Borja Bartolomé

Purification and characterization of Pla a 1, a major allergen from Platanus acerifolia pollen

Background: Plane trees, as Platanus acerifolia, are an important source of airborne allergens in cities of the United States and Western Europe. Little is known about the relevant allergens of this pollen. The aim of this study was to identify relevant allergens from P. acerifolia pollen and purify and characterize a major allergen of 18 kDa.

Sensitization to Sunflower Pollen: Only an Occupational Allergy?

Sunflower (Helianthus annuus) pollen sensitization has been reported as an occupational allergy. In this report, the sensitization of the general population living in sunflower-growing areas to Helianthus pollen was studied. Both RAST results in 32 adults with summer symptoms previously diagnosed as allergic to Artemisia pollen, and cross-reactivity studies between H. annuus and other Compositae suggested that H. annuus pollen was the main allergen involved in the hypersensitivity reaction of those patients. Good correlation was found between RAST and SPT to Helianthus and between RAST and conjunctival provocation test to Helianthus. Bronchial challenge tests performed on 8 of the 32 patients confirmed the clinical implication of Helianthus pollen in suspected subjects. Five workers, handling sunflower pollen, who suffered from related symptoms were subjected to the same study, showing lesser wheal areas and lesser specific IgE levels than a non-worker group. Thirteen patients with RAST values > or = class 2 showed 2 IgE-binding fractions at 34.0 and 42.8 kD in 65% of sera and 3 IgE-binding fractions at pI 4.9, 9.6 and 10.2 in 54% of sera. By means of micropreparative high-resolution chromatography, it was possible to purify a 34-kD major allergen. Analysis performed by RAST inhibition with sera from atopic patients and ELISA inhibition with experimental anti-Helianthus rabbit sera demonstrated a cross-reactivity between Helianthus and other Compositae, but low affinity of specific anti-Helianthus antibodies for heterologous antigens. Taking into account the above-mentioned data, and the high prevalence of Helianthus pollen in the atmosphere during harvesting (in spite of its entomophilous character), Helianthus pollen should be considered as an allergenic source to be investigated in the general population living in sunflower-growing regions suffering from seasonal summer allergy.

Anaphylaxis to a pine caterpillar

• A 41-year-old male pine-forest worker was admitted with a history of a sudden onset of itching rash that started in the neck and then extended to the whole body, with tongue edema, shortness of breath, weakness, blurred sight, and nausea. He said he had disturbed caterpillar nests while knocking pine cones down with a long pole. ——^^^^^———^-^— The patient was nonatopic although he had complained of rash and conjunctivitis for several years, but these symptoms appeared only in association with his work. On physical examination, his blood pressure was 8(V5(J, and the patient had tongue edema and a generalized rash, although it mainly involved the neck. Chest radiography, electrocardiogram, and physical examination were normal. He received 1 mg of subcutaneous epinephrine, as well as tnethylprednisolone and dexchlorpheniramine, both intravenously. A case of severe, IgE-

Glucoamylase: another fungal enzyme associated with baker's asthma.

BACKGROUND Aspergillus-derived enzymes are widely used as dough additives in the baking industry. These enzymes may give rise to immunoglobulin (Ig)E-mediated sensitization and occupational asthma. Glucoamylase (or amyloglucosidase) is an important industrial enzyme obtained from Aspergillus niger and used to provide fermentable sugars for yeast to improve loaf volume and texture. OBJECTIVE The aim of our study was to investigate the potential allergenic role of glucoamylase in bakers asthma. METHODS We report four subjects with work-related allergic respiratory symptoms who were exposed to glucoamylase and other starch-cleaving enzymes used as baking additives. The causative role of glucoamylase in work-related asthma was investigated by immunologic tests and specific inhalation challenges (SIC). Glucoamylase allergenic components were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. RESULTS Skin prick tests to glucoamylase (10 mg/mL) gave a positive response in all patients. Further, a positive skin prick test to alpha-amylase was obtained in the four patients and to hemicellulase in two of them. SIC to glucoamylase elicited isolated early asthmatic responses in the three patients tested, and SIC to alpha-amylase elicited early asthmatic responses in two patients and a dual asthmatic response in another patient. Immunoblotting with glucoamylase showed several IgE-binding bands with molecular masses between 33 and 96 kD. IgE-inhibition assays showed scarce to moderate allergenic cross-reactivity between glucoamylase and alpha-amylase. CONCLUSIONS These bakers had developed IgE-mediated occupational asthma to glucoamylase and alpha-amylase. Fungal glucoamylase is widely used as a baking additive and this enzyme may give rise to allergic respiratory reactions among exposed workers.

Allergy to pomegranate (Punica granatum).

We present the case of a 7-year-old IgE-dependent asthmatic child who, moments after ingesting several pomegranate seeds, showed a clinical condition of bronchospasm which responded to treatment with inhaled salbutamol. Cutaneous tests using the prick-by-prick techniques with extract of the fresh fruit were positive and the RAST for pomegranate was 0.8 PRU/ml. In the literature studied, we have found few specific references to allergy to pomegranate.

Anaphylactic reaction after artificial insemination

BACKGROUND Bovine seroalbumin is known as an allergen for human beings, but reactions to it in an artificial insemination procedure are much rarer. We report a case of anaphylaxis after intrauterine insemination (IUI) in which sensitization to bovine serum albumin (BSA) is demonstrated. OBJECTIVE Report the allergy evaluation performed in a patient who suffered a severe reaction immediately after an IUI procedure. METHODS A 33-year-old woman was referred because of an anaphylactic reaction after a second trial of IUI. She developed pruritus, abdominal pain, nausea and vomiting, bronchospasm, and generalized urticaria. She had an atopic medical history of pollen allergy and sensitization to cat epithelium. She had never had trouble with minor surgery and she usually uses latex material. She had never received heterologous sera before. Her husbands semen for the IUI was processed in a standard fluid medium called upgraded INRA B 2 (Laboratoires CCD, Paris, France), which contains amino acids, lipids, vitamins, BSA, penicillin, and streptomycin in addition to inorganic salts. RESULTS Skin prick tests with the medium and BSA 10 mg/mL were positive. In vitro studies demonstrated an immunoglobulin E binding protein of 60 to 65 kDa and mast cells and basophil activation (CD63 expression) against BSA contained in the medium. Cutaneous and challenge tests with penicillin and streptomycin were negative. CONCLUSIONS We consider the BSA in the semen culture medium to be the factor which triggered the anaphylactic reaction. This case supports the authors who state that media free from heterologous proteins should be used for human application, especially on atopic patients, to avoid sensitization.

Sequence polymorphism and structural analysis of timothy grass pollen profilin allergen (Phl p 11)

Three cDNA clones encoding timothy grass pollen profilin (Phl p 11) were newly isolated. Comparison of the sequences of four cDNA clones, including a previously isolated clone, showed a low level of polymorphism. Isoelectrofocusing of highly purified timothy grass profilin indicated the existence of at least five isoforms. One recombinant profilin showed similar immunological properties to natural timothy grass profilin. Tertiary structure of Phleum pratense profilin was obtained by homology-based molecular modeling.

Occupational allergy caused by carnation (Dianthus caryophyllus)

BACKGROUND Occupational respiratory symptoms caused by decorative flowers are seldom reported in the literature. In our area a large portion of the population works in carnation (Dianthus caryophyllus) winter quarters, and many workers have symptoms of rhinitis and asthma related to exposition. OBJECTIVE The purposes of this study were to investigate whether the symptoms induced by carnation were IgE-mediated and to study the possible allergens involved. METHODS A total of 16 subjects employed in indoor carnation cultivation with symptoms during exposition time were studied along with 15 patients with allergic asthma who were not exposed to carnations and 15 healthy carnation workers used as control subjects. Skin prick tests with carnation extract and RASTs were performed. Protein bands were isolated by SDS-PAGE, and afterwards immunoblotting was performed to characterize the extract. Specific nasal provocation and nonspecific bronchial provocation tests were performed for all the asthmatic patients. Diurnal variation in peak expiratory flow was also measured. RESULTS Skin prick test responses with carnation extract were positive in 15 of the 16 patients and negative in all control subjects. Nasal provocation test responses with carnation extract were positive in 13 of 16 patients. A significant correlation was seen between RAST and nasal provocation results (P <.01). Immunoblotting of sera from 13 patients showed 2 major IgE-binding fractions of 34 and 35 kd in most of the patients, which could constitute the major allergen. Methacholine PD20 showed a variable degree of nonspecific bronchial hyperresponsiveness in all asthmatic subjects. CONCLUSION Data demonstrate the involvement of carnation in occupational allergy, mediated by an IgE-dependent mechanism.

Involvement of Can f 5 in a Case of Human Seminal Plasma Allergy

Background: The existence of IgE binding to dog dander extract without IgE antibodies against the described dog allergens (Can f 1, 2, 3 and 4) implies the presence of other dog allergens yet to be identified. Recently, an IgE-binding protein was isolated from dog urine and identified as prostatic kallikrein; it has been named Can f 5. Cross-reactivity between a dog dander allergen and human prostate-specific antigen (PSA) has been described. The aim of this study was to identify the dog dander allergen that presents cross-reactivity with PSA and demonstrate its clinical relevance in our patient with human seminal plasma allergy. Methods: SDS-PAGE immunoblotting and inhibition tests were performed. Mass spectrometry was carried out to identify the protein involved in the allergy reactions. Results: SDS-PAGE immunoblotting-inhibition with an IgE-binding protein from dog prostatic secretion showed total IgE binding inhibition to a 28-kDa IgE-reactive band identified as PSA. The electroeluted protein from dog prostatic secretion was identified by mass spectrometry as Can f 5. IgE immunoblotting of human seminal plasma incubated with the serum of the patient revealed two IgE-binding bands (28 and 32.7 kDa). Both SDS-PAGE immunoblotting inhibition assays, with human seminal plasma or purified PSA in solid phase, showed complete IgE binding inhibition when the serum of the anaphylactic patient was preincubated with dog dander extract or recombinant Can f 5. Conclusions: The dog dander allergen that shows cross-reactivity with human PSA has been characterized and turns out to be the recently described Can f 5. We demonstrated the clinical relevance of this cross-reactivity in a patient.

Specific sublingual immunotherapy with peach LTP (Pru p 3). One year treatment: a case report.

IntroductionFood allergy is an increasing problem with limited therapeutic approaches apart from to the eviction diet.Case presentationA 40-year-old female patient with food allergy symptoms was polysensitized to almost all vegetable food since the age of 36; the onset of symptoms was during pregnancy. The allergological study demonstrated positive skin prick tests (SPT) to nuts, legumes, cereals, spices, several fresh fruits including peach, and other groups of vegetable foods however, it was negative to common aeroallergens. Serum specific IgE levels were negative (<0.35 kU/L) to profilin and carbohydrate determinants, but positive to Pru p 3 (3.5 kU/L). Positive double-blind placebo-controlled food challenge to peach confirmed the allergic disease. She received specific sublingual immunotherapy with native Pru p 3 at a concentration of 40 μg/ml with 5 administrations per week and a cumulative dose of 200 μg of nPru p 3 per month. After an ultra-rush build-up phase concluded in one day she continued therapy during a year with 5 administrations per week. The clinical evolution and laboratory studies demonstrated an early reduction on SPT reactions with no relevant changes on serum specific IgE, IgG, IgG1 and IgG4 to Pru p 3 during the immunotherapy period. The challenge test was negative 4 months after the beginning of the SLIT. Regarding clinical response she markedly improved after the first month of treatment, and by the 3th month she had no major vegetable dietary restrictions, except for nuts and pepper.ConclusionThese results demonstrate the excellent efficacy and safety of sublingual specific protein immunotherapy developed according to the patient specific sensitivity profile to Pru p3.