Evaluation of the efficacy of isopathic immunotherapy in the treatment of allergic asthma in BALB/C mice.

Introduction: Homeopathy is a therapeutic method based on the fundamental principle of "like cures like." Homeopathic remedies are extremely dilute but involve vigorous shaking at each dilution. Isopathy is one approach of homeopathy, in which the causative agents or products of a disease are used to treat the same disease. Allergen immunotherapy is the only potential disease-modifying treatment for allergic patients. Subcutaneous immunotherapy is more effective than sublingual immunotherapy. However, subcutaneous immunotherapy is ineffective at a low dose, whereas at high doses it can result in an unacceptably high frequency of systemic reactions. In the current study, we evaluated the efficacy of isopathic immunotherapy with highly diluted ovalbumin (HD OVA) in the treatment of OVA-induced allergic asthma in BALB/c mice.Methods: BALB/c mice were sensitized with OVA and alum. Two weeks later, the mice received HD OVA on days 21, 22, 32 and 41 (8 h after the last challenge) of the treatment. The mice were challenged with OVA (5%) aerosols on days 35, 38 and 41 for 20 minutes using an ultrasonic nebulizer and sacrificed the next day.Results: Isopathic immunotherapy significantly reduced lung tissue inflammation, the number of eosinophils in bronchoalveolar fluid, allergen-specific IgE and interleukin-4 production. It also insignificantly increased the production of transforming growth factor-beta and proliferation of regulatory T cells against the allergen.Conclusion: Isopathic immunotherapy may be a good candidate treatment for allergic asthma.

Utility of opium seed extract tests in preventing hypersensitivity reactions during surgery.

BACKGROUND: Anaphylaxis during anaesthesia is fatal in 3-9% of patients and analgesics, including opioids, and is the second most common medicament-related cause, although the prevalence is underestimated. We recently found that patients may generate IgE antibodies to opium seeds. OBJECTIVES: To determine the diagnostic accuracy of specific antibodies to morphine, codeine, rocuronium and oil body and aqueous fractions of Papaver somniferum seeds in the diagnosis and prevention of allergy to opioids. METHODS: Patients with hypersensitivity reactions during surgery, and severe clinical allergy (pollen, tobacco), and illicit heroin users were selected. The sensitivity, specificity and predictive values of in vivo and in vitro diagnostic techniques including oil body and aqueous fractions of P. somniferum seeds were measured. RESULTS: We studied 203 patients, with mean age 35.1±17.1 and 200 healthy controls. Patients sensitised to heroin or with hypersensitivity reactions during surgery responded to P. somniferum seed tests. Of patients not known to be sensitised to opioids, the highest positivity was in patients sensitised to tobacco (p<0.001). Opium seed skin tests and IgE, especially the oil body fraction, were more sensitive (64.2%) and specific (98.4%) than morphine, codeine and rocuronium tests for opioid sensitivity. Pollen allergy was not a risk factor for sensitisation to morphine. CONCLUSIONS: Sensitivity to opioids and intraoperative anaphylaxis can be diagnosed by routine tests. IgE and skin tests for the oil body fraction of P. somniferum had the highest sensitivity for sensitisation to opioids.

Clinical value of morphine, pholcodine and poppy seed IgE assays in drug-abusers and allergic people.

BACKGROUND: The diagnosis of anaphylactic reactions due to opiates during anaesthesia can be difficult, since in most cases various drugs may have been administered. Detection of specific IgE to poppy seed might be a marker for sensitisation to opiates in allergic people and heroin-abusers. This study assessed the clinical value of morphine, pholcodine and poppy seed skin-prick and IgE determination in people suffering hypersensitivity reactions during anaesthesia or analgesia and drug-abusers with allergic symptoms. METHODS: We selected heroin abusers and patients who suffered severe reactions during anaesthesia and analgesia from a database of 23,873 patients. The diagnostic yield (sensitivity, specificity and predictive value) of prick and IgE tests in determining opiate allergy was analysed. RESULTS: Overall, 149 patients and 200 controls, mean age 32.9 ± 14.7 years, were included. All patients with positive prick to opiates showed positive prick and IgE to poppy seeds, but not to morphine or pholcodine IgE. Among drug-abusers, 13/42 patients (31%) presented opium hypersensitivity confirmed by challenge tests. Among non-drug abusers, sensitisation to opiates was higher in people allergic to tobacco (25%), P<.001. Prick tests and IgE against poppy seed had a good sensitivity (95.6% and 82.6%, respectively) and specificity (98.5% and 100%, respectively) in the diagnosis of opiate allergy. CONCLUSIONS: Opiates may be significant allergens. Drug-abusers and people sensitised to tobacco are at risk. Both the prick and specific IgE tests efficiently detected sensitisation to opiates. The highest levels were related to more-severe clinical profiles.

Clinical and immunological effects of a treatment with desentizating low-dose multicomponents in IgE mediated and non IgE mediated food allergies: observational retrospective pilot study.

BACKGROUND: Alimentary allergy has high impact on the quality of life (Qol) of patients and their families: it represents an economic burden for individuals and National Health System. The disease, particularly frequent in pediatric age, recognizes different pathogenetic mechanisms and expresses itself through the production of IgE (IgE mediated form) antibodies or through cell-mediated immunune responses (non IgE mediated forms). The aim of this clinical observational retrospective study is to evaluate the effect of a long-term treatment with Low Dose Medicine (LDM) drugs in pediatric patients affected by IgE and non IgE mediated food allergy. OBJECTIVE: The purpose of the study is to determine the efficacy of the treatment with Allergy Plex (Guna Laboratory, Milan, Italy) to induce clinical and/or immunological tolerance both to IgE mediated and non IgE mediated food allergy; the secondary endpoint is to investigate the treatment tolerability, the reduction of positivity to Skin Prick test and Patch test to food allergens and the decrease on the peripheral blood of the specific IgE to food allergens. The treatment efficacy was measured through a clinical score. METHODS: In this study the immunomodulant activity of Allergy Plex 13, Allergy Plex 7 and Allergy Plex 10 (Guna S.p.A., Milano, Italy) was evaluated. In every patient the state of allergical clinical responses and the immuno-allergological state were evaluated by means of specific parameters letting know the regulatory response to the allergical Th fenotype. RESULTS: Data about Clinical tolerance to food, Symptomatological clinical score, ECP, ACTH, Cortisol; IL-4, IL-10 was collected. There was evidence of improvement of clinical score, reduction of the diameter of cutaneous pomphus obtained through the Prick test and a decrease of IgE specifics values. CONCLUSIONS: The data issued from this study seem to confirm the efficacy of treatment with Allergy Plex in allowing the restoration of immune tolerance and the definite reduction of the clinical score.

[Effect of dongchong xiacao capsule on airway inflammation of asthmatic patients].

OBJECTIVE: To investigate the effect of dongchong xiacao capsule on the airway inflammation of asthmatic patients and to explore the relevant mechanism of therapeutic effect of Dongchongxiacao capsule. METHOD: Sixty patients with moderate persistent asthma were randomized into the treatment group (n=30) and the control group (n=30). Inhaled corticosteroid and as-needed beta-agonist were used in the treatment group while this therapy plus dongchong xiacao capsule were used in the control group for two months. Serum IL-4, IFN-gamma, sICAM-1, MMP-9, IgG, IgE level were assessed at randomization and 2 months after randomization. RESULT: The serum level of IgE, sICAM-1, IL-4 and MMP-9 of the treatment group was lowered to a greater degree than that of the control group (P < 0.05 or P < 0.01 ). CONCLUSION: Dongchong xiacao capsule can reduce the serum markers of airway inflammation, which suggests this therapy bares the anti-inflammation effects probably through regulating the balance of TH1/TH2, inhibiting the activity of adherence molecule and reducing IgE production. It may also have the effect of reversing airway remodeling, which needs further research to determine.

Sensitive flow cytometric method to test basophil activation influenced by homeopathic histamine dilutions.

OBJECTIVE: In an experimental setting, human basophil degranulation was triggered by anti-IgE to measure the effects from homeopathic solutions in an in-vitro cell system. A 3-color flow cytometric method with enhanced accuracy was established. As an example we looked at the influence of histamine on anti-IgE activation of basophils. METHODS: Basophils were identified in the flow cytometer by their physical properties in the forward and side scatter light depiction and by gating on CD2(-), CD14(-), CD16(-), CD19(-), HLA-DR(-) negative and CD123-positive cells. CD63 expression on the cell surface of the anti-IgE-activated basophils served as an activation marker. RESULTS: With this method we were able to study basophil function of the 0.6-3.9% basophils out of the mononuclear blood cell fraction and to document their activation status upon anti-IgE activation. Optimal activation occurs at 0.6 microg/ml final anti-IgE concentration; not less than 10,000 basophils have to be counted per batch to reduce the variation of the measurement. The fixation method was able to stabilize activation for two days. After investigation and reduction of the source of measurement variability, an unequivocally inhibited basophil activation was documented in a partly optimized system with homeopathic dilutions of histamine (10(-22)M, 10(-23)M, 10(-24)M, and 10(-25)M histamine). Dilutions greater than 10(-20)M histamine (Avogadro's number 6.02 x 10(23)) account for less than 1.36 molecules of histamine in the test sample, indicating a true homeopathic effect. CONCLUSIONS: This test system is adequate for studying the effects of highly diluted mediators on basophil activation by anti-IgE. The systematic application of this experimental arrangement is recommended to study the effects of homeopathic dilutions on basophils.

Effects of pancuronium and vecuronium on plasma levels of immunoglobulins, IgG, IgM and IgE, in patients undergoing minor gynaecological surgery.

1. The effects of two competitive neuromuscular blocking agents, pancuronium and vecuronium, on plasma levels of immunoglobulins, IgG, IgM and IgE, were studied and compared, using the ELISA assay, in patients undergoing minor gynaecological surgery. 2. The results showed that neither pancuronium (0.1 mg/kg) nor vecuronium (0.1 mg/kg) had any significant effect on IgG, IgM and IgE levels in these patients. However, the underlying trend was that both pancuronium and vecuronium slightly increased the IgG and IgM levels, (by 19.5%, 22.1%, of control IgG, and by 17.3%, 28.8% of control IgM, respectively), with vecuronium being more effective than pancuronium). In addition, pancuronium, but not vecuronium, also slightly increased the IgE levels (by 5.9% of control IgE). Vecuronium, on the other hand, slightly decreased the IgE level (by 4.9% of control IgE). However, these increases or decreases were not statistically significant. 3. The significance of the present results are discussed in relation to changes in immunoglobulins and adverse effects induced by drugs, such as muscle relaxants.

[Severe anaphylactic reactions after administration of rocuronium]. / Accidents anaphylactiques graves après administration de rocuronium.

The authors report four cases of severe anaphylactic reactions (grade III or IV) to rocuronium bromide. In three of them, it was the first contact with a muscle relaxant. In three patients the reaction was mediated by IgE anti-bodies. A cross-reactivity with other muscle relaxants was existing in two cases (suxamethonium, vecuronium and atracurium in one patient, suxamethonium, vecuronium and pancuronium in the other.

Histaminoid responses to atracurium, vecuronium and tubocurarine.

Sixty patients scheduled for elective surgery underwent intradermal testing with 0.1 ml of the following solutions diluted in 0.9% saline: vecuronium and tubocurarine (1 in 1,000), atracurium (1 in 1,000 and 1 in 10,000), thiopentone (1 in 100) and also a 0.9% saline control. Thirty minutes later, an area of erythema of greater than 1.5 cm, or a wheal exceeding 1.0 cm in diameter, was recorded as a positive reaction. The patients then randomly received equipotent doses of atracurium, vecuronium or tubocurarine during a standardized anaesthetic induction. Any cutaneous reaction and the percentage fall in systolic pressure three minutes after administration of the relaxant were recorded. In 51 patients plasma IgE levels were measured. The incidence of positive cutaneous reactions to intradermal and intravenous relaxants was significantly different with each agent (p less than 0.01). The percentage fall in systolic pressure after tubocurarine was significantly different relative to the other two agents (p less than 0.01). This was regarded as reflecting potency in releasing histamine and placed the relaxants in the same order: tubocurarine, atracurium and vecuronium. The response to intradermal administration was no guide to the subsequent response after intravenous administration of the three relaxants. IgE levels below 15 IU X ml-1 occurred significantly more often in females and were associated with a significantly higher incidence of cutaneous reactions after intradermal atracurium (1 in 1,000 and 1 in 10,000) (p less than 0.05 and 0.001 respectively) and tubocurarine (1 in 1,000). With these two agents, generalized flushing after intravenous administration was also more common in this group, relative to the normal/high IgE group.

[Therapy of cantharides extract for perennial allergic rhinitis and its effect on total IgE in serum].

The therapy of 10% Cantharides extract in treating 50 cases of perennial allergic rhinitis (PAR) was studied. The extract was plastered and blistered on Dazhui, Neiguan point. It was observed by nasal mucosa provocative test, cells in nasal secretion test and serum total IgE test. The results showed that its effective rate was 88%, the allergic nasal mucosa provocative test of treated group alleviated obviously after the treatment (P < 0.01), the number of eosinophil and basophil in nasal secretion decreased (P < 0.01, P < 0.05); and the serum total IgE also reduced significantly (P < 0.01).

Anaphylaxis following administration of papaveretum. Case report: Implication of IgE antibodies that react with morphine and codeine, and identification of an allergenic determinant.

IgE antibodies that reacted with morphine and codeine were detected in the serum of a subject who experienced a life-threatening anaphylactic reaction following the administration of Omnopon-Scopolamine (papaveretum-hyoscine). Hapten inhibition studies with morphine and a number of structurally-related analogues revealed that morphine and codeine were the most potent inhibitors of IgE binding to a morphine-solid phase. Nalorphine, meperidine, and methadone were also good inhibitors of IgE binding, but naltrexone, buprenorphine, and fentanyl proved to be poor inhibitors. From a detailed examination of structure-activity relationships, the authors conclude that the important structural features of the morphine allergenic (that is, IgE binding) determinant comprises the cyclohexenyl ring with a hydroxyl group at C-6 and, most important of all, a methyl substituent attached to the N atom. The authors' findings suggest that morphine analogues administered to such a patient may provoke clinical anaphylaxis. Hyoscine reacted weakly with IgE antibodies in the subject's serum, but this was thought to be due to weak cross-reaction between this compound and morphine.

Allergenic potential of gonadotrophic preparations in experimental animals: relevance of purity.

Local reactions have been frequently reported following repeated injections of human menopausal gonadotrophins (HMG) for the treatment of infertility. Also immunoglobulin (Ig) E-mediated systemic reactions have sporadically been observed. Since most HMG preparations contain significant amounts of non-hormonal urine-derived proteins, it was suggested that these contaminating proteins are responsible for the various allergic reactions. In order to verify this hypothesis, different human follicle stimulating hormone (HFSH) and HMG preparations (Metrodin and Pergonal from Ares-Serono, and Humegon from Organon), were compared with a highly purified preparation (Metrodin HP from Ares-Serono) for the frequency and severity of allergic reactions induced in laboratory animals. The occurrence of anaphylactic shock or related symptoms was studied in sensitized guinea-pigs. The production of specific IgE was evaluated in serum from mice sensitized with the test drugs by the induction of passive cutaneous anaphylaxis in rats. In both models, two different schedules of sensitization were used. Severe allergic reactions were found in 20 of 7% of the guinea-pigs receiving highly purified FSH (Metrodin HP) in the two schedules, respectively, compared to 90 and 88% with the other preparations. Similarly significantly lower IgE titres were induced by highly purified FSH in respect to the other preparations. It can be concluded that the elimination of contaminating proteins significantly reduces the allergenicity of urine-derived HFSH preparations.

Cross-reactivity of metocurine, atracurium, vecuronium and fazadinium with IgE antibodies from patients unexposed to these drugs but allergic to other myoneural blocking drugs.

An inhibition assay was used to determine quantitatively the allergenic cross-reactivity of some myoneural blocking drugs not yet released for use in Australia, in the sera of patients who had experienced anaphylactic reactions to neuromuscular blocking drugs. Two of the compounds, metocurine and atracurium were highly cross-reactive with the currently used myoneural blockers; fazadinium was weakly cross-reactive and vecuronium intermediate in potency between these two extremes. From these results, we predict that anaphylactic reactions to these compounds, and particularly to metocurine and atracurium, will occur in some patients allergic to the currently used neuromuscular blocking agents.

In vitro immunological degranulation of human basophils is modulated by lung histamine and Apis mellifica.

1. The effect of high dilutions of two homeopathic drugs Lung histamine (Lung his) and Apis mellifica (Apis mel) used for the treatment of allergic diseases has been assessed on in vitro human basophil degranulation. Experiments were conducted blind. 2. Basophil degranulation induced by 1.66 X 10(-9) M anti-IgE antibody was significantly inhibited in the presence of 5 Lung his (5th centesimal dilution of Lung his) and 15 Lung his (15th centesimal dilution of Lung his) by 28.8% and 28.6% respectively and by 65.8% in the presence of 9 Apis mel (9th centesimal dilution of Apis mel). Basophil degranulation induced by 1.66 X 10(-16) to 1.66 X 10(-18) M anti-IgE antibody was also inhibited by high dilutions of Lung his and Apis mel with an inhibition of nearly 100% with 18 Lung his (18th centesimal dilution of Lung his) and 10 Apis mel (10th centesimal dilution of Apis mel). An alternance of inhibition, inactivity and stimulation was observed when basophils were incubated in the presence of serial dilutions of Lung his and Apis mel. 3. The investigation of the clinical efficacy of high dilutions of Lung his and Apis mel should be envisaged in allergic diseases in parallel with in vitro and ex vivo biological assays.

Allergy to suxamethonium: persisting abnormalities in skin tests, specific IgE antibodies and leucocyte histamine release.

Twenty-one patients, who had previously experienced an anaphylactic reaction to suxamethonium during general anaesthesia, were selected for this study. Initially, skin tests with muscle relaxants were carried out in the twenty-one patients, detection of specific anti-choline IgE in nineteen, and leucocyte histamine release in seventeen. These three tests were then repeated between 1 year and 4 years after the initial evaluation. In the majority of patients, sensitization to the muscle relaxants persisted for more than 1 year after the anaphylactic reaction. Only three patients out of twenty-one (4%) had negative skin tests when retested 1-4 years later. A reduction in leucocyte histamine release was noticed in one of the seventeen retested patients (6%). Modifications of anti-choline IgE were observed in five of nineteen patients (26%). The persistence of sensitization to suxamethonium may result from repeated stimulation by occasional contacts with quaternary ammonium compounds. This study demonstrates the reliability of skin tests, leucocyte histamine release and detection of anti-choline IgE to diagnose allergic reactions to suxamethonium, even when they are performed a long time after the initial anaphylactic reaction.

Anaphylaxis to muscle relaxants: cross-sensitivity studied by radioimmunoassays compared to intradermal tests in 34 cases.

Thirty-four patients (31 female and three male patients) with a previous anaphylactoid shock to muscle relaxants were investigated. The seric antimyorelaxant IgE was detected by radioimmunoassay (RIA), and the results were compared to intradermal test (IDR) reactions to dilutions of the commercial drugs. The RIA was carried out with a Sepharose-myorelaxant solid phase and anti-IgE 125I-labeled IgG. The results corresponded to the percentage of labeled anti-IgE bound on the solid phase. The RIA with Sepharose-alcuronium and Sepharose-choline was estimated positive from determination with normal sera (n = 12) when bound IgE was greater than 1.0% and 1.5%, respectively. The RIA and IDR were positive in 43.5% and 75%, respectively, of the cases, with a concordance of 66%. One test at least was positive in 79.4% of the cases. No correlation was found between IgE seric levels and the RIA nor between the cutaneous sensitivity and the RIA. Cross-reactivity with Sepharose-choline and Sepharose-alcuronium was observed in 50%, and it was demonstrated by IDR in only 34.2%. The RIA demonstrated the specificity of IgE to quaternary ammonium compounds as myorelaxant drugs. The positive IDR revealed the bridging of mast cell-bound specific IgE, depending on structural conditions, such as the flexibility of the molecules or the variable specificity of the antibodies, restricted to quaternary ammonium ions or enlarged to a broader part of the incriminated molecules.