{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T05:06:24Z","timestamp":1770699984031,"version":"3.49.0"},"reference-count":32,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2005,3,8]],"date-time":"2005-03-08T00:00:00Z","timestamp":1110240000000},"content-version":"vor","delay-in-days":6521,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry"],"published-print":{"date-parts":[[1987,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>In RBL\u20102H3 rat mucosal mast cells, the crosslinking of cell\u2010surface IgE\u2010receptor complexes by multivalent antigens initiates a sequence of responses leading to degranulation. We have developed a family of dinitrophenol (DNP)\u2010conjugated fluorescent antigens to study dynamic membrane events associated with these responses. Lysyl groups on the phycobiliproteins, B\u2010phycoerythrin and C\u2010phycocyanin, were labelled with DNP, yielding fluorescent conjugates that cause the release of [<jats:sup>3<\/jats:sup>H]serotonin from anti\u2010DNO\u2010IgE\u2010primed RBL\u20102H3 cells. The binding of these antigens to IgE\u2010receptor complexes was observed by fluorescence microscopy and quantified by flow cytometry. Incubation with 1 \u03bcg\/ml DNP<jats:sub>42<\/jats:sub>\u2010B\u2010phycoerythrin stimulates maximum degranulation from IgE\u2010saturated cells. Under these conditions, approximately 26 \u00d7 10<jats:sup>3<\/jats:sup> molecules of DNP<jats:sub>42<\/jats:sub>\u2010B\u2010phycoerythrin are bound per cell at equilibrium. The rate and extent of antigen binding and of antigenstimulated mediator release decrease in parallel as the concentration and DNP:protein ratio of the fluorescent conjugates is reduced. Secretion stops immediately when the nonfluorescent monovalent antigen, DNP\u2010lysine, is added to degranulating cell suspensions. DNP\u2010lysine also displaces surface\u2010bound antigen when added during the first minutes after multivalent antigen. However, the ability of DNP\u2010lysine to displace surface\u2010bound DNP<jats:sub>42<\/jats:sub>\u2010B\u2010phycoerythrin from IgE\u2010receptor complexes decreases progressively with time. Treatment with dihydrocytochalasin B and several analogs that prevent antigen\u2010stimulated F\u2010actin assembly enhances secretion and delays the transition of antigen to its DNP\u2010lysine\u2010resistant form. Cytochalasin treatment also permits the long\u2010range movement of antigen into surface caps. Based on these data, we propose that secretion is triggered by the act of IgE\u2010receptor crosslinking or by a shortlived excited state of the crosslinked antigen\u2010IgE\u2010receptor complex. We propose further that antigen\u2010stimulated F\u2010actin assembly contributes to the transition of antigen\u2010IgE\u2010receptor complexes to a DNP\u2010lysine\u2010resistant form that does not trigger secretion. Two posible mechanisms for the transition to DNP\u2010lysine resistance are discussed.<\/jats:p>","DOI":"10.1002\/cyto.990080309","type":"journal-article","created":{"date-parts":[[2005,8,8]],"date-time":"2005-08-08T14:00:02Z","timestamp":1123509602000},"page":"287-295","source":"Crossref","is-referenced-by-count":31,"title":["DNP\u2010phycobiliproteins, fluorescent antigens to study dynamic properties of antigen\u2010IgE\u2010receptor complexes on RBL\u20102H3 rat mast cells"],"prefix":"10.1002","volume":"8","author":[{"given":"Jean Clare","family":"Seagrave","sequence":"first","affiliation":[]},{"given":"Grace G.","family":"Deanin","sequence":"additional","affiliation":[]},{"given":"John C.","family":"Martin","sequence":"additional","affiliation":[]},{"given":"Bruce H.","family":"Davis","sequence":"additional","affiliation":[]},{"given":"Janet 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