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124 Characterization of oilseed rape allergens
J ALLERGYCLIN IMMUNOL VOLUME 97, NUMBER1, PART3 121
122
Abstracts
A n a l y s i s o f I g E a n t i b o d y r e s p o n s e s to D. pteronyssinus a n d Blomia tropicalis u s i n g r e c o m b i n a n t G r o u p 5 a l l e r g e n s . LD Yailes MS, MC ~.izzo MD, C K Naspitz MD,..F. Montealegre Phi), E Fernandez.Caldas Phl), TAE Platte-Mills MD PhD, MD Chapman PhD.~ LK Arruda MD. Charlottesville VA, Tampa FL, Ponce PR, USA; Sac Paulo, Brazil. • A B. tropicalis allergen, Blo t 5, has been identified by molecular cloning, and shows -40% sequence homology to the 14kd D. pteronyssinus allergen Der p 5. Recombinant(r) 131o t 5 expressed in E. coli gave positive immediate prick test at 51zg/ml or intzadermal test at a concentration as low as 103t~g/ml, in selected B. tropicalis allergic children in Brazil. lgE ab responses to rBlo t 5 and rDer p 5 (Linet al, JACI 1994;94:989), were compared in sera from 138 mite allergic patients living in different areas of the world, using an antigen binding RIA. The prevalence of IgE ab to BIot 5 among patients from Brazil, Puerto Rico (PR) and Florida, exposed to both D. pteronyssinus and B. tropicalis, was 45%. On the other hand, among patients from Charlottesville and England the prevalence was only 20% (p<0.05). In addition, the levels of lgE ab were lower in the second group (geom. mean 29.2 units]el versus 106 units/nil, p<0.005). The results suggested that IgE ab to Blot 5 in patients not exposed to B. tropicalis reflect antigenic crossreativity with the equivalent antigen from D. pteronyssinus. IgE ab to rDer p 5 was detected in 55% and 54% of patients (geom. mean 389 units/el and 436 units/el, p=0.38) in these groups. The prevalence of IgE ab to Group 5 allergens combined was 71%. To further study eXlX~ore to B. tropicalis, a panel of Blot 5-specific monoclonal abe has been raised. Recombinant Blot 5 allergen has a major role in the study of sensitization and exposure to 11. tropicalis.
123
Allergic rhinoconjunctivitis by lilac flowers. Bainza T,MD. Barrio de M.MD. Montoro A,MD, Ordouui E.MD, Baeza ML,MD. Madrid. Spain.
124
Extrinsic rhlnoconjunotlvitis caused by ornamental flowers is an uncommon disorder and extremely rare in non floral industry workers. We report an atoplc patient who developed allergic rhinoconjunctivitie to lilac flowers exposure. A 17 yrs old female consulted by ocular itching, tearing, sneezing and watery discharge when exposed to lilac flowers. She had previously been diagnosed of allergic rhinocon~unctivitie and asthma by olive and grass pollen. Skin prick tests were positive to olea, fraxinus and lolium pollens and slightly positive to a crude lilac extract. Intradermal tests were positive to lilac extract (0.2 ~g/ml) in our patient and negative in controls. Specific serum IgE was detected to olea pollen (0.61 kU/L, Pharmacia CAP-System) and to crude lilac extract by indirect ELISA (patient's O.D.~ 0,798, control= 0.009). Immunoblottlng from lilac extract SDS-PAGE showed IgE recognition of an apparent MW of 45 kD band. In conclusionr we report a nonprofessional sensitization to lilac flowers in an atopic patient.
21:3
IMMUNOLOGIC RESPONSES TO CHIRONOMUS FLAVIPLUMUS IN ATOPIC CHILDREN Hye Youn~ KanQ. Ki Youn~l Lee. DonQ Soo Kim. Kvu Eam Kim. Bvuna Ju JeonQ Yonsei University College of Medicine, Seoul, Korea
To evaluate the role of chironomidae(non-biting midge) as an inhalant allergen in childhood atopic patients, we performed allergy skin test to crude extracts of Chironomus flaviplumus(CF). Thirty-five out of 456 atopic patients(7.8%) showed positive skin reactions. All of the positive reactors also showed positive responses to other inhalant allergens, especially house dust mite. Specific IgE to CF antigens could be detected 3 out of 8 patients by ELISA. Eight patients showed specific IgG and IgG subclasses with very low O.D. SDS-PAGE analysis of CF extracts contains at least 16 bands and 15 IgE-binding bands were noted by immunoblotting analysis. Among these bands 3 bands of 32KD, 27KD, 19KD were bound strongly with the sera of atopic patients.
C h a r a c t e r i z a t i o n o f oilseed r a p e a l l e r g e n s . M Focke P ~ W Hemmer Phi3, R Bracun MD~ S_ Wolf MD1 F Wantke MD~ I~ O6tz MD~ R Jarisch MD, Vienna, Austria. Oilseed rape (OSR) cultureshave increased twcntyfold in the last decade in Austria.8 % of pollen allergicpatientsare also sensitizedto O S R in our area and cases of isolated O S R allergy cxlst.As O S R pollen antigens have not yet been identified, sera of scnsitiz.ed patients were investigatedby immunoblot. An aqueous extract of O S R pollen (AIIergon)was prepar,~d. Proteins were loaded onto SDS-PAGE and ¢lectroblotted to nitroo~Jlulose. Nitrocelluloses w e e probed with see from >50 patientsshowin~ O S R sensitization.Blot resultswere compared with results from patzontswith isolatedsensitizationto eitherbirch, grass and mugwort, and control blood samples drawn from healthy nonatopic individuals.Furthermore, blotted O S R pmtaina were screened for cross-reacting properties by incubation with monoclonal
antibodies (moAbs) raised against birch (B1P 1,3,4, anti-Bet v 2) and ross'won pollen (MUP 6,9,12,15). IgE-binding proteins with a molecular mass of 70, 61/58, 51, 42, and 33kD, and double bands with 14/12kD and 8/6kD were detected. 70% of OSR positive patients reacted to proteins within the high molecular weight range. In about 50%, IgE-binding to low molecular weight proteins (6/8, 12/14kD) was observed. The 6/BkD proteins usually showed high lgE-binding capacity and are considered OSR unique proteins. Cross-reactivity with birch and ross'won is indicated by positive signals with moAbs BIP3 and MUP6 at 26/28/30 and 27kD, respectively. Intense binding of the anti-Bet v 2 moAb to the 12/14kD double band suggests this protein to be a profilin. The data demonstrate that immunoblotted proteins of oilseed rape pollen are capable to bind lgE in patients diagnosed by skin prick test or RAST to be sensitized, and thus prove that oilseed rape is an allergen.
122
Abstracts
A n a l y s i s o f I g E a n t i b o d y r e s p o n s e s to D. pteronyssinus a n d Blomia tropicalis u s i n g r e c o m b i n a n t G r o u p 5 a l l e r g e n s . LD Yailes MS, MC ~.izzo MD, C K Naspitz MD,..F. Montealegre Phi), E Fernandez.Caldas Phl), TAE Platte-Mills MD PhD, MD Chapman PhD.~ LK Arruda MD. Charlottesville VA, Tampa FL, Ponce PR, USA; Sac Paulo, Brazil. • A B. tropicalis allergen, Blo t 5, has been identified by molecular cloning, and shows -40% sequence homology to the 14kd D. pteronyssinus allergen Der p 5. Recombinant(r) 131o t 5 expressed in E. coli gave positive immediate prick test at 51zg/ml or intzadermal test at a concentration as low as 103t~g/ml, in selected B. tropicalis allergic children in Brazil. lgE ab responses to rBlo t 5 and rDer p 5 (Linet al, JACI 1994;94:989), were compared in sera from 138 mite allergic patients living in different areas of the world, using an antigen binding RIA. The prevalence of IgE ab to BIot 5 among patients from Brazil, Puerto Rico (PR) and Florida, exposed to both D. pteronyssinus and B. tropicalis, was 45%. On the other hand, among patients from Charlottesville and England the prevalence was only 20% (p<0.05). In addition, the levels of lgE ab were lower in the second group (geom. mean 29.2 units]el versus 106 units/nil, p<0.005). The results suggested that IgE ab to Blot 5 in patients not exposed to B. tropicalis reflect antigenic crossreativity with the equivalent antigen from D. pteronyssinus. IgE ab to rDer p 5 was detected in 55% and 54% of patients (geom. mean 389 units/el and 436 units/el, p=0.38) in these groups. The prevalence of IgE ab to Group 5 allergens combined was 71%. To further study eXlX~ore to B. tropicalis, a panel of Blot 5-specific monoclonal abe has been raised. Recombinant Blot 5 allergen has a major role in the study of sensitization and exposure to 11. tropicalis.
123
Allergic rhinoconjunctivitis by lilac flowers. Bainza T,MD. Barrio de M.MD. Montoro A,MD, Ordouui E.MD, Baeza ML,MD. Madrid. Spain.
124
Extrinsic rhlnoconjunotlvitis caused by ornamental flowers is an uncommon disorder and extremely rare in non floral industry workers. We report an atoplc patient who developed allergic rhinoconjunctivitie to lilac flowers exposure. A 17 yrs old female consulted by ocular itching, tearing, sneezing and watery discharge when exposed to lilac flowers. She had previously been diagnosed of allergic rhinocon~unctivitie and asthma by olive and grass pollen. Skin prick tests were positive to olea, fraxinus and lolium pollens and slightly positive to a crude lilac extract. Intradermal tests were positive to lilac extract (0.2 ~g/ml) in our patient and negative in controls. Specific serum IgE was detected to olea pollen (0.61 kU/L, Pharmacia CAP-System) and to crude lilac extract by indirect ELISA (patient's O.D.~ 0,798, control= 0.009). Immunoblottlng from lilac extract SDS-PAGE showed IgE recognition of an apparent MW of 45 kD band. In conclusionr we report a nonprofessional sensitization to lilac flowers in an atopic patient.
21:3
IMMUNOLOGIC RESPONSES TO CHIRONOMUS FLAVIPLUMUS IN ATOPIC CHILDREN Hye Youn~ KanQ. Ki Youn~l Lee. DonQ Soo Kim. Kvu Eam Kim. Bvuna Ju JeonQ Yonsei University College of Medicine, Seoul, Korea
To evaluate the role of chironomidae(non-biting midge) as an inhalant allergen in childhood atopic patients, we performed allergy skin test to crude extracts of Chironomus flaviplumus(CF). Thirty-five out of 456 atopic patients(7.8%) showed positive skin reactions. All of the positive reactors also showed positive responses to other inhalant allergens, especially house dust mite. Specific IgE to CF antigens could be detected 3 out of 8 patients by ELISA. Eight patients showed specific IgG and IgG subclasses with very low O.D. SDS-PAGE analysis of CF extracts contains at least 16 bands and 15 IgE-binding bands were noted by immunoblotting analysis. Among these bands 3 bands of 32KD, 27KD, 19KD were bound strongly with the sera of atopic patients.
C h a r a c t e r i z a t i o n o f oilseed r a p e a l l e r g e n s . M Focke P ~ W Hemmer Phi3, R Bracun MD~ S_ Wolf MD1 F Wantke MD~ I~ O6tz MD~ R Jarisch MD, Vienna, Austria. Oilseed rape (OSR) cultureshave increased twcntyfold in the last decade in Austria.8 % of pollen allergicpatientsare also sensitizedto O S R in our area and cases of isolated O S R allergy cxlst.As O S R pollen antigens have not yet been identified, sera of scnsitiz.ed patients were investigatedby immunoblot. An aqueous extract of O S R pollen (AIIergon)was prepar,~d. Proteins were loaded onto SDS-PAGE and ¢lectroblotted to nitroo~Jlulose. Nitrocelluloses w e e probed with see from >50 patientsshowin~ O S R sensitization.Blot resultswere compared with results from patzontswith isolatedsensitizationto eitherbirch, grass and mugwort, and control blood samples drawn from healthy nonatopic individuals.Furthermore, blotted O S R pmtaina were screened for cross-reacting properties by incubation with monoclonal
antibodies (moAbs) raised against birch (B1P 1,3,4, anti-Bet v 2) and ross'won pollen (MUP 6,9,12,15). IgE-binding proteins with a molecular mass of 70, 61/58, 51, 42, and 33kD, and double bands with 14/12kD and 8/6kD were detected. 70% of OSR positive patients reacted to proteins within the high molecular weight range. In about 50%, IgE-binding to low molecular weight proteins (6/8, 12/14kD) was observed. The 6/BkD proteins usually showed high lgE-binding capacity and are considered OSR unique proteins. Cross-reactivity with birch and ross'won is indicated by positive signals with moAbs BIP3 and MUP6 at 26/28/30 and 27kD, respectively. Intense binding of the anti-Bet v 2 moAb to the 12/14kD double band suggests this protein to be a profilin. The data demonstrate that immunoblotted proteins of oilseed rape pollen are capable to bind lgE in patients diagnosed by skin prick test or RAST to be sensitized, and thus prove that oilseed rape is an allergen.