Introduction of a novel proliferation assay for pharmacological studies allowing the combination of BrdU detection and phenotyping.
A recently developed technology for the non-enzymatic detection of the thymidine analog 5-bromo-2-deoxyuridine (BrdU) has been evaluated. In contrast to previous enzymatic approaches, Ultraviolet-Induced Detection (UVID) of halogenated pyrimidines allows for a mild detection procedure which enables the simultaneous detection of cellular markers and DNA-synthesis without enzyme-specific disadvantages. Superantigen-stimulated peripheral blood mononuclear cells (PBMNCs) have been treated with two different inhibitors of proliferation and the cell cycle of different lymphocyte subsets has been analysed. Both pentoxifylline (POF) and 2-methoxyestradiol (2ME2) exhibited strong antiproliferative activity, but led to distinctive changes in the cell cycle distribution. This study shows that the UVID technology is a simple and viable method which should find a wide range of applications in immunological and pharmacodynamic assays.
Institute of Immunology and Transfusion Medicine, Medical University of Luebeck School of Medicine, Ratzeburger Allee 160, Lübeck, Germany.
H J
HJ
Hammers
M
M
Saballus
S
S
Sheikzadeh
P
P
Schlenke
D000077584:2-Methoxyestradiol; D018941:Antigens, CD19 / Q000096:biosynthesis; D001973:Bromodeoxyuridine / Q000032:analysis* / Q000528:radiation effects; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000187:drug effects / Q000276:immunology / Q000528:radiation effects; D002478:Cells, Cultured; D004247:DNA / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D016193:G1 Phase / Q000187:drug effects / Q000528:radiation effects; D006131:Growth Inhibitors / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D018655:Lymphocyte Count; D016131:Lymphocyte Subsets / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects; D010431:Pentoxifylline / Q000494:pharmacology; D016192:Resting Phase, Cell Cycle / Q000187:drug effects / Q000528:radiation effects; D014466:Ultraviolet Rays
D018941:Antigens, CD19; D006131:Growth Inhibitors; D004958:Estradiol; D000077584:2-Methoxyestradiol; D004247:DNA; D001973:Bromodeoxyuridine; D010431:Pentoxifylline