node cell proliferative responses in BALB/c mice has been examined. Conventional contact sensitization with 4-ethoxymethylene-2-phenyloxazol-5-one (oxazolone) has been shown to induce a rapid and systemic suppression of subsequent proliferative responses to topically applied chemical which can be adoptively transferred to recipient mice with immune lymph node cells. In contrast to some previous reports in which such suppression was found to be largely antigen-specific in nature, we report that, at least initially, the inhibition of lymphocyte proliferation induced by skin sensitization is hapten-non-specific. The relevance of this phenomenon to the regulation of contact sensitization is discussed.
Endotoxin-Induced Auto-Immunity in Mice
Authors: Bloembergen Peter , Hofhuis Frans M.A. , van Dijk Hans , Verhagen Cora , Rademaker Pieternel M. , Willers Jan M.N.
Published: August 2009

Injection of mice with endotoxin results in the formation of auto-antibodies and the appearance of soluble immune complexes in the blood. In this study the relationship between the production and serum levels of autohaemolysins and circulating immune complex titres was investigated. Immune complexes were detected by a solid-phase C1q-binding assay and found to contain antibodies of the IgM, but not of the IgG class. Individual mice showed marked differences as to their splenic plaque-forming cells, serum autohaemolysin, and circulating immune complex responses, both in kinetic studies and dose-response experiments. The dissociation between production and serum levels of auto-antibodies was ascribed to extrasplenic synthesis or a disproportionate production per plasma cell. The independent behaviour of the circulating immune complex response could, at least partially, be attributed to differential complement-dependent clearance from the circulation. The implications of our findings for the laboratory diagnosis of auto-immunity at the blood level are being discussed.
Thymus Dependence of Compound 48/80-Induced Mucosal Mast Cell Proliferation
Authors: Aldenborg Frank
Published: August 2009

The mucosal and connective-tissue mast cells (MMC and CTMC, respectively) of the rat are histochemically and functionally distinct. MMC, unlike CTMC, are insensitive to the degranulating action of the mast cell secretagogue Compound 48/80 and instead increase in number after treatment with this drug. T cell-derived growth factors are necessary for the growth of MMC-like cells from hemopoietic tissues in vitro, as well as for nematode-induced MMC proliferation in vivo. We therefore examined the possible role of the thymus in the pharmacologically induced MMC expansion. Mast cell numbers and histamine content after treatment with Compound 48/80 were determined in the skin, tongue and gut of athymic male LEW/MOL-rnu/ rnu rats and their normal littermates at the age of 6–7 weeks. The influence of the strain of rat and the mode of administration of the drug was assessed by studying age- and sex-matched Sprague-Dawley rats as well. Injections of Compound 48/80 for 5 days resulted in a statistically significant increase in the number of intestinal MMC and the histamine content of all thymus-bearing rats. The increase was more pronounced in the Sprague-Dawley rats, however, and unrelated to the mode of administration of the drug, indicating that the MMC-stimulating action of Compound 48/80 is at least partly controlled by genetic factors. Athymic rats showed no statistically significant MMC expansion. This suggests that the MMC response after Compound 48/80 treatment may be at least partly dependent on the thymus. The mast cell numbers and histamine content in the tissues of athymic control rats were higher than in their thymus-bearing littermates. Subepidermal skin mast cells, especially frequent in the athymic rats, contained little histamine and a large fraction of these cells did not degranulate after Compound 48/80 treatment.
Use of the Cutaneous Anaphylactoid Test to Detect Differences in Mast Cell Mediator-Releasing Activity among LHRH Peptides
Authors: Nekola Mary V. , Morgan Jane E. , O’Neil Carol E. , Hocart Simon J. , Coy David H.
Published: August 2009

The cutaneous anaphylactoid test was evaluated as a screen for assessing mast cell mediator-releasing (MCMR) activity of luteinizing hormone-releasing hormone (LHRH) peptides. As expected, LHRH and D-Trp6-LHRH, an agonist of LHRH, were nonreactive at concentrations of antagonist which induced pronounced skin lesions. Differences between LHRH and antagonists were obvious, but it was difficult to detect differences among antagonists using arithmetic means. However, when the geometric means of the highest positive dilution were calculated, the MCMR activities of the antagonists differed and these differences appeared to be related to the hydrophobicities of their N-termini. Thus, it appears that the cutaneous anaphylactoid test is an appropriate screen to assess the MCMR activity of LHRH peptides and should assist in the development of clinically useful inhibitory analogs of LHRH.
Isolation of a Histamine-Releasing Factor from Two-Cell Human Embryo
Authors: Cocchiara Roberta , di Trapani Giovanna , Azzolina Antonina , Albeggiani Giuseppe , Geraci Domenico
Published: August 2009

An embryo-derived histamine-releasing factor (EHRF) was identified and partially purified from media in which two-cell human embryos were cultured. The EHRF at 5 μg/ml was capable of inducing 22 ± 7% release of histamine from sensitized human leukocytes, reaching a maximum of 56 ± 4% over an EHRF concentration range of 1–30 μg/ml. The EHRF was not detected in media where unfertilized oocytes were cultured or in medium alone. The effect of EHRF was not due to cytotoxicity since unsensitized leukocytes were unreactive. Histamine release did not occur when the assay was performed at 4°C or in presence of EDTA.
Antibodies to Food and Environmental Antigens in Serum, Respiratory Tract and Intestine of Pigs
Authors: Morgan K.L. , Patel D.P. , Bourne F.J.
Published: August 2009

Serum antibodies to food, bedding and dust were detected in pigs by precipitation and ELISA. There was an age-related decrease in the number of animals with precipitating antibodies and in the specific activity of IgG and IgM antibodies. Antibodies to food, bedding and dust were detected in respiratory and intestinal secretions, but there were differences in the distribution and isotype of antibodies to these three antigens.
Lack of IgA Antibody Response in Secretions of Rat Dams during Long-Term Ovalbumin Feeding
Authors: Wold Agnes E. , Dahlgren Ulf I.H. , Ahlstedt Staffan , Hanson Lars Å.
Published: August 2009

Antigen uptake, serum and secretory immune response was studied in rat dams fed pellets substituted with ovalbumin during three consecutive pregnancies and lactation periods. The concentration of ovalbumin was somewhat higher in milk than in serum, and both serum and milk concentrations showed a parallel decrease with feeding time. IgA antibodies did not develop in the milk during the 7-month feeding period and no antibody response could be detected in saliva or bile. By the time of the first lactation about half of the dams had developed low titres of IgG antibodies in serum and milk and the number of IgG responders, as well as the mean antibody titre, increased with each lactation. Also when injected directly into the Peyer’s patches ovalbumin induced solely IgG antibodies in the milk, while immunization with ferritin in addition resulted in both IgA and IgM antibodies in the milk against ferritin. The presence or absence of immunological tolerance to ovalbumin was examined in three groups of rats: (1) dams which had been fed ovalbumin-substituted pellets for one pregnancy and the first 14 days of the lactation period, i.e. for approximately 5 weeks altogether; (2) pups that were born by and kept together with these mothers, and (3) pups that were born by and kept together with mothers that were fed ovalbumin-substituted pellets during the pregnancy and until weaning at day 21 (altogether for 6 weeks). The rat dams in group 1 were not tolerant to a subcutaneous injection of ovalbumin 4 weeks after the termination of feeding with ovalbumin-substituted food. The pups (group 2) from these mothers also responded in a normal way to an injection of ovalbumin at the age of 2 months. In contrast, the pups in group 3 that were kept together with dams fed ovalbumin-substituted pellets for the whole lactation period, thus having access to the mothers’ food during 3 weeks, were tolerant to ovalbumin when tested at 2 or 3 months of age.
Age-Dependent Bone Marrow Mast Cell Maturation in Specific Pathogen-Free Rats
Authors: Ahlstedt S. , Håård J.
Published: August 2009

In the present study we have investigated an age-dependent mast cell increase. The mast cell numbers in bone marrow from (BN × Wi/Fu)F1 rats were determined using Alcian blue staining of cells in solution. From the 6th to the 19th week of age the number of mast cells in the bone marrow increased 6-fold with a marked peak around the 12th week of age. This increase in mast cell numbers was accompanied with increased histamine levels. Immunization increased the numbers further, also when the age-dependent mast cell increase was corrected for. The results should be considered in relation to previous studies revealing increased mast cell numbers in the bone marrow after immunization and also increasing numbers in the lungs of rats immunized with protein antigens or exposed to environmental antigens.
Lung Nonresponsiveness Produced in Guinea Pigs by Aerosolized Antigen
Authors: Blanchard G.C. , Pathak S.K. , Taylor Gercel , Berman L.D.
Published: August 2009

Nonresponsiveness to aerosolized antigen induced by repeated exposure of guinea pigs to either bovine serum albumin (BSA) or ragweed (RW) aerosols was not antigen specific, since induction of lung sensitization with the same, as well as heterologous antigen, was prevented. However, prior lung sensitization to an antigen was maintained in spite of subsequent treatment with aerosolized antigen. In this case treatment with aerosolized antigen could abrogate sensitization of the lung to all subsequent antigens. Chopped lung pieces could not be passively sensitized by antisera to homologous or heterologous antigens unless they were previously treated at low pH. High levels of conventional and homocytotropic antibody were demonstrated in nonresponder animals. Nonresponsiveness induced by fed antigen, however, was antigen specific, and elevated homocytotropic antibody levels were not present. Aerosol-induced nonresponsiveness appears to be a local phenomenon which involves a blocking of the pulmonary mast cells by immune complexes or other blocking factors so that they can no longer respond to antigen challenge.
Induction of Photoallergy in Guinea-Pigs by Injection of Photoallergen-Protein Conjugates
Authors: Barratt M.D. , Goodwin B.F.J. , Lovell W.W.
Published: August 2009

Photoconjugates were prepared by ultraviolet irradiation of guinea-pig albumin (GPA) with the photoallergens tetrachlorosalicylanilide (T4CS) and fentichlor. Injections of T4CS-GPA induced photoallergy to T4CS in 11 of 12 guinea-pigs whereas injections of fentichlor-GPA induced photoallergy in 5 of 12 guinea-pigs. Thus the fentichlor-GPA photoconjugate, which contained a molar ratio of hapten to protein 3 times higher than the corresponding T4CS conjugate, produced a significantly lower response. The results demonstrate the importance of protein conjugate formation in the induction of photoallergy, i.e. the role of carrier protein in contact sensitivity. The high potency of the T4CS-GPA photoconjugate in inducing photoallergy suggests that albumin may have a special role as a carrier protein in T4CS photoallergy.
Inhibition of Antigen-Induced Contraction of Guinea Pig Isolated Tracheal Muscle with 2-n-butyl-3-dimethylamino-5,6-methylenedioxy indene (MDI-A), indane (MDI-B) and 8-(diethylamino)octyl-3,4,5-trimethoxy benzoate hydrochloride (TMB-8)
Authors: Nagai Hiroichi , Yakuo Ikuhisa , Arimura Akinori , Hara Satoshi , Inagaki Naoki , Koda Akihide
Published: August 2009

The effects of three intracellular calcium antagonists on antigen-induced contraction of sensitized guinea pig tracheal muscle were investigated. The agents employed were: 2-n-butyl-3-dimethylamino-5,6-methylenedioxy indene hydrochloride (MDI-A), 2-n-butyl-3-dimethylamino-5,6-methylenedioxy indane hydrochloride (MDI-B) and 8-(diethylamino)octyl-3,4,5-trimethoxy benzoate hydrochloride (TMB-8). Each of these agents caused a concentration-dependent inhibition of the antigen-induced contraction of sensitized tracheal smooth muscle. Moreover, in histamine and leukotriene D4 (LTD4) incited contractions of guinea pig tracheal muscle, they showed antagonistic action. The induced tracheal muscle contraction, achieved through the addition of compound 48/80 to a solution of ethylene glycol bis (β-aminoethylether)-NNN1, N1-tetraacetic acid (EGTA) in a contained Ca-free medium, was completely inhibited by individual pretreatment with MDI-A, MDI-B and TMB-8. In the case of tracheal muscle contraction induced by CaCl2 in a high potassium, Ca-free medium, only TMB-8 inhibited contraction. Lastly, none of these three intracellular calcium antagonists affected the antigen-induced release of histamine and slow reacting substance of anaphylaxis (SRS-A). These results suggest that MDI-A and MDI-B inhibit the antigen-induced contraction of sensitized guinea pig tracheal muscle by interfering with the contractile responses caused by histamine and LTD4.
Platelet-Activating Factor in the Inflammatory Exudate in the Anaphylactic Phase of Allergic Inflammation in Rats
Authors: Watanabe Masako , Ohuchi Kazuo , Sugidachi Atsuhiro , Hirasawa Noriyasu , Hayashi Yasuhiro , Tsurufuji Susumu
Published: August 2009

Using a model of allergic inflammation of air pouch type in rats, the platelet-activating factor (PAF) in the pouch fluid in the anaphylactic phase was analyzed. Anaphylactic reaction was induced by injecting an antigen (azobenzene-arsonate-conjugated acetyl bovine serum albumin) solution into a subcutaneous air pouch preformed on the dorsum of immunized rats. The pouch fluid was collected 30 min after the antigenic challenge, and chloroform extract was subjected to normal phase high-performance liquid chromatography to isolate two fractions, PAF and lyso-PAF. In the pouch fluid, however, there was little activity of PAF as examined by the aggregation of guinea pig platelets. The lyso-PAF fraction obtained was acetylated to PAF chemically with pyridine and acetic anhydride. This acetylated lyso-PAF fraction induced the aggregation of guinea pig platelets, which was inhibited dose-dependently by a PAF antagonist, CV-3988. The amount of lyso-PAF in the pouch fluid of the immunized group in the anaphylactic phase was significantly higher than that of the nonimmunized group. When (3H-)PAF was incubated with the supernatant fraction of the pouch fluid it was metabolized into lyso-PAF time-dependently. The significance of the higher level of lyso-PAF in the pouch fluid in the anaphylactic phase of allergic inflammation is discussed.
Characterization of Trypanosoma cruzi Antigens Recognized by Sera from Patients with Chronic Chagas’ Disease
Authors: Gea S. , Rodriguez P. , Vottero-Cima E.
Published: August 2009

The aim of this work was to study the Trypanosoma cruzi antigens more reactive with antibodies from chagasic patients. For this purpose, the polypeptidic spectrum of cytosol was analyzed by isoelectric focusing. It showed about thirty bands distributed along a pH gradient of 3–10. In order to study the antigenicity of these polypeptides, four fractions from the focused cytosol were eluted and then analyzed by ELISA with chagasic and control sera. The more acidic and homogeneous fraction (FIV, pI 4–5,5) was the most reactive with antibodies from Chagas’ disease patients. The 125I-labeled FIV was immunoprecipitated with chagasic and control human sera and subjected to SDS-PAGE analysis. The autoradiography revealed the presence of 4 major antigens, with apparent molecular weights of 49, 80, 86 and 110 kilodaltons only reactive with sera from chagasic patients. Two antigens with molecular weights of 34 and 21 kilodaltons were reactive with both chagasic and control sera.
Circulating Basophil Counts in Atopic Individuals
Authors: Reilly K.M. , Yap P.L. , Dawes J. , Barnetson R.S.C. , MacKenzie F. , Allan T.L.
Published: August 2009

Absolute numbers of circulating basophils were measured in 55 atopic and 35 non-atopic individuals, using a flow cytometer for automated cell counting. They were significantly elevated in the atopic group (p < 0.001), but the total leucocyte count was not significantly different between the two groups. Eosinophil counts, which correlated with the number of basophils for both populations, were also significantly raised in the atopic group (p < 0.001). Individual variation in absolute basophil counts was not detected in sequential samples taken at daily intervals over 5 days, in 5 atopies and 4 non-atopies. In addition, no significant variation was detected in 6 atopies and 5 non-atopies over a period of up to 20 months.
Demonstration of Allergen Components in the Storage Mite Lepidoglyphus destructor by an Immunoblotting Technique
Authors: Johansson Eva , van Hage-Hamsten Marianne , Johansson S.G.O.
Published: August 2009

The allergen composition of crude extracts of the storage mite Lepidoglyphus destructor were investigated by sodium dodecylsulphate polyacrylamide gel electrophoresis and immunoblotting. The allergen components were detected by sera from allergic farmers with a positive radioallergosorbent test to L. destructor. The allergen-antibody complexes were visualized by rabbit anti-IgE and β-galactosidase-labelled sheep antirabbit IgG in a chromogenic substrate. Six out of the forty-three sera also had IgE antibodies to the culture medium, which were neutralized by addition of medium antigens before the demonstration of mite allergen components. A total of 11 allergen components were identified. Two of them were found to be major allergen components, identified by more than 50% of the 43 sera used in the study. These 2 IgE-binding components had molecular weights of approximately 18,000 and 16,000 daltons, respectively.
Involvement of the Outer Wall Layer of Cladosporium cladosporioides in an IgG-Mediated Hypersensitivity
Authors: Bergen M.S. , Yang T.J. , Collins R.P.
Published: August 2009

The serum of an individual hypersensitive to the fungus Cladosporium cladosporioides reacted with the outer wall layer of germinating spores, immature hyphae, and, to a lesser extent, mature mycelium, as detected by the indirect immunofluorescence assay when goat anti-human IgG, but not goat anti-human IgE, was employed. The outer wall layer of ungerminated spores, however, did not react with the patient’s serum and the anti-human IgG probe. When ungerminated spores were vortexed in physiological saline for 1 min, approximately 8 pg of protein/spore was released; this rapid release of protein may have been the cause of the loss of antigenic activity from the outer wall layer of ungerminated spores during the immunoassay process. Immunoblotting revealed the presence of a high molecular weight antigen in all the extracts of the fungus at various stages of the life cycle, including ungerminated and germinating spores, hyphae in the logarithmic phase, mature mycelium, and a culture filtrate of C. cladosporioides.
A Comparison of the Antigenic and Allergenic Components of Birch and Alder Pollens in Scandinavia and Australia
Authors: Hemmens V.J. , Baldo B.A. , Bass D. , Vik H. , Florvaag E. , Elsayed S.
Published: August 2009

Allergens in birch (Betula) pollens from B. pendula grown in Australia and Norway, B. davurica and B. populofolia and from alder (Alnus incana) were identified by electroblotting, following separation by SDS-PAGE, transfer to nitrocellulose membranes and incubation with sera from birch pollenallergic subjects. Of 42 antigenic components detected by protein staining in the pollen extract from B. pendula grown in Norway, 17 bound IgE. The allergenic components included those already reported in the literature at MWs of 40, 29, 25, 17 and 10–12 kd, as well as previously undescribed components at MWs of 90, 79, 60, 50, 38, 35, 31, 27, 23, 16, 15 and 14 kd. The major IgE-binding components were located in the low MW region 10–17 kd for all species of birch and alder pollen proteins studied. Results reported here provide the first evidence of birch and alder pollen allergies in Australia. Extensive heterogeneity was observed amongst the sera from birch pollen-allergic subjects in both Norway and Australia. Cross-reactivity appears to exist among the proteins present in pollen from the various birch species and from alder.
Dissection of Adjuvant and Suppressive Effects of Mycobacteria in Experimental Allergic Encephalomyelitis Production
Authors: Mostarica-Stojković Marija , Vukmanović Stanislav , Petrović Milica , Ramić Zorica , Lukić Miodrag L.
Published: August 2009

Dark August rats exhibit clinically and histologically verified experimental allergic encephalomyelitis (EAE) when immunized with appropriate antigen (nervous tissue, myelin basic protein) emulsified in complete Freund’s adjuvant (CFA). We provide evidence that 6,6′-trechalose dymicolate (TDM) incorporated in incomplete Freund’s adjuvant replaces CFA in EAE induction. The animals that recovered from EAE were resistant to the reinduction of the disease irrespectively whether Mycobacterium tuberculosis or TDM was used as an adjuvant. Finally, pretreatment with CFA alone was sufficient for prevention of disease elicited by challenge with encephalitogen + CFA. However, TDM, despite its adjuvant capacity when applied prior to the induction of the disease with encephalitogen + CFA, did not exhibit any protective effect. Thus, our study implicates that adjuvant and suppressive capacities of M. tuberculosis may be related to the different determinants of the microorganisms, TDM possessing the adjuvanticity only.
Decreased T-Colony Formation by Lymphocytes from Patients with Focal Glomerular Sclerosis
Authors: Matsumoto Koichi , Okano Kikuo , Yoshizawa Nobuyuki , Hatano Michinobu
Published: August 2009

The T-colony-forming capacity was examined in 13 normal subjects and 8 patients with biopsy-proven focal glomerular sclerosis (FGS). Fifteen patients with chronic mesangial proliferative glomerulone-phritis (CGN) without renal insufficiency were studied as a disease control. The two main assays used were the counting of T colonies formed by the patients’ lymphocytes, and the measurement of T-colony-forming activity in conditioned medium from cultures of patients’ lymphocytes. The levels of T-colony-forming cells (TCFC) and T-colony-stimulating factor (TCSF) were decreased in patients with FGS compared with those in normal controls and lower in FGS patients with the nephrotic syndrome (NS) than in those without NS. In contrast, these parameters in CGN patients with or without NS did not differ from normal subjects. TCSF activity for TCFC in both normal individuals and FGS patients was removed from media conditioned by phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes (PHA-LCM) with interleukin-2 (IL-2) receptor bearing cultured T cells. These in vitro findings suggest that IL-2 is the essential factor contained in PHA-LCM. Thus, depressed TCFC in FGS ptients with NS may result in part from impaired generation of TCSF by lymphocytes.
Induction of Allergen-Specific T Cells by Conjugates of N-Formyl-Methionyl-Leucyl-Phenylalanine and Rye Grass Pollen Extract
Authors: Cook R.M. , Wheeler A.W. , Spackman V.M. , Musgrove N.R.J. , Dave Y.K. , Moran D.M.
Published: August 2009

A conjugate of the biologically active peptide N-formyl-methionyl-leucyl-phenylalanine and rye grass pollen extract (F-MLP/rye), previously shown to react with rye grass pollen extract-specific T cells, induced the formation of allergen-specific T cells in mice. Lymph node cells prepared from mice immunized with either native extract or F-MLP/rye gave an enhanced response to unmodified rye grass pollen allergens in vitro. Syngeneic spleen macrophages were able to present the unmodified allergens to T cells obtained from both groups of mice causing their proliferation in vitro. Conjugation of the peptide into the extract brought about an extensive reduction in its reactivity with grass pollen-specific human IgE, and a loss of its ability to induce specific IgG antibody in guinea-pigs. A state of delayed hypersensitivity specific for rye grass pollen extract was produced in guinea-pigs by immunization with either the F-MLP/rye or unmodified extract. It is concluded that conjugates such as F-MLP/rye or other T′ allergoids could be used as probes to investigate whether changes in T cell activity are important in immunotherapy.
Medium Conditioned by Spleen Cells of Nematospiroides dubius-lnfected Mice Does Not Support Development of Cultured Mast Cells
Authors: Reed N.D. , Dehlawi M.S. , Wakelin D.
Published: August 2009

The ability of different conditioned media to support mast cell development from precursors in normal bone marrow was evaluated. Many mast cells developed in bone marrow cultures containing medium conditioned by concanavalin-A-stimulated spleen cells of normal mice, Trichinella spiralis-infectedmice, or mice infected for 6 days by Nematospiroides dubius. By contrast, medium conditioned by concanavalin-A-stimulated spleen cells of mice having 18-day infections of the nematode N. dubius failed to support mast cell development. The difference between medium conditioned by spleen cells of mice having 6-day versus 18-day infections of N. dubius may be due to the life cycle stage of the parasite, larval or adult, present at those times. These results from cultures are consistent with those from in vivo studies in which mice given primary infections of N. dubius failed to develop the intestinal mastocytosis characteristic of nematode infections.
Cloning and Expression of DNA Coding for the Major House Dust Mite Allergen Der p 1 in Escherichia coli
Authors: Thomas W.R. , Stewart G.A. , Simpson R.J. , Chua K.Y. , Plozza T.M. , Dilworth R.J. , Nisbet A. , Turner K.J.
Published: August 2009

A cDNA clone coding for the major house dust mite allergen Der p 1 was isolated from a lambda gt 11 library. Its sequence correlates with known amino acid sequences of Der p 1 and it produces a fusion protein which reacts with rabbit anti-Der p 1 antiserum.
Functional and Phenotypic Characterization of Human B Lymphocyte Subsets Isolated by Unit Gravity Sedimentation
Authors: Sherr Elliott H. , Saxon Andrew , Wells John R.
Published: August 2009

We report the use of unit gravity sedimentation with a CelSep® apparatus to generate two volumetrically similar but functionally and phenotypically distinct subsets of human peripheral blood B cells. One subset, comprised of small B lymphocytes, underwent a significant size change in response to anti-mu, proliferated synergistically to low concentrations of anti-mu plus B cell growth factor (BCGF) or phorbol myristate acetate plus BCGF, and could be induced to produce immunoglobulin in response to pokeweed-mitogen-derived T-lymphocyte-replacing factors. These cells were primarily slg+, B1+, B2 + , and were virtually free of monocytes ( < 0.01 %). Unlike these resting B lymphocytes, the large cells proliferated directly to BCGF, without displaying synergy with anti-mu. These cells displayed very little B2 ( < 7%), did not increase in volume in the presence of anti-mu, and made more immunoglobulin in response to TRF than the small resting B lymphocytes. However, neither population synthesized immunoglobulin spontaneously. This technique, which is highly reproducible, not equipment intensive, and produces high cell recovery ( > 90%), allows for a precise analysis of the steps involved in the maturation of a resting B lymphocyte to an immunoglobulin-secreting cell.
Basidiospore Extracts: Evidence for Common Antigenic/Allergenic Determinants
Authors: O’Neil Carol E. , Hughes Janet M. , Butcher Brian T. , Salvaggio John E. , Lehrer Samuel B.
Published: August 2009

Spore extracts, prepared from Armillariella tabescens, Pleurotus ostreatus, Coprinus quadrifidus, Amanita muscaria, Ganoderma lucidum, Psilocybe cubensis, Pisolithus tinctorius, Scleroderma sp. and Calvatia cyathiformis, were examined for antigenic/allergenic relationships by Ouchterlony and radioallergosorbent testing (RAST) inhibition, respectively. Ouchterlony, using hyperimmunized rabbit sera, demonstrated a high degree of cross-antigenicity among the extracts tested; however, some unique antigens were also present. RAST inhibition, evaluated by comparing extract concentrations which inhibited the RAST by 50% (IC-50), varied with the allergen tested. P. cubensis was the most potent inhibitor (IC-50 ranged from 0.034 mg/ml for A.tabescens RAST to 0.29 mg/ml for G. lucidum RAST). P. tinctorius was the least potent inhibitor, failing to reach IC-50 at 10 mg/ml for any basidiospore extract. Evaluation of slopes and intercepts of the dose-response lines demonstrated qualitative and quantitative differences among allergens in these extracts. These results indicate the presence of shared allergenic epitopes, and suggest that representative extract panels could be developed for future use in diagnosis and treatment of basidiospore-sensitive individuals.
Cancer-Induced Alterations in T-Cell Subsets in Normal and Cyclophosphamide-Treated Mice
Authors: Tonner Elizabeth , Sewell Herb F. , Thomson Angus W.
Published: August 2009

The influence of an ascites carcinoma on splenic T-cell subsets in normal or cyclophosphamide (Cy)-pretreated mice immunized with sheep erythrocytes was determined by immunophenotypic analysis, using an ‘EPICS C’ flow cytometer. As suggested by our previous studies on lymphokine production, the results obtained were consistent with direct impairment of Th-cell function and/or with tumour-induced suppression of cell-mediated immunity by Cy-insensitive suppressor cells, such as the ‘Ts effector’ (Ts3) or those of non-T-cell lineage.
Allergen Nomenclature
Authors: Marsh David G. , Goodfriend Lawrence , King Te Piao , Løwenstein Henning , Platts-Mills Thomas A.E.
Published: August 2009

This article presents a nomenclature system for allergens which has been officially recommended by the International Union of Immunological Societies (IUIS). The nomenclature is based on proposals of the IUIS Sub-Committee for Allergen Nomenclature and is applicable to highly purified, well-characterized allergens and to nonpurified or partially purified allergenic extracts.
Induction of Secondary IgE Antibody Response in Rats Immunized with X-Irradiated Metacercariae of Paragonimus ohirai
Authors: Ikeda Teruaki , Tani Shokichi
Published: August 2009

Rats reinfected with Paragonimus ohirai (P.o.) elicited little secondary IgE response to adult P.o. antigen. In contrast, rats immunized with X-irradiated (2–10 krad) metacercariae elicited not only a marked primary IgE response comparable to that in normally infected rats, but also a marked secondary IgE response after challenge infection. Rats immunized with 2 krad X-irradiated metacercariae yielded a higher secondary IgE response than with 5 or 10 krad irradiated ones, and elicited a secondary response lasting at least 1 year. An IgE response to antigen of newly excysted juvenile parasites (NEJ) was clearly different from the IgE response to adult P.o. antigen. The former IgE response was weak after infection or immunization, but after challenge infection a marked secondary response occurred at a level similar to the IgE response to adult P.o. antigen. The secondary IgE responses to adult P.o. and NEJ antigens were also elicited by reinoculation with X-irradiated metacercariae. Cross-absorption experiments confirmed that there were adult type and NEJ type allergens.
Inhibition by Nedocromil Sodium of IgE-Mediated Activation of Human Mononuclear Phagocytes and Platelets in Allergy
Authors: Thorel T. , Joseph M. , Tsicopoulos A. , Tonnel A.B. , Capron A.
Published: August 2009

The demonstration of a specific receptor for IgE on nonmast cell or basophil leukocytes, such as mononuclear phagocytes, eosinophils, and platelets, suggests that these cells may participate directly in immunological disorders of allergy. Thus, a full understanding of the mode of action of antiallergic or antiasthma drugs must take into account their activity on these nonmast cell leukocytes. Consequently, inhibition by nedocromil sodium of IgE-dependent activation of human alveolar macrophages, blood monocytes and platelets, was investigated. This compound induced an inhibition of the IgE-mediated generation of cytotoxic molecules from monocytes and platelets, together with a concomitant inhibition of their oxidative metabolism, measured by chemiluminescence, and a reduction of the potential ability of alveolar macrophages to synthesize and release mediators, estimated by lysosomal beta-glucuronidase activity. These observations confirm the hypothesis that nedocromil sodium acts on a cell compartment other than the classical mast cell population, in IgE-dependent allergy and, more particularly, in asthma.
Heteroclitic Antibodies: Differences in Fine Specificities between Monoclonal Antibodies Directed against Dinitrophenyl and Trinitrophenyl Haptens
Authors: Furusawa Shuichi , Ovary Zoltan
Published: August 2009

The inhibition of binding of monoclonal antibodies by different haptens was studied using the 50% antibody binding assay. The binding of antidinitrophenyl and antitrinitrophenyl antibodies to dinitrophenylated or trinitrophenylated bovine serum albumin could be inhibited by monovalent dinitrophenyl or trinitrophenyl ε-aminocaproic acid. Some of the antibodies could be inhibited to a greater degree with the cross-reacting haptens than with the haptens homologous to the immunizing antigen, therefore these antibodies were heteroclitic.