title: Identification of compounds that enhance the anti-lymphoma activity of rituximab using flow cytometric high-content screening.
abstract: In this report, we describe a new flow cytometry technique termed flow cytometric high-content screening (FC-HCS) which involves semi-automated processing and analysis of multiparameter flow cytometry samples. As a first test of the FC-HCS technique, we used it to screen a 2000-compound library, called the National Cancer Institute (NCI) Diversity Set, to identify agents that would enhance the anti-lymphoma activity of the therapeutic monoclonal antibody rituximab. FC-HCS identified 15 compounds from the Diversity Set that significantly enhanced the ability of rituximab to inhibit cell cycle progression and induce apoptosis in lymphoma cells. The validity of the screening results was confirmed for several compounds using additional assays of cell proliferation, apoptosis and cell growth. The FC-HCS technique was relatively simple and reliable and could process up to 1000 samples/day on a single flow cytometer. The FC-HCS technique may be useful for a variety of applications including drug discovery, immunologic monitoring of patients, functional genomics studies and tissue engineering efforts.
authors.affiliation: Duke University Medical Center, Duke University, Durham, NC, USA.
authors.forename: Maura
authors.initials: M
authors.lastname: Gasparetto
authors.forename: Tracy
authors.initials: T
authors.lastname: Gentry
authors.forename: Said
authors.initials: S
authors.lastname: Sebti
authors.forename: Erica
authors.initials: E
authors.lastname: O'Bryan
authors.forename: Ramadevi
authors.initials: R
authors.lastname: Nimmanapalli
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Blaskovich
authors.forename: Kapil
authors.initials: K
authors.lastname: Bhalla
authors.forename: David
authors.initials: D
authors.lastname: Rizzieri
authors.forename: Perry
authors.initials: P
authors.lastname: Haaland
authors.forename: Jack
authors.initials: J
authors.lastname: Dunne
authors.forename: Clay
authors.initials: C
authors.lastname: Smith
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D058846:Antibodies, Monoclonal, Murine-Derived; D016590:Aphidicolin / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D008223:Lymphoma / Q000188:drug therapy* / Q000473:pathology; D010618:Phenanthrolines / Q000494:pharmacology; D000069283:Rituximab; D019772:Topotecan / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D010618:Phenanthrolines; D016590:Aphidicolin; D000069283:Rituximab; D019772:Topotecan