Identification of compounds that enhance the anti-lymphoma activity of rituximab using flow cytometric high-content screening.
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.
Gasparetto
Maura
M
Duke University Medical Center, Duke University, Durham, NC, USA.
Gentry
Tracy
T
Sebti
Said
S
O'Bryan
Erica
E
Nimmanapalli
Ramadevi
R
Blaskovich
Michelle A
MA
Bhalla
Kapil
K
Rizzieri
David
D
Haaland
Perry
P
Dunne
Jack
J
Smith
Clay
C
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
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D010618:Phenanthrolines; D016590:Aphidicolin; D000069283:Rituximab; D019772:Topotecan