Donald Barnett

Allergenic cross-reactions among legume foods—An in vitro study

The specific IgE binding by protein extracts of 11 food legumes, including soybean, was examined by RAST and RAST inhibition. Sera from 15 peanut-sensitive patients were, with very few exceptions, positive in the RAST to all the legumes. RAST-inhibition testing of each extract against RAST discs of the other legumes indicated considerable cross-reactivity of IgE binding between the legumes. Cross-allergenicity was demonstrated to be most marked between the extracts of peanut, garden pea, chick pea, and soybean. The results have important implications for selection of effective hypoallergenic diets and for the diagnosis of patients hypersensitive to foods.

Multiplicity of allergens in peanuts

Crude peanut protein fractions from raw and roasted peanuts were examined in the RAST with 10 sera from patients showing clinical peanut sensitivity. The radioactive uptake results, which were generally high, did not reveal any distinguishable pattern. Two commercially available peanut proteins, peanut lectin and phospholipase D, gave poor RAST responses. Three purified peanut proteins, alpha-arachin, conarachin I, and concanavalin A-reactive glycoprotein, all gave significant RAST results that were generally lower than those obtained with the crude extracts. The extent of RAST inhibition obtained with these materials was inversely related to their abundance in the total peanut protein. Crossed immunoelectrophoresis with extracts from raw and roasted peanut indicated the presence of 22 and 10 anodically migrating antigens, respectively. Sixteen IgE binding antigens were revealed for raw peanut and seven for roasted peanut after incubation with a mixed serum from the 10 patients in crossed radioimmunoelectrophoresis (CRIE) using 125I-labeled anti-IgE. CRIE plates treated with individual serum samples showed that all the patients had specific IgE for the major antigen peak, which has been tentatively identified as alpha-arachin. This major storage protein of peanut, which is known to be particularly heat resistant; may be of greater clinical significance than its apparently low RAST activity would seem to indicate.

Partial characterization of an allergic glycoprotein from peanut (Arachis hypogaea L.)

A concanavalin A-reactive glycoprotein allergen has been isolated from peanut (Arachis hypogaea). The allergen was separated by affinity chromatography and purified by gel permeation and ion-exchange chromatography. The monomeric molecular weight is 65,000 and the pI is 4.6. The presence of one cysteine residue per molecule results in some dimer formation. Concanavalin A-reactive glycoprotein is a potent allergen for peanut-sensitive patients in both in vivo and in vitro tests. It is allergenically stable, on in vitro examination, at temperatures of up to 100 degrees C and over the pH range 2.8-10. Removal of the carbohydrate moiety failed to eliminate the allergenicity. Concanavalin A-reactive glycoprotein is identified in the crossed immunoelectrophoretic pattern as a major antigen of peanut protein extract but its structural characteristics indicate that it is probably not a component of the major storage-protein complex, arachin.

New allergens from hen's egg white and egg yolk. In vitro study of ovomucin, apovitellenin I and VI, and phosvitin.

Three hen egg yolk proteins, apovitellenins I and VI and phosvitin, and one egg white protein, ovomucin, were purified and tested for their ability to bind IgE in the sera of patients hypersensitive to egg. All of the proteins bound IgE from the sera of egg-allergic individuals in the radioallergosorbent test, and they also inhibited binding of IgE to the parent fractions-either egg yolk (apovitellenins I and VI and phosvitin) or egg white (ovomucin). It appears that apovitellenins I and VI are major allergens for some of the individuals tested. This is the first report of the in vitro allergenicity of these proteins.

Isolation and amino acid sequence of a new long-chain neurotoxin with two chromatographic isoforms (Aa el and Ae e2) from the venom of the australian death adder (Acanthophis antarcticus)

The amino acid sequence of a previously undescribed toxin from Australian death adder venom (Acanthophis antarcticus) has been elucidated. It appears to exist in two forms which are separated by reverse-phase high-performance liquid chromatography, but which have the same sequence and mol. wt. It has 79 amino acid residues and is therefore longer than other long postsynaptic neurotoxins. It shows homology with the conserved regions of the other long postsynaptic neurotoxins except for three unique substitutions of conserved residues, which are Arg-29 instead of Trp or Phe, Leu-33 instead of Arg and Thr-43 instead of Ala.

Immunologic and clinical responses to parenteral immunotherapy in peanut anaphylaxis ­ a study using IgE and IgG4 immunoblot monitoring

BACKGROUND Specific desensitisation to food allergens, which produce anaphylaxis after ingestion, has not been considered as a treatment for food allergy until recently. The purpose of this study was to assess if a parenteral immunotherapy program, using a partially characterised crude peanut extract, could induce a state of immunological tolerance in a patient who exhibited anaphylaxis, asthma and urticaria on exposure to peanut and other legumes. A further aim was to measure the serum antibody responses to the immunotherapy. METHODS AND RESULTS We report the successful desensitisation towards all of the legumes tested of a male patient on parenteral immunotherapy using a partially characterised peanut extract. The immunologic parameters measured during treatment included specific IgE and IgG4 for peanut, soybean, pea and lentil extracts. Immunoblots of specific IgE and IgG4 were made before and after therapy. The antibody response followed the same pattern seen in successful desensitisation of patients with bee venom anaphylaxis. The IgG4 levels increased strongly from a low pre-treatment level in proportion to the antigen dose received. The antigen-specific IgE levels gradually fell from a high pretreatment level, but remained significantly elevated. Immunoblotting for specific IgE and IgG4 demonstrated that acquisition of clinical tolerance after therapy was associated with declines in the number and intensity of bands in IgE blots and the development of more bands of increasing density in the IgG4 blots. CONCLUSIONS Parenteral immunotherapy may offer an alternative treatment to lifelong dietary restriction and epinephrine injections in patients who exhibit life-threatening IgE-mediated anaphylaxis to peanut. Cross desensitisation to other legumes appears to have occurred in this study. The quality and potency of the extract used is an important factor in achieving the desired acquisition of clinical tolerance. In our patient this tolerance correlated with his ability to maintain high levels of specific IgG4, which acted as a marker of protection against anaphylaxis. The use of IgG4 immunoblotting may provide an improved level of discrimination in the assessment of correlation of clinical efficacy with the immunologic response.

Lectins and the radioallergosorbent test

An investigation into the possible role of lectin binding of IgE in RAST of legume and wheat extracts is reported. Lectins from pea, broad bean, lentil, jack bean, soybean, peanut, and wheat germ were coupled to RAST discs. The discs were pretreated with lectin-specific sugars in an attempt to inhibit RAST with sera from 11 sensitive patients. In all cases, RAST was almost unaffected by the inhibitory sugars, indicating that nonimmune binding of IgE by lectins in legume or wheat RAST was not significant when RAST was carried out with allergens bound to the usual paper discs. IgE contains binding sites for all the lectins examined, and five of the sera had high total IgE greater than 300 IU/ml. It is suggested that competition by IgG and other serum glycoproteins may explain the lack of effect by the added sugars. All sera contained endogenous glucose at a level of about 3 mmol/L that may have accounted for some self-inhibition of the lectin binding by pea, broad bean, lentil, and jack bean lectins. There was, however, significant immune binding of some of the lectins by specific IgE, and it is concluded that these lectins may be important in expression of IgE-mediated allergic responses.

Neural Network Processing of Responses to Odorants by a Biological Nose and a Sensor Array

A neural network is a processing device, either an algorithm or actual hardware, whose design was motivated by biological neural functions. It can be trained to operate as a classifier. The application of neural networks to the study of olfactory processing in vivo and to identifying and classifying complex chemical mixtures from the outputs of chemical sensor arrays is the subject of this study.