bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of Parietaria judaica Pollen Allergens
Authors: Ford S.A. , Baldo B.A. , Geraci D. , Bass D.
Published: August 2009

Parietaria judaica pollen allergens, fractionated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes, were identified using 52 sera collected in Australia and Sicily from P. judaica pollen-allergic patients. IgE-binding pollen components transferred to nitrocellulose were detected by reaction with 125I-anti-human IgE and autoradiography. Nine pollen components, ranging in molecular weight (MW) from approximately 10,000 to 80,000 daltons, bound IgE antibodies but the two fastest migrating components sometimes each separated into two very closely migrating bands. The faster of the two components exhibiting doublet formation (MW approximately 10,000 daltons) showed by far the highest frequency of IgE binding, being recognised by 50 of the 52 sera examined. Although patients’ IgE reaction patterns to P. judaica allergens were heterogeneous, the degree of heterogeneity was much less than that observed with house dust mite and other pollen extracts studied by electrophoretic transfer analysis. Results with gradient gel-nitrocellulose transfer experiments which showed no IgE-binding components with MWs less than 70,000 daltons, and comparisons of our electroblotting results with crossed radioimmunoelectrophoresis results of others, suggested that the doublet proteins with MWs of approximately 10,000 probably bind to higher MW proteins in P. judaica pollen extracts.
Autoantibodies of the IgM Class against a Human Myeloma Protein IgE(DES). II. Specificity
Authors: Magnusson C.G.M. , Vaerman J.P.
Published: August 2009

Five different monoclonal IgE proteins, papain and pepsin fragments of myeloma protein IgE(DES) and particle-counting immunoassay were used to study in detail the epitope(s) of IgE(DES) involved in the agglutination with IgM anti-IgE(DES) antibodies. The specificity of these autoantibodies was restricted to IgE(DES) as they did not react with latex particles coated with four other IgE myeloma proteins. Antibodies reactive with latex-IgE other than latex-IgE(DES), when present, were also of restricted specificity as shown by criss-cross absorption experiments. These differences between IgE myeloma proteins could not be attributed to the light chain type nor to the Em(l)-allotype. The epitope(s) of IgE(DES) participating in the reaction was heat-resistant (56 °C, 2 h) and localized in the pepsin F(ab′)2ε fragment, but was absent in the papain Fcε and Fabε fragments. Further degradation by pepsin of the F(ab′)2ε fragment (molecular weight 145,000 daltons) to 5Sε (90,000 daltons) and FcεXXX fragments (30,000 daltons) destroyed the reacting epitope. These results indicate that the IgM antibodies were not directed against k chain or idiotypic determinants of IgE(DES), but to some epitope(s) in the undegraded hinge region. Therefore, it seems that some kind of antigenic heterogeneity, perhaps related to the carbohydrate moieties, is present in IgE myeloma proteins besides the known idiotypic, allotypic and isotypic ones.
Induction of IgE Antibody Production to Aerosolized Ovalbumin in Mice Treated Intratracheally with Aluminum Silicate
Authors: Fujimaki Hidekazu , Ozawa Masashi , Ashikawa Tetsuo , Kubota Kentaro , Watanabe Naohiro
Published: August 2009

Induction of IgE antibody production was studied in mice with aerosolized antigen exposure and intratracheal instillation of adjuvants. BALB/c mice were immunized intratracheally with 10 μg ovalbumin (OA) adsorbed with or without aluminum silicate as an adjuvant. A primary IgE antibody production was observed in mice immunized with OA plus adjuvant, but not with OA alone. In mice immunized with OA plus adjuvant, a secondary IgE antibody production was observed after a booster of OA aerosol exposure on day 28. Mice were instilled intratracheally with aluminum silicate or aluminum hydroxide (alum) and subsequently exposed to aerosolized OA for 30 min. Primary and secondary anti-OA IgE antibody productions were obtained in the groups instilled with the adjuvants. After the intratracheal instillation of alum, aluminum remained in the mice for a long period. These results suggest that the intratracheal instillation of adjuvants is effective in the induction of IgE antibody production by aerosolized antigen exposure.
Tolerance to the Trinitrophenol Ligand in Guinea Pigs: Studies on the Role of the Solvent Used in Feeding
Authors: Pomeranz Jerome R.
Published: August 2009

Guinea pigs fed picryl sulfonic acid in aqueous solvents did not become unresponsive to contact sensitization with picryl chloride. In contrast, a substantial proportion of those fed picryl sulfonic acid, emulsified with triglyceride oil, were tolerant to subsequent immunization with picryl chloride. Feeding a related hapten (picryl chloride) in aqueous solvents was also considerably less effective in the induction of unresponsiveness to contact sensitization than feeding this hapten in vegetable oil. Animals fed picryl sulfonic acid in either aqueous or emulsified triglyceride solvents developed picryl-specific antibody after the initial feeding. However, those fed picryl chloride did not develop antibody and its role, if any, in the mechanism of unresponsiveness is unclear.
Plasma Protease Inhibitor and Anaphylatoxin Inactivator Levels in Chronic Urticaria/Angioedema and in Patients Experiencing Anaphylactoid Reactions to Radiographic Contrast Media
Authors: Mathews Kenneth P. , Pan Pauline M. , Amendola Marco A. , Lewis Fred.H.
Published: August 2009

Since a protease inhibitor or anaphylatoxin inactivator deficiency might explain why certain individuals are prone to develop chronic urticaria/angioedema or anaphylactoid reactions to radiographic contrast media, serum αj-protease inhibitor, α1-antichymotrypsin, α2-macroglobulin, inter-α-inhibitor, anti-thrombin III, α2-plasmin inhibitor, C1 inhibitor, and serum carboxypeptidase N were assessed by immunologic or functional methods. These values all were within normal limits in both groups of patients except for a low mean α1-protease inhibitor level in chronic idiopathic urticaria/angioedema and cold urticaria patients and marginal decreases of α1-protease and inter-α-inhibitor levels in radiographic contrast medium reactors. However, these abnormalities were not thought to be of pathogenetic significance.
The Antiallergic Agent Amoxanox Suppresses SRS-A Generation by Inhibiting Lipoxygenase
Authors: Saijo T. , Makino H. , Tamura S. , Kuriki H. , Ashida Y. , Terao S. , Maki Y.
Published: August 2009

Amoxanox has potent antiallergic activity because it inhibits the release of chemical mediators such as histamine and leukotrienes. We studied the in vitro effect of amoxanox on arachidonic acid metabolism, including the lipoxygenase and cyclooxygenase pathways. Amoxanox inhibited calcium ionophore A23187-induced formation of 5-HETE, LTB4, SRS-A (LTC4, LTD4 and LTE4), and 12-HETE in rat peritoneal resident monocytes. These results indicate that amoxanox inhibits 5- and 12-lipoxygenases. The compound, however, did not affect the formation of TXB2 or 6-keto-PGF1α in guinea pig lung fragments and PGE2 or PGF2α in bovine seminal vesicles, suggesting that it did not inhibit cyclooxygenase. These results show that the antiallergic action of amoxanox is associated, at least in part, with the reduction of leukotrienes due to the inhibition of lipoxygenases.
Allergic Bronchopulmonary Aspergillosis in Asthmatic Patients Presenting with Allergic Rhinitis
Authors: Grammer Leslie C. , Greenberger Paul A. , Patterson Roy
Published: August 2009

The early diagnosis of allergic bronchopulmonary aspergillosis (ABPA) is important because it may allow prevention of evolution to the fibrotic lung disease stage (stage V). In two immunotherapy studies of 62 patients recruited for allergic rhinitis, we conducted studies to exclude ABPA in any subject with asthma and positive immediate skin reactivity to Aspergillus fumigatus. The evaluation included serum precipitating antibodies against A. fumigatus, total serum IgE, blood eosinophilia, serum IgE and IgG antibody indices against A. fumigatus, and chest X-rays. Three of the 62 patients (5%) had features indicative of ABPA. This finding underscores the importance of excluding the diagnosis of ABPA in all asthmatics with skin reactivity to A. fumigatus, even if the patients major initial complaint is allergic rhinitis.
IgE Antibodies against Piperazine and N-Methyl-Piperazine in Two Asthmatic Subjects
Authors: Welinder Hans , Hagmar Lars , Gustavsson Cecilia
Published: August 2009

The presence of specific IgE antibodies against conjugates between human serum albumin and piperazine, a compound previously known to cause occupational asthma, and N-methyl-piperazine, respectively, was demonstrated by RAST, RAST inhibition and skin prick tests in 2 asthmatic subjects occupationally exposed to these agents.
A Partially Purified Glycoprotein Antigen from Aspergillus fumigatus
Authors: Kurup V.P. , John K.V. , Resnick A. , Fink J.N.
Published: August 2009

Culture filtrate antigens of Aspergillus fumigatus were fractionated by isoelectric focusing using a pH gradient of 4–6.5. Three fractions, namely, 18, 19 and 20 were pooled and subjected to immunochemical analysis. It contained a concanavalin A binding glycoprotein. Antibody raised against this component was used to prepare an affinity column of IgG-Sepharose and was used to purify crude culture filtrate antigens. This component produced three precipitin arcs in crossed immunoelectrophoresis using anti-A. fumigatus rabbit serum. This fraction has an isoelectric point of 6.5 and showed three components in two-dimensional electrophoresis with approximate molecular weights of 20, 40 and 80 kilo daltons. This antigen reacted with patient sera in the biotinavidin linked immunosorbent assay and showed high levels of anti-A. fumigatus IgG and IgE antibodies in allergic bronchopulmonary aspergillosis and IgG antibodies in aspergilloma. Both controls and Aspergillus skin test positive asthmatics showed only low levels of specific antibodies. Because of the purity of this antigen, its potential use as a standardized antigen in the detection of antibody is discussed.
Extensive Cross Reactivity between Treponema pallidum and Cultivable Treponemes Demonstrated by Sequential Immunoadsorption
Authors: Wos Susan M. , Wicher Konrad
Published: August 2009

Extensive cross reactivity between Treponema pallidum and three nonpathogenic species of treponemes was demonstrated by the Western blot technique. Rabbit antiserum produced by adjuvant immunization with solubilized T. pallidum antigens reacted with 34 T. pallidum antigens and with approximately 30 antigens each of T. phagedenis biotype Reiter, T. noguchii and T. vincentii. Adsorption of the antiserum with T. phagedenis Reiter removed only about half of the cross-reacting antibodies. Sequential adsorption with all three nonpathogenic treponemes removed antibodies to all but three polypeptides of 36,000, 34,000 and 27,000 daltons.
Induction of IgE-Secreting Cells in the Lymphatic Drainage of the Lungs of Rats following Passive Antigen Inhalation
Authors: Sedgwick J.D. , Holt P.G.
Published: August 2009

Repeated challenge of high-IgE-responder BN rats with an antigen aerosol in the absence of adjuvant, elicited both specific IgE and IgG responses in serum. Employing the recently developed ELISA plaque assay to detect individual IgE and IgG secreting cells, the lymph nodes draining the lower respiratory tract were pinpointed as the site for initiation of this response, and the sole location of specific IgE-secreting cells. Subsequent analysis of the tissue distribution of total IgE-secreting cells revealed that ≥50% were located in the lymph nodes draining the upper and lower respiratory tracts.
Detection of an IgM Antiglobulin in the Sera of Atopic Patients Using Insolubilised IgG4, and its Capacity to Release Histamine from Leucocytes
Authors: Shakib F. , Brown Morrow , Phelps A. , Redhead R.
Published: August 2009

ELISA plates coated with highly pure IgG4 were employed to detect IgM antiglobulin in atopic sera. The use of IgG4, rather than whole IgG, on the solid-phase was to provide direct evidence for the IgG4 reactivity of the antiglobulin. Bound IgM was shown to be antiglobulin in that binding can be inhibited by pre-absorption of serum with IgG. Some 75% of asthmatic patients and 29% of eczema patients were found to have significantly raised level of IgM antiglobulin. This antiglobulin resembles rheumatoid factor in that it appears to be directed against antigenic determinants common to human and rabbit IgG. Isolated antiglobulin-enriched IgM fractions released histamine from leucocytes of 7 out of 12 atopic patients. The histamine-releasing capacity of the IgM antiglobulin was shown to be operating via basophil-bound IgG in that the process can be blocked by pre-absorption of the antiglobulin with IgG. Furthermore, heating the antiglobulin-enriched IgM fraction did not affect its histamine-releasing capacity. We conclude that the IgM antiglobulin detectable in our atopic patients may contribute to the pathological changes in them.
Route-Dependent Immunomodulation: Local Stimulation by a Surfactant and Systemic Stimulation by a Polyanion
Authors: Hilgers L.A.T. , Snippe H. , Jansze M. , Willers J.M.N.
Published: August 2009

Immunomodulatory activity of the two synthetic adjuvants dimethyldioctadecylammonium bromide (DDA) and dextran sulfate (DXS) in relation to route and time of injection was investigated in mice. Humoral responses to sheep red blood cells (SRBC) were measured as the number of direct anti-SRBC plaque-forming cells (PFC) in the spleen 5 days after immunization. Both adjuvants stimulated the anti-SRBC response if adjuvant and antigen were injected simultaneously via the same route (either intraperitoneally or intravenously). Administration of adjuvant and antigen via different routes (intraperitoneally or intravenously, respectively or vice versa) resulted in enhanced humoral responses after DXS, but not after DDA. Intraperitoneal immunization of mice which were injected intraperitoneally with either adjuvant 4 days earlier resulted in diminished humoral responses. Immune responses in pretreated mice were not suppressed when the antigen was injected intravenously instead of intraperitoneally. In conclusion, DDA and DXS differ in iramuno-stimulating properties as DDA enhanced only a response to antigen injected via the same route whereas DXS induced a systemic state of increased immunoresponsiveness. The immunosuppressive state induced by intra-peritoneal injection of either adjuvant prior to immunization is restricted to the peritoneal compartment. Mechanisms underlying differences between both adjuvants and aspects of systemic immunopotentiation are discussed.
Cytotoxic Effect of Heterologous and Autologous Serum Factor on Guinea Pig Thymocytes Assayed by Electronic Cell Volume Distribution Analysis
Authors: Sandberg G. , Stenvinkel C.
Published: August 2009

The cytotoxic effect of serum on guinea pig thymocytes was studied by dye exclusion and by analysis of cell volume distribution curves obtained by means of a computer-assisted electronic cell volume distribution analyzer. Guinea pig, rabbit and human sera were shown to be cytotoxic for guinea pig thymocytes (incubation for 1 h at 37°C or overnight at 4°C). A similar, but less potent, activity was found in autologous guinea pig serum (4°C). The activity was dialysable and lyophilizable but was destroyed by heating at 56°C for 30 min. It was recovered after ammonium sulphate precipitation followed by dialysis. The effect of this dialysate was dose-dependent. A tentative definition of viable and dead cells was made on the basis of ‘electronic’ cell volume. The following results indicate that by this definition electronic cell volume distribution allowed detection of the cytotoxic serum factor: (1) sera and dialysates produced marked distortion of cell volume distribution curves, not seen in controls treated with heat-inactivated preparations; (2) the dose-response relationship was similar when dye exclusion and cell volume distribution was used as assay; (3) both methods detected similar differences in sensitivity of different thymocyte subpopulations to serum; (4) the electronic method could be used on separate thymocyte subpopulations although they differed in cell volume. This rapid, objective and reproducible method for studying the cytotoxic effect of sera may be useful during attempts to purify and identify the active factor(s).
Regulation of Ongoing IgE Synthesis by Human T-Cell Supernatants Derived from Atopic and Nonatopic Donors
Authors: Kanowith-Klein Susan , Saxon Andrew
Published: August 2009

Supernatants derived from T cells of donors with and without atopic disease were assessed to determine whether they could regulate ongoing IgE, IgG, or IgA synthesis of three different human B-cell lines. The results were compared with the ability of T-cell supernatants from the atopic donors to specifically induce IgE synthesis of nonatopic peripheral blood B lymphocytes (PBL-B cells). We found that, when tested on the B-cell lines, the T-cell supernatants from atopic donors doubled the IgE as well as the IgG and IgA synthesis. Although the T-cell supernatants from nonatopic donors had no effect on Ig synthesis of PBL-B cells or on the IgG of the IgG-secreting cell line, the supernatants doubled the IgE and moderately enhanced IgA synthesis of the IgE- and IgG-secreting cell lines. These results demonstrate that T-cell supernatants derived from atopic and nonatopic donors differ in their regulatory effects, not only on PBL-B cells, as had been shown previously by others, but also in their effects on ongoing Ig production.
IgG2 Antibodies Block IgE Antibody-Induced Asthma in Guinea Pigs
Authors: Yamauchi Nobukazu , Ito Koji , Suko Matsunobu , Ishii Akira , Miyamoto Terumasa
Published: August 2009

In order to examine the blocking activity of IgG2 antibodies to guinea pig for IgE antibodies-induced guinea pig asthma, experiments were carried out as follows. Guinea pigs were passively sensitized intravenously with guinea pig serum containing IgE antibodies to ovalbumin (OA). 8 days after sensitization, IgG2 purified from guinea pigs hyperimmunized with OA was intravenously injected. One hour later, the guinea pigs were challenged by inhalation of OA solution. Asthma attacks were not observed in the guinea pigs, whereas the attacks were observed in guinea pigs passively sensitized with the IgE antibodies but injected IgG2 fraction from normal guinea pigs 1 h before inhalation. These observations suggested that IgG antibodies that increased after immunotherapy might block asthma caused by inhalation of allergens in humans.
Allergens in Pollen from Mugwort (Artemisia vulgaris L.)
Authors: Nilsen B.M. , Paulsen Smestad
Published: August 2009

A crude and a partly purified extract of mugwort pollen were characterized with particular emphasis on the glycoprotein allergen Ag7. Crossed immunoaffinoelectrophoresis with immobilized Con A in the intermediate gel demonstrated binding of Ag7 to this lectin, indicating that Ag7 contains terminal α-D-mannopyranosyl and/or α-D-glucopyranosyl residues. Crossed immunoaffinoelectrophoresis with free Con A in the first-dimension gel demonstrated microheterogeneity in the carbohydrate moiety of the allergen. Crossed radio-immunoelectrophoresis analysis using sera from 26 mugwort-allergic patients indicated that Ag7 is an important allergen in mugwort pollen. Heat stability experiments demonstrated the presence of at least three heat-resistant allergens, one of these being Ag7. Crossed immunoelectrofocusing indicated a pi value for Ag7 of 4.3.
Enhanced IgE Antibody Production in Mice Injected with Fly Ash
Authors: Fujimaki Hidekazu , Hirano Seishiro , Takenaka Shinji , Murakami Masataka , Watanabe Naohiro
Published: August 2009

The adjuvant effect of fly ash injected via various routes on IgE antibody production was examined in mice. Primary anti-ovalbumin (OA) IgE antibody production was induced by intratracheal and intraperitoneal injections with fly ash and OA, but not by the intragastric route. Both the intratracheal and the intraperitoneal routes showed similar efficiency in inducing IgE antibody production. High secondary anti-OA IgE antibody response by exposure to aerosolized OA was also observed in mice primed with intratracheal instillation of fly ash plus OA. The activity to induce IgE antibody production by fly ash instillation was almost the same as that by aluminum silicate, studied previously. Primary and secondary anti-OA IgG antibody production was also stimulated by intratracheal instillation of fly ash. These results suggest that fly ash has an adjuvant activity in the induction of IgE and IgG antibody production in mice.
Binding of Compound 48/80 by Isolated Rat Mast Cells in Connection with Histamine Release
Authors: Grosman Nina
Published: August 2009

The binding of compound 48/80 in connection with histamine release from isolated rat mast cells was investigated by a two-step incubation procedure. After incubation of mast cells with known concentrations of compound 48/80, the supernatants were collected and subsequently exposed to fresh mast cells. The response in the second incubation step provided a measure of the amount of compound 48/80 remaining in the supernatants. At noncytotoxic concentrations of the releasing agent, binding capacities for compound 48/80 of up to 4 μg/l05 mast cells (4% w/v) were observed. The binding of compound 48/80 was of high affinity, giving mast cell concentrations of the compound exceeding that in the incubation medium by four orders of magnitude. The binding occurred rapidly with a substantial proportion bound within the first minute. These findings indicate that quantitation of exocytotic events by means of basic dyes may be compromised by competition for granular binding sites by basic releasing agents
Tumor Immunity in Rats Immunized with Rat Carcinoembryonic Antigen
Authors: Diakun Kate R. , Wilhelm Sally A. , Abeyounis John , Milgrom Felix
Published: August 2009

Active immunization of rats with an emulsion consisting of Freund’s complete adjuvant (FCA) and an extract of rat tumor containing carcinoembryonic antigen (CEA) induced clear-cut protection from growth of the syngeneic CEA-positive tumor, RCA-1. No protection was observed in rats treated with FCA alone nor was there protection against a tumor that had no serologically detectable CEA. The results suggested that the tumor immunity exhibited by the immunized rats was mediated by an immune response specific for rat CEA. It was shown further that multiparous rats were more resistant to growth of RCA-1 tumor than nulliparous rats. This suggested that immunization against rat CEA, which is an oncofetal antigen, may occur during pregnancy.
Labile Disulfide Bonds and Free Thiol Groups in Human IgG II.
Authors: Schauenstein E. , Reiter M. , Gombotz H. , List W.
Published: August 2009

As recently shown, the measurement of labile disulfide bonds and free thiol groups of human IgG using 5,5′-dithio(2,2′-dinitro)benzoic acid (DTNB) provides direct information about the relative concentrations of IgG1 and IgG2, respectively. Furthermore, using this technique data have been obtained that show a decrease in labile S-S (corresponding to IgG1) and increase of free SH (corresponding to IgG2) in IgG from patients afflicted with various malignant diseases. In the present study a total of 280 sera from patients with malignancies of various tissue origin was investigated. Significantly reduced ΣS values, i.e. the sum of free SH and labile S-S, were obtained with IgG from cases of carcinoma of the mammary gland, urinary bladder, skin and prostate, as compared to cases with benign diseases of the respective organs. Most pronounced differences were observed between benign and malignant disorders of the prostate. It was confirmed with this group of patients that the reduction of ΣS was due to the decrease of S-S bonds, whereas the number of free SH groups remained unchanged or was increased. The cases of mammary gland carcinoma were grouped according to clinical stage. A clear correlation between reduction of ΣS and tumor size was observed. The possible theoretical and clinical implications of these results are discussed.
Are Cross-Reacting Natural Antibodies Multispecific?
Authors: Milgrom Felix , Swierczynska Zdzislawa
Published: August 2009

Human natural antibodies to antigens of Escherichia coli and Serratia marcescens were studied for cross-reactivity. The organisms were grown on synthetic media and extracted at lOO°C. The extracts were precipitated three times at 71% ethanol concentration and redissolved at the desired concentration. These preparations were referred to as Escherichia antigen (Ea) and Serratia antigen (Sa). They readily coated red blood cells (RBC), which then could be used for passive agglutination tests. Human serum selected for this study had strong agglutinins combining with both antigens (cross-reacting antibody) and rather weak agglutinins combining with Ea only or Sa only, a property that became obvious from absorption experiments. Absorption of the serum with RBC coated by Ea removed all activity for Ea and most of the activity for Sa, and the opposite effect was noted after absorption of the serum by RBC coated by Sa. The activity against both antigens could be recovered by elution of antibodies at 56 °C from RBC coated by either Ea or Sa. Significantly, inhibition of the serum or the eluate with soluble antigens gave strictly specific results in that Ea abolished only the reaction with RBC coated by Ea and did not influence the reaction of RBC coated by Sa, and the opposite result was obtained upon inhibition of the serum with Sa. These results strongly indicated to us that the cross-reacting antibodies under study were multispecific, i.e., had different antigen-reactive sites (haptophore groups) for Ea and Sa. Further evidence supporting this contention was obtained from a study in which the cross-reacting antibody was ‘labelled’ by a bacterial antigen. To this end, the tested serum was neutralized with Sa and then reacted with RBC coated by Ea. From these RBC, antibodies were eluted from which Sa was recovered. A ‘mirror image’ experiment was also conducted in which Ea was recovered from antibodies that were blocked by Ea and reacted with RBC coated by Sa.
Superoxide Anion Generation during Airway Anaphylaxis
Authors: Weiss Earle B. , Bellino Joseph R.
Published: August 2009

Extracellular release of superoxide anion (O2) during in vitro anaphylaxis in guinea pig trachealis was found to exhibit both a time- and a concentration-dependent generation of O2 Trachealis O2 release was unaffected by treatment with diphenhydramine HC1 (10––5M, 10––6M). While indomethacin (10––5M) augmented anaphylaxis tension it did not enhance O2 release. However, the semiselective SRS-A antagonist FPL 55712 (10––5M) resulted in essentially complete inhibition of O2 generation. These data indicate that O2 generation occurring during acute airway anaphylaxis is associated with the activation of SRS-A products.
Identification of Binding Site for Heparin and Other Polysulfated Glycosaminoglycans on Human Thrombocytes
Authors: Wolf H. , Glassl H. , Nowack H. , Wick G.
Published: August 2009

Heparin and other polysulf ated glycosaminoglycans (GAGPS) can evoke an immune response. when complexed to the surface of platelets in vivo. We have previously reported on the detection and characteristics of such antibodies leading to thrombocytopenia. Here we report data concerning the biochemical characterization of the heparin-/GAGPS-binding site on human thrombocytes. By means of heparin affinity chromatography, GAGPS chromatography and subsequent ‘Western blotting’ of specifically eluted proteins we were able to detect two proteins of apparent molecular mass of 207 and 170 kilodaltons, which could be reduced to 57 and 142 kilodaltons, respectively. The 170-kilodalton glycoprotein was stained by the carbohydrate specific PAS-reagent and by surface labeling with 3H. The second molecule binding to heparin (GAGPS)-Sepharose was detected by platelet surface labeling with 125I, protein-specific staining Coomassie brilliant blue and PAS staining. Binding studies using enzyme-labeling techniques with five monoclonal antibodies (MCA) against platelet surface proteins GPI, GPIIb/IIIa complex, GPIIIa and CRI (C3b receptor) revealed specific interaction of the 170-kilodalton heparin-binding glycoprotein with an MCA to GPI, suggesting identity or immunological cross-reactivity of these two moieties. No specific reactions were observed between the 207-kilodalton glycoprotein and any of the MCA tested. For further evaluation of the antigenicity of separated platelet proteins, an ELISA in microtiter plates was developed, and a cellular sandwich ELISA for serological detection of antibodies against whole native thrombocytes or GAGPS-derivatized proteins was set up. The identification of two thrombocytes surface glycoproteins as major binding sites for heparin (GAGPS) may provide the means for the elucidation of the molecular basis of the development of the immunologic mechanisms leading to thrombocytopenia in patients treated with these substances. Furthermore, a simple and sensitive ELISA can now be applied for the diagnosis of this severe complication of heparin (GAGPS) therapy.
Function of Phagocytes in Experimentally Induced Colitis in Rabbits I.
Authors: Hermanowicz A. , Hermel A. , Śliwiński Z. , Nawarska Z. , Borys D. , Banaś B.
Published: August 2009

Adhesion, random and chemotactic migration, phagocytosis and nitroblue tetrazolium (NBT) dye reduction of peritoneal exudate polymorphonuclear leukocytes (PMN) and macrophages were examined in 110 rabbits with immune complex-mediated colitis and in controls. We may conclude that Arthus-like colitis in rabbits is accompanied by an impairment of adhesion and phagocytosis, and an increase in chemotactic and random migration of both PMN and macrophages. The examined phagocytes were obtained from the peritoneal cavity, and the changes in their function are related much more directly to the presence of circulating immune complexes than to the colitis. The mentioned impairment in their function may be responsible for prolonged persistence of immune complexes in the serum of rabbits with colitis; however, it does not prolong the time of healing of the ulcers.
Function of Phagocytes in Experimentally Induced Colitis in Rabbits II.
Authors: Hermanowicz A. , Hermel A. , Nowak A. , Śliwiński Z. , Nawarska Z. , Borys D. , Banaś B.
Published: August 2009

Adhesion, phagocytosis, chemotactic and random migration, nitroblue tetrazolium dye reduction of peritoneal exudate neutrophils and macrophages, fibrinogen level, gelation of soluble fibrin and serial dilution protamine sulfate test were investigated in 115 New Zealand white rabbits with experimentally induced Shwartzman phenomenon in the colon, and in control animals. The results presented in this report demonstrated impairment of chemotactic migration of phagocytes in the presence of endotoxin. The depression was dose-dependent and less marked when neutrophils were stimulated with monocyte-derived chemotactic factor or with casein, than with complement-derived chemotactic factor. The prolonged depression of chemotactic migration of neutrophils and macrophages in rabbits with colitis, however, did not affect the healing time of the ulcers in the colon.
Enhancement of the Bronchial Reactivity in Immunized Rats by Neonatal Treatment with Capsaicin
Authors: Ahlstedt S. , Alving K. , Hesselmar B. , Olaisson E.
Published: August 2009

Rats were injected with capsaicin at 1–2 days of age to abolish the content of substance P (SP) in nerve terminals. At 6 weeks of age the capsaicin-treated and control rats were sensitized daily for 1 or 2 subsequent weeks Monday through Friday with ovalbumin (OA). The OA was given without adjuvant as 300 ng subcutaneous (s.c.) injections in the neck region or as 1% aerosol for 30 min. The capsaicin-treated animals which were sensitized s.c. for 2 weeks reacted moderately with increased transpulmonary pressure (TPP) to airway challenge with OA, and strongly to intravenous (i.v.) challenge with OA or serotonin. The capsaicin-untreated animals, which were sensitized with OA, reacted weakly to the challenge. In the animals sensitized with aerosolized OA, slightly lower reactivity was seen compared with those sensitized s.c. Untreated and unsensitized control rats reacted only to serotonin challenge. No animal had any detectable serum or bronchial IgE antibodies. Aerosol-sensitized animals had IgG antibodies in both serum and bronchial lavage. Histologically, the animals treated with capsaicin in contrast to the untreated controls demonstrated a pronounced increase of lymphoid tissue around their bronchi. Their mast cell numbers were increased around vessels and in the pleura and their mucous cell numbers were increased in the epithelium of the bronchi and bronchioli. The sensitization did not add much to this histological picture.
IgG Anti-IgA1 and Anti-IgA2 Antibodies: Their Measurement by an Enzyme-Linked Immunosorbent Assay and Their Relationship to Disease
Authors: Petty R.E. , Sherry D.D. , Johannson J.M.
Published: August 2009

IgG anti-IgA antibodies were measured by an enzyme-linked immunosorbent assay using one IgA1 and one IgA2 (ml) myeloma and a pooled IgA protein preparation as antigens. Class-specific anti-IgA antibodies occurred in 0.8% of non-IgA-deficient sera and 24.3% of IgA-deficient sera. Antibodies reacting with IgA1 only occurred in 2.6% of non-IgA-deficient sera and 6.7% of IgA-deficient sera. Antibodies reacting with IgA2 only occurred in 0.6% of non-IgA-deficient sera and 2.7% of IgA-deficient sera. The prevalence of anti-IgA in IgA deficients with inflammatory disease was higher (81.8%) than in IgA deficients without disease (24.1 %) and was accounted for by class-specific antibodies.
Suppression of Murine IgE Responses with Amino Acid Polymer/Allergen Conjugates
Authors: Cook R.M. , Henderson D.C. , Wheeler A.W. , Moran D.M.
Published: August 2009

Conjugates of poly-N-methylglycine (polysarcosine) and grass pollen allergen extracts, which have been previously shown to suppress murine IgE responses, were examined for their ability to modify lymphocyte activity in vitro. Allergen-specific T lymphocytes obtained from Balb/c mice gave a reduced response to syngeneic accessory cells pulsed with conjugates of polysarcosine-allergen compared with the response found using equivalent concentrations of native extract. Pretreatment of accessory cells with either polysarcosine or polysarcosine-allergen conjugates did not impair their subsequent ability to present grass pollen extract to immune T cells. Incubation of allergen-specific spleen cells with polysarcosine-allergen conjugates, but not with polysarcosine or allergen alone, resulted in specific cell-mediated suppression which significantly reduced proliferation in vitro. This activity was sensitive to treatment of cells with anti-T-lymphocyte antisera plus complement. Spleen cells obtained from animals immunised with allergen and taken 21 days after intravenous treatment with polysarcosine-allergen conjugates, a regimen that suppressed IgE antibody production, did not proliferate in the presence of grass pollen extract and failed to suppress a secondary lymphoproliferative response in vitro. Spleen cells obtained from similarly treated animals 3 days after the final polysarcosine-allergen injection responded to pollen extract in culture and, additionally, impaired a secondary response. The results suggest that the reduced IgE response found in animals treated with polysarcosine-allergen conjugates may be due, in part, to the generation of a short-lived antigen-specific T cell suppression.
Study on the E-Rosette-Blocking Effects of E Receptors
Authors: Gu S.-Q. , Wasserman J.
Published: August 2009

Lymphocytes were purified from 10 samples of normal peripheral blood. E receptors were detached by heating to +45°C. The sheep red blood cells (SRBC) and 2-aminoethylisothiouronium bromide (AET)-treated SRBC (AET-SRBC) were incubated at +4°C for 2 h, with E receptors. E receptors detached by heating blocked E-rosette formation with SRBC, but not with AET-SRBC. The results suggest that E receptors may exhibit different conformational structures or different molecules which are responsible for E-rosette formation and E-AET-rosette formation, respectively.
Enhancement of Antibody Production by Mercury and Platinum Group Metal Halide Salts
Authors: Murdoch R.D. , Pepys J.
Published: August 2009

The effects of halide salts of mercury, platinum and palladium on the synthesis of total and specific serum IgE and of total IgG were studied in groups of Hooded Lister rats immunised with antigen (ovalbumin) in the presence and absence of an adjuvant (Bordetella pertussis vaccine or aluminium hydroxide). Repeated intraperitoneal injections of mercuric chloride alone rapidly enhanced total IgE levels in control rats, independent of adjuvant. Injections of the platinum salt, however, elevated total IgE levels more slowly and then only in the B. pertussis-treated group. The halide salt of palladium was ineffective. In rats immunised with antigen and adjuvant, mercury treatment rapidly produced enhancement of the titre-specific IgE antibodies, whereas treatment with platinum again raised these levels more slowly. The palladium salt had no such effect.
Chemotactic Reactivity of Eosinophils Obtained from Bone Marrow and Peritoneal Cavity of Cyclophosphamide-Treated, Toxocara canis-Infected Mice
Authors: Nawa Y. , Owhashi M. , Imai J. , Abe T.
Published: August 2009

Chemotactic reactivity of eosinophils obtained from the bone marrow (BM-Eo) of cyclophosphamide-treated, Toxocara canis-infected mice was compared to that of eosinophils obtained from the peritoneal cavity (PEC-Eo). BM-Eo responded well to alanyl-tetrapeptide, a synthetic eosinophil chemotactic factor of anaphylaxis (ECF-A), whereas PEC-Eo did not. Both BM-Eo and PEC-Eo showed almost equally high chemotactic reactivity to parasite-derived ECF, ECF lymphokine and complement-derived ECF. Chemotactic reactivity of BM-Eo to synthetic ECF-A was deactivated by preincubation with ECF-A. Unresponsiveness of PEC-Eo to synthetic ECF-A could be explained by chemotactic deactivation by ECF-A, because an ECF-A-like substance was detected in the ascitic fluid; this substance could deactivate the chemotactic reactivity of BM-Eo to synthetic ECF-A. From these results, BM-Eo are naive and seem to be a good indicator for eosinophilotaxis and its modulation.
Induction of in vitro Antigen-Specific Antibody Production against NIP-KLH in Nonadherent Cells Derived from Human Peripheral Blood Mononuclear Cells
Authors: Komatsu Hirotsugu , Shimada Shigetoshi , Terashima Masazumi , Osawa Toshiaki
Published: August 2009

A primary in vitro antibody response to 4-hydroxy-3-iodo-5-nitrophenylacetyl keyhole limpet hemocyanin (NIP-KLH) by plastic dish nonadherent human peripheral blood mononuclear cells (PBMC) was examined. Nonadherent cells of most donors produced an anti-NIP antibody when they were cultured with the antigen at a cell density of 2.5 × 106/ml in the presence of Staphylococcus aureus Cowan I (SAC) at a concentration of 0.003% for 5 days, washed and further cultured in the absence of the antigen and SAC for an additional 5 or 6 days. Nonadherent cells of ‘low-responders’ responded to NIP-KLH plus SAC and produced anti-NIP antibody when they were cultured with not only NIP-KLH plus SAC but also with the culture supernatant obtained on phytohemagglutinin (PHA) stimulation of a mixture of PBMC from 2 donors. Furthermore, it was found that the PHA supernatants could be very effectively substituted for by human recombinant interleukin-2.
Genetic Control of Delayed-Type Hypersensitivity in Mice to Salmonella Antigen
Authors: Cho Norio , Saito-Taki Tatsuo , Nakano Masayasu
Published: August 2009

Genetic restriction on the expression of delayed-type hypersensitivity (DTH) to Salmonella typhimurium in mice transferred passively with immune spleen cells was studied. After the intravenous transfer of immune C3H/HeJ (H-2Ik) cells into A.TL (H-2Ik) or A.TH (H-2P) mice, footpad DTH responses could be evoked in the A.TL recipients, but not in the A.TH mice. When the immune cells of BALB/c or C3H/He mice were intravenously-transferred into F1 hybrids produced by mating BALB/c and C57BL/6 or C3H/He and C57BL/6, respectively, no DTH response could be evoked in these F1 hybrids that received immune parental cells. Local transfer as well as systemic intravenous transfer of immune parental cells to F1 haplotype recipients did not cause any DTH. Previous treatment of the F1 hybrid recipients with cyclophosphamide did not result in the expression of the DTH response. Transfer of immune F1 spleen cells into parental strains also did not induce DTH. When the immune cells of parental strains were transferred into F2 mice and into back-cross mice, examination of the DTH response in these mice showed that some of them did not have any obvious footpad swelling, while others revealed various magnitudes of swelling. The resistance of F1 hybrids to transfer of DTH is discussed.
Identification of HLA-Linked Antigens by Pregnancy-Associated Non-Cytotoxic Alloantisera
Authors: MacLeod A.M. , Mather A.J. , Mason R.J. , Stewart K.N. , Power D.A. , Innes A. , Catto G.R.D.
Published: August 2009

Non-cytotoxic sera obtained from post-partum primiparous and multiparous women were examined by a rosette inhibition technique for the presence of antibodies mediating blockade of human B lymphocyte Fc receptors. Selective activity was demonstrated against a panel of normal human B lymphocytes and lymphocytes from patients with chronic lymphocytic leukaemia (CLL). A pattern of specific activity was found in sera and in their IgG fractions, which was not accounted for by antibodies directed to known HLA-A, -B or -DR antigens. Several sera were identified with selective activity in this assay. As the results of testing sera in a direct binding assay correlated with those of the EA inhibition assay, and since EA inhibitory activity occurred in F(ab′)2 fractions of sera, it is possible that these non-cytotoxic antibodies bind directly to B cell surface antigens. Sera may therefore have been identified which possess antibodies to hitherto undefined HLA antigens.
Development of a Competitive Radioimmunoassay for Human Plasma Fibronectin
Authors: Ouaissi M.A. , Neyrinck J.L. , Capron A.
Published: August 2009

A competitive radioimmunoassay (CRIA) for quantitating human plasma fibronectin levels has been developed and compared with a conventional immunoturbidimetry assay (IMTA). The assay ranges for CRIA and IMTA were 0.05–5 μg/ml and 100–1,000 μg/ml, respectively, and the lowest detectable amounts of fibronectin that differed significantly from zero were 17.6 ng and 50 μg. The correlation coefficient between CRIA and IMTA was r = 0.953 (p <0.001). We consider the CRIA as potentially useful in the identification and study of fibronectin in certain biological fluids where it may be present in low concentrations.
Purification of Serum Proteins with Inhibitory Activity on the Histamine Release in vitro and/or in vivo
Authors: Theobald K. , Gross-Weege W. , Keymling J. , König W.
Published: August 2009

Recently, we described a factor in rat and human serum with inhibitory activity on the 2-hour passive cutaneous anaphylactic (PCA) reaction in rat skin. The Ca-ionophore-induced histamine release from rat peritoneal mast cells was inhibited as well. In human serum, inhibitory activity for the in vitro histamine release from rat mast cells was shown to be present within two serum fractions. One molecule was purified by 50–70% ammonium sulfate precipitation and subsequent DEAE chromatography at pH 5.4. The main component showed complete identity with transferrin. Commercially available transferrin was active in vitro as well. The dose-response plot revealed two inhibition maxima at transferrin concentrations of 0.05–5 ng/ml and at more than 1 mg/ml, respectively. Subsequently it was shown that the degree of iron saturation was critical for the decrease in histamine release. The mediator release in vivo (PCA) could not be inhibited by the iron transport protein. Inhibitory activity in vivo was however mediated by the second serum fraction, which was characterized by a molecular weight of approximately 150,000 daltons and an IEP within the alkaline range. These data suggest that exogenous factors may be potent modulators of inflammatory reactions.
Interleukin-1 Induces Mucus Secretion from Mouse Intestinal Explants
Authors: Han V. , Resau J. , Prendergast R. , Scott A. , Levy D.A.
Published: August 2009

Secretion of goblet cell (GC) mucus occurs during immune reactions in the gut. As human macrophages produce a substance that induces mucus secretion from lung explants, we tested the effect of macrophage-derived factor(s) on mucus secretion from intestinal explants. Fragments of mouse duodenum were incubated with macrophage culture supernatants and purified interleukin-1 (IL-1) preparations, and the amount of mucus released was estimated by an enzyme-linked lectin assay. Both the culture supernatants and the IL-1 preparations induced dose- and time-dependent mucus release. Lipopolysaccharide-induced culture fluids were shown to contain IL-1. Thus, stimulation of mucus secretion from GC can be added to the list of biological activities attributable to IL-1.
Percutaneous Sensitisation to House Dust Mite May Occur Naturally in Eczema
Authors: Carswell F. , Thompson S.J.
Published: August 2009

The concentrations of serum IgE with specificity for the faecal and body allergens of the house dust mite, Dermatophagoides pteronyssinus, were compared in 69 children with various combinations of asthma, eczema and/or rhinitis. The concentration of IgE antibodies to the mite body was higher in those with eczema than those with asthma but the concentrations of anti-faecal antibodies were similar. Our results suggest that anti-mite body antibodies are characteristic of eczema and that sensitization to the body allergen may occur because of its passage through the skin.
The Role of Leukotriene-inducing and -Metabolizing Enzymes in Inflammation
Authors: König W. , Bremm K.D. , Brom H.J. , Köller M. , Knöller J. , Raulf M. , Schönfeld W. , Stüning M.
Published: August 2009

Leukotrienes are potent mediators of allergy and inflammation. Polymorphonuclear granulocytes which participate in acute and chronic disease processes can be activated by immunological and nonimmunological stimuli to generate leukotrienes. It appears that on activation of the cells 5-lipoxygenase is released into the supernatant and thus may exert a potent role with regard to interdependent cellular interactions. The release of leukotriene-metabolizing enzymes (e.g., γ-glutamyl-transpeptidase, dipeptidase) is stimulus-dependent. Polymorphonuclear granulocytes alter their release profile once the cells have been prestimulated. Thus, a precise knowledge on the leukotriene-inducing and -metabolizing enzymes is of utmost importance for future immunopharmacological interventions. It appears that the enzyme activity reflects the state of cellular activation.
Effect of a New Selective H1 Receptor Antagonist (Levocabastine) in a Nasal and Conjunctival Provocation Test
Authors: Pécoud A. , Zuber P. , Kolly M.
Published: August 2009

Levocabastine is a new selective H1 receptor antagonist. The effect of the drug administered locally was compared to placebo in a quantified nasal and conjunctival provocation test with allergens performed in patients allergic to grass pollen. In the nasal provocation test, levocabastine was able to increase the ‘reaction threshold’ (dose of allergen necessary to trigger allergic symptoms) in 9 out of 12 patients; the drug inhibited rhinorrhea and sneezing, but not nasal obstruction. In the conjunctival provocation test, the ‘reaction threshold’ clearly increased in 10 out of 11 patients. The local administration of levocabastine might be useful in allergic rhinitis and conjunctivitis.