bluuebunny/crossref_metadata_2025 huggingface dataset:
Beta-2-Microglobulin of Rat Major Histocompatibility Complex Class I Antigens
Authors: Misra D.N. , Kunz H.W. , Gill, III T.J.
Published: August 2009

Detection of the β2-microglobulin (β2m) component of the rat MHC class I antigens has been difficult. In the present report, we have addressed this issue by a systematic study of rat class I antigens from red blood cells or from lymphocytes that were freshly isolated or cultured in the presence of autologous or heterologous sera and surface-labeled with 125I or intrinsically labeled with radioactive amino acids. First, specific radioiodination of rat β2m in association with the antigen heavy chain on red blood cells or lymphocytes is minimal, resulting in its poor identification by SDS-PAGE. Second, labeling with radioactive methionine or lysine gives a more intense β2m band with respect to the heavy chain than labeling with arginine or tyrosine. Third, the β2m component shows a large increase in intensity compared to the heavy chain when the antigens are isolated from lymphocytes that are cultured in the presence of fetal bovine serum prior to 125I-labeling. This increase is due to exchange of endogenous rat β2m with bovine β2m and to a much higher level of radioiodination of the latter. Fourth, rat red blood cells and lymphocytes contain free surface β2m molecules in addition to those associated with the antigen heavy chains. The free molecules show a much higher level of radioiodination than those associated with the heavy chains, and there is little exchange between the antigen-bound and the free β2m after radioiodination. Thus, intrinsic labeling with suitable radioactive amino acids rather than surface labeling with 125I is the method of choice for unambiguous detection of the endogenous β2m component of the rat MHC class I antigens.
Detection and Characterization of IgE-Producing Cells in Patients with Clonorchiasis
Authors: Yanagihara Yukiyoshi , Kiniwa Mamoru , Kajiwara Keiichi , Joh Kosuke , Kobayashi Akio , Watanabe Naohiro
Published: August 2009

The ability of peripheral B cells of patients with Clonorchis sinensis infections to secrete IgE spontaneously was investigated in vitro. The de novo synthesis of IgE was observed in unstimulated B-cell cultures of patients. There was a significant relationship between the serum IgE level and the amount of IgE spontaneously secreted by B cells. Pretreatment of patients’ B cells with 10 μg/ml of rabbit anti-human IgE resulted in the clear suppression of spontaneous IgE synthesis without affecting the IgG synthesis. Their B cells capable of spontaneously secreting IgE were partially sensitive to irradiation with 1,000 rad. The results obtained suggest that such IgE-forming cells may be responsible for at least part of the persistent IgE formation in patients with helminthic infections as well as in those with atopic disease.
Modulation of Human Peripheral Blood Eosinophil Function by Tumor Necrosis Factor-Alpha
Authors: Whitcomb Elizabeth A. , Wesolek John H. , Pincus Stephanie H.
Published: August 2009

Eosinophils are important effectors in helminthic parasitic infection. Tumor necrosis factor (TNF-α) has been implicated as a mediator in the host response to parasitic infection and enhances eosinophil-mediated helminthotoxicity. We have examined the direct effects of recombinant human (rh)TNF on eosinophil functions of degranulation and oxidative metabolism. This report describes the minimal effects of rhTNF-α on eosinophil superoxide anion generation and enzyme secretion, which do not satisfactorily explain the observed increases in helminthotoxicity. In contrast to other cell types, eosinophils are unique in their differential responses to interleukin-1β and TNF.
IgE-Positive Cells in Human Intestinal Mucosa Are Mainly Mast Cells
Authors: Rognum T.O. , Brandtzaeg P.
Published: August 2009

Nine biopsy specimens from the jejunum of patients with a clinical history of food allergy and 10 from the rectal mucosa of patients with presumed ‘allergic proctitis’ were fixed in cold ethanol and further processed for paraffin embedding. Serial tissue sections were stained for IgE by direct (polyclonal antibody) and indirect (monoclonal antibody) immunofluorescence methods. Adjacent sections were subjected to conventional mast cell staining (astra blue). In all mucosal specimens from the jejunum and in 8 rectal ones, numerous cells were found to be positive both for astra blue by transmission microscopy and for IgE by fluorescence microscopy of the same section. With the monoclonal antibody all astra-blue-positive cells were IgE-positive; however, this was not always the case with the polyclonal reagent, probably because the fluorescence was weaker with the direct technique. Detection of IgE-positive mucosal mast cells may turn out to be of diagnostic interest in patients with adverse reactions to food.
Antibody Specificity in the Immune Response to Insulin
Authors: Mirza Intisar H. , Wilkin Terence J.
Published: August 2009

The immune response to insulin in Balb/c mice has been investigated with regard to the fine specificity of antibodies present during the primary stage of immune responsiveness. Hybridoma technology was used to raise monoclonal antibodies and a solid phase enzyme-linked immunoassay to test their specificity for human, bovine and porcine insulins. Heteroclitic antibodies were observed. These showed greater binding to the cross-reactive bovine and porcine insulins than to the inducing human insulin.
Responses of Antigen-Specific Long-Term Murine T Cell Lines to Wheat Gliadin Fractions
Authors: Howdle P.D. , Hanson D.G. , Trejdosiewicz L.K. , Ciclitira P.J. , Smart C.J. , Walker W.A.
Published: August 2009

Recent evidence suggests that the four electrophoretically defined gliadin subfractions (α, β, γ and ω) of wheat can induce the typical pathological finding of coeliac disease. We have prepared long-term murine T cell lines to gliadin and its four major subfractions. The cell lines were tested in proliferative assays with each homologous gliadin subfraction, and to the other gliadin subfractions. There was some cross-reactivity, with unfractionated gliadin and its α-subfraction being the most antigenic, while ω-gliadin was the least. These data demonstrate that gliadin components are effective stimuli for specific T cell responses, and further suggest that the α-gliadin subfraction generates the highest specific responses. This accords with observations in man that all four gliadin subfractions exacerbate coeliac mucosa, but that the α-subfraction is the most active.
Monitoring of Active Groups of Cyanogen-Bromide-Activated Paper Discs Used as Allergosorbent
Authors: Jaggi Kuldeep S. , Gangal Sharad V.
Published: August 2009

The amount of active groups of cyanate esters and imidocarbamates present on cyanogen-bromide-activated paper discs were estimated. The protein-binding capacity of activated discs was found to be dependent on the amount of these active groups. The results suggested that monitoring of active groups on activated discs can help in preparing consistent allergosorbent for immunoassays.
Inhibition by Histamine of Platelet-Activating-Factor-Induced Neutrophil Chemotaxis in Bronchial Asthma
Authors: Rabier Mireille , Damon Marcelle , Chanez Pascal , Mencia-Huerta Jean-Michel , Braquet Pierre , Bousquet Jean , Michel François-Bernard , Godard Philippe
Published: August 2009

Circulating human polymorphonuclear neutrophils are involved in asthma after their migration into the lung by local chemotactic factors. Investigation of the locomotion of neutrophils in Boyden chambers, showed that the chemotactic intensity of the platelet-activating factor (PAF) was similar in cells from healthy subjects and allergic asthmatics, although the optimal effect of the mediator was observed at 10––6M and 10––8M, respectively. Histamine had no direct chemoattractant effect on neutrophils but inhibited PAF-induced chemotaxis of neutrophils from healthy subjects and allergic asthmatics. This study provides additional evidence that neutrophils are involved in asthma, and points out the interaction between PAF and histamine in the migration of neutrophils to the lung.
Tetrandrine and Transmembrane Signal Transduction: Effect on Phosphoinositide Metabolism, Calcium Flux and Protein Kinase C Translocation in Human Lymphocytes
Authors: Ioannoni B. , Chalmers A.H. , Seow W.K. , McCormack J.G. , Thong Y.H.
Published: August 2009

Time-course and dose-dependent studies showed consistent suppression of phosphoinositide turnover in Con A-stimulated human lymphocytes in the presence of the plant alkaloid, tetrandrine. Significant inhibition of Con A-stimulated calcium flux was also observed. Furthermore, protein kinase C activity was also significantly inhibited by tetrandrine irrespective of whether Con A or phorbol myristate acetate was the stimulant. These results suggest that the immunosuppressive properties of tetrandrine are in part mediated by the capacity of tetrandrine to interfere with transmembrane signalling.
Human Immune Response against Timothy Grass Pollen Allergens
Authors: Malley Arthur
Published: August 2009

Enzyme-linked immunoassays were used to examine the binding specificity of affinity-purified anti-idiotypic antibody [against the idiotypic determinant on timothy grass pollen antigen B (AgB)-specific IgE] with murine AgB-specific IgG and IgE antibodies, and the serum from timothy-sensitive patients. In addition, the proliferative response of peripheral blood lymphocytes from timothy-sensitive patients with either antigen or anti-idiotypic antibody was examined. These studies suggest that the idiotypic determinant expressed on murine AgB-specific IgE is highly conserved on human AgB-specific IgE and T cells of timothy-sensitive patients.
Identification of a Mouse Anti-Rat IgE Monoclonal Antibody, 44.7b, Which Inhibits IgE Binding to RBL Cells
Authors: Rup Bonita J. , Kahn Larry E.
Published: August 2009

We have identified a monoclonal antibody specific for rat IgE, 44.7b, which blocks binding of rat IgE to the mast cell-like line RBL. Other monoclonal anti-rat IgE antibodies (MARE1 and B5) did not inhibit IgE binding to these cells. Furthermore, 44.7b did not react with IgE previously bound to these cells. These results indicate that 44.7b binds to an epitope on the IgE molecule which is within the binding site of the mast cell IgE Fc receptor (FcERI). The 44.7b monoclonal antibody did recognize IgE bound to a B lymphocyte cell line indicating that mast cell and B lymphocyte FcERs recognize different regions of the IgE molecule.
Experimental Asthma in Guinea Pigs Sensitized with Mites (Dermatophagoides farinae)
Authors: Ishii Akira , Ito Koji , Ino Yoshitaka , Miyamoto Terumasa
Published: August 2009

Guinea pigs were intraperitoneally immunized with 1, 10, 100 or 1,000 μg protein of an extract from mites (Dermatophagoide farinae) incorporated with aluminium hydroxide gel 3 times every 28 days. Ten days after the last injection, a bronchial provocation test was performed by the use of the mite extract. The group immunized with 1 μg protein produced very low levels of IgE-, IgG1- and IgG2 antibodies, and did not show asthmatic response upon the challenge. Although the group immunized with 10 μg protein showed higher rates of asthmatic onset than other groups, the sera of this group contained lower levels of IgE antibodies than the group immunized with 100 or 1,000 μg protein. However, the sera of the group immunized with 10 μg protein showed higher ratios of IgE/IgG1, IgE/IgG2 and lower ratios of IgG1/IgG2 antibodies than the sera of the group immunized with 100 or 1,000 μg protein.
Cellular Communication in Leukotriene C4 Production between Eosinophils and Neutrophils
Authors: Kloprogge Ed , de Leeuw Anna J. , de Monchy Jan G.R. , Kauffman Henk F.
Published: August 2009

Eosinophilic granulocytes as well as neutrophilic granulocytes produced leukotriene C4 (LTC4) on stimulation with 1 μM A23187 (Ca2+ ionophore). In healthy volunteers, the LTC4 production in eosinophils was about 3 times the production in neutrophils. Within 15 min >90% of the LTC4 was released into the supernatant. Stimulation of eosinophils and neutrophils together resulted in a synergistic increase in LTC4 production of 306 ± 40%. LTC4 synthesis by hypodense eosinophils was also enhanced if stimulated in the presence of neutrophils. These findings suggest a communication between eosinophils and neutrophils, which may play a role in bronchial asthma.
Modulation of Human Neutrophil Adherence by Periodontopathic Bacteria: Reversal by Specific Monoclonal Antibodies
Authors: Seow Kim , Bird Philip S. , Seymour Gregory J. , Thong Y.H.
Published: August 2009

Previous investigations by us have shown that direct interaction of Fusobacterium nucleatum with polymorphonuclear neutrophils (PMNs) results in the stimulation of PMN adherence whereas direct interaction with Bacteroides gingivalis results in PMN suppression. In the present study, panels of monoclonal antibodies (MAbs) raised against cell wall antigens of F. nucleatum and B. gingivalis were tested to determine their ability to block the modulatory effects of the bacteria in their interactions with PMNs. While no activity was demonstrated for any of the 9 MAbs raised against F. nucleatum, it was found that 2 of 16 MAbs raised against B. gingivalis were able to reverse the suppression of PMN adherence induced by the bacteria. Further studies on these 2 reactive MAbs showed that the effect of MAbs were abrogated by heat treatment as well as by trypsin proteolysis. Investigations into the nature of the reactive epitopes on the bacterial surface showed that they probably contain protein components susceptible to proteolytic attack by subtilisin. In addition, β-galactose may be a component of the reactive epitopes for one of the MAbs, but sialic acid residues on the bacterial surface are probably not involved as their elimination by neuraminidase did not affect the binding of both MAbs. The results of the present study strongly validate our previous observations that direct specific interaction of B. gingivalis with human PMNs occurs, resulting in the suppression of PMNs.
The Biological Response Modifier OK-432 (a Streptococcal Preparation) Inhibits the Development of Autoimmune Kidney Disease in NZB/W F1 Hybrid Mice: Possible Involvement of Tumor Necrosis Factor
Authors: Mihara Masahiko , Ohsugi Yoshiyuki
Published: August 2009

OK-432 (a streptococcal preparation) has been widely used for cancer immunotherapy in Japan. It is a potent immunostimulator, activating macrophages and T lymphocytes, and increasing the production of TNF and several other cytokines in both humans and animals. In the present study, we evaluated the prophylactic effect of OK-432 on the development of autoimmune kidney disease in NZB/W F1 (BWF1) mice. The mice were given 0.5 or 2.0 KE (‘klinische Einheit’; clinical unit) of OK-432 intraperitoneally every week from 21 weeks of age to the time of death. The control group received the same volume of saline (vehicle). OK-432 delayed the development of proteinuria and prolonged the survival of these mice dose dependently. At 49 weeks, 33.3% of control mice were alive, whereas 55.6% in the 0.5-KE- and 75% in the 2.0-KE-treated mice were alive. In the control group, the serum cholesterol level increased due to the development of glomerulonephntis. In contrast, mice treated with OK-432 had significantly lower levels of serum cholesterol. The serum levels of anti-DNA and anti-TNP antibodies were not affected by OK-432 administration. OK-432 induced the production of tumor necrosis factor (TNF)-α in the peritoneal fluid in the BWF1 mice. These results indicate that the effect of OK-432 in preventing the development of autoimmune disease in the mice may result from the stimulation of the endogenous TNF-α production.
Intrathyroidal Mast Cells Express Major Histocompatibility Complex Class-II Antigens
Authors: Banovac K. , Ghandur-Mnaymneh Latifa , Leone J. , Neylan D. , Rabinovitch A.
Published: August 2009

We studied the expression of major histocompatibility complex (MHC) class-II antigen in mast cells of rat thyroid glands. In the normal rat thyroid, mast cells express MHC class-II antigen. In BioBreeding Wistar (BB/W) rats, the strain of animals prone to develop spontaneous autoimmune thyroiditis, the number of MHC class-II-positive mast cells was significantly higher than in normal rats (p <0.01). Furthermore, intrathyroidal mast cells in BB/W rats showed an increased MHC class-II expression before the appearance of circulating thyroid antibodies or the infiltration of tissue with mononuclear cells. These data suggest that mast cells in the rat thyroid gland may have a function as antigen-presenting cells.
Human Antibodies against Trypanosoma cruzi Exoantigens Recognizing Parasite Surface Antigens and Heart Tissue Components
Authors: Gruppi A. , Gea S. , Moretti E.R.A. , Vottero-Cima Elsa
Published: August 2009

This work describes the occurrence of antibodies against exoantigens of Trypanosoma cruzi in a high percentage of patients with chronic Chagas’ disease. When analyzing selected groups of patients, it was observed that sera from individuals with electrocardiographic alterations showed a greater number of precipitin systems and higher antibody titers than sera from patients with positive serology only. Partial characterization of exoantigens of T. cruzi was performed by isoelectric focusing and immunoblotting. Two distinct groups of antigens at pi around 5 and 6, respectively, were identified by these methods. It was also shown in absorption experiments that exoantigens of lower pi share epitopes with components of the cellular surface of epimastigotes of T. cruzi, whereas exoantigens of higher pi share epitopes with normal human heart tissue.
Immunoregulatory Effect of Antibody on Delayed-Type Hypersensitivity in Mice
Authors: Morikawa Y. , Kuribayashi K. , Saito K.
Published: August 2009

We analyzed the conditions for in vivo toleration of delayed-type hypersensitivity (DTH). The intravenous injection of a high dose of keyhole-limpet hemocyanin (KLH) into BALB/c mice did not induce DTH in vivo, but the serum titers of the anti-KLH antibody were significantly elevated. This lack of DTH response was antigen-specific, and the intravenous injection of the antigen induced effector-phase suppressor T cells. The findings suggested that the lack of a DTH response brought about by the intravenous injection of a high dose of antigen was not immunological tolerance. Treatment with a high dose (250 mg/kg) of cyclophosphamide – but not a low dose (50 mg/kg) – enhanced the DTH, but suppressed antibody production. These results indicate that humoral immune responses participate in the regulation of DTH. In addition, the transfer of serum or immunoglobulin from mice that were injected intravenously with a high dose of the antigen suppressed the DTH. We concluded that the antibodies regulate DTH in the antigen-specific manner.
Anti-Asthmatic Activity of Newly Synthesized Calcium Antagonists: 2-n-Butyl-1{N-methyl-N-[2-(N′, N′-dimethylamino)ethyl]amino}-5,6-methylenedioxy-Indene and -Indane
Authors: Nagai Hiroichi , Yakuo Ikuhisa , Arimura Akinori , Choi Soo Hyung , Inagaki Naoki , Koda Akihide
Published: August 2009

The anti-asthmatic activity of the newly synthesized methylenedioxyindene (MDI) derivatives, 2-n-butyl-1{N-methyl-N-[2-(N′, N′-dimethylamino)ethyl]amino}-5,6-methylenedioxy-indene (MDI-C) and -indane (MDI-D), were investigated in vitro and in vivo in guinea pigs. The in vitro pharmacological activity of both derivatives was compared to that of 8-(diethylamino)octyl-3,4,5-trimethyoxybenzoate hydrochloride. All three agents caused a concentration-dependent inhibition of the antigen-induced contraction of sensitized guinea pig tracheal smooth muscle. In histamine and leukotriene D4-induced contractions of guinea pig tracheal smooth muscle, each agent showed clear antagonistic actions. Additionally, all three agents demonstrated potent calcium antagonistic actions via inhibition of guinea pig tracheal smooth muscle contractions caused by CaCl2 in a high potassium and Ca-free medium and by compound 48/80 in a solution of ethylene glycol bis(β-aminoethylether)-N, N, N′, N′-tetraacetic acid in a contained Ca-free medium. MDI-C and MDI-D inhibited the antigen-induced release of histamine and slow reacting substance of anaphylaxis from sensitized guinea pig lung tissue. Lastly, both MDIs clearly inhibited asthmatic respiratory disorders without affecting blood pressure in guinea pigs.
Antigen-Specific T-Cell Factors Induce Isotype-Like Suppression of Mast Cell and Eosinophil-Rich T-Cell-Dependent Inflammation in the Intestine of Mice Infected with Trichinella spiralis
Authors: Parmentier H.K. , Dijkstra J.W. , Wissink A. , Ruitenberg E.J. , Askenase P.W. , van Loveren H.
Published: August 2009

The recent identification of a T-cell-derived antigen-binding molecule (TABM), Trichinella spiralis factor (Tric-F), isolated from culture supernatants of lymphoid cells from mice infected with the intestinal helminth T. spiralis, has led to investigation of the ability of Tric-F to induce a T-cell-dependent feedback circuit that ultimately suppresses the production of other TABMs with similar (isotype-like) features. This form of regulation that has been identified in contact hypersensitivity and in delayed-type hypersensitivity (DTH) responses to tumor cells, was shown not to be antigen-specific but to be DTH-specific. Injection of mice with the TABM called picryl chloride factor (PCl-F) induced suppression of the production of DTH-initiating TABMs of other antigenic specificities. In this study, we report that intravenous injection of mice with Tric-F or PC1-F, 8 days before an oral infection with T. spiralis, induced suppressor cells that inhibited the T-cell-dependent influx into the gut of inflammatory cells, comprising mast cells and eosinophils. Similar results were obtained when the mice were skin sensitized with PC1 8 days prior to a T. spiralis infection, i.e. in a system where TABMs are known to be produced. The phenotype of these suppressor cells was Lyt-1––2+. This suppression preferentially affected the parasite-induced DTH-like response in the gut. In contrast, increased levels of IgA plasma cells in the gut, and worm expulsion were not affected by these treatments. In reciprocal experiments, intravenous injection of Tric-F, or PC1-F, or an oral infection with T. spiralis (that results in the production of TABMs) given 8 days before contact sensitizing mice with PC1, resulted in a suppression of elicitation of cutaneous DTH, as measured by ear swelling. In contrast, pretreatment with anti-dinitrophenyl IgE antibody did not interfere with intestinal inflammation to T. spiralis nor with DTH to PC1. Our results suggest that similar to cutaneous DTH, T. spiralis-specific T-cell factors are involved in the initiation and regulation of the DTH-like mast cell and eosinophil-rich intestinal inflammation that accompanies T. spiralis infections in the gut. Since both Tric-F and PCl-F induce suppression of cellular immune responses in vivo, independent of antigen specificity, it is concluded that Tric-F belongs to the same isotype of TABMs as PC1-F that therefore can be regulated by a non-antigen-specific, isotype-like, T-cell-dependent feedback mechanism.
Immunological Studies of the Glycoprotein Allergen Ag-54 (Cla h II) in Cladosporium herbarum with Special Attention to the Carbohydrate and Protein Moieties
Authors: Swärd-Nordmo Marit , Paulsen Berit Smestad , Wold Jens Kristian
Published: August 2009

The glycoprotein allergen Ag-54 (Cla h II) isolated from Cladosporium herbarum was split into its carbohydrate and protein moieties by using alkaline-borohydride treatment and deglycosylation, respectively. The native Ag-54, the deglycosylated protein and the protein-free carbohydrate moieties were tested for IgE-binding activity by radioallergosorbent test inhibition using selected sera from individuals with C. herbarum allergy. The deglycosylated material had a stronger allergenic activity than the native Ag-54 with all the sera tested. The Ag-54 carbohydrate moieties were not found to possess any IgE-binding activity. The galactoglucomannan part of the carbohydrate moiety was isolated and it was demonstrated to precipitate IgG rabbit antibody. Removal of the galactofuranose units by mild acid hydrolysis did not alter the IgG-binding capacity of the polysaccharide, indicating that galactofuranose was not immunodominant.
Measurement of Allergens Associated with Dust Mite Allergy
Authors: Yasueda Hiroshi , Mita Haruhisa , Yui Yasuo , Shida Takao
Published: August 2009

Sensitive radioimmunoassays were developed to measure the two groups of major allergens (Der I and Der II) of Dermatophagoides pteronyssinus and Dermatophagoides farinae, in which radiolabeled protein A served as a general tracer. Glass rods covalently coupled with F(ab′)2 fragments of rabbit antiserum IgG were incubated first with mite or house dust extracts, and then with affinity-purified rabbit antibodies. The bound allergen-antibody complex was detected with a 125I-labeled protein A. The two Der I allergens, Der p I and Der f I, were measured separately with the use of immunoabsorbed rabbit antibodies directed against species-specific determinants, while the two Der II allergens, Der p II and Der f II, were measured as ‘Der II’ with the use of antibodies directed against common determinants on both allergens. Each assay demonstrated consistently parallel dilution curves with mite and house dust extracts. The mean intraassay and interassay coefficient of variation for each assay ranged from 4.3 to 7.7% and from 3.6 to 9.0%, respectively. The Der p I and Der f I assays were shown to be highly species-specific so that the ratio of Der p l:Der f I would provide a good index of the distribution of the two mite species in a dust sample. The concentrations of the Der I and Der II allergens in different types of mite extract and dust samples from houses were compared using these assays. The results demonstrated that the ratio of Der I: Der II in the mite body extracts varied markedly from those in the whole culture extracts, while the ratio in individual dust samples was nearly constant and was close to that in the mite body extracts. These assays will be useful in standardizing mite and house dust extracts and for assessments of mite allergen exposure. Furthermore, the high assay sensitivity will make it possible to use these assays to measure mite allergens associated with airborne particles.
Measurement of Allergens Associated with Dust Mite Allergy
Authors: Sakaguchi Masahiro , Inouye Sakae , Yasueda Hiroshi , Irie Tatehisa , Yoshizawa Susumu , Shida Takao
Published: August 2009

Assays of mite allergens (Der p I, Der f l and Der II) in the air of houses became feasible with the use of a low-noise air sampler and a sensitive radioimmunoassay described previously. The levels of the airborne allergens Der I (Der p I + Der f l)and Der II in the living room of 10 houses during usual domestic life were very low, 29.5 and 6.3 pg/m3, respectively, with a Der I: Der II ratio of 4.7:1. At the time of bedmaking, they greatly increased, about 1,000-fold, to 30,900 and 12,600 pg/m3, respectively, with a Der I: Der II ratio of 2.5:1. The amounts of Der I and Der II in the floor dust of the living room were 2,040 and 2,690 ng/g of fine dust, respectively, with a Der I: Der II ratio of 0.8:1. Der I seemed more prone to become airborne than Der II.
Cyclic AMP Inhibits Chemotactic-Peptide-Induced but not Ca2+-Ionophore- or Tetradecanoylphorbol-Acetate-Induced Enzyme Secretion in Guinea Pig Neutrophils
Authors: Wada Yasutaka
Published: August 2009

cAMP-increasing agents such as prostaglandin E1, dibutyryl cAMP and 8-bromo-cAMP inhibited N-acetyl-β-D-glucosaminidase secretion induced by the chemotactic peptide formylmethionyl-leucyl-phenylalanine in guinea pig neutrophils. However, they could not inhibit but rather potentiated the secretion significantly when secretion was induced by the Ca2+ ionophore A23187. Similarly, these agents did not inhibit tetradecanoylphorbol acetate (TPA)-induced secretion. Moreover, the secretion induced by combined treatment with Ca2+ ionophore A23187 and TPA was hardly blocked by cAMP-increasing agents. These results strongly suggest that cAMP inhibits the secretion through suppression of a signal transduction from receptor activation to events such as Ca2+ mobilization and protein kinase C activation, probably of the inositol phospholipid metabolism.
Plasma Neuropeptide Pattern in Acute Idiopathic Urticaria
Authors: Hernanz A. , Muelas G. , Borbujo J.
Published: August 2009

Immediate hypersensitivity responses, as acute urticaria, produce a release of neuropeptides by nerve endings, which present specificity of recognition by mast cells, basophils and other target cells. We have measured vasoactive intestinal peptide (VIP), somatostatin, bombesin, neurotensin and β-endorphin by radio-immunoassay in plasma extracts of 20 patients with acute idiopathic urticaria and of 20 healthy subjects. VIP-and β-endorphin-like immunoreactivities were found to be significantly decreased with respect to controls (p < 0.001 and p < 0.01, respectively). On the contrary, somatostatin- and bombesin-like immunoreactivities were significantly increased (p < 0.001 and p < 0.05, respectively). These findings could be a reflection in blood of a raised release of somatostatin and bombesin by nerve endings in the urticaria process. Moreover, the decreased plasma levels of VIP- and β-endorphin-like immunoreactivities could be explained by a raised specific metabolism of these peptides in the urticaria process.
Enhanced Leukotriene C4 Production by Peripheral Eosinophilic Granulocytes from Children with Asthma
Authors: Schauer U. , Eckhart A. , Müller R. , Gemsa D. , Rieger C.H.L.
Published: August 2009

Granulocytes and mononuclear cells were isolated from the blood of asthmatic and healthy children. Stimulation with ionophore A 23187 induced a significantly higher leukotriene C4 (LTC4) generation from granulocytes of asthmatic children than from granulocytes of healthy controls. In contrast, mononuclear cells from patients and controls did not differ in their ability to produce LTC4. Additional analysis showed that the difference in LTC4 generation of granulocytes was due to increased formation but not to decreased oxidative degradation of LTC4. Analysis of LTC4 generation of purified neutrophils and eosinophils revealed that LTC4 was generated almost exclusively by eosinophils and, in particular, the hypodense population. Granulocytes from patients with a history of severe asthma displayed a higher LTC4 formation than granulocytes from patients with less severe disease.
Polymorphonuclear Leukocyte Stimulation Measured by Phage Inactivation
Authors: Ferrini U. , Mileo A.M. , Nista A. , Mattei E. , Orofino A.
Published: August 2009

A quantitative assay for determining the polymorphonuclear neutrophil (PMN) stimulation is described. The method is based on the inactivation of lambda vector phages that occurs after a brief exposure to stimulated PMNs. The determination of the number of residual plaque-forming units on the appropriate bacterial host allows a reproducible and sensitive quantitative assay for measuring the stimulation level of the PMN. In comparison with other methods that employ bacteria or eukaryotic cells, this assay provides several advantages and can be used for investigating the biochemical and physiological processes responsible for PMN stimulation.
Investigation of the Allergenic Activity of Isolated Fractions of a Standardized Partly Purified Whole Mite Body (Dermatophagoides pteronyssinus) Extract
Authors: Wahl R. , Meineke D. , Maasch H.J. , Schultze-Werninghaus G.
Published: August 2009

Twelve fractions of a molecular weight range of 1.35–670.00 kilodaltons (kD) were isolated from abiologically standardized partly purified whole mite body extract (Dermatophagoidespteronyssinus) by preparative size exclusion high-performance liquid chromatography. The allergenic activity and the antigen and allergen patterns of the isolated fractions were investigated by RAST, RAST inhibition and crossed (radio)immunoelectrophoresis (CIE/CRIE). By CRIE, each of the fractions showed allergen patterns, mostly of different compositions. Each fraction showed allergenic activity of different degrees by RAST inhibition. The highest allergenic activity could be measured by RAST inhibition with fractions which contained predominantlythe major allergens Der p I and PY as detected by CRIE. Also proteins of higher molecular weights (> 158 kD) showed IgE-binding capacities. Nearly all antigens detected by CIE could be identified as allergens using CRIE.
Identification and Partial Characterization of a T-Cell-Derived Antigen-Binding Factor from Mice Infected with the Intestinal Helminth Trichinella spiralis
Authors: Parmentier H.K. , Dijkstra J.W. , Wissink A. , Ruitenberg E.J. , Askenase P.W. , Van Loveren H.
Published: August 2009

Immunochemical and biological characterization was performed of an antigen-binding factor derived from culture supernatants of T cells from mice infected 4 days previously with the intestinal helminth Trichinella spiralis. Affinity chromatography with T. spiralis antigen resulted in the purification of a protein, provisionally designated Trichinella factor (Tric-F), that shared antigenic and other properties with a known T-cell-derived antigen-binding factor of different antigenic specificity, picryl chloride factor, which mediates an early 2-hour component of contact sensitivity. Tric-F lacked determinants of immunoglobulins and possessed determinants shared by other antigen-specific T cell factors, as determined by ELISA and antibody affinity chromatography. Biological activity of Tric-F was assayed in vivo and in vitro. Mice injected intravenously with Tric-F developed an antigen-specific early 2-hour ear swelling response following local challenge with T. spiralis antigen. These results corresponded to delayed-type hypersensitivity responses in the ears of T. spiralis-infected mice that comprised early 2-hour and late classical 24-hour responses. In vitro, Tric-F induced serotonin release by mast cells in the presence of T. spiralis antigen. Mast cells sensitized with Tric-F formed rosettes with antigen-coated sheep erythrocytes. It is suggested that Tric-F, an antigen-binding molecule that is T-cell-derived, mediates the early 2-hour component of delayed-type hypersensitivity and is involved in the initiation and regulation of T-cell-mediated intestinal inflammation during a T. spiralis infection in mice.
Direct Modulation of Human Neutrophil Behaviour by Candida albicans
Authors: Vuddhakul V. , Seow W.K. , McCormack J.G. , Thong Y.H.
Published: August 2009

We examined the direct effect of unopsonized yeast particles of Candida albicans on two aspects of neutrophil behaviour, namely adherence and 3H-deoxyglucose uptake. The data show that brief exposure of C. albicans to human neutrophils resulted in decreased ability of the neutrophils to adhere to Dacron fibre and take up deoxyglucose. This inhibitory effect was further shown to be dependent on yeast concentration and on the integrity of the yeast cell wall. Additional experiments indicate that this effect was direct rather than indirect through soluble mediators in the supernatant. Interference experiments with glucan and mannan, the two major polysaccharide components of the yeast cell wall, suggest that the ligand in direct interaction between C. albicans and neutrophil contains glucan. Finally, it was shown that nonpathogenic species of Candida such as C. krusei, C. parapsilosis and C. guilliermondii did not display neutrophil-modulatory properties while the occasionally pathogenic C. tropicalis did. These results indicate that C. albicans has the ability to circumvent neutrophil defence mechanisms, and may in part explain the propensity of this fungus to cause infection.
Anti-IgE- and Complement-Induced Histamine Release from Peripheral Leukocytes in Normals and Atopics
Authors: Stephan V. , Kühr J. , Urbanek R.
Published: August 2009

Anti-IgE- and complement-induced histamine release from isolated basophils was compared in normal and atopic subjects. Activated complement was generated by incubation of a serum pool from normal donors with zymosan. Histamine was measured by a single-isotope radioenzymatic assay. In atopies, the mean histamine release induced by heterologous complement was significantly higher than in normals (p ≤0.01). Anti-IgE-mediated histamine release showed no differences between atopies and normals. Regarding the histamine release as a function of the age of the donor, we observed significantly higher complement-induced release values in normal children aged 1–10 years compared to those aged 10–20 and >20 years (p ≤0.05). Anti-IgE-induced histamine release showed no age-related differences in normals or atopic patients. As a high complement-mediated leukocytic histamine release was more commonly found in atopies with heterologous complement, this phenomenon appears not to be due to a higher rate of complement activation in the serum of atopies, but rather to an intrinsic cell abnormality. Moreover, these results suggest that age-related releasability patterns might be considered when studying complement-induced histamine release. Differences between IgE-and complement-mediated histamine release indicated that parameters of releasability must be defined with respect to each stimulus.
Passive Air-Pouch Anaphylaxis in Rats
Authors: Konno-Amano Satoshi , Hirao Tetsuji , Kohno Hiroyuki , Mori Hiroshi , Sato Yoshihisa
Published: August 2009

Induction of an experimental passive anaphylaxis of the air-pouch type, passive air-pouch anaphylaxis, was carried out in an attempt to induce a reproducible anaphylaxis model suitable for quantitative studies. Rats were injected subcutaneously with 10 ml of air into the dorsal skin to make an air-pouch and with 2 ml of antiserum at an appropriate dilution for passive sensitization, and then 5 ml of air was removed. The challenge with 5 ml of antigen solution into the air-pouch 48 h later provoked mast cell degranulation and increased vascular permeability induced by released histamine. Treatment with monovalent hapten prior to the antigen challenge almost completely inhibited histamine release and plasma exudation to levels similar to those in the nonsensitized group. In this model, mast cell-dependent late-phase allergic reaction, such as leukocyte migration or the increase of plasma exudation following mast cell degranulation, was not observed.
Involvement of Tumor Necrosis Factor in Human Granulocyte-Mediated Killing of WEHI 164 Cells
Authors: Mándi Yvette , Degré M. , Béládi I.
Published: August 2009

Human polymorphonuclear leukocytes (PMNLs) kill WEHI 164 clone 13 cells in an 18-hour 51Cr release assay. Antibody to human tumor necrosis factor (TNF) blocks the lysis of targets mediated by human granulocytes. PMNLs triggered by sensitive targets not only displayed cytotoxic activity, but also released a soluble factor capable of selectively lysing WEHI 164 cells. The killing of these cells by supernatants of triggered granulocytes was totally inhibited by anti-TNF antibody. These experiments suggest that the killing of WEHI 164 sarcoma cells by human PMNLs involves TNF or TNF-like molecules.
Regulatory Role of Cytokines and CD23 in the Human IgE Antibody Synthesis
Authors: Pène Jérôme
Published: August 2009

IL-4 specifically induces IgE antibody production by mononuclear cells (MNC) from healthy nonatopic donors. At least part of the effect of IL-4 is indirect since IgE-induced synthesis also requires CD4+ T cells and, to a lesser extent, monocytes. The IL-4-mediated IgE synthesis is enhanced by IL-2, IL-5 and IL-6 and suppressed by IFN-γ, IFN-α and PGE2. In vitro IFN-γ plays a central role in the down-regulation of IL-4-induced IgE synthesis. Interestingly, studies performed with atopic donors indicated that IL-4 and IFN-γ may also be operational in vivo. Moreover, IL-4 inhibits the IFN-γ synthesis, which indicates that these two cytokines regulate IgE synthesis by inhibiting each other’s activities and synthesis. In addition, it was demonstrated that IgE production is associated with the expression of the FcεRII/CD23 antigen and its soluble part (sCD23). However, the expression of CD23 and the release of sCD23 does not strictly correlate with the production of IgE since also highly purified B cells express CD23 to the same extent as MNC but without producing IgE. Production of IgE can only be achieved by addition of CD4+ T cells, suggesting that sCD23 also acts indirectly on B cells and may have a possible activity on T cells. The addition of sCD23 to MNC does not induce IgE production in the absence of IL-4. These data indicate that the release of sCD23 in itself, even in the presence of T cells, is not sufficient for triggering the B cells to produce IgE antibodies. In conclusion, our findings demonstrate that IL-4-induced IgE production in human involves complex interactions between B cells and at least T cells and monocytes and is positively modulated by IL-2, IL-5, IL-6 and sCD23 and down-regulated by IFN-γ, IFN-α, and PGE2.
Low-Affinity Receptor for IgE (FcERII, CD23) and Its Soluble Fragments
Authors: Delespesse G. , Hofstetter H. , Sarfati M.
Published: August 2009

The low-affinity receptor for IgE (FcERII or CD23) is a membrane 45-kD glycoprotein which is cleaved by an autoproteolytic mechanism into soluble 37-, 33- and 25-kD fragments that are capable of binding to IgE (IgE-binding factors, IgE-BFs). FcERIIa (which is expressed only on fresh B cells) differs from FcE-RIIb (which is expressed on IL4-stimulated B cells, monocytes, eosinophils and T cells) by a few intracytoplas-mic amino acids. The only function of FcERII which is clearly demonstrated is the IgE-dependent cytotoxicity of FcERIIb on monocytes and eosinophils. We here review our recent observations indicating that 37-kD IgE-BFs regulate the synthesis of human IgE. Recombinant 37-kD IgE-BFs increase the IL4-induced synthesis of IgE by peripheral blood lymphocytes, as well as the IL4-independent, ongoing synthesis of IgE by either in vivo activated B cells from allergic patients or by in vitro IL4-preactivated B cells.
Establishment of a Memory in vitro Murine IgE Response to Benzylpenicillin and its Resistance to Suppression by Anti-lL-4 Antibody
Authors: Heusser Ch.H. , Wagner K. , Brinkmann V. , Severinson E. , Blaser K.
Published: August 2009

Regulation of a memory IgE antibody response may be different from the induction of a primary response and may, therefore, be more relevant to the study of allergic diseases and the therapeutic manipulation of IgE antibody formation. In this paper a murine hapten-specific in vitro memory IgE antibody response to benzylpenicilloyl(BPO)-KLH is described. The response was analyzed by determining the number of antibody-producing cells (APC) in an ELISA spot assay. Of the total number of BPO-specific APC (10,000 APC/ 106 cultured spleen cells), about 1 % were IgE-producing cells (100/106 cultured cells), as detected on day 6 of culture. The level of the antibody response is antigen dose-dependent, and the detected APC are BPO specific. The memory IgE response is not inhibited by the addition of anti-IL-4 antibody (11B11), even at a high excess. In the presence of the mitogen lipopolysaccharide, it has been shown that switch of B cells to IgE is induced by IL-4, a process which can be inhibited by anti-IL-4 antibody. Because the antigen-induced IgE response cannot be inhibited by anti-IL-4 antibody, in vitro responding cells derived from BPO-KLH-preimmunized mice may, therefore, have already switched in vivo to IgE. On the other hand, B cells switching to IgE in a situation of cognate T-B cell interaction might receive IL-4 in a transsynaptical way from T cells which might not be accessible to inhibition by anti-IL-4 antibody. The identification of the two possibilities in situations of established allergic disorders will be decisive for determining whether pharmacological inhibition of IL-4 (or IL-4-induced switch) – e.g., by putative low molecular weight compounds – will ever be a meaningful approach to suppress allergic diseases.
Studies of Chemotactic, Chemotactic Movement-Inhibiting and Random Movement-Inhibiting Effects of Interleukin-1 Alpha and Beta, Tumor Necrosis Factors Alpha and Beta and Interferon Gamma on Human Neutrophils in Assays Using ‘Sparse-Pore’ Polycarbonate (Nuclepore) Membranes in the Boyden Chamber
Authors: Bignold Leon P. , Ferrante Antonio , Haynes David R.
Published: August 2009

Interleukin-1 alpha and beta (IL-1α and β), tumour necrosis factors alpha and beta (TNFα and β) and interferon gamma (IFNγ) were tested for their chemotactic effects, their effects on chemotactic movement towards N-formyl-methionyl-leucyl-phenylalanine (FMLP) and their effects on random locomotion of human peripheral blood neutrophils through polycarbonate membranes in Boyden-type chambers. Both IL-1α and β, but no other cytokine tested were chemotactic for neutrophils using ‘sparse-pore’ polycarbonate membrane. Both TNFs, but no other cytokine, inhibited neutrophil chemotactic movement towards FMLP using the same membrane. No cytokine influenced random migration of neutrophils through polycarbonate membrane of standard pore density. These results suggest that IL-1 may have a role as a chemotactic mediator of inflammation, but that TNFs may inhibit chemotactic migration of neutrophils into inflammatory lesions.
Endogenous Histamine in Immune Inflammation in 6-Day-Old Air Pouch of Facsimile Synovium
Authors: Sin Y.M. , Ang L.N.
Published: August 2009

Immune inflammation was induced by injecting bovine serum albumin (BSA) into 6-day-old air pouches of mice presensitized with 2 weekly injections of an emulsion containing BSA and complete Freund’s adjuvant. Control mice were also similarly pretreated with the same emulsion without BSA. The results show that the numbers of exudate leucocytes in the air pouches of both test and control groups increased and peaked at 4 h and then declined after the antigen challenge. However, the values of exudate leucocytes in the test animals at 4- and 24-hour intervals were significantly lower than those of the control. On the other hand, exudate histamine of the test group peaked at 1 h, and this was significantly higher than that of the control. Injection of exogenous histamine or histaminase with the challenging antigen increased the number of exudate leucocytes in both test and control animals. The findings thus suggest that endogenous histamine released in immune inflammation most probably plays the same role as in non-immune inflammation by enhancing the vascular permeability at the inflammatory site in the early phase of the inflammatory reaction.
Immunogenicity of Amodiaquine in the Rat
Authors: Clarke J.B. , Maggs J.L. , Kitteringham N.R. , Park B.K.
Published: August 2009

Amodiaquine is an antimalarial drug that has been associated with adverse reactions which may be immune mediated. Specific IgG anti-amodiaquine antibodies were detected after administration of the drug to rats (269 μmol/kg for 4 days), using an enzyme-linked immunosorbent assay employing amodiaquine conjugated to metallothionein as an antigen. A positive immune response was observed regardless of the route of administration, but the magnitude of the response in terms of antibody titre was in the order intraperitoneal administration > intramuscular administration > oral administration. Hapten inhibition experiments with structurally related drugs defined the specificity of the antibody which appears to recognize a conjugate of amodiaquine quinone imine and cysteine residues present in protein. Amodiaquine was converted to a protein-reactive species by activated human polymorphonuclear leucocytes in vitro, and this may provide a mechanism for immunogen formation in vivo. A humoral immune response was observed with doses of amodiaquine that did not produce either direct hepatotoxicity or leucopenia. Thus an animal model has been developed with which to investigate the toxicological consequences of amodiaquine immunogenicity.