Fg. Blankenberg et al., QUANTITATIVE-ANALYSIS OF APOPTOTIC CELL-DEATH USING PROTON NUCLEAR-MAGNETIC-RESONANCE SPECTROSCOPY, Blood, 89(10), 1997, pp. 3778-3786
Quantification of apoptotic cell death in vivo has become an important
area of investigation in patients with acute lymphoblastic leukemia (
ALL). We have devised a noninvasive analytical method to estimate the
percentage of apoptotic lymphoblasts in doxorubicin-treated Jurkat T-c
ell ALL cultures, using proton nuclear magnetic resonance spectroscopy
(H-1 NMR). We have found that the ratio of the methylene (CH2) resona
nce (at 1.3 ppm) to the methyl (CH3) resonance (at 0.9 ppm) signal int
ensity, as observed by H-1 NMR, is directly proportional to the percen
tage of apoptotic lymphoblasts in vitro. The correlation between the C
H2/CH3 signal intensity ratio and the percentage of apoptotic lymphobl
asts was optimal 24 to 28 hours after doxorubicin treatment (r(2) = .9
47, N = 27 samples), There was also a direct temporal relationship bet
ween an increase in the CH2/ CH3 signal intensity ratio and the onset
of apoptosis as detected by nuclear morphologic analysis, fluorescein-
annexin V flow cytometry, and DNA gel electrophoresis. Thin-layer chro
matography confirmed that a dynamic and/or compositional change of the
plasma membrane, rather than increases in lipase activity or fatty ac
id production, appears to account for the increase in the CH2/CH3 sign
al intensity ratio during apoptosis. H-1 NMR may have clinical utility
for the early noninvasive assessment of chemotherapeutic efficacy in
patients with ALL. (C) 1997 by The American Society of Hematology.