Class I chitinases as potential panallergens involved in the latex-fruit syndrome Carlos Blanco, PhDa, Araceli Diaz-Perales, BSb, Carmen Collada, PhDc,

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Class I chitinases as potential panallergens involved in the latex-fruit syndrome Carlos Blanco, PhDa, Araceli Diaz-Perales, BSb, Carmen Collada, PhDc, Rosa Sánchez-Monge, PhDb, Cipriano Aragoncillo, PhDc, Rodolfo Castillo, MDa, Nancy Ortega, MDa, María Alvarez, MDa, Teresa Carrillo, PhDa, Gabriel Salcedo, PhDb Journal of Allergy and Clinical Immunology Volume 103, Issue 3, Pages 507-513 (March 1999) DOI: 10.1016/S0091-6749(99)70478-1 Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 1 CAP inhibition assays of chestnut by crude extract (CT) , chitinase-enriched preparation (CS) , purified chitinases class I (CI) and class II (CII ) from chestnut, and the natural latex extract (Hb ). A pool of sera from patients with latex-fruit allergy was preincubated with different 1:10 dilutions of each inhibitor solution (initial concentrations: 250 μg/mL for CT, CS, and Hb and 30 μg/mL for the purified chestnut chitinases). D pteronyssinus commercial extract was used as negative-inhibition control. Data represent means ± SEM of duplicate determinations. Journal of Allergy and Clinical Immunology 1999 103, 507-513DOI: (10.1016/S0091-6749(99)70478-1) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 2 CAP inhibition assays of avocado by crude extract (AT) , chitinase-enriched preparation (AS ), purified chitinases class I (PI ) and class II (PII ) from avocado, and natural latex extract (Hb ). A pool of sera from patients with latex-fruit allergy was preincubated with different 1:10 dilutions of each inhibitor solution (initial concentrations: 250 μg/mL for AT, AS, and Hb and 30 μg/mL for the purified avocado chitinases). D pteronyssinus commercial extract was used as negative-inhibition control. Data represent means ± SEM of duplicate determinations. Journal of Allergy and Clinical Immunology 1999 103, 507-513DOI: (10.1016/S0091-6749(99)70478-1) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 3 SDS-PAGE fractionation of extracts (15 μg of protein) from chestnut (Cs ), avocado (Pa ), and natural latex (Hb ), as well as from the purified chitinases (2 μg of protein) CI , CII , PI , and PII . Replica gels were stained with Coomassie Blue (A) or electrotransferred to PVDF membranes and immunodetected with a pool of sera from patients with latex-fruit allergy (B) or from control subjects allergic to latex but not fruit (C) . Journal of Allergy and Clinical Immunology 1999 103, 507-513DOI: (10.1016/S0091-6749(99)70478-1) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 4 Immunoblot inhibition assays with latex extract as inhibitor. Crude extracts (15 μg of protein) from chestnut, (Cs) and avocado (Pa), and the purified allergens (2 μg of protein) Cl and Pl were separated by SDS-PAGE and electrotransferred to PVDF membranes. Subsequently, they were immunodetected with the pool of sera from the patients with latex-fruit allergy and preincubated with 250 μg of the natural latex extract (+Hb) or with 250 μg of BSA (+BSA) as negative inhibition control. Journal of Allergy and Clinical Immunology 1999 103, 507-513DOI: (10.1016/S0091-6749(99)70478-1) Copyright © 1999 Mosby, Inc. Terms and Conditions