Shida T

Isolation and partial characterization of the major allergen from Japanese cedar (Cryptomeria japonica) pollen

A purified allergen, antigen SBP, was obtained from the pollen of the Japanese cedar (Cryptomeria japonica, Sugi in Japanese) by ammonium sulfate precipitation, ion-exchange chromatography on diethylaminoethyl and carboxymethyl cellulose, and gel chromatography on Sephadex G-150. Antigen SBP was a heat-sensitive basic glycoprotein of approximately 40,000 molecular weight. By preparative isoelectric focusing and gel chromatography on Sephadex G-100, antigen SBP could be further separated into four subfractions, differing in both isoelectric point and molecular weight. By immunodiffusion analysis, direct skin testing, and radioallergosorbent test inhibition, it was shown that antigen SBP was the major allergen of Japanese cedar pollen, and the four subfractions were seen to be antigenically and allergenically identical.

Allergenicity of acid protease secreted by Candida albicans

We have previously reported the cases of Candida albicans (C. alb) acid protease (CAAP)‐induced atopic asthma. In this study, the allergenicity of the released enzyme CAAP was examined among asthmatic patients with positive immediate skin response to crude C. alb antigen. Among 49 patients with positive skin response to crude C. alb, anti‐crude C. alb IgE antibodies were detected in 40 and anti‐CAAP IgE antibodies were detected in 18. Moreover, anticrude C. alb IgE antibodies were detected in all of the patients in whom anti‐CAAP IgE antibodies were detected. No correlations between IgG antibodies to both antigens or between IgE and IgG antibodies to CAAP were observed. CAAP induced significant T‐cell proliferation in 20/28 patients showing positive T‐cell proliferation response to crude C. alb antigen. Most of the patients showing positive conjunctival response to crude C. alb antigen also showed positive response to CAAP. Most of the patients showing high levels of serum IgE antibody and positive histamine‐release response of peripheral blood leukocytes to CAAP showed positive conjunctival response. The results indicate that CAAP is an important allergen in C. alb‐related mucosal allergy.

Anti-Allergic Activity of Formoterol, a New Beta-Adrenoceptor Stimulant, and Salbutamol in Human Leukocytes and Human Lung Tissue

Formoterol and salbutamol were compared for in vitro inhibition of allergen‐induced histamine release from allergic leukocytes and human lung tissue passively sensitized with allergic serum. Formoterol inhibited the release of histamine from leukocytes but salbutamol showed little inhibiting effect. When combined with theophylline, formoterol was a more potent inhibitor of the release of histamine from leukocytes in allergic patients than salbutamol. In fragments of human lung sensitized with allergic serum, the concentration required to inhibit histamine release by 50% was 2 × 10−11 M for formoterol and 8.5 × 10−9 M for salbutamol. The potency of salbutamol and formoterol in blocking specific 3 H‐dihydroalprenolol binding to beta‐adrenoceptors on guinea pig lung membranes revealed that formoterol had higher affinity for beta‐adrenoceptors than salbutamol, and the concentration required for half‐maximum stimulation of adenylate cyclase was approximately 200‐fold higher for salbutamol than for formoterol.

Effect of AA‐861, a 5‐Lipoxygenase Inhibitor, on Leukotriene Synthesis in Human Polymorphonuclear Leukocytes and on Cyclooxygenase and 12‐Lipoxygenase Activities in Human Platelets

AA‐861, a selective inhibitor of 5‐lipoxygenase of arachidonic acid, was tested for ability to inhibit leukotriene C4 and leukotriene B4 synthesis in human polymorphonuclear leukocytes after calcium ionophore stimulation. AA‐861 dose‐dependently inhibited leukotriene B4 and leukotriene C4 generation in human polymorphonuclear leukocytes; the concentration required to inhibit generation by 50 % (IC50) was 3 × 10−7 M for leukotriene B4 and 1 × 10−8 M for leukotriene C4. BW‐755C inhibited the generation of leukotriene C4 with an IC50 of about 10−5 M, indicating that AA‐861 is about 1000 times more potent than BW‐755C. AA‐861 did not affect the activity‐ of either cyclooxygenase or 12‐lipoxygenase at a concentration up to 10−5 M in human platelets. AA‐861 did not inhibit histamine release from human basophils. These results indicate that AA‐861 selectively inhibits 5‐lipoxygenase but not cyclooxygenase or 12‐lipoxygenase in human specimens.

Changes of Alpha1- and Beta-Adrenergic and Cholinergic Muscarinic Receptors in Guinea Pig Lung Sensitized with Ovalbumin

The effect of immunization of guinea pigs with ovalbumin on the number and affinity of alpha 1- and beta-adrenergic and cholinergic muscarinic receptors was studied in lung membranes by direct binding techniques using 3H-prazosin, 1-3H-dihydroalprenolol and 1-3H-quinuclidinyl benzilate. After immunization by intraperitoneal injection of ovalbumin to guinea pigs, the number and affinity of each receptor in sensitized animals were not significantly different from those of control animals. Sensitization of guinea pigs by an aerosol exposure with the antigen resulted in a decreased number of beta-adrenergic receptor (458 +/- 29 vs. 687 +/- 56 fmol/mg protein; p less than 0.01), and an increased number of alpha 1-adrenergic receptor (36 +/- 2 vs. 25 +/- 2 fmol/mg protein; p less than 0.01), but no change was observed in the number of muscarinic receptors, as compared with control animals. On the other hand, following a prolonged sensitization of guinea pigs with a low dose of the aerosolized antigen, the number of muscarinic receptors was significantly increased in the lung of sensitized animals (50 +/- 2 vs. 42 +/- 2 fmol/mg protein; p less than 0.01); however, we found no significant differences between sensitized and normal animals in the number of alpha 1- and beta-adrenergic receptors. There were no different changes in the affinity of these receptors in all experiments.

An attempt to produce an antibody of histamine and histamine derivatives

AbstractThree conjugates of different histamine derivatives to bovine serum albumin were prepared, and an attempt was made to determine whether antibody against histamine and histamine derivatives, could be produced in rabbits by immunization with these conjugates.1.Antibody produced by immunization with bovine serum albumin-succinylhistamine conjugate could not recognize the hapten moiety of the immunogen.2.Immunization of rabbits with conjugate of bovine serum albumin andp-[2-(Nα-trifluoroacetyl-histamine)azo]benzoic acid resulted in the production of antibody toNα-trifluoroacetylhistamine. The antibody had a specificity for histamine metabolites and their derivatives. However, the antibody also showed a small but not negligible affinity for trifluoroacetylated serotonin, norepinephrine and aniline, indicating that the antibody specificity was directed mainly to the trifluoroacetamide group.3.After immunization of rabbits with bovine serum albumin andp-[2-(Nα-propionylhistamine)azo]benzoic acid conjugate, antibody againstNα-propionylhistamine could be produced. However, the antibody had a very low affinity even forNα-propionylhistamine.nSpecific antibody to histamine and histamine derivatives could thus not be produced by immunization with the present hapten-carrier conjugates.

Enhancement of IgE synthesis and histamine release by T cell factors derived from atopic patients with bronchial asthma

Culture supernatants of unstimulated T cells (TCS) derived from normal donors or from atopic patients with bronchial asthma were tested for their ability to regulate the spontaneous IgE synthesis by B cells of normal and atopic subjects. The same TCS were also tested for their influence on the histamine release from leukocytes of house dust mites-sensitive patients. Addition of TCS to B cell cultures from allergic donors induced a dose-dependent increase of the spontaneous IgE production without affecting the synthesis of IgG, IgM, and IgA. The potentiating activity of TCS was observed only in B cell cultures spontaneously producing IgE; TCS were still active on irradiated B cells. The maximal IgE-enhancing activity was observed when TCS were added at the onset of B cell cultures. The supernatants of T cells lysed at day 0 did not contain IgE-potentiating factors. The antigen-induced but not the spontaneous histamine release from leukocytes of house dust mite-sensitive patients was enhanced by pretreatment with TCS from allergic donors. The enhancing activities of TCS on IgE synthesis and on histamine release could be removed by absorption with IgE-Sepharose and subsequently recovered by elution with glycine buffer. The results indicate that T cells of patients with asthma spontaneously release IgE-binding factors capable of increasing both the spontaneous IgE synthesis by B cells and the antigen-induced histamine release.

Effect of Bordetella pertussis on α1and β-Adrenergic and Cholinergic Muscarinic Receptors in Guinea Pig Lung Membranes

After intraperitoneal injection of Bordetella pertussis vaccine to guinea pigs, the α 1 - and β -adrenergic and chol

Changes of Beta-Adrenergic Receptor Number and Catecholamine-Sensitive Adenylate Cyclase in Guinea Pig Lung after Inhalation of Histamine Aerosol

After guinea pigs were exposed to histamine or acetylcholine aerosol for 1 week, alpha 1- and beta-adrenergic and cholinergic muscarinic receptors on the lung membranes were measured by direct binding techniques using 3H-prazosin, l-3H-dihydroalprenolol, and l-3H-quinuclidinyl benzilate, respectively, and adenylate cyclase responses to l-isoproterenol (10(-5) to 10(-8) M) and NaF (20 mM) were also examined on the membranes. After the inhalation, the number of beta-adrenergic receptors was decreased by about 25% without a significant change in binding affinity for l-3H-dihydroalprenolol, while the number and the affinity of alpha 1-adrenergic and muscarinic receptors on the membranes were not significantly different from those in the control animals. When compared with the control animals, the lung membranes showed a reduced adenylate cyclase response to l-isoproterenol, and the response to 10(-5) M of l-isoproterenol was significantly decreased by 11% in the lung membranes. The histamine inhalation showed no significant effect on basal adenylate cyclase activity and adenylate cyclase response to NaF.

Anti-mite IgE Antibody Production in vitro by Peripheral Blood Lymphocytes from Mite-Sensitive Atopic Patients

Anti-mite IgE antibody production in vitro was investigated using peripheral blood lymphocytes (PBL) from mite-sensitive patients with bronchial asthma. Preculture of PBL with the mite antigen, which induced the remarkable cell proliferation, resulted in a marked decrease of the IgE antibody production directed to mite but not to unrelated antigens. Anti-mite IgE antibody formation was not inhibited by autologous or allogeneic T cells stimulated with the mite antigen. The spontaneous production of IgE antibody was observed when fractionated B cells were cultured alone or with pokeweed mitogen (PWM), but was suppressed when they were precultured with the antigen. Coculture of autologous T cells with B cells showed a considerable augmentation of anti-mite IgE antibody formation. The enhancing activity of autologous T cells was not observed when B cells were irradiated at 1,000 rad. The spontaneous IgE antibody production by B cells alone was radioresistant. Both mitotic inhibitors and 1,000 to 2,000 rad of gama-irradiation did not significantly suppress the spontaneous anti-mite IgE antibody production by B cells.