Kinetics of cell death and disintegration in human lymphocyte cultures.
In order to quantitate lymphocyte proliferative responses, we explored the role of cell death in the kinetics of phytohemagglutinin-stimulated cultures. Unless the disintegration time (tDIS) of nonviable lymphocytes in culture is known, the rate of cell death cannot be calculated. To obtain tDIS, we determined the time interval between total and viable cell population decay after various killing events. Two subpopulations of lymphocytes were observed, the major (80%) with a mean (+/-SEM) tDIS of 16+/-2 hr and the minor (20%) with a tDIS of 45+/-7 hr. Kinetic balance sheets were constructed predicting total culture DNA content (cells plus medium), as calculated both from proliferation rates and from observed death and disintegration rates. In an experiment characterized by extensive cell death, the two tallies were well-matched when the above data were utilized. The large discrepancy between predicted and observed DNA contents of the medium indicates that the DNA of disintegrated lymphocytes is extensively degraded. We conclude that cell death explains proliferation deficits in stimulated lymphocyte cultures. Our approach to quantitation of cell death may have general applicability to kinetic studies of cultured cells.
D002455:Cell Division; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002478:Cells, Cultured; D003486:Cyanides / Q000494:pharmacology; D003561:Cytarabine / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D037102:Lectins; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects
D003486:Cyanides; D037102:Lectins; D003561:Cytarabine; D003703:Demecolcine
Ann N Y Acad Sci. 1961 Nov 13;95:743-57
14039747
Biochem J. 1967 Nov;105(2):679-84
5584011
J Biol Chem. 1958 Jul;233(1):184-8
13563465
Clin Exp Immunol. 1977 Jan;27(1):111-7
849644
Proc Natl Acad Sci U S A. 1976 Sep;73(9):3211-5
1067614
J Clin Invest. 1971 Apr;50(4):818-26
4100684
Cell Immunol. 1975 Apr;16(2):237-50
46788
Proc Natl Acad Sci U S A. 1972 Jul;69(7):1685-9
4505646
J Exp Med. 1974 May 1;139(5):1037-48
4825240
Immunol Commun. 1973;2(3):277-85
4742972
J Exp Med. 1969 Aug 1;130(2):327-43
4893888
J Exp Med. 1976 May 1;143(5):1249-64
1262786
Science. 1975 Dec 12;190(4219):1093-5
1188385
Blood. 1975 Oct;46(4):509-18
1174688
J Reticuloendothel Soc. 1975 Jan;17(1):47-56
1113269
Int Arch Allergy Appl Immunol. 1970;37(4):393-408
5411051
Clin Exp Immunol. 1968 Jul;3(6):571-84
5700693
Virchows Arch Pathol Anat Physiol Klin Med. 1960;333:421-65
13765553