bluuebunny/crossref_metadata_2025 huggingface dataset:
Histamine and Sulfidoleukotriene Release from Human Basophils: Different Effects of Antigen, Anti-IgE, C5a, f-Met-Leu-Phe and the Novel Neutrophil-Activating Peptide NAF
Authors: Dahinden Clemens A. , Kurimoto Joshiyuki , Baggiolini Marco , Dewald Beatrice , Walz Alfred
Published: August 2009

Human blood leukocytes were challenged by IgE-dependent stimuli (antigens or anti-IgE antibodies) and the chemotactic agonists C5a, N-formyl-Met-Leu-Phe (fMLP) or NAF, a novel neutrophil-activating peptide produced by stimulated human monocytes. IgE-dependent stimuli induced abundant release of histamine and sulfidoleukotrienes (LTC, LTD and LTE). The effects of the chemotactic peptides differed considerably: C5a induced high percentages of histamine release already at concentrations as low as 10––9M, but no leukotriene formation; NAF was completely inactive up to 10––6M, and fMLP induced moderate release of both products at relatively high concentrations only (1–2.5 × 10––6M). All three chemotactic peptides, however, induced a concentration-dependent release of elastase from cytochalasin-B-treated human neutrophils. The EC50 was approximately 0.3, 2 and 5 × 10––9M for C5a, NAF and fMLP, respectively. NAF showed activity over a wider concentration range and its dose-response curve was less steep than that of the two other agonists. Our results suggest that NAF, that may be generated at the sites of hypersensitivity reactions, is likely to induce the immigration of neutrophils, but not the direct activation of basophils and mast cells. In this respect, NAF differs from less selective chemotactic peptides like the complement product C5a or the N-formyl-methionyl peptides.
Interleukin-2 Receptor Expression in Human Mast Cells and Basophils
Authors: Maggiano Nicola , Colotta Francesco , Castellino Flora , Ricci Riccardo , Valitutti Salvatore , Larocca Luigi M. , Musiani Piero
Published: August 2009

Surgical human thymus, upper respiratory tract, lung and small and large bowel specimens were analyzed for the presence of interleukin 2 receptor (IL2-R)-positive cells. Histochemical (toluidine) and immunologic (anti-IL2-R monoclonal antibody) staining procedures revealed a distinct anti-IL2-R positivity in most of metachromatically staining cells. These positive cells were observed not only in tissues showing strong inflammatory reaction and mast cell hyperplasia, as in Crohn’s disease, but also in those not histologically affected by pathologic conditions. This finding suggested that human mast cells, like T cell blasts, express the p55 chain of IL2-R on their surface. To see whether IL2-R was being actively synthesized, a cell preparation rich in peripheral blood basophils (PBB), which are cells closely related to mast cells, was obtained. Ultrastructural analysis of PBB after indirect immunogold procedure revealed that the vast majority expressed the IL2-R. Moreover, the presence of intracellular reaction products in the cytoplasm of most membrane-positive PBB was indicative of active antigen synthesis. Furthermore, Northern blot analysis evidenced specific IL2-R mRNA in PBB, while its expression was augmented several times when PBB were cultured in the presence of stimulated T cell supernatant.
Allergen Entrapped in Liposomes Reduce Allergenicity and Induce Immunogenicity on Repeated Injections in Mice
Authors: Arora Naveen , Gangal S.V.
Published: August 2009

Liposomes which are nontoxic, biodegradable and biocompatible lipid vesicles are known to act as adjuvants and can be used to formulate sustained release preparation by encapsulation. In the present study, allergen entrapped in liposomes were injected at different time intervals in Swiss mice (made responders to IgE by injecting cyclophosphamide) and Balb/C mice (high IgE responders). Tissue distribution studies after intra-peritoneal injection of allergen (entrapped and untrapped) revealed that liposome-entrapped allergen was retained for a longer time in all the tissues except kidney as compared to the free allergen given similarly. It was observed that serum specific IgG antibody levels were higher and specific IgE levels were lower in animals given repeated injections of liposome-entrapped allergen as compared to those animals injected free allergen in the same manner. The entrapment of allergen in liposomes somehow suppressed the specific IgE response on repeated injections. The immunomodulatory effect of liposomes may be useful in the immunotherapy of Respiratory allergic disorders.
Correlation between Atopy and Gm Allotypes
Authors: Oxelius Vivi-Anne
Published: August 2009

In 50 consecutive atopic Caucasian patients with increased IgE > 600 kU/l, the phenotypic Gm allotype constellation deviated from that to be expected, with significantly increased frequency of patients with the phenotype Gm(fnb). There was an increased frequency of the G2m(n) allotype, more frequent in patients with IgE > 1,000 kU/l, and in patients with IgG4 > 1 g/l. In patients with IgE > 1,000 kU/l, the phenotype Gm(afnb) was significantly increased and in patients with IgG4 > 1 g/l the phenotype Gm(fnb) was significantly increased. Those atopic patients with increased IgE and increased IgG4, according to earlier studies known to have the most severe forms of the disease, were thus mainly found to have the m(fnb) phenotype.
Immunogenicity Testing of Food Proteins: in vitro and in vivo Trials in Rats
Authors: Steinmann Jörg , Wottge Hans-Ulrich , Müller-Ruchholtz Wolfgang
Published: August 2009

The immunogenicity of an industrially produced bacterial food protein (single cell protein, SCP) was analyzed in a comparative study. SCP, casein and ovalbumin were injected into or fed to rats. The systemic response was tested in vitro with a lymphocyte transformation test and in vivo with a footpad swelling assay. Feeding of casein induced cell-mediated reactivity against casein, while feeding of SCP or ovalbumin had no effect. One injection of casein led to sensitization compared with three injections of SCP. Prior feeding of SCP did not abrogate the response to injected SCP. The footpad swelling test is suggested as a particularly effective model for helping to determine the immunological risk of foodstuffs to man.
Endothelial Cells Modulate Both T-Cell-Dependent and T-Cell-Independent Plaque-Forming Cell Generation in vitro
Authors: Teitel J.M. , Shore A. , McBarron J.M. , Leary P.L. , Schiavone A.
Published: August 2009

The effects of live endothelial cells (EC), paraformaldehyde fixed EC, and EC supernatant were measured on pokeweed mitogen (PWM)-induced T-cell-dependent plaque-forming cell (PFC) generation by peripheral blood mononuclear cell (PBM). At low doses (≤ 2 × 104 cells/culture) live EC helped PFC generation. At higher doses (≥ 10 × 104 cells/culture) the effect of live EC was always marked suppression (< 10% of baseline PFC). In contrast both fixed EC and EC supernatant provided help exclusively over a wide dose range. The EC-helper effect enhanced the sensitivity of PBM to suboptimal PWM doses and also accelerated the rate of PFC generation during culture. EC influences on PFC could not be modified by gamma-interferon induction of surface DR which is known to modify EC accessory cell ability. There was also only minimal helper activity of live EC and fixed EC on the PFC generation by Epstein-Barr virus-induced cultures of purified B cells (which had been depleted of both T cells and monocytes). In contrast, suppression (> 97%) of PFC in isolated B-cell cultures was found even when EC constituted < 1 % of cultured cells. These results imply EC have the potential of providing multiple regulatory signals which modulate in vitro antibody production. EC-derived mechanisms are independent of their accessory cell function and require interaction with non-B cells for help, but suppression may occur directly at the B-cell level. EC are thus not only capable of providing an accessory cell signal which can initiate immune responses, they also may profoundly influence on-going immune processes initiated by other immunocytes.
Biological and Immunological Properties of Sugi Basic Protein-Pullulan Conjugate
Authors: Usui Mitsuko , Taniguchi Yoshifumi , Ando Shunsaku , Kurimoto Masashi , Matuhasi Tyoku
Published: August 2009

Ability of Sugi basic protein (SBP)-pullulan conjugate to elicit the Arthus reaction was found to be markedly reduced, about 100 times lower than that of native SBP. To analyze this reduced ability, activation of complement by immune complex consisting of SBP-pullulan and anti-SBP antibodies was studied. Tests for complement consumption, C3 conversion and cleavage of factor B revealed that immune complex formed with SBP-pullulan is incapable of supporting efficient activation of the complement system. Previously, we have shown data suggesting that SBP-pullulan conjugate would be a good candidate for desensitization therapy against cedar pollinosis. The results presented in this paper provide additional support for the suggestion.
Activation of Kallikrein-Kinin System in Human Plasma with Purified Serine Protease from Dermatophagoides farinae
Authors: Takahashi Katsunobu , Aoki Takashi , Kohmoto Shoichi , Nishimura Hiroyuki , Kodera Yoh , Matsushima Ayako , Inada Yuji
Published: August 2009

An aqueous extract from a mite culture, of Dermatophagoides farinae, activated prekallikrein to kallikrein in normal plasma. Crude protein preparation, obtained by ammonium sulfate precipitation (95 % saturation) from the extract, exhibited high activity (0.81 units/mg protein) towards a synthetic substrate of coagulation factor XIIa, Boc-Gln-Gly-Arg-MCA, and had also activity to form kallikrein in human plasma deficient in coagulation factor XII. Treatment of the protein preparation with phenylmethylsulfonyl fluoride (PMSF), an inhibitor of serine enzyme, gave rise to inactivation of both activities. Thus, the serine protease specific for Boc-Gln-Gly-Arg-MCA in mite cultures of D.farinae was purified by ammonium sulfate precipitation and chromatographies on p-aminobenzamidine-sepharose CL-4B, DEAE-Toyopearl 650M, Sephadex G-75 superfine and Sephacryl S-200. The purified protease was homogeneous electrophoretically, and its molecular weight was estimated to be 30,000. The optimum pH and temperature were around 7.5 and 50 °C, respectively. The specific activity was 36 units/mg protein at pH 7.4 and 37 °C. The activity was completely inhibited by PMSF. The serine protease had the activity to activate the kallikrein-kinin system in normal human plasma.
Regulation of in vivo Expression of Fc Receptors for IgE (FcεR) on Murine Lymphocytes
Authors: Kiniwa Mamoru , Yanagihara Yukiyoshi , Watanabe Naohiro , Tasaka Kenji
Published: August 2009

Homologous monomeric IgE was employed in a flow cytometric assay for the detection of IgE Fc receptors (FcεR) on mouse lymphocytes. The expression of FcεR in normal BALB/c mice was detected on splenic and circulating lymphocytes, but not on bone marrow cells. The FcεR expression was observed in B cells with B220, surface IgM, and IgD, but not in T cells. Infection of mice with Nippostrongylus brasiliensis resulted in a marked increase in the expression of FcεR on splenic B cells. T cells, however, did not express FcεR even after N. brasiliensis infection. On the other hand, the FcεR expression on normal B cells decreased after a simple incubation at 37 °C for 24 h, while in the presence of IgE this decrease was inhibited. In contrast, B cells stimulated with interleukin 4 display FcεR with high densities. Interestingly, IgE enhanced the FcεR expression induced by interleukin 4, suggesting that both interleukin 4 and IgE may be responsible for an increase in the expression of FcεR on B cells of N. brasiliensis infected mice.
Enhancing Effect of C1q on IgG Monoclonal Antibody Binding to Hapten
Authors: Hamada Atsuo , Watanabe Naohiro , Azuma Takachika , Kobayashi Akio
Published: August 2009

We have previously demonstrated that IgG antibody binding to microfilariae of Dirofilaria immitis increased in the presence of purified C1q. The present study was designed to examine the mechanism of the C1q effect using a system with an antihapten monoclonal antibody (MoAb) and a hapten as an antigen. Microtiter plates were coated with 4-hydroxy-3-nitrophenyl-acetyl (NP)-bovine serum albumin (BSA), and mouse anti-NP MoAb (IgG) was added in the presence of C1q. The amount of IgG which bound to NP-BSA increased with the addition of C1q (p < 0.01) when the antibody had both specificity to the antigen and ability to fix C1q. The C1q effect, examined using two anti-NP MoAbs with different affinities, was more apparent with the low-affinity antibody (LAMoAb) than with the high-affinity (HAMoAb; percent enhancement of IgG binding was 19 vs. 12%). The C1q effect on LAMoAb binding was doubled when a small amount of HAMoAb was incubated with LAMoAb. The C1q effect on IgG binding might be operative in the early phase of infection, where a small amount of high-affinity antibody and a relatively large amount of low-affinity antibody are produced in the host.
Basophil Releasability in Human Hydatidosis
Authors: Aceti Antonio , Celestino Domenico , Caferro Marta , Teggi Antonella , Pennica Alfredo , Adriani Emilio , Grilli Anastasio , Paparo Samuele , Leri Oriana , De Rosa Franco , Sebastiani Antonio
Published: August 2009

We studied immunoglobulin E (IgE)- and non-IgE-mediated releasability in basophils from 31 patients with hydatidosis. Histamine release to non-IgE-dependent stimuli did not differ significantly between normal individuals and patients with hydatidosis. On the contrary, an increased histamine liberation was obtained by challenging basophils from hydatid patients with anti-human IgE. It is concluded that Echinococcus granulosus infection induces an enhanced sensitivity of basophils to IgE-dependent stimuli.
Expression of Dermatophagoides pteronyssinus Allergen, Der p II, in Escherichia coli and the Binding Studies with Human IgE
Authors: Chua K.Y. , Dilworth R.J. , Thomas W.R.
Published: August 2009

Lambda gt11 clones expressing the major house dust mite allergen, Der p II, have been reported to react with IgE in the serum of a high proportion of allergic patients. The clones described, however, only produced small quantities of protein which was not fused to the β-galactosidase of the vector. A construct of the Der p II is described which produces a fusion of Der p II, minus its leader sequence, with the glutathione-S transferase in the pGEX vector. This could be readily isolated and was shown to react with IgE in 22 of 24 patient sera showing reactivity to native Der p II, the sera not reacting having low reactivity to native protein. Absorption analysis showed that the recombinant material removed most of the IgE reactivity of patients to native Der p II. The construct described, therefore, should be valuable for quantitative studies of a pure mite allergen.
Characterization of Allergenic Components in Sap Extract from the Weeping Fig (Ficus benjamina)
Authors: Axelsson I.G.K. , Johansson S.G.O. , Larsson P.H. , Zetterström O.
Published: August 2009

The allergen composition of crude extract from sap (latex) of the weeping fig (Ficus benjamina) was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting. The allergenic components were detected by sera from 11 occupationally exposed plant keepers, of whom 7 were non-atopic, and from 9 non-occupationally exposed atopic patients with a positive radio-allergosorbent test to weeping fig. The allergen-antibody complexes were visualized by rabbit anti-IgE and β-galactosidase-labelled sheep anti-rabbit IgG, using a chromogenic insoluble substrate. A total of 11 allergenic components were identified. Three of them were found to be major allergenic components, identified by more than 50% of the investigated sera. These 3 IgE-binding components had molecular weights of approximately 29,000, 28,000 and 25,000 daltons, respectively. The major allergenic components were denaturated by heat in the temperature range of 60–90 °C.
Blood Histamine Levels in HIV-1-Infected Infants and Children
Authors: Burtin C. , Blanche S. , Galoppin L. , Merval R. , Griscelli C. , Scheinmann P.
Published: August 2009

Blood histamine levels (BHL; ng histamine base/ml blood, mean ± SEM) were determined in 118 infants born to HIV-1-infected mothers and in children infected after blood transfusion. In asymptomatic infected infants, BHL were not significantly different from HIV-1-negative infants born to HIV-infected mothers (54.115.6 vs. 56.4 + 3.5). BHL progressively decreased in the group with polyadenopathies (40.0 ± 4.0), in the group with AIDS-related complex (28.8 ± 2.9), and in patients with AIDS (20.4 ± 0.9). The decrease in BHL was associated with a decrease in blood basophil number.
Immunopathological Response of C57BL/6 and C3H/HeN Mice to Aspergillus fumigatus Antigens
Authors: Kurup Viswanath P. , Choi Hongyung , Resnick Abe , Kalbfleisch John , Fink Jordan N.
Published: August 2009

C3H/HeN and C57BL/6 mice were exposed to culture filtrate (CF) and mycelial extracts (ME) of Aspergillus fumigatus (Af) intranasally. Animals received 6, 8 or 10 biweekly doses and were sacrificed 2 weeks after the last dose was administered. Specific antibodies against Af were detected in their sera by biotin-avidin-linked immunosorbent assay (BALISA). Antibodies against Af belonging to all isotypes showed an increase in both strains of mice. A progressive increase in IgG and IgA antibody isotypes against both CF and ME antigens was detected in C3H/HeN mice during the entire experimental period, whereas most antibody levels peaked after the 8th dose and remained steady or decreased slightly in the C57BL/6 strain. Lung lavage studies showed a relative decrease in the number of macrophages and an increase in the number of lymphocytes after the 6th and 8th instillation of Af antigens in both strains of mice. Histology of the lung demonstrated a progressive inflammatory reaction in C57BL/6 mice during the experimental period. On the other hand, the C3H/ HeN mice showed a negligible inflammatory pulmonary reaction. The antibody responses and inflammatory changes detected in the lungs of mice exposed to Af antigens are comparable to allergic bronchopulmonary aspergillosis (ABPA) in humans and hence this model will be of value in understanding the disease mechanism in ABPA and related diseases.
Influence of Glutathione Conjugation on the Immunogenicity of Dinitrophenyl Derivatives in the Rat
Authors: Tingle M.D. , Clarke Janet B. , Kitteringham N.R. , Park B.K.
Published: August 2009

The role of metabolic detoxication (glutathione conjugation) in the humoral response to three model haptens, dinitrofluorobenzene (DNFB), dinitrochlorobenzene (DNCB) and dinitrobenzenesulphonate (DNBS), was investigated in male Wistar rats. All three haptens produced a measurable anti-dinitrophenyl IgG antibody response over a wide dose range (2.7 nmol/kg to 0.27 mmol/kg) given for 4 days. A significant difference in antibody titre was only observed at the highest dose (DNFB > DNCB = DNBS), despite a marked difference in reactivity towards a protein carrier (albumin) and N-acetyl-lysine in vitro. These observations can be partly explained by the fact that the most reactive hapten (DNFB) conjugates most rapidly with glutathione in vitro and in vivo.
Effect of Age, Culture Medium and Lymphocyte Presence on Ascorbate Content of Peritoneal Macrophages from Mice and Guinea Pigs during Phagocytosis
Authors: Hernanz A. , Collazos M.E. , de la Fuente M.
Published: August 2009

Changes in ascorbate content have been measured by high-performance liquid chromatography in peritoneal macrophages from mice and guinea pigs stimulated by latex particles, in the presence and absence of peritoneal lymphocytes. There was a significant decrease in ascorbate content in murine macrophages 5 min after stimulation that did not occur in controls. The rate of ascorbate consumption was nearly identical with murine macrophages incubated in three different ascorbate-free culture media (phosphate-buffered saline, Hanks’ solution and RPMI) studied, as well as with macrophages from young and old mice. In contrast to mice, both control and stimulated peritoneal macrophages from guinea pigs showed significant decreased contents of ascorbate in the absence of lymphocytes compared with those in the presence of lymphocytes. Moreover, stimulated macrophages showed significantly decreased ascorbate contents with respect to controls. These results indicate that ascorbic acid plays an important role in macrophages and that peritoneal lymphocytes also seem to play a significant role in the maintenance of ascorbate content in macrophages during phagocytosis in guinea pigs.
Reactivity of Mast-Cell-Bound IgE Idiotypes with Anti-Idiotypic Antibody: Mediator Release or Inhibition of Antigen-Induced Mediator Release?
Authors: Wheeler A.W. , Deards M.J. , Hickman B.E. , Spackman V.M. , Johansson S.G.O.
Published: August 2009

When anti-idiotypic antibodies specific for idiotopes on IgE antibodies react with mast-dell-bound IgE antibody, the reaction may be expected to lead either to mediator release or to inhibition of allergen-induced release. This study was carried out to determine which would occur. Anti-idiotype antiserum (anti-Ids) was raised in syngeneic mice by immunisation with affinity-purified DNP-specific mouse monoclonal IgE. Sera from these immunised mice and from a similarly immunised rabbit inhibited the binding of the monoclonal IgE to radio-iodinated dinitrophenylated (DNP) ovalbumin, whereas a high concentration of affinity-purified rabbit antimouse ε-chain-specific antibody was unable to inhibit even though it was shown to bind to the monoclonal IgE. The mouse anti-Ids did not give a positive passive cutaneous anaphylactic reaction in rats sensitised with high-titre grass-pollen-specific IgE-containing antiserum, whereas it gave a positive result in rats sensitised with the DNP-specific monoclonal IgE. Rabbit antimonoclonal antiserum and rabbit anti-ε-chain antibody both gave positive results in these two assays. The results indicated that anti-idiotypic antibodies specific for idiotopes on IgE antibody can react and release mediators from IgE-sensitised mast cells. The significance of this in view of the increased levels of anti-idiotypes that can occur during immunotherapy is discussed.
Further Characterization of Surface Membrane Structures Expressed on Human Basophils and Mast Cells
Authors: Valent Peter , Majdic Otto , Maurer Dieter , Bodger Michael , Muhm Manfred , Bettelheim Peter
Published: August 2009

Recently, we were able to establish the immunologic surface marker profile of human basophils and mast cells. In the present study, the characterization of these cell types was extended by the use of monoclonal antibodies (mAbs) to hemopoietic differentiation antigens. Basophils and mast cells were enriched by mAbs and complement from chronic myeloid leukemia blood (n = 5) and dispersed lung tissue (n = 4), respectively. A panel of 80 mAbs was tested for being reactive with purified cell populations using flow cytometry and/or a combined toluidine blue-immunofluorescence staining procedure. In addition to previous findings, basophils were found to react with mAbs directed against the 126-kilodalton dipeptidylpeptidase IV (CD26), platelet glycoprotein IIa (CD31), CD40 antigen known to share sequence homology with nerve growth factor receptor, leukosialin (CD43), CD44 antigen, the ICAM-1 antigen (CD54) and VIM2-reactive gangliosides involving the sialofucooligosaccharide sequence (CDw65L). Bsp-1 was found to be a specific marker for human basophils, whereas mast cells were not stained by this reagent. Basophils apparently lack CD22 antigen, gangliosides detected by CDw65 mAbs (except CDw65L) and CD71 antigen (transferrin receptor). Mast cells were found to express CD43 and CD44 antigen. In contrast, mast cells lack CD22, CD26, CD31, CD40 and CDw65 antigen. These results provide further evidence that both blood basophils and mast cells express a unique immunologic surface marker profile including binding sites for a variety of immunomodulating ligands and adhesion molecules.
Interleukin-1 Release by Alveolar Macrophages in Asthmatic Patients and Healthy Subjects
Authors: Pujol Jean-Louis , Cosso Béatrice , Daurès Jean-Pierre , Clot Jacques , Michel François-Bernard , Godard Philippe
Published: August 2009

A thymocyte proliferative response assay was used to compare spontaneous and lipopolysaccharide (LPS)-induced interleukin-1 (IL-1) release by alveolar macrophage (AM) in asthmatic patients and normal subjects. Twelve asthmatic patients and seven nonsmoking healthy subjects underwent a bronchoalveolar lavage (BAL). All asthmatic patients had a reversible airway obstruction and 7/12 were allergic. BAL AM were separated by adherence on tissue culture plates in medium RPMI-1640 supplemented with antibiotics and fetal calf serum, and were incubated with or without 10 μg/ml LPS for 20 h. Free-cell supernatants were tested by C3H/HeJ mice thymocyte proliferative assay. Unstimulated AM supernatant IL-1 activity was significantly higher in asthmatic patients (mean ± SEM: 47.8 ± 11.9 units/106 AM) in comparison with healthy subjects (4.8 ± 2.3 units/106 AM; p < 0.05, Mann-Whitney U test) but did not significantly differ between allergic (42.2 ± 15.5 units/106 AM) and intrinsic asthmatic patients (55.8 ± 20.7 units/106 AM). For healthy subjects, IL-1 activity was significantly higher in LPS-stimulated AM supernatants (85 ± 20 units/106 AM, p < 0.05; Mann-Whitney U test) in comparison with unstimulated ones; for asthmatic patients, unstimulated and LPS-stimulated AM supernatant IL-1 activity did not significantly differ. This finding is in accordance with previous work suggesting that AM from asthmatic patients have a weak suppressive activity upon lymphocyte proliferation and emphasize the enhanced AM releasability in asthma.
Sequential Analysis of Histamine Release and Intracellular Ca2+ Release from Murine Mast Cells
Authors: Tasaka Kenji , Sugimoto Yukio , Mio Mitsunobu
Published: August 2009

Stimulation of murine peritoneal mast cells with compound 48/80 at a concentration of 1 μg/ml elicited rather slow histamine release; the onset of release was observed 5 s after stimulation, and it reached a plateau at about 60 s. Both inositol-1,4,5-trisphosphate (IP3) and inositol-1,4-bisphosphate (IP2) contents increased to their maximum 5 s after stimulation. The IP3 content decreased to the control level more rapidly than that of IP2. Changes in the intracellular Ca2+ concentration of the quin 2 loaded mast cells were determined using a video-intensified microscopy system. The fluorescence intensity due to Ca-quin 2 complex increased rapidly after 48/80 stimulation in a Ca-free medium and reached the maximum at about 6–7s. It became clear that the increase in IP3 content and the resulting Ca2+ release from the intracellular Ca store precede histamine release from murine mast cells.
Further Thoughts on Mast Cells, Calcium Channels and Histamine Release
Authors: West G.B.
Published: August 2009

Calcium movements across membranes of mast cells represent a major target for effective anti-allergic drugs, as these are essential events linking stimulation to secretion. Using isolated peritoneal mast cellsfrom two types of genetically different rats and a blocker of the entry of calcium ions into cells, histaminerelease induced by antigen, calcium ionophore and potassium (in a lowered calcium medium) has been followed. Lower concentrations of the calcium entry blocker were required to inhibit potassium-induced release than those needed for release induced by allergic and other non-allergic stimuli, suggesting that different specialised calcium channels are involved in histamine release from rat mast cells.
Recombinant Neutral Endopeptidase Attenuates Substance P-Induced Plasma Extravasation in the Guinea Pig Skin
Authors: Rubinstein Israel , Iwamoto Itsuo , Ueki Iris F. , Borson Benjamin , Nadel Jay A.
Published: August 2009

To determine whether exogenously administered neutral endopeptidase (NEP; enkephalinase, EC 3.4.24.11) inhibits the substance P-induced increase in vascular permeability in the skin, we examined the effects of recombinant human NEP on plasma extravasation induced by intradermal injection of substance P in guinea pig skin. Injection of substance P (2.5 × 10––8M) induced significant plasma extravasation in the skin (53 ± 4 mm2 of Evans blue extravasation; mean ± 1 SEM). In vitro incubation of substance P with recombinant human NEP prior to injection prevented the substance P-induced plasma extravasation in the skin in a dose-dependent fashion. Intradermal preinjection of recombinant human NEP partially inhibited plasma extravasation induced by subsequent injection of substance P (52 ± 9% of the control without NEP). The H1 and H2 histamine antagonists pyrilamine and cimetidine, and a muscarinic antagonist, atropine, had no effects on substance P-induced responses. Two products of substance P degradation by NEP containing the carboxy-terminal portion, substance P7–11 and substance P8–11, were also without effect. These findings suggest that recombinant human NEP can attenuate substance P-induced increases in vascular permeability in guinea pig skin and, therefore, may be useful in treating dermatologic disorders in which abnormal responses to substance P or other neuropeptides cleaved by NEP may occur.
Antigen-Induced Mucosal Damage and Restitution in the Small Intestine of the Immunized Rat
Authors: D’lnca Renata , Ramage John K. , Hunt Richard H. , Perdue Mary H.
Published: August 2009

Intestinal mucosal damage and restitution were examined following antigen-induced systemic anaphylaxis in Nippostrongylus brasiliensis immunized rats. The rats were injected intravenously with N. brasiliensis antigen or saline. At 60 min, morphological and biochemical parameters were determined in jejunum and ileum, and the epithelial permeability was assessed by measuring recovery of 51Cr-ethylenediaminetetraacetic acid in the blood after injecting it into a ligated segment. Antigen challenge resulted in significant abnormalities: (1) villus damage with sloughing of enterocytes; (2) decreased activities of brush border enzymes; (3) decreased levels of mucosal histamine and rat mast cell protease II (mast cell mediators), and (4) increased uptake of 51Cr-ethylenediaminetetraacetic acid. Progression of the injury was examined by taking consecutive biopsies at 15-min intervals for 60 min and then at 5 h. At 15 min, an abnormality was present in all sections which ranged from minor oedema and enterocyte detachment at villus tips to virtual complete destruction of the apical region. Restitution occurred by villus contraction with migration of the epithelium over the damaged regions. At 5 h, the epithelium had resealed, but the villi were significantly reduced in height.
Antigenic and Allergenic Characterization of the Enzymes Alcalase and Savinase by Crossed Immunoelectrophoresis and Crossed Radioimmunoelectrophoresis
Authors: Arlian L.G. , Vyszenski-Moher D.L. , Merski J.A. , Ritz H.L. , Nusair T.L. , Wilson E.R.
Published: August 2009

Alcalase and savinase, produced by Bacillus species, are proteolytic enzymes that are used in laundry products and are known to cause respiratory allergy. Antigenic and allergenic characteristics of alcalase and savinase and their potential cross-reactivity were evaluated using crossed immunoelectrophoresis and crossed radioimmunoelectrophoresis. Alcalase exhibited two distinct antigens; one electropositive and one electronegative. The electropositive antigen exhibited some retrograde anodic mobility when coupled with antiserum components. Savinase exhibited one electropositive and two electronegative antigens. The antigens of the two enzymes were clearly different from each other, the three savinase antigens exhibiting greater electrophoretic mobility than the two alcalase antigens. In crossed radioimmunoelectrophoresis studies, only the electropositive antigen of alcalase, its retrograde complex, and the electropositive antigen of savinase bound IgE from the sera of individuals who were skin test positive to one or both enzymes. No evidence of cross-reactivity was observed in heterologous and tandem crossed immunoelectrophoresis studies and heterologous microimmunodiffusion reactions.
Relationship between Alterations in Atrial and Ventricular Histamine Content and Cardiac Function during Cardiac Anaphylaxis of Isolated Guinea Pig Hearts
Authors: Heller Lois J. , Regal Jean F.
Published: August 2009

Antigen-induced histamine release from sensitized cardiac tissue has been shown to contribute significantly to the increases in atrial rate, atrioventricular nodal conduction, and ventricular contractile force that occur during cardiac anaphylaxis. These findings suggest that there might be a strong negative correlation between changes in these variables and the residual histamine content in the tissue after the anaphylactic reaction. In the present study using isolated hearts of passively sensitized guinea pigs, antigen challenge evoked transient increases in atrial automaticity, P-R intervals, vascular resistance, and left ventricular pressure. The histamine content of atrial and ventricular tissue from antigen-challenged hearts (1,475 ± 296 and 4,543 ± 360 ng/g, respectively) was significantly less than that of nonchallenged hearts (2,652 ± 335 and 6,298 ± 251 ng/g, respectively). Significant correlations found were between the residual histamine content of the ventricular tissue or the total histamine released and the magnitude of the antigen-induced increase in left ventricular systolic pressure. These findings suggest that antigen-induced increases in left ventricular systolic pressure can be used as an index of the local ventricular histamine release. The lack of significant correlations between local residual histamine levels and changes in atrial rate, P-R intervals, or coronary vascular resistance suggests that complicated interactions between histamine and other mediators or factors may be involved in the antigen-induced chronotropic, dromotropic, and vasoconstrictive alterations in these preparations.
Antibodies from Patients with Rheumatoid Arthritis and Juvenile Chronic Arthritis Analyzed with Core Histone Synthetic Peptides
Authors: Tuaillon Nadine , Muller Sylviane , Pasquali Jean-Louis , Bordigoni Pierre , Youinou Pierre , Van Regenmortel Marc H.V.
Published: August 2009

Antihistone antibodies were detected in the sera of a randomly selected group of patients with rheumatoid arthritis (RA) and juvenile chronic arthritis (JCA) by an enzyme-linked immunosorbent assay (ELISA) with the five individual histones and by immunoblotting. In ELISA, the overall frequency of antihistone antibodies in RA and JCA was 51 and 44%, respectively. Antibodies present in these serum samples were further studied by ELISA by means of 17 core histone synthetic peptides. The fragment 1–21 of H3 was recognized by 60% of RA sera and by 62% of JCA sera. In addition, at least four terminal or internal peptides of H3 and H4 were recognized by more than a third of JCA sera, while only two of these peptides reacted with 20% of RA sera. Many of the sera that did not show any reactivity with the whole histone reacted with various histone peptides. This finding demonstrates the usefulness of synthetic peptides for identifying autoantibodies.
T Cell Receptor Variable Gene Expression: Analysis in Ragweed-Sensitive Patients during Allergen Exposure
Authors: Burton Daniel A. , Shepherd Gillian M. , Siskind Gregory W. , Posnett David N.
Published: August 2009

Four monoclonal antibodies (MAb) to V region determinants of the α/β-chain of the T cell antigen receptor (TCR) were used, by cytofluorography, to detect discrete populations of peripheral blood T cells (PBT). Together they identify 10–15% of circulating CD3+ T cells. Each MAb is known to detect specific V regions of the β-chain. Thus Vβ5 gene products are recognized by MAb C37, Vβ6 by OT145, Vβ8 by Ti3a, and Vβ12 by MAb S511. In previous studies, we found that the percentages of PBT detected by these MAb show little variation over time in normal individuals. In order to determine if there is a change in TCR V gene usage during an immune response to an environmental antigen, 12 atopic patients with known ragweed sensitivity by history and skin test were followed for a 6-month period encompassing the ragweed season. No shifts in V gene usage that could be correlated with the ragweed season were consistently observed. The patients could be arbitrarily divided into two groups: in group I little variation over time was observed in the T cell populations identified by the MAb used, while group II was characterized by marked variation of the same T cell populations over time. In group II individuals, the population of Ti3a+ T cells showed the most variation over time. Failure to observe shifts in PBT subpopulations, identified by expression of different TCR V regions, during exposure to an allergen to which an IgE response has been made may mean that such shifts do not occur or that they occur primarily at the tissue site of antigen exposure and not in the peripheral circulation or that they occur in T cell subpopulations not identified by the reagents used.
Identification of a ‘Disease-Associated’ Antigen in Pigeon Breeder’s Disease by Western Blotting
Authors: de Beer P.M. , Bouic P.J. , Joubert J.R.
Published: August 2009

Sera from 9 symptomatic and 7 asymptomatic pigeon breeders were analyzed for their reactivity to pigeon serum by western blotting. All 9 symptomatic sera (9/9; 100%) and only 4 of 7 (57%) asymptomatic sera revealed specific antibodies. The immunoreactivity patterns of the sera varied: the majority of the sera reacted to antigens having molecular weights of 220 kD or more (9/16 sera), 98 kD (8/16 sera), and 86 kKD (8/16 sera). However, only sera from symptomatic breeders recognized an antigen of approximate molecular weight of 29–32 kD (9/9; 100% of symptomatics). We conclude that this antigen is ‘disease associated’ and may be useful in the diagnosis of pigeon breeder’s disease.
Human Recombinant Lymphokines and Cytokines Induce Pulmonary Eosinophilia in the Guinea Pig which Is Inhibited by Ketotifen and AH 21–132
Authors: Kings M.A. , Chapman I. , Kristersson A. , Sanjar S. , Morley J.
Published: August 2009

Subcutaneous or intraperitoneal injection of recombinant human granulocyte-macrophage colony-stimulating factor, interleukin 3, or mouse tumour necrosis factor alpha, but not recombinant human interferon gamma, platelet-derived growth factor, or transforming growth factor beta caused selective eosinophilia of the pulmonary airways in the guinea pig. Unlike responses to latelet-activating factor, there was no attendant detectable airway hyperreactivity, but in common with responses to platelet-activating factor, eosinophilia of the airways was prevented by pretreatment with ketotifen or AH 21–132. Cytokines or lymphokines may contribute to pulmonary eosinophilia in diseases such as asthma.
Isolation and Characterization of a cDNA Clone Encoding an IgE-Binding Protein from Kentucky Bluegrass (Poa pratensis) Pollen
Authors: Mohapatra Shyam S. , Hill Robert , Astwood Jim , Ekramoddoullah Abul K.M. , Olsen Egil , Silvanovitch Andre , Hatton Tom , Kisil Fred T. , Sehon Alec H.
Published: August 2009

We reported previously on the isolation and characterization of several allergens from Kentucky bluegrass (KBG) (Poa pratensis L.) pollen with the aid of the corresponding murine monoclonal antibodies (Mabs). In the present study, (1) an analysis of various tissues of this grass revealed that the allergenic components recognized by these Mabs were confined to the pollen; (2) intact translatable mRNA was isolated from the KBG pollen, and (3) a cDNA library was constructed with this mRNA in the λgt11 expression vector. Screening of this library with a pool of six sera from KBG-allergic patients, in combination with enzyme-labeled antibodies to human IgE, led to the isolation of a cDNA clone, referred to as KBG7.2. The nick-translated cDNA probe of KBG7.2 hybridized to a 1.5-kbp RNA transcript from KBG pollen. Moreover, transcripts corresponding to KBG7.2 were found in pollens of eight other grasses, indicating that the proteins similar to the one encoded by this cDNA may be present in these grasses. The nucleotide sequence of KBG7.2 was determined; interestingly, the corresponding derived amino acid sequence did not match any other sequence recorded in the protein data banks. The peptide encoded by KBG7.2 was expressed as a fusion protein utilizing the plasmid vector pWR590.1. Whereas none of the above allergen-specific Mabs bound to the fusion protein, all the 15 individual sera from grass pollen allergic patients recognized the fusion protein. It is, therefore, concluded that the cDNA clone KBG7.2 encodes a polypeptide of a major IgE-binding protein of KBG pollen and that the murine Mabs used in this study do not recognize the epitopes responsible for induction of human IgE antibodies.
Rhesus Monkey Airway Responses to Substance P
Authors: Patterson Roy , Harris Kathleen E.
Published: August 2009

The tachykinin, substance P (SP), was used as a model to study airway and cutaneous responsesin a group of normal rhesus monkeys or animals with IgE-mediated cutaneous reactions to Ascaris antigen (AA) alone or animals with both IgE-mediated cutaneous and airway responses to AA. Aerosolized SP (10 mg/ml) resulted in airway responses qualitatively similar in duration and pulmonary function abnormalities to AA. These SP airway responses were most closely associated with the presence of IgE antibody to AA and were not increased in animals with airway reactivity to AA. A dose response to aerosolized SP can be established. An enkephalinase inhibitor, thiorphan, did not potentiate SP airway responses but appeared to potentiate SP cutaneous responses in some animals. SP cutaneous reactions could be demonstrated and did not correlate with the IgE-mediated cutaneous dilution titers to AA.
Histamine Release from Human Basophils by the Insect Allergen Chi t I
Authors: Baur Xaver , Mazur Gertraud , Jarosch Barbara
Published: August 2009

Hemoglobins of the Diptera species Chironomus thummi thummi (Chi t I) are potent inhalant allergens. Chi t I-specific histamine release was measured by a new radioimmunoassay in whole human blood taken from 20 sensitized patients, 11 exposed nonsensitized probands, and 11 nonexposed controls. The sensitized patients, who all had positive skin tests and radioallergosorbent test results with Chi t I, showed a significantly higher histamine release than the two other groups. However, within the patient group, the percentage of released histamine did not correlate with the intensity of the skin test response or the concentration of Chi t I-specific IgE antibodies. Our results demonstrate that this method is a sensitive and specific in vitro test for evaluation of IgE-mediated sensitization.
The Airway Epithelial Lining in Guinea Pigs Is Intact Promptly after the Mucosal Crossing of a Large Amount of Plasma Exudate
Authors: Luts A. , Sundler F. , Erjefält I. , Persson C.G.A.
Published: August 2009

In the present study of anaesthetized guinea pigs, we have confirmed by chemical analyses that 10 min after topical mucosal provocation with capsaicin 0.4 nmol of platelet-activating factor 8 nmol plasma macromolecular tracers (FITC-dextran, molecular weight 156 kD, previously injected intravenously) have been significantly exuded into airway tissue and lumen. We have also examined the airway mucosa and the mucosal surface material using fluorescence, light, and electron microscopy. Thus we have demonstrated by histological techniques that promptly after the induction of large exudative responses plasma macromolecules are abundant in the lumen as well as in the lamina propria. Furthermore, the exudate is seen on the surface of a mucosa that has an intact epithelial lining. These observations are evidence in favour of the possibility that plasma exudation into the lumen is a first-line defense mechanism of the normal airway mucosa.
Induction of Interleukin-1-Beta Release from Human Monocytes by Cotton Bract Tannin
Authors: Vuk-Pavlović Zvezdana , Rohrbach Michael S.
Published: August 2009

The human T lymphocyte proliferative response to cotton bract tannin was shown to be dependent upon the presence of monocytes. Since monocytes support the T cell mitogenic response by interleukin-1 (IL-1) production, it was anticipated that tannin has IL-1-inducing ability. To examine this possibility, human monocytes were cultured alone or with peripheral blood T lymphocytes, and stimulated with tannin. Control cultures included unstimulated cells, and cells challenged with other IL-1 inducers: concanavalin A (Con A) and lipopolysaccharide from Escherichia coli or Enterobacter agglomerans. IL-1β was measured in culture super-natants 24 h after initiation of the culture by the use of an ELISA or an RIA. The results showed that tannin stimulated monocytes to secrete IL-1β in a manner similar to Con A, i.e. substantially more cytokine was measured in the supernatants of monocyte-T-lymphocyte co-cultures than in the cultures of monocyte alone. Endotoxin from E. coli was less effective than the endotoxin from E. agglomerans in IL-1 induction. Contaminating endotoxin present in the tannin preparation accounted for the majority of IL-1β released from monocytes alone stimulated with tannin, but only 20% of the IL-1β released from tannin-stimulated monocyte-T-lymphocyte co-cultures. These results show that tannin itself has IL-1-inducing ability. The dose-response studies show that the extent of IL-1β release is dependent on tannin dose and that increased levels of monocyte-produced IL-1β precede the increase in T lymphocyte proliferation. These findings suggest that tannin stimulates T cell proliferation by a mechanism similar to that in lectin-induced T lymphocyte proliferation, namely by providing the second signal by stimulating monocytes for IL-1 production.
Sequential Appearance of Basophils and Mast Cells from Human Bone Marrow in Long-Term Suspension Culture
Authors: Bressler Robert B. , Friedman Marc M. , Kirshenbaum Arnolds.K. , Irani Anne-Marie A. , Schwartz Lawrence B. , Metcalfe Dean D.
Published: August 2009

Human bone marrow cells grown in liquid culture in the presence of conditioned medium from concanavalin A stimulated peripheral blood mononuclear cells, from mixed lymphocyte reactions, or from an osteogenic sarcoma tumor line gave rise to basophils which were maximal in number at 1–3 weeks. Basophils had a multilobed nucleus, contained large cytoplasmic granules of variable size which stained metachromatically with acid, but not neutral toluidine blue, were negative for chloroacetate esterase, and did not contain human mast cell tryptase. In long-term cultures, mast cells were detected after 5–6 weeks. Mast cells had cytoplasmic granules that stained with acid and neutral toluidine blue, were positive for chloroacetate esterase, and contained human mast cell tryptase. It, therefore, appears that in liquid cultures even in the absence of feeder layers, human bone marrow cell cultures can give rise initially to basophils, and then to mast cells.
Monoclonal Antibodies to Proteins from Cocksfoot Grass (Dactylis glomerata) Pollen: Isolation and N-Terminal Sequence of a Major Allergen
Authors: Walsh David J. , Matthews Jayne A. , Denmeade Russell , Maxwell Paula , Davidson Matthew , Walker Matthew R.
Published: August 2009

Monoclonal antibodies (Mabs) raised to an aqueous extract of cocksfoot grass (Dactylis glomerata) pollen have been characterised. Mab 1B9 was demonstrated by SDS-PAGE and Western blotting to recognise a major allergen of an approximate molecular weight of 28 kD in this extract, termed DG3, and a component with a molecular weight of between 35 and 40 kD in an extract of Secale cereale (cultivated rye) pollen. The 28 kD component of cocksfoot grass pollen isolated by affinity chromatography using Mab 1B9 was recognised by IgE antibodies in 80% (8 of 10) atopic sera, but only weakly by 25% (1 of 4) non-atopic sera tested. N-terminal sequencing of DG3 purified by affinity chromatography, 2 D electrophoresis and electroblotting to polyvinylidenedifluoride revealed significant homology with a group-V allergen (Phi p V) from timothy grass (Phleum pratense).