The JAM Test and its daughter P-JAM: simple tests of DNA fragmentation to measure cell death and stasis.
Cytotoxic T lymphocytes, and other death-inducing agents, have at least two different ways of killing their targets: drilling holes in the target cell membrane, or triggering the targets to commit suicide. The JAM Test is a method that measures the DNA fragmentation that accompanies cell suicide. We label target cells with radioactive DNA-precursor nucleotides and harvest them onto fiberglass filters, which trap large pieces of DNA but pass smaller fragments of apoptotic cells. As a general measure of apoptosis, the JAM Test described here is faster (can be completed in 4 h [or less if labeling is done the night before]), more quantitative, easier, more sensitive, more flexible and cheaper than most other current assays of apoptosis. The P-JAM, also discussed, additionally allows for assessment of death in cells that don't fragment their DNA, and allows for assays of agents that induce cell stasis rather than death.
Usharauli
David
D
Laboratory of Cellular and Molecular Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 4, Room 111, Bethesda, Maryland 20892, USA.
Perez-Diez
Ainhoa
A
Matzinger
Polly
P
D000818:Animals; D016923:Cell Death; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D053938:DNA Fragmentation; D051379:Mice; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*