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116 Cross-reactivity studies of Lepidoglyphus destructor and Blomia tropicalis
J ALLERGYCLIN IMMUNOL VOLUME 97, NUMBER 1, PART 3
113
114
Abstracts
Allergenic Components in Three Different Species of PenciUium: Cross-reactivity among Major Allergens HD Shen MS, WL Lin BS, JJ Tsai MD, PhD., SS Han MD, PhD, Taipei, Taiwan, Republic o f China For analyzing allergens of the nmuld Penicillium, IgE-binding components of 3 prevalent air-borne Penicillium species (P. citrinum, P. notatum and P. breviconlpactum) were identified by immunoblottiug using sera from 67 asthnmtic patients. Among the 67 serum samples tested, 15, 14 and I1 showed lgE reactivity, to components of P. citrinum, P. notatum and P. brevicompactum respectively. All 15 P. citriuum-positive senml samples showed lgEbinding to the 32.5-kD component of this Penicillium species. Thirlecn (93%) of the 14 P. notatum-positive serum samples reacted with the 34-kD component of P. notatum. Tea (91%) of the 11 P. brevicompactu,n-positive sera showed IgE reactivity to the 33-kD colnpoueut of_P. brevicompactum All of the 10 P. brovi.compaclum 33 kD component-positive serum samples showed IgE reactivity to the 32.5 and 34 kD components of the other 2 different Penicillium species tested. Dose-dependent inhibition of lgE-binding to these major allergens was observed when the positive serum sample was absorbed with different amounts of individual allergenic extract as well as with different anaounts of the other 2 different Penicillium extracts. In conclusion, although different allergenic profiles were observed in 3 different Penicillium species tested, results obtained in the present study showed that there was lgE-crossreactivity among the 33 kD-group major allergens of P. citrinum, P. not~qum and P. brevicompactum. The ild'ormation obtained will be useful in cliuic~ diagnosis and treatment of Penicillinm allergy.
115
Cross-allergenicity of the House Dust Mites
116
Euroglyphus maynei a n d Blomia tropicalis. L G Arlian PhD. MS Morgan PhD. E Fernandez-Caldas PhD, Dayton, OH and Madrid, Spain The domestic mites Euroglyphus maynei (EM) and Blomia tropicalis (BT) frequently co-inhabit homes in subtropical/tropical regions around the world. Both species are the source of substances that cause allergic reactions in patients. The purpose of this study was to examine the immunologic cross-reactivity between E M and BT. Sera of 19 mite-sensitive patients who were skin test positive to B T and/or R A S T positive to E M were used. Western blotting showed that individual sera had IgE that bound to 0-17 and 2-15 proteins in E M and BT extracts, respectively. Corresponding IgE-binding proteins of 105, 75, 57, 18 and <10 kD were detected in both EM and B T extracts. The majority of IgE-binding proteins did not show corresponding bands in both extracts. Heterologous CRIE showed IgE binding to 6 of the 9 E M antigens precipitated by a n t i - B T s e r u m with individual sera recognizing 0-4 of the 6 allergens. In the reciprocal reaction, 10 of the 12 proteins of B T that were precipitated by anti-EM serum bound IgE with individual sera binding to 0-5 proteins. This study indicated that E M and B T are the source of both species-specific and cross-reactive allergens, but most allergens in each extract were species-specific.
211
AUTOLOGOUS WESTERN BLOTTING WITH HOUSE DUST IN ATOPIC SUBJECTS. Pearson DJ. and Lees M., Manchester, UK. W e have investigated the specificity of IgE responses to house dust in atopie patients from the N o r t h West of the UK. Western blots were performed using sera from a random group o f atopic patients using house dust collected from their own homes, a reference dust and a panel o f 14 extracts o f common allergens (including D. Pteronyssinus, cat, dog, pollens and molds). 9 o f 20 subjects reacted to one or more mite allergen bands, 10 to cat or dog, 17 to one or more molds. 4 subjects demonstrated binding to a high molecular weight component o f dust which did not correlate with any band seen in any o f the purified allergen extracts. W e conclude that Western blotting with patients serum demonstrates a currently unidentified allergen.
Cross-reactivity studies of Lepidoglj,phus destructor and Blomia tropicalis. E. Johansson PhD, L. Machado MD, PhD'. SGO Johanss.on MD. PhD, M van Hage-Hamsten MD, Ph_._.DD.Stockholm. Sweden. *Sao Paulo. Brazil Tile mite Lepidog(~phusdestructor (Ld) occurs abundantly in stored farm products but has also been obsen,ed in dwcllings. It is the most common cause of sensitization among Swedish farmers. Blomiatropicalis(Bt). a common mite in house dust, is an important allergen in tropical and subtropical areas. Earlier. we have reported allergenic cross-reactivity between Ld and other mite species and now, we demonstrate cross-reactivity also between Ld and Bt. The allergens of the mite cultures were detected by immunoblotting, after separation by SDS-PAGE, using non-reducing conditions. Sera which were positive in RAST to Ld and BI were used. Twenty-one Swedish farmers sensitized to Ld. reacted with 26 IgE-binding components of Bt and 23 patients attending the Allergy Clinic at Hospital das Clinicas in Sao Paulo. reacted with 23 components of Ld. Pools were made from the Swedish and the Brazilian sera respectively, containing lgE to the most frequent allergens and used in the cross-reaclivity stud),. A serial dilution of each mite extract was preincubalcd with each serum pool before probing to the mite allergens on blots. The IgE-binding to several allergens of each species were i,ahibited by the heterologous mite. Each mile species contains a major allergen of appro:,:imately 15 kDa that is inhibiled by the heterologous mite extract. We will fiwther specie' the cross-reactivity to these two allergens by using tile 15 kDa recombinant Lep d 2 allergen of Ld produced in our labotators'.
113
114
Abstracts
Allergenic Components in Three Different Species of PenciUium: Cross-reactivity among Major Allergens HD Shen MS, WL Lin BS, JJ Tsai MD, PhD., SS Han MD, PhD, Taipei, Taiwan, Republic o f China For analyzing allergens of the nmuld Penicillium, IgE-binding components of 3 prevalent air-borne Penicillium species (P. citrinum, P. notatum and P. breviconlpactum) were identified by immunoblottiug using sera from 67 asthnmtic patients. Among the 67 serum samples tested, 15, 14 and I1 showed lgE reactivity, to components of P. citrinum, P. notatum and P. brevicompactum respectively. All 15 P. citriuum-positive senml samples showed lgEbinding to the 32.5-kD component of this Penicillium species. Thirlecn (93%) of the 14 P. notatum-positive serum samples reacted with the 34-kD component of P. notatum. Tea (91%) of the 11 P. brevicompactu,n-positive sera showed IgE reactivity to the 33-kD colnpoueut of_P. brevicompactum All of the 10 P. brovi.compaclum 33 kD component-positive serum samples showed IgE reactivity to the 32.5 and 34 kD components of the other 2 different Penicillium species tested. Dose-dependent inhibition of lgE-binding to these major allergens was observed when the positive serum sample was absorbed with different amounts of individual allergenic extract as well as with different anaounts of the other 2 different Penicillium extracts. In conclusion, although different allergenic profiles were observed in 3 different Penicillium species tested, results obtained in the present study showed that there was lgE-crossreactivity among the 33 kD-group major allergens of P. citrinum, P. not~qum and P. brevicompactum. The ild'ormation obtained will be useful in cliuic~ diagnosis and treatment of Penicillinm allergy.
115
Cross-allergenicity of the House Dust Mites
116
Euroglyphus maynei a n d Blomia tropicalis. L G Arlian PhD. MS Morgan PhD. E Fernandez-Caldas PhD, Dayton, OH and Madrid, Spain The domestic mites Euroglyphus maynei (EM) and Blomia tropicalis (BT) frequently co-inhabit homes in subtropical/tropical regions around the world. Both species are the source of substances that cause allergic reactions in patients. The purpose of this study was to examine the immunologic cross-reactivity between E M and BT. Sera of 19 mite-sensitive patients who were skin test positive to B T and/or R A S T positive to E M were used. Western blotting showed that individual sera had IgE that bound to 0-17 and 2-15 proteins in E M and BT extracts, respectively. Corresponding IgE-binding proteins of 105, 75, 57, 18 and <10 kD were detected in both EM and B T extracts. The majority of IgE-binding proteins did not show corresponding bands in both extracts. Heterologous CRIE showed IgE binding to 6 of the 9 E M antigens precipitated by a n t i - B T s e r u m with individual sera recognizing 0-4 of the 6 allergens. In the reciprocal reaction, 10 of the 12 proteins of B T that were precipitated by anti-EM serum bound IgE with individual sera binding to 0-5 proteins. This study indicated that E M and B T are the source of both species-specific and cross-reactive allergens, but most allergens in each extract were species-specific.
211
AUTOLOGOUS WESTERN BLOTTING WITH HOUSE DUST IN ATOPIC SUBJECTS. Pearson DJ. and Lees M., Manchester, UK. W e have investigated the specificity of IgE responses to house dust in atopie patients from the N o r t h West of the UK. Western blots were performed using sera from a random group o f atopic patients using house dust collected from their own homes, a reference dust and a panel o f 14 extracts o f common allergens (including D. Pteronyssinus, cat, dog, pollens and molds). 9 o f 20 subjects reacted to one or more mite allergen bands, 10 to cat or dog, 17 to one or more molds. 4 subjects demonstrated binding to a high molecular weight component o f dust which did not correlate with any band seen in any o f the purified allergen extracts. W e conclude that Western blotting with patients serum demonstrates a currently unidentified allergen.
Cross-reactivity studies of Lepidoglj,phus destructor and Blomia tropicalis. E. Johansson PhD, L. Machado MD, PhD'. SGO Johanss.on MD. PhD, M van Hage-Hamsten MD, Ph_._.DD.Stockholm. Sweden. *Sao Paulo. Brazil Tile mite Lepidog(~phusdestructor (Ld) occurs abundantly in stored farm products but has also been obsen,ed in dwcllings. It is the most common cause of sensitization among Swedish farmers. Blomiatropicalis(Bt). a common mite in house dust, is an important allergen in tropical and subtropical areas. Earlier. we have reported allergenic cross-reactivity between Ld and other mite species and now, we demonstrate cross-reactivity also between Ld and Bt. The allergens of the mite cultures were detected by immunoblotting, after separation by SDS-PAGE, using non-reducing conditions. Sera which were positive in RAST to Ld and BI were used. Twenty-one Swedish farmers sensitized to Ld. reacted with 26 IgE-binding components of Bt and 23 patients attending the Allergy Clinic at Hospital das Clinicas in Sao Paulo. reacted with 23 components of Ld. Pools were made from the Swedish and the Brazilian sera respectively, containing lgE to the most frequent allergens and used in the cross-reaclivity stud),. A serial dilution of each mite extract was preincubalcd with each serum pool before probing to the mite allergens on blots. The IgE-binding to several allergens of each species were i,ahibited by the heterologous mite. Each mile species contains a major allergen of appro:,:imately 15 kDa that is inhibiled by the heterologous mite extract. We will fiwther specie' the cross-reactivity to these two allergens by using tile 15 kDa recombinant Lep d 2 allergen of Ld produced in our labotators'.