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Biomedical subjects

R Ferrándiz

Publications and source records attributed to R Ferrándiz.

10 recordsLinked to original sources

Biologic activity of Dermatophagoides siboney and Blomia tropicalis allergens in exposed and unexposed mite-allergic individuals. Effect of patient selection on the biologic standardization of mite extracts.

BACKGROUND: This study aimed to investigate the influence of patient selection criteria, i.e., mite-allergic individuals exposed and not exposed to Dermatophagoides siboney and Blomia tropicalis, on the biologic activity of mite extracts. Determination of the potency of mite extracts in vivo requires selection of patients with a clinical history of mite allergy. In Scandinavia, there are some anamnestic criteria for mite allergy, whereas in the tropics, where patients are continuously exposed to high levels of mites, selection of patients with mite allergy by clinical history is difficult. METHODS: A total of 210 Cuban asthmatics with continuous symptoms, and 43 Swedes with a clinical history of mite allergy were investigated. Skin prick tests were performed with D. siboney, D. pteronyssinus, D. farinae, B. tropicalis, Acarus siro, Lepidoglyphus destructor, and Tyrophagus putrescentiae extracts. For analysis of the biologic activity of mite extracts, Cuban patients were divided into four groups: 1) all patients skin-test-positive to mites 2) patients positive to mites, but not to other inhalant allergens 3) patients reacting most to the mite species analyzed 4) patients reactive only to mites and reacting most to the mite species analyzed. The biologic potency was calculated according to the Nordic Guidelines. RESULTS: Due to cross-reactivity between mites, Swedish mite-sensitive patients, with a clear clinical history of mite allergy, but not exposed to D. siboney and B. tropicalis, were more skin reactive to these mites than were Cubans. The estimated potency increased gradually to >200% in group 4. In group 1 Cubans, the reactivity to all mites but B. tropicalis was lower than that in mite-sensitive Swedes. CONCLUSIONS: According to the influence of patient selection criteria on the estimation of the potency of mite extracts, the determination of the biologic activity of allergenic extracts in subjects without a clear-cut clinical history should be replaced by new methods when available.

Adolescent↗

Circulating cat allergen and immune complexes in cat-allergic children.

BACKGROUND: The first encounters with allergens seem to influence the development of allergy. Food antigens have been detected in sera as free antigens and in complexes with IgG but less is known about the presence of inhalant allergens. OBJECTIVE: To investigate the presence of the major cat allergen Fel d 1, either as free allergen and/or in complexes with IgG and IgE antibodies in sera from atopic children. METHODS: Serum samples from 33 cat allergic asthmatic children, 7-17 years old, and 15 non-allergic controls were investigated for the presence of Fel d 1 by ELISA (detection limit 0. 13 microg/L). To detect immune complexes (IC), the IgG fraction from Fel d 1 positive sera was purified by affinity chromatography. Purified and non-absorbed material was then analysed for allergen content and specific IgG antibody levels. Immune complexes with Fel d 1 IgE were detected by coupling anti-Fel d 1 MoAb to paramagnetic particles. RESULTS: Fel d 1 was detected (0.15-1.8 microg/L) in 23 of the 33 patients (70%) but not from any of the controls. Eighteen samples contained IgE-Fel d 1 IC and two of four tested samples contained Fel d 1 in the IgG fraction. Electrophoresis and Western blotting of IgG purified material using anti-Fel d 1 MoAb corroborated the presence of IgG-Fel d 1 IC. CONCLUSION: Free-circulating inhalant allergen and IC with allergens may contribute to maintaining immune responsiveness and sensitivity.

Adolescent↗

Cross-reactivity between Dermatophagoides siboney and other domestic mites. II. Analysis of individual cross-reacting allergens after SDS-PAGE and Western blotting inhibition.

BACKGROUND: Dermatophagoides siboney (Ds) cross-reacted with other mite allergens in mite-allergic patients. The aim of this study was to investigate the cross-reactivity between individual allergens responsible for this cross-reactivity. METHODS: The inhibition of IgE binding to Ds allergens was investigated by non-reducing SDS-PAGE and Western blotting. Nitrocellulose membranes were incubated with a pool of sera from mite-sensitive asthmatics, after the addition of serial dilutions of D. farinae (Df), D. pteronyssinus (Dp), D. microceras (Dm), Lepidoglyphus destructor (Ld), Tyrophagus putrescentiae (Tp), Acarus siro (As) and Blomia tropicalis (Bt). N-terminal amino acid sequences and amino acid analysis of the purified major allergens, Der s 1, 2 and 3, were performed after transfer to polyvinylidene difluoride membranes. RESULTS: The inhibition was higher with Df(86%), Dp (54%) and Dm (49%) extracts than with Ld (20%), Tp (11%), As (18%) and Bt (6%). The dose-response inhibition showed a diverse pattern for the individual allergens. Despite the high cross-reactivity between the pyroglyphid mites, some proteins of Ds were less inhibited, e.g. by the Df and Dp 80-kD protein, and by the Dm and Dp 52-, 37-, 30- and 14-kD allergens. The 65-, 62-, 37- and 30-kD proteins were always inhibited more than 50% by all the mite extracts at the maximum concentration used. The 80-, 52-, 43-, 27- and 14-kD proteins cross-reacted to a lesser extent. Individual allergens of Ds were much less inhibited by non-pyroglyphid mites. However, at the highest concentration, Ld also inhibited most of the Ds allergens. All the ten selected allergens were inhibited to some extent by the heterologous mite extracts. The N-terminal sequences of Der s 1, 2 and 3 allergens showed higher homology to Df and Dm than to Dp. The homology of the group 2 allergens was higher than that of the group 1 allergens. CONCLUSION: The individual allergens of Ds were more similar to Df and Dm than to Dp. There was a limited and variable cross-reactivity with non-pyroglyphid mites. No single allergen was unique for Ds.

Allergens↗

Purification and IgE binding capacity of Der s 3, a major allergen from Dermatophagoides siboney.

BACKGROUND: Sensitization to the house dust mite Dermatophagoides siboney has been demonstrated in asthmatic patients. Previously, Dermatophagoides siboney group 1 and group 2 allergens, named Der s 1 and Der s 2, respectively, have been purified. OBJECTIVES: The aim of this study was to purify and to study the IgE reactivity of 30 kDa component, suspected to correspond to group 3 allergens. METHODS: The protein was purified by affinity chromatography using anti-Der f 3 monoclonal antibodies and semi-preparative SDS-PAGE. The IgE binding capacity of the purified fractions was tested with sera from 106 mite-sensitive asthmatic patients using a modified chemiluminiscent method. RESULTS: Affinity chromatography resulted in fractions containing the 30 kDa component which was further purified to homogeneity by SDS-PAGE. Seventy-three per cent of the sera showed IgE reactivity to this protein, indicating that it is a major allergen. The protein also reacted with anti Der f 3 polyclonal antibodies and had tryptic activity. There were differences in the reactivity to Der s 3 according to the age of the patients. CONCLUSION: Based on the frequency of IgE reactions and the reactivity with antibodies directed to Der f 3, it is proposed to name this 30 kDa allergen from D. siboney, Der s 3.

Adolescent↗

Allergens of horse dander: comparison among breeds and individual animals by immunoblotting.

Some patients who are allergic to horses have reported that they can tolerate certain breeds, and the presence of breed-specific allergens has been suggested. Breeders and patients with asthma have claimed that Bashkir horses are nonallergenic. We used sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting to determine IgE-binding profiles of extracts of dander obtained from horses of several breeds. We found considerable inter-breed and within-breed variation but no breed-specific allergens. Danders from all breeds investigated contained the most important allergens, and the allergenic content of dander from Bashkir horses was similar to that of other breeds. Difference in scale production could account for differences in sensitivity to breeds and individual horses.

Allergens↗

Sensitization to Dermatophagoides siboney, Blomia tropicalis, and other domestic mites in asthmatic patients.

Mite species adapted to warm, humid climates are commonly found in house dust in the tropics. In Cuba, Dermatophagoides pteronyssinus, D. siboney, and Blomia tropicalis are the most common and abundant mite species in house dust. To investigate the pattern of sensitization of Cuban asthmatic patients to common mite species, we skin-prick-tested (SPT) 148 patients with a clinical history of asthma and possible mite allergy, and determined specific IgE antibodies against mite allergens (D. pteronyssinus, D. farinae, D. siboney, B. tropicalis, Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, and Glycyphagus domesticus). The prevalence of positive SPT was high to D. siboney (88%), D. pteronyssinus (87%), A. siro (85%), B. tropicalis (85%), and D. farinae (83%). The largest skin reactions were obtained with D. siboney and B. tropicalis extracts. The skin test response to the D. siboney extract correlated to those of D. farinae, D. pteronyssinus, B. tropicalis, and A. siro. The highest IgE levels were found to Dermatophagoides species and B. tropicalis. IgE to D. siboney and B. tropicalis were found in 97% and 96% of the patients, respectively. The prevalence of specific IgE to the other mites studied varied from 46 to 65%. D. siboney and B. tropicalis are important sensitizers among asthmatic patients in Cuba.

Adolescent↗

Characterization of allergenic components from house dust mite Dermatophagoides siboney. Purification of Der s 1 and Der s 2 allergens.

BACKGROUND: Allergic reactions to house dust mites of the genus Dermatophagoides play an important role in the pathogenesis of asthma and other atopic diseases. Dermatophagoides siboney has been described as a species from Cuba. Together with D. pteronyssinus and Blomia tropicalis, it is frequently found in house dust from homes of asthmatics. OBJECTIVES: The aim of this study was to investigate the allergenic composition from the house dust mite D. siboney. METHODS: The characterization of D. siboney extract was performed by SDS-gPAGE and immunoblotting. Purification of individual components was performed by affinity chromatography. RESULTS: At least 16 components between 13 and 98 kDa stained by Coomassie Blue were found. Using a panel of 35 sera from atopic mite sensitive patients 13 components reacted to different extent with patient IgE. Two components, 25 and 14 kDa, bound to specific IgE strongly and frequently, i.e. 80 and 91% of the patients, respectively. Affinity chromatography using crossreacting monoclonal antibodies to group 1 and 2 allergens resulted in purified preparations of 25 and 14 kDa proteins, which showed IgE-binding with the majority of the human sera when tested by immuno-dot. CONCLUSION: Based on the IgE binding profile of D. siboney and on the capacity to react with crossreacting monoclonal antibodies for groups 1 and 2, it is proposed to name these two allergens, 25 and 14 kDa, Der s 1 and Der s 2, respectively.

Adult↗

Crossreactivity between Dermatophagoides siboney and other house dust mite allergens in sensitized asthmatic patients.

BACKGROUND: The role of Pyroglyphid mites in IgE-mediated sensitization has been well established. Although mites belonging to the genus Dermatophagoides dominate in the acarofauna of domestic dust, non-pyroglyphid mites might also be of clinical importance. In Cuba, Dermatophagoides siboney is found in dust coexisting with D. pteronyssinus and Blomia tropicalis. Dermatophagoides farinae is not found. Storage mites, such as Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae, might also be present. OBJECTIVES: The aim of this study was to investigate the allergenic relationships among the mites present in the environment of allergic patients in Cuba. METHODS: The crossreactivity between an extract of D. siboney and the above mentioned six mites was studied by specific IgE inhibition, using sera of mite-sensitive patients after incubation with serial dilutions of D. siboney. RESULTS: The inhibitory effect of D. siboney extract was more pronounced against D. farinae, 91%, than against D. pteronyssinus, 62%. Specific IgE to B. tropicalis, A. siro, L. destructor and T. putrescentiae were inhibited to a much lesser extent, i.e. between 47 and 58%. The correlation between IgE to D. siboney and IgE to D. pteronyssinus and D. farinae was of 0.92 and 0.90, respectively. A weaker correlation was found between D. siboney and L. destructor (0.7), A. siro (0.67), B. tropicalis (0.54) and T. putrescentiae (0.51). CONCLUSION: Thus, D. siboney seems to be most closely related to D. farinae.

Adolescent↗