A quantitative approach for studying IgE-FcεRI aggregation

Richard G. Posner, Paul B. Savage, Adam S. Peters, Angelica Macias, Jamael DelGado, Gordon Zwartz, Larry A. Sklar, William S. Hlavacek

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Aggregation of cell surface receptors is a ubiquitous means of initiating signal transduction in many cellular systems. In this manuscript, we describe a combined theoretical and experimental approach based on multiparameter flow cytometry for measuring the time course of ligand induced aggregation of IgE-FcεRI on RBL cells. By fluorescently labeling both the ligand and surface IgE (sIgE), we have developed an assay that permits us to simultaneously measure both occupancy of sIgE combining sites and association of antigen with the cell surface. This allows for a direct calculation of the degree of receptor aggregation present on the cell. By employing new mixing technologies developed for flow cytometry, we are able to look at aggregation in the sub second time domain. To extend our work, we have synthesized a new set of chemically well defined ligands (of valences 1-3) to use as probes in our studies. We show that the magnitude of the cellular response is dramatically increased as the valence of our ligand is raised from two to three.

Original languageEnglish (US)
Pages (from-to)1221-1228
Number of pages8
JournalMolecular Immunology
Volume38
Issue number16-18
DOIs
StatePublished - Nov 12 2002

Keywords

  • IgE-FcεRI aggregation
  • Ligand-receptor binding
  • Receptor aggregation

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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    Posner, R. G., Savage, P. B., Peters, A. S., Macias, A., DelGado, J., Zwartz, G., Sklar, L. A., & Hlavacek, W. S. (2002). A quantitative approach for studying IgE-FcεRI aggregation. Molecular Immunology, 38(16-18), 1221-1228. https://doi.org/10.1016/S0161-5890(02)00067-6