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57 Immunochemical characterization of parietaria judaica antigens
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CHARACTERIZATIONOF PARIE!WUA -CAL JuDAIcAAiW1GENs. D.Geraci, K.Billesbolle, R. Cocchiara, H.Igsen, H.Lawenstein. Is'cituto di
Biologia dell0 Sviluppo,CNR,Paler,Italy & The Protein Laboratory,University of Copenhagen, Denmark. A crude extract was obtained
of Parietaria judaica pollen by means of extraction, centrifuga-
tion and dialysis and studied by means of gumtitative imtiun~lectrophoresis. Crossed inmnoelectrophoresis (CIE), using a high-titer purified rabbit antibody fraction, shmed that the pollen extract contained at least26 antigens, of which 18movedtomrds the anode, 6rmved towards thecathode andlmvedbothtomrds the cathode and the anode. The allergens in the extract were identified by mans of crossed radio irmnunoelectrophoresis (CRIE). Seven of the 26 antigens were able to bind specific human IgE to their corresponding imnunoprecipitates, and 4 of these antigens can be classified as major allergens. The apparent mleculac weight of 17 antigens was determined by a combination of size exclusion chrmatography and immm&mical analysis. Ten antigens had K(av) values corresponding to molecular weights in the range 10 to 40 kD, 1 antigen possessed anappamantmolecular weight of less than 10 kD and six antigens had n-olecular weights above 40 kD. Most of the antigens had pI values between 4 to 7 .
ALLERGENICITY OF THE AUSTRALIAN PINE POLLEN G. Bucholtz, M.D., (cASUARINA F.~~I~ETIF~LIA). A. Hensel III, M.D., R. Lackey, M.D., D. Serbousek, B.A., D. Ledford, M.D., Tampa, FL Australian pine (AP) is the source of the fourth most common tree pollen in Tampa and controlled studies are lacking proving its allerDouble-blind nasal challenges (NC) genicity. were done on 14 patients (Pt) with allergic rhinitis and positive skin tests (wheal L 8mmwith 0.02 ml I.D. 1:500 w/v) to AP pollen extract (AR+) and on 22 control Pt (8 with allergic rhinitis, 8 with vasomotor rhinitis, 6 normals) with negative skin test to AP pollen extract. The NC was done by instilling 50 ul of increasing concentrations of AP pollen extract l:lO,OOO, l:l,OOO and 1:lOO w/v into one naris Pt were oband 50 ul diluent into the other. served for pallor, discharge, edema, sneezing Bronchial challenges (BC) were and itching. done (Chai et al, JACI 56:323, 1975) on 10 Pt with extrinsic asthma and positive skin tests to AP pollen extract (EA+) and on 20 control Pt (11 extrinsic asthmatics, 3 intrinsic asthmatics, 6 normals) with negative skin tests to AP pollen RAST was done on all Pt sera. A RAST extract. score of 2+ or greater was considered positive. Nasal Challenge Results p P&ST+ RAST- p Pt Groups AR+ Controlc.001 6/10 4126 c.01 Positive lo/14 O/22 (60%) (15%) NC/tPt (71%) (0%) Bronchial Challenge Results p RAST+ RAST- P Pt Groups EA+ Controll/24 c.001 o/20 c.001 416 Positive 5/10 (67%) (4%) (50%) (0%) BC/tPt The Australian pine pollen is an important aeroallergen.
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of cross-reacting and partially identical allergens by inhibition of crossed radioimmunelectrophoresis (CRIE-inh,). H.J. Maasch, W. Geissler, PhD R Wahl 7sFio -9 1-. ._ .L-...- .^ -.ein e ) PhD, .._.
Analysis
Cross-reactivity and identity of allergens were studied on a molecular basis by CRiE-inh. In this modification the patient sera were preincubated with an allergen extract to deplete sera of allergen specific IgE. The pretreated sera were employed in a normal CRIE. An uninhibited serum served as control. By this we studied the degree of crossreactivity and identity of allergens from birch-, timothy-, barleyand rye-pollen extracts. oat-, wheat-, Moreover th'e composition of extracts from purified bodies and whole cultures of Dermatophagoides pteronyssinus were investigated. By self-inhibition of the birch system and RAST-inhibition we established the optimal concentration for the inhibiting extract. The fading of the allergens in CRXE-Bnh, correlated well with the increasing inhibition capacity in RAST. Individual allergens of timothy were inhibited to different degrees by cereal pollen extracts. However, the different cereal pollen extracts show similar inhibition patterns, Extracts from purified mite bodies inhibited allergens from whole mite cultures completely. On the other hand, some allergens of purified mites were not completely inhibited by extracts from whole mite cultures. This indicates a partially heterogeneity of the two preparations. CRIE-inh. appears to be a useful tool for analysis of allergen extracts.
119
CHARACTERIZATIONOF PARIE!WUA -CAL JuDAIcAAiW1GENs. D.Geraci, K.Billesbolle, R. Cocchiara, H.Igsen, H.Lawenstein. Is'cituto di
Biologia dell0 Sviluppo,CNR,Paler,Italy & The Protein Laboratory,University of Copenhagen, Denmark. A crude extract was obtained
of Parietaria judaica pollen by means of extraction, centrifuga-
tion and dialysis and studied by means of gumtitative imtiun~lectrophoresis. Crossed inmnoelectrophoresis (CIE), using a high-titer purified rabbit antibody fraction, shmed that the pollen extract contained at least26 antigens, of which 18movedtomrds the anode, 6rmved towards thecathode andlmvedbothtomrds the cathode and the anode. The allergens in the extract were identified by mans of crossed radio irmnunoelectrophoresis (CRIE). Seven of the 26 antigens were able to bind specific human IgE to their corresponding imnunoprecipitates, and 4 of these antigens can be classified as major allergens. The apparent mleculac weight of 17 antigens was determined by a combination of size exclusion chrmatography and immm&mical analysis. Ten antigens had K(av) values corresponding to molecular weights in the range 10 to 40 kD, 1 antigen possessed anappamantmolecular weight of less than 10 kD and six antigens had n-olecular weights above 40 kD. Most of the antigens had pI values between 4 to 7 .
ALLERGENICITY OF THE AUSTRALIAN PINE POLLEN G. Bucholtz, M.D., (cASUARINA F.~~I~ETIF~LIA). A. Hensel III, M.D., R. Lackey, M.D., D. Serbousek, B.A., D. Ledford, M.D., Tampa, FL Australian pine (AP) is the source of the fourth most common tree pollen in Tampa and controlled studies are lacking proving its allerDouble-blind nasal challenges (NC) genicity. were done on 14 patients (Pt) with allergic rhinitis and positive skin tests (wheal L 8mmwith 0.02 ml I.D. 1:500 w/v) to AP pollen extract (AR+) and on 22 control Pt (8 with allergic rhinitis, 8 with vasomotor rhinitis, 6 normals) with negative skin test to AP pollen extract. The NC was done by instilling 50 ul of increasing concentrations of AP pollen extract l:lO,OOO, l:l,OOO and 1:lOO w/v into one naris Pt were oband 50 ul diluent into the other. served for pallor, discharge, edema, sneezing Bronchial challenges (BC) were and itching. done (Chai et al, JACI 56:323, 1975) on 10 Pt with extrinsic asthma and positive skin tests to AP pollen extract (EA+) and on 20 control Pt (11 extrinsic asthmatics, 3 intrinsic asthmatics, 6 normals) with negative skin tests to AP pollen RAST was done on all Pt sera. A RAST extract. score of 2+ or greater was considered positive. Nasal Challenge Results p P&ST+ RAST- p Pt Groups AR+ Controlc.001 6/10 4126 c.01 Positive lo/14 O/22 (60%) (15%) NC/tPt (71%) (0%) Bronchial Challenge Results p RAST+ RAST- P Pt Groups EA+ Controll/24 c.001 o/20 c.001 416 Positive 5/10 (67%) (4%) (50%) (0%) BC/tPt The Australian pine pollen is an important aeroallergen.
60
of cross-reacting and partially identical allergens by inhibition of crossed radioimmunelectrophoresis (CRIE-inh,). H.J. Maasch, W. Geissler, PhD R Wahl 7sFio -9 1-. ._ .L-...- .^ -.ein e ) PhD, .._.
Analysis
Cross-reactivity and identity of allergens were studied on a molecular basis by CRiE-inh. In this modification the patient sera were preincubated with an allergen extract to deplete sera of allergen specific IgE. The pretreated sera were employed in a normal CRIE. An uninhibited serum served as control. By this we studied the degree of crossreactivity and identity of allergens from birch-, timothy-, barleyand rye-pollen extracts. oat-, wheat-, Moreover th'e composition of extracts from purified bodies and whole cultures of Dermatophagoides pteronyssinus were investigated. By self-inhibition of the birch system and RAST-inhibition we established the optimal concentration for the inhibiting extract. The fading of the allergens in CRXE-Bnh, correlated well with the increasing inhibition capacity in RAST. Individual allergens of timothy were inhibited to different degrees by cereal pollen extracts. However, the different cereal pollen extracts show similar inhibition patterns, Extracts from purified mite bodies inhibited allergens from whole mite cultures completely. On the other hand, some allergens of purified mites were not completely inhibited by extracts from whole mite cultures. This indicates a partially heterogeneity of the two preparations. CRIE-inh. appears to be a useful tool for analysis of allergen extracts.
119