{"id":{"repo_id":"bologna","oai_identifier":"oai:amsdottorato.cib.unibo.it:1613"},"canonical_url":"https://search.dev.ndltd.org/etd/bologna/oai:amsdottorato.cib.unibo.it:1613","repository":{"repo_id":"bologna","name":"Università di Bologna","base_url":"https://amsdottorato.unibo.it/cgi/oai2"},"display":{"title":"Development of screening assays to test novel integrin antagonists in allergic inflammation","abstract":"Aim of the research: to develop a prototype of homogeneous high-throughput screening (HTS) for identification of novel integrin antagonists for the treatment of ocular allergy and to better understand the mechanisms of action of integrin-mediated levocabastine antiallergic action. Results: This thesis provides evidence that adopting scintillation proximity assay (SPA) levocabastine (IC50=406 mM), but not the first-generation antihistamine chlorpheniramine, displaces [125I]fibronectin (FN) binding to human a4b1 integrin. This result is supported by flow cytometry analysis, where levocabastine antagonizes the binding of a primary antibody to integrin a4 expressed in Jurkat E6.1 cells. Levocabastine, but not chlorpheniramine, binds to a4b1 integrin and prevents eosinophil adhesion to VCAM-1, FN or human umbilical vein endothelial cells (HUVEC) cultured in vitro. Similarly, levocabastine affects aLb2/ICAM-1-mediated adhesion of Jurkat E6.1 cells. Analyzing the supernatant of TNF-a-treated (24h) eosinophilic cells (EoL-1), we report that levocabastine reduces the TNF-a-induced release of the cytokines IL-12p40, IL-8 and VEGF. Finally, in a model of allergic conjunctivitis, levocastine eye drops (0.05%) reduced the clinical aspects of the early and late phase reactions and the conjunctival expression of a4b1 integrin by reducing infiltrated eosinophils. Conclusions: SPA is a highly efficient, amenable to automation and robust binding assay to screen novel integrin antagonists in a HTS setting. We propose that blockade of integrinmediated cell adhesion might be a target of the anti-allergic action of levocabastine and may play a role in preventing eosinophil adhesion and infiltration in allergic conjunctivitis.","abstract_html":"Aim of the research: to develop a prototype of homogeneous high-throughput screening (HTS) for identification of novel integrin antagonists for the treatment of ocular allergy and to better understand the mechanisms of action of integrin-mediated levocabastine antiallergic action. Results: This thesis provides evidence that adopting scintillation proximity assay (SPA) levocabastine (IC50=406 mM), but not the first-generation antihistamine chlorpheniramine, displaces [125I]fibronectin (FN) binding to human a4b1 integrin. This result is supported by flow cytometry analysis, where levocabastine antagonizes the binding of a primary antibody to integrin a4 expressed in Jurkat E6.1 cells. Levocabastine, but not chlorpheniramine, binds to a4b1 integrin and prevents eosinophil adhesion to VCAM-1, FN or human umbilical vein endothelial cells (HUVEC) cultured in vitro. Similarly, levocabastine affects aLb2/ICAM-1-mediated adhesion of Jurkat E6.1 cells. Analyzing the supernatant of TNF-a-treated (24h) eosinophilic cells (EoL-1), we report that levocabastine reduces the TNF-a-induced release of the cytokines IL-12p40, IL-8 and VEGF. Finally, in a model of allergic conjunctivitis, levocastine eye drops (0.05%) reduced the clinical aspects of the early and late phase reactions and the conjunctival expression of a4b1 integrin by reducing infiltrated eosinophils. Conclusions: SPA is a highly efficient, amenable to automation and robust binding assay to screen novel integrin antagonists in a HTS setting. We propose that blockade of integrinmediated cell adhesion might be a target of the anti-allergic action of levocabastine and may play a role in preventing eosinophil adhesion and infiltration in allergic conjunctivitis.","abstract_has_math":false,"creators":["Spartà, Antonino Maria <1977>"],"institution":"Alma Mater Studiorum - Università di Bologna","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Spampinato, Santi Mario"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2009,"date_issued":"2009-05-05","date_published":"2009-05-05","updated_at":"2026-07-24T01:12:16Z","subjects":["BIO/14 Farmacologia"],"languages":["en"],"rights":["info:eu-repo/semantics/openAccess"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["urn:nbn:it:unibo-1363"],"render_values":[{"text":"urn:nbn:it:unibo-1363","href":null,"code":true}]}]},"links":{"outbound_url":"https://doi.org/10.6092/unibo/amsdottorato/1613.","outbound_label":"DOI","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Spampinato, Santi Mario"]},{"key":"dc:creator","label":"Author","values":["Spartà, Antonino Maria <1977>"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2009-05-05"]},{"key":"dc:publisher","label":"Institution","values":["Alma Mater Studiorum - Università di Bologna"]},{"key":"dc:relation","label":"Dc Relation","values":["https://amsdottorato.unibo.it/id/eprint/1613/"]},{"key":"dc:type","label":"Dc Type","values":["Doctoral Thesis","PeerReviewed"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["BIO/14 Farmacologia"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["info:eu-repo/semantics/openAccess"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://amsdottorato.unibo.it/id/eprint/1613/1/FILE_COMPLETO_CON_INDICE_E_FRONTESPIZIO_130309.pdf","urn:nbn:it:unibo-1363","Spartà, Antonino Maria (2009) Development of screening assays to test novel integrin antagonists in allergic inflammation, [Dissertation thesis], Alma Mater Studiorum Università di Bologna. Dottorato di ricerca in Biotecnologie cellulari e molecolari <https://amsdottorato.unibo.it/view/dottorati/DOT210/>, 21 Ciclo. DOI 10.6092/unibo/amsdottorato/1613."]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Aim of the research: to develop a prototype of homogeneous high-throughput screening (HTS) for identification of novel integrin antagonists for the treatment of ocular allergy and to better understand the mechanisms of action of integrin-mediated levocabastine antiallergic action. Results: This thesis provides evidence that adopting scintillation proximity assay (SPA) levocabastine (IC50=406 mM), but not the first-generation antihistamine chlorpheniramine, displaces [125I]fibronectin (FN) binding to human a4b1 integrin. This result is supported by flow cytometry analysis, where levocabastine antagonizes the binding of a primary antibody to integrin a4 expressed in Jurkat E6.1 cells. Levocabastine, but not chlorpheniramine, binds to a4b1 integrin and prevents eosinophil adhesion to VCAM-1, FN or human umbilical vein endothelial cells (HUVEC) cultured in vitro. Similarly, levocabastine affects aLb2/ICAM-1-mediated adhesion of Jurkat E6.1 cells. Analyzing the supernatant of TNF-a-treated (24h) eosinophilic cells (EoL-1), we report that levocabastine reduces the TNF-a-induced release of the cytokines IL-12p40, IL-8 and VEGF. Finally, in a model of allergic conjunctivitis, levocastine eye drops (0.05%) reduced the clinical aspects of the early and late phase reactions and the conjunctival expression of a4b1 integrin by reducing infiltrated eosinophils. Conclusions: SPA is a highly efficient, amenable to automation and robust binding assay to screen novel integrin antagonists in a HTS setting. We propose that blockade of integrinmediated cell adhesion might be a target of the anti-allergic action of levocabastine and may play a role in preventing eosinophil adhesion and infiltration in allergic conjunctivitis."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Development of screening assays to test novel integrin antagonists in allergic inflammation"]}]}],"canonical_facts":{"dc:contributor":["Spampinato, Santi Mario"],"dc:creator":["Spartà, Antonino Maria <1977>"],"dc:date":["2009-05-05"],"dc:description":["Aim of the research: to develop a prototype of homogeneous high-throughput screening (HTS) for identification of novel integrin antagonists for the treatment of ocular allergy and to better understand the mechanisms of action of integrin-mediated levocabastine antiallergic action. Results: This thesis provides evidence that adopting scintillation proximity assay (SPA) levocabastine (IC50=406 mM), but not the first-generation antihistamine chlorpheniramine, displaces [125I]fibronectin (FN) binding to human a4b1 integrin. This result is supported by flow cytometry analysis, where levocabastine antagonizes the binding of a primary antibody to integrin a4 expressed in Jurkat E6.1 cells. Levocabastine, but not chlorpheniramine, binds to a4b1 integrin and prevents eosinophil adhesion to VCAM-1, FN or human umbilical vein endothelial cells (HUVEC) cultured in vitro. Similarly, levocabastine affects aLb2/ICAM-1-mediated adhesion of Jurkat E6.1 cells. Analyzing the supernatant of TNF-a-treated (24h) eosinophilic cells (EoL-1), we report that levocabastine reduces the TNF-a-induced release of the cytokines IL-12p40, IL-8 and VEGF. Finally, in a model of allergic conjunctivitis, levocastine eye drops (0.05%) reduced the clinical aspects of the early and late phase reactions and the conjunctival expression of a4b1 integrin by reducing infiltrated eosinophils. Conclusions: SPA is a highly efficient, amenable to automation and robust binding assay to screen novel integrin antagonists in a HTS setting. We propose that blockade of integrinmediated cell adhesion might be a target of the anti-allergic action of levocabastine and may play a role in preventing eosinophil adhesion and infiltration in allergic conjunctivitis."],"dc:format":["application/pdf"],"dc:identifier":["https://amsdottorato.unibo.it/id/eprint/1613/1/FILE_COMPLETO_CON_INDICE_E_FRONTESPIZIO_130309.pdf","urn:nbn:it:unibo-1363","Spartà, Antonino Maria (2009) Development of screening assays to test novel integrin antagonists in allergic inflammation, [Dissertation thesis], Alma Mater Studiorum Università di Bologna. Dottorato di ricerca in Biotecnologie cellulari e molecolari <https://amsdottorato.unibo.it/view/dottorati/DOT210/>, 21 Ciclo. DOI 10.6092/unibo/amsdottorato/1613."],"dc:language":["en"],"dc:publisher":["Alma Mater Studiorum - Università di Bologna"],"dc:relation":["https://amsdottorato.unibo.it/id/eprint/1613/"],"dc:rights":["info:eu-repo/semantics/openAccess"],"dc:subject":["BIO/14 Farmacologia"],"dc:title":["Development of screening assays to test novel integrin antagonists in allergic inflammation"],"dc:type":["Doctoral Thesis","PeerReviewed"]},"updated_at":"2026-07-24T01:12:16Z"}