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Notes: The H1 antagonist loratadine has the capacity to inhibit histamine release from human basophils. The aim of this study was to investigate whether loratadine can also inhibit leukotriene C4 (LTC4) release from human leucocytes. Basophil-enriched mononuclear cell suspensions were prepared by centrifugation of peripheral venous blood (n= 10) on discontinuous Percoll gradients. Leucocytes were stimulated with anti-IgE, N-formylmethionyl-leucyl-phenylalanine (FMLP) and Ca2+ ionophore A23187; immunoreactive (i) LTC4 release in the cell supernatant was measured by a competitive radioimmunoassay and histamine release was evaluated by an automated fluorometric technique. Loratadine, in the concentration range of 1–50μM, exerted a dose-dependent inhibitory effect on IgE-mediated and IgE-independent histamine and iLTC4 release. The concentrations inhibiting 50% of histamine release were 30 μM (anti-IgE), 27μM (FMLP) and 19μM (Ca2+ ionophore A23187). The concentrations inhibiting 50% of iLTC4 release were 2–3 μM (anti-IgE). 11 μM (FMLP) and 1.7μM (Ca2+ ionophore A23187). The inhibitory activity on iLTC4 release was optimal after preincubation for 2h at 37°C, and was no longer evident when leucocytes were stimulated 2h after cell washing. Increased extracellular Ca2+ concentrations reduced the inhibitory activity of loratadine. These results indicate that loratadine has the capacity to inhibit the release of preformed and newly generated mediators from human basophil-enriched mononuclear cell suspensions.

Notes: Background Inflammatory alterations of respiratory airways have been found in patients with non-allergic asthma, but the triggering event has not been defined. An autoimmune activation of inflammatory cells has been hypothesized.Objective To evaluate whether histamine-releasing factors are present in sera from non-allergic asthmatics.Methods Twenty-four patients with non-allergic asthma underwent in vivo autologous serum skin test (ASST) and in vitro basophil histamine release assay using autologous basophils as well as basophils from normal donors. Twenty-seven subjects with respiratory allergy and three normal subjects were chosen as control.Results ASST was positive in 14/24 non-allergic asthmatics (58%) whereas it was negative in all 30 control subjects (P〈0.001). The serum of only one ASST-positive patient out of 12 (8.4%) induced in vitro histamine release from autologous basophils. The serum from another ASST-positive patient induced histamine release from membrane IgE-stripped autologous basophils. Sera from either non-allergic asthmatics or from control subjects did not provoke significant histamine release from basophils from three normal donors.Conclusion Skin reactivity to autologous serum is common among non-allergic asthmatics, indicating the presence of circulating histamine-releasing factors. However, only in a minority of patients in vitro functional evidence of histamine-releasing autoantibodies (anti-FcɛRI or anti-IgE) was obtained. The presence of circulating histamine-releasing factors might contribute to initiation/maintenance of inflammation in respiratory airways of non-allergic asthmatics.

Notes: Background All previous studies agree that only a proportion of sera from patients with chronic urticaria (CU) positive on the autologous serum skin test (ASST) are able to induce histamine release in vitro. A non-specific release of bradykinins during clotting of blood samples has been suggested; however, ASST seems rather specific and some data point to the existence of a mast cell-specific histamine-releasing factor.Objective To assess whether, and to what extent, the use of both human basophils and mast cells increases the sensitivity of in vitro histamine release assays (HRAs) in ASST-positive patients with CU.Methods The histamine-releasing activity of sera from 93 patients with CU selected on the basis of strong skin reactivity on ASST was assessed in vitro on basophils from 1 (n=86), 2 (n=31), or 3 (n=20) normal donors, and on mast cells from 1 (n=3), 2 (n=3), or 3 (n=87) normal donors.Results Sera from 88/93 (95%) patients induced significant histamine release from mast cells or basophils on at least one HRA. 76/93 (82%), 45/90 (50%), 22/80 (28%), and 6/12 (50%) sera were able to induce significant histamine release from cells of 2/5, 3/5, 4/5 and 5/5 donors, respectively.Conclusion Sera from nearly all ASST-positive patients with CU are able to induce histamine release in vitro. However, the serum from each single patient seems to show its maximal activity on autologous mast cells in vivo, and functional in vitro tests show much variability and seem less sensitive than ASST in the detection of patients with histamine-releasing factors in their blood.

Notes: Background Recently, distinct studies have shown that: (a) chronic idiopathic urticaria (CIU) is autoimmune in 30–50% of cases; (b) in patients with CIU the autologous serum skin test is inhibited by heparin; and (c) basophil histamine release induced in vitro by CIU sera maybe complement-dependent.Objective To carry out a comprehensive clinical and serological study on CIU based upon these observations.Methods Three hundred and six adults with CIU underwent intradermal (ID) test with autologous serum; 57 of them with autologous heparinized plasma as well. Sera from 121 patients (plasmas from 17) were employed to induce in vitro histamine release from basophils of normal donors. The effects of heating (56 °C, 60 min), filtration through membrane, and preincubation with heparin were evaluated as well.Results Autologous serum and plasma induced a weal and flare reaction in 205 out of 306 (205/306; 67%) and in 8/57 (14%) patients, respectively. Positive plasma skin tests were observed only in patients showing strongly positive serum skin tests. Plasma did not elicit any skin reaction in 3/3 patients with dermatographism who showed a positive intradermal test with saline. Sera from 20/121 (16.5%) patients induced significant histamine release from basophils of normal donors. 19/20 sera were from patients with a positive intradermal test; thus, basophil histamine release assay was positive in 19/87 (21.8%) patients with a positive serum skin test. Heating at 56 °C × 1 h markedly reduced the histamine-releasing activity of both serum and plasma from in vitro reactors. Ultrafiltered fractions 〉 100 kDa of both sera tested retained the histamine-releasing activity, whereas fractions 〈 100 kDa were not able to induce any histamine release. Heparin dose-dependently inhibited histamine release induced by sera and plasma, and by basophil agonists such as anti-IgE, formyl-methionyl-leucyl-phenilalanine, and interleukin (IL)-3.Conclusions 67% of our patients with CIU showed a positive autologous serum skin test. Sera from about 20% of those positive on autologous serum skin test induced histamine release from normal basophils in vitro probably as a consequence of the presence of functional autoantibodies. The marked difference between in vivo and in vitro findings could reflect the existence of a mast cell-specific histamine-releasing factor which does not release histamine from basophils of healthy blood donors. However, it might be also the result of in vivo priming of patients' cutaneous mast cells or of heterogeneity of basophil donors. At least in some cases complement seems essential for histamine-releasing activity of serum from patients with CIU. Heparin inhibits histamine release from both basophils (in vitro) and mast cells (in vivo), probably acting directly at a cellular level.

Notes: A novel class of histamine receptors (H3, controlling histamine synthesis and release, was described in rat and human brain and peripheral nerve endings. The present study was undertaken to evaluate whether H3, receptors contribute to the regulation of histamine release from human basophils. Basophil leucocytes were incubated with a H., antagonist (thioperamide; concentrations ranging from 1 nM to 10 μM) or with a H3 agonist ((R)αmethyl-histamine; concentrations ranging from I to 100 μM), and subsequently were stimulated with optimal doses of anti-IgE and formyl-methionyl-leueyl-phenyl-alanine (f-met peptide). No significant modifications of histamine release were observed after incubation either with the H3 agonist or with the H3 antagonist. By contrast, a H3 antagonist (cimetidine; concentrations ranging from 1 to 100 μM) exerted a dose-dependent enhancing effect on anti-IgE- and, to a lesser extent, on f-met peptide-induced histamine release. A H, antihistamine (chlorpheniramine; concentrations ranging from 100 nM to 1μM), at the highest concentration employed, displayed an inhibitory activity on IgE-dependent and IgE-independent histamine release. Exogenous histamine was shown to exert a dose-dependent inhibitory effect on two-staged anti-IgE-induced histamine release. Taken as a whole, these results suggest that H3 receptors are not involved in the regulation of histamine release from human basophils; by contrast, H2 receptors participate in controlling histamine release from human basophils, as previously demonstrated by other authors.

Notes: In 6 normal subjects and 6 patients with allergic rhinitis, nasal response to insufflation of paf-acether (paf, platelet-activating factor), lyso-paf and histamine was evaluated. Nasal challenge with paf, at doses of 300 and 600 nM, induced nasal obstruction, associated with an increase in nasal airway resistances, measured by antherior passive rhinomanometry. Maximum increase in nasal airway resistance was observed at 30 min after challenge (mean percent change + 481 with 600 nM paf; P 〈 0.05). Other symptoms induced by paf insufflation were rhinorrhea (6 out of 12 subjects), itching (8 out of 12), sneezing (4 out of 12) and a burning sensation (6 out of 12). No differences were observed between normal and rhinitic subjects, concerning nasal sensitivity to paf. Neither nasal symptoms nor changes in nasal airway resistance were observed after nasal challenge with lyso-paf (300 and 600 nM); by contrast, histamine (100 nM) induced sneezing, nasal obstruction, itching and rhinorrhea in all the studied subjects, associated with an increase in nasal airway resistance (maximum 5 min after challenge: percent change + 358; P 〈 0.02). Nasal effects of paf were not mediated by histamine, since no increase in histamine levels was observed in nasal washings following paf insufflation. We conclude that paf may have pathogenetic relevance in allergic rhinitis.

Notes: The effect of the new calcium antagonist nitrendipine on in vitro basophil activation was evaluated in 10 subjects. The histamine release induced by calcium ionophore A23187, f-met peptide and anti-IgE was inhibited, in a dose-dependent fashion, by nitrendipine in the concentration range of 1–100 μM. The activity of this calcium antagonist seems complex and related to an interference with calcium at multiple sites. At concentrations higher than 200 μM, nitrendipine causes histamine release from basophil leukocytes. This histamine secretion is likely to be due to a cytotoxic effect, since it is associated with an increase in LDH levels in the cell supernatant.

Notes: The neuropeptide substance P is a possible candidate as histamine-releasing factor in some patients with chronic ordinary urticaria (COU), particularly in those with evidence of a nonautoantibody circulating histamine-releasing factor. In this study, serum substance P levels were measured by enzyme immunoassay in 117 COU patients, 40 atopic subjects and 24 normal subjects. In vivo and in vitro assays for histamine-releasing factors, autologous serum skin test (ASST) and basophil histamine release (BHR) assay, respectively, were performed in all COU patients. Mean serum substance P concentration was not significantly different in COU patients and in normal subjects; however, significantly higher levels were detected in atopic subjects than in COU patients (P 〈 0.003). ASST and BHR assays allowed us to distinguish different subsets of COU patients. Mean serum substance P concentration did not vary significantly in the different subsets of patients. Nevertheless, interestingly three patients with positive ASST and negative BHR assay showed very high substance P levels. These results suggest that substance P does not play an important role as histamine-releasing factor in COU in general but only in occasional patients in whom it might act as a trigger of urticarial symptoms.