In vitro assay for the quantitative measurement of apoptotic lymphocytes phagocytosis by peripheral blood monocytes.
Currently used assays for the quantification of apoptotic cells uptake by phagocytes have several methodological problems. Our assay overcomes some of these problems. As a source of apoptotic cells we used peripheral blood lymphocytes obtained from the patients with chronic lymphoblast leukaemia. Apoptosis was induced by incubating cells with cycloheximide for up to 24 h. The assay was performed in suspension of peripheral blood mononuclear cells. For the visualisation of the phagocytes and phagocyted cells and discrimination of phagocyted from bound apoptotic cells we used Acridine orange/Ethidium bromide double staining. Here we offer a simple test which enables reliable measurement and it can show the difference of phagocytic potential between different individuals.
Popovic
S
S
Institute of Microbiology and Immunology, Faculty of Medicine, University of Kragujevac, Svetozara Markovica 69, 34000 Kragujevac, Serbia. pop21@ptt.yu
Arsenijevic
N
N
Baskic
D
D
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology; D003584:Cytological Techniques / Q000379:methods*; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000502:physiology*; D008214:Lymphocytes / Q000166:cytology* / Q000187:drug effects; D008856:Microscopy, Fluorescence; D010587:Phagocytosis / Q000502:physiology*; D013997:Time Factors
D003513:Cycloheximide