{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T13:10:01Z","timestamp":1789132201443,"version":"build-2803163510"},"reference-count":57,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T00:00:00Z","timestamp":1786924800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002428","name":"Austrian Science Fund","doi-asserted-by":"publisher","award":["I3946-B33"],"award-info":[{"award-number":["I3946-B33"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>The major birch pollen allergen Bet v 1 primarily affects the nose and eyes of allergic patients. After crossing the epithelial barrier, it causes IgE-mediated effector cell release, triggering inflammatory responses. We sought to develop bispecific nanobodies that bind Bet v 1 on mucosal surfaces to prevent allergen penetration and subsequent activation of effector cells. Intercellular adhesion molecule 1 (ICAM-1), highly upregulated on epithelial cells of allergic individuals, was chosen as an anchor.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Genetically fused nanobodies targeting Bet v 1 and ICAM-1 were generated and characterized for specificity, affinity, Bet v 1 immobilization on the surface of the human bronchial epithelial cell line, 16HBE14o-, and their potential to inhibit Bet v 1 penetration through a cell monolayer.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Bispecific nanobodies recognized ICAM-1, Bet v 1 and related tree pollen allergens (Aln g 1, Cor a 1). While we observed a clear co-localization of nanobodies and Bet v 1 on the 16HBE14o-cell surface after 30\u00a0min, Bet v 1 was slightly internalized after 4h at 32\u00a0\u00b0C. Dissociation rate constants between bispecific nanobodies and antigens were 3\u20136 times faster at 32\u00a0\u00b0C compared to 25\u00a0\u00b0C. Bispecific nanobodies reduced Bet v 1 penetration and hence decreased basophil activation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>We generated bivalent nanobodies specific for ICAM-1 and Bet v 1. These nanobodies diminished trans-epithelial transport of Bet v 1 by ICAM-1-mediated immobilization at the cell surface. Reduced Bet v 1 penetration resulted in decreased basophil activation, demonstrating the potential of local administration of nanobodies for the prevention of pollen allergy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1864741","type":"journal-article","created":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T05:25:49Z","timestamp":1786944349000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Bispecific nanobodies targeting Bet v 1 and ICAM-1 decrease respiratory epithelial penetration of Bet v 1"],"prefix":"10.3389","volume":"17","author":[{"given":"Ines","family":"Zettl","sequence":"first","affiliation":[{"name":"Institute of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna","place":["Vienna, 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