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Figure 2

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Kinetics for γ-H2AX foci loss in macaque lymphocytes after total body irradiation.

(A) Representative images of macaque lymphocytes in Sham-irradiated (Sham) and 0.3, 1, 2, 3, 7 and 10 days (d) after 8.5 Gy-TBI. As previously observed with blood samples irradiated ex-vivo, the γ-H2AX foci were fewer and fainter in neutrophils than in lymphocytes. Green, γ-H2AX; red, DNA stained with PI. Arrowheads indicate neutrophils. (B) γ-H2AX foci distribution in lymphocytes taken from individual animals subjected to TBI. The rows from top to bottom show the focal distributions from individual animals after 8.5 Gy-TBI (6 macaques E25 to E30) and Sham-TBI (2 macaques E23 and E24, noted S), 6.5 Gy-TBI (6 macaques E15 toE20) and Sham-TBI (2 macaques E21 and E22, noted S), 3.5 Gy-TBI (6 macaques E01to E06) and Sham-TBI (2 macaques E07and E08, noted S), and 1 Gy-TBI (6 macaques E09 to E14) and sham-TBI (2 macaques E21 and E22, noted S). Results from the sham-IR are shown with red lines (noted S). The columns from left to right correspond to the different sampling times post-TBI, 0.3 d (1 and 3.5 Gy), 1 d (1 , 3.5 , 6.5 and 8.5 Gy), 2 d (1 , 3.5 , 6.5 and 8.5 Gy), and 4 d (3.5 , 6.5 and 8.5 Gy). (C) Values for γ-H2AX foci per cell vs days after the indicated TBI doses. The data for the TBI treated cohorts (blue bars) and sham treated controls for each cohort (red bars) are plotted on a log scale to bring out the details concerning differences between the TBI and sham treated animals. The data are shown as mean numbers of γ-H2AX foci per lymphocyte from each cohort ± standard deviations (n = 2 for Sham irradiated samples; n = 6 for TBI-irradiation, except 16 days after 3.5 Gy, n = 3 and 11 days after 8.5 Gy; no SD was determined 14 days after 8.5 Gy as data were prepared from 1 animal). (D) Foci per cell values in lymphocytes taken at the noted time (Pre = the day before TBI) and plotted vs TBI dose. Data are means ± standard deviations (n = 6). Equations for trendlines and R-squared values are y = −0.005x+0.096 / R2 = 0.225 for Pre-IR; y = 3.167x−0.578 / R2 = 0.980 for 0.3 d; y = 0.672x−0.034 / R2 = 0.996 for 1 d; y = 0.286x+0.014 / R2 = 0.993 for 2 d and y = 0.158x+0.097 / R2 = 0.956 for 4 d. Asterisks in (C) denote statistically difference between TBI-and sham-irradiated macaques (P<0.05, t-test). Detailed statistical analysis is shown in Table 2. Numbers in (C) for the 6.5 and 8.5 Gy panels correspond to the numbers of animals alive after TBI at the indicated time (no number indicates that all 6 TBI-irradiated animals are alive).

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