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This study was undertaken to investigate the effect of estrogens on the histamine release mediated by IgE in rat peritoneal mast cells (PMC) and in sensitized human basophils. The estrogens were found to enhance the histamine release of either rat PMC and sensitized human basophils upon stimulation with anti-IgE. The enhancement was estrogens dose-dependent reaching the maximum value of 23% for rat PMC and 41% for sensitized human basophils stimulated with anti-IgE upon preincubation with 10(-8) M estrogens. Moreover, when purified PMC were used, the enhancing effect was still detected, suggesting a direct interaction between estrogens and mast cells. The enhancing effect took place quite rapidly reaching plateau levels in about 60 min. Basophils preincubated at 4 instead of 37 degrees C did not give any appreciable enhancement, suggesting that it was temperature-dependent and that the effect observed was not due to cytotoxicity. Incubation of PMC or human basophils with estrogens alone, without challenge with anti-IgE, did not give any detectable histamine release. The enhancement of histamine release by estrogens is probably mediated by IgE molecules present on the cell membrane, since this effect was not observed on challenge with substance P or compound 48/80, two segretagogues known to induce histamine release not via IgE.

An embryo-derived histamine-releasing factor (EHRF) was identified and partially purified from media in which two-cell human embryos were cultured. The EHRF at 5 micrograms/ml was capable of inducing 22 +/- 7% release of histamine from sensitized human leukocytes, reaching a maximum of 56 +/- 4% over an EHRF concentration range of 1-30 micrograms/ml. The EHRF was not detected in media where unfertilized oocytes were cultured or in medium alone. The effect of EHRF was not due to cytotoxicity since unsensitized leukocytes were unreactive. Histamine release did not occur when the assay was performed at 4 degrees C or in presence of EDTA.

A crude extract of Parietaria judaica pollen was obtained by means of extraction, centrifugation and dialysis, and studied by means of quantitative immunoelectrophoresis. Crossed immunoelectrophoresis, using a high-titer purified rabbit antibody fraction, showed that the pollen extract contained at least 26 antigens of which 18 moved towards the anode, 6 moved towards the cathode and 1 moved both towards the anode and the cathode. The allergens in the extract were identified by means of crossed radio immunoelectrophoresis. Nine of the 26 antigens were able to bind specific human IgE to their corresponding immunoprecipitates, and 4 of these antigens can be classified as major allergens. The apparent molecular weights of 17 antigens were determined by a combination of size exclusion chromatography and immunochemical analysis. Ten antigens had Kav values corresponding to molecular weights in the range of 10-40 kilodaltons, 1 antigen possessed an apparent molecular weight of less than 10 kilodaltons, and six antigens had molecular weights above 40 kilodaltons. Most of the antigens, as analysed by column isoelectric focusing, had pI values between 4 and 7. Crossed line immunoelectrophoresis using an extract of Parietaria officinalis shows that the antigens of this extract exhibit a high degree of identity with those of P. judaica.

Parietaria judaica pollen allergens, fractionated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes, were identified using 52 sera collected in Australia and Sicily from P. judaica pollen-allergic patients. IgE-binding pollen components transferred to nitrocellulose were detected by reaction with 125I-anti-human IgE and autoradiography. Nine pollen components, ranging in molecular weight (MW) from approximately 10,000 to 80,000 daltons, bound IgE antibodies but the two fastest migrating components sometimes each separated into two very closely migrating bands. The faster of the two components exhibiting doublet formation (MW approximately 10,000 daltons) showed by far the highest frequency of IgE binding, being recognised by 50 of the 52 sera examined. Although patients' IgE reaction patterns to P. judaica allergens were heterogeneous, the degree of heterogeneity was much less than that observed with house dust mite and other pollen extracts studied by electrophoretic transfer analysis. Results with gradient gel-nitrocellulose transfer experiments which showed no IgE-binding components with MWs less than 70,000 daltons, and comparisons of our electroblotting results with crossed radioimmunoelectrophoresis results of others, suggested that the doublet proteins with MWs of approximately 10,000 probably bind to higher MW proteins in P. judaica pollen extracts.

The data obtained in this study suggest that eating Euparipha pisana (snail), a common food in Mediterranean countries, could give serious allergic reaction such as asthma. We describe here the identification and partial characterization of allergenic molecules form this new source. An aqueous extract of snail was obtained by homogenization in distilled water, centrifugation, dialysis and defatting with ethyl ether. Skin prick test (SPT) performed with the snail extract on 70 subjects allergic to the more common allergens of the Mediterranean area gave a SPT positivity in 61% of the subjects tested, with a mean value of histamine-equivalent prick (HEP) equal to 0.81 +/- 0.25 (n = 43), while no SPT-snail-positive reactions were obtained by using the same extract on 30 not allergic subjects. To ascertain if such a sensitivity was IgE-mediated, sera from SPT-snail-positive subjects were analyzed by RAST, coupling the snail extract to polystyrene balls and to paper discs. 19% of the sera tested were RAST-positive, mean value of binding 4.8 +/- 2.8% (n = 13), while when using sera from SPT-snail-negative subjects, the RAST mean value was 0.49 +/- 0.18% (n = 27). Histamine release (HR) was also performed. Basophils prepared from SPT-snail-positive subjects were incubated with a snail extract. All of the SPT-snail-positive subjects gave a significant value of HR, mean value 21.8 +/- 7% using 1 micrograms of snail extract (n = 16), while 1.41 +/- 1.1% (n = 10) was the mean value obtained when SPT-snail-negative subjects were analyzed.(ABSTRACT TRUNCATED AT 250 WORDS)

A major allergen, the Parj I, was purified to homogeneity from Parietaria judaica pollen by means of ultrafiltration dialysis, preparative polyacrylamide gel chromatography and affinity chromatography through a column of Sepharose-monoclonal antibody specific for Parj I. The homogeneity of the Parj I was assessed by one single arc of immunoprecipitation both in cross immunoelectrophoresis (CIE) and crossed radioimmunoelectrophoresis, by one single band of radiostaining after a sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transfer to nitrocellulose and by one single peak after a size exclusion chromatography on high-performance liquid chromatography (HPLC). The homogeneity was further supported by crossed Laurell immunoelectrophoretic analysis, in that only one arc of precipitation was magnified in CIE after addition of the purified allergen. The purified Parj I allergen was capable of interacting in vitro with 70% of the human IgE specific for a crude P. judaica extract, as determined by radioallergosorbent test inhibition. The purified Parj I was capable of inducing positive reactions in vivo in skin prick tests, and of inducing release of histamine from blood containing basophils as determined by a histamine release assay. The amino acid analysis of the Parj I showed 118 amino acid residues per monomer analyzed and, among other residues, three methionine residues were detected. The molecular weight of the Parj I estimated by HPLC and amino acid composition was 26 kilodaltons.