Page 348 - 2014 Printable Abstract Book
P. 348
evaluated for 60 days (d) for hematology, body weight, core temperature, food consumption, and clinical
observations. Supportive care included nutritional support, analgesics, rehydration fluids, antibiotics,
antidiarrheals, antipyretics, antiemetics, antiulceratives, and blood transfusions. The primary end-point
was survival at 60 d post-irradiation. The survival curve demonstrated a dose-dependent effect of
irradiation. Twenty-five of 48 animals (52%) succumbed over the dose range 6.25 - 8.75 Gy. The LD30/60,
LD50/60, LD70/60 and LD 90/60 [95% CI] were estimated at 6.88 [5.98, 7.31], 7.43 [6.92, 7.90], 7.98 [7.56,
8.79], and 8.77 [8.20, 10.36]. The slope of the DRR was 0.96 (±0.51) probits per linear dose. The respective
overall mean survival time (MST) for each cohort ranged from 15 - 20 d with a MST of decedents at all
doses of 16.7 d (range 11 - 23 d). No deaths were observed following d24 post TBI. There was a reduction
-1
-1
in absolute neutrophil count (ANC) to <500 µL 6 - 7 d after irradiation and to <100 µL within 8 – 13 d.
-1
The ANC for all animals fell to <100 µL (grade 4 neutropenia). The 7.25 Gy cohort, evaluated for its
-1
proximity to the LD50/60 of 7.43 Gy on the probit plot, had an average nadir of 3 µL on day 13. The ANC
-1
of survivors recovered to ≥100 µL between days 14 – 22. Only surviving animal’s attained recovery to
-1
ANC ≥500 µL (d 17– 24). The first day the PLT count decreased to <20,000 µL was similar between all
-1
-1
dose cohorts and ranged from days 10 – 13. The duration of thrombocytopenia (PLT <20,000 µL ) at 7.25
Gy was 8.6 d (range, 4 - 14 d). The DRR established at SNBL is consistent with the previously published
DRR in rhesus macaques administered medical management (Health Physics 103, 2012 pg.367). The
comparable LD50/60 and slope provides definitive validation of the radiation physics and medical
management administered in the study.
1
1
1
(PS6-41) X-ray irradiation of mice and blood. David J. Sandgren ; Sereyratana Som ; Cielita L. Page ;
1
1
1
1
1
Norbert Makori ; Steven M. Glaza ; Ronald G. Manning ; Thomas W. Beck ; Koichiro Fukuzaki ; and Ryoichi
Nagata , SNBL USA, Ltd., Everett, WA and Shin Nippon Biomedical Laboratories, Ltd., Kagoshima, Japan
1
2
2
Radiation research capabilities at SNBL USA, Ltd. have expanded to include two total-body
irradiation (TBI) mouse models and ex vivo blood irradiation. SNBL USA can support drug development
efforts around mitigation of and treating radiation injury specifically related to Acute Radiation Syndrome
(ARS) with mouse survival studies and mouse time course biosampling studies. We have successfully
established and herein present a 30 day dose response curve for TBI (0-14 Gy, X-Ray, 160 kV at 25 mA)
male C57BL/6 mice, including LD10 through LD90. During the study, it was essential to ensure objectivity
among staff, better animal welfare, and consistent endpoint recording. We accomplished this by
establishing an ARS severity scoring system based on animal appearance and behavior. Individual scores
for posture, coat, and behavior, as well as combined scores, increased as dose increased and time
progressed. Differences in total scores (out of 12 possible) compared to control mice from 7, 9, and 14 Gy
show up as early as 10, 3, and 2 days post-TBI, respectively. At day 5 post-TBI, 14 Gy mice mean total
scores were 6 counts higher than 8 Gy mean total scores. A daily plasma and biomarker collection post-
TBI is another established male C57BL/6 mouse model. The volume of blood and plasma was inversely
proportional to radiation dose as time progressed, correlating well with loss in body weights. Neutrophil
counts show a significant increase (R < 0.01) over control at 6 h post-TBI (4.3-, 5.6-, and 6.2-fold for 3, 8,
and 14 Gy, respectively) followed by the classic fall in concentration. Lymphocyte counts show a 4.2-, 4.7-
, and 4.1-fold decrease at 6 h for 3, 8, and 14 Gy vs control, respectively (R < 0.001). Both neutrophils and
lymphocytes show a dose dependant change through day 6. Select histopathological results on 2 d, 6 d,
30 d, or last day of survival post-TBI will also be presented. Part of SNBL USA’s full supportive care
paradigm in TBI nonhuman primate studies includes blood transfusions. On-site X-ray irradiation
346 | P a g e
observations. Supportive care included nutritional support, analgesics, rehydration fluids, antibiotics,
antidiarrheals, antipyretics, antiemetics, antiulceratives, and blood transfusions. The primary end-point
was survival at 60 d post-irradiation. The survival curve demonstrated a dose-dependent effect of
irradiation. Twenty-five of 48 animals (52%) succumbed over the dose range 6.25 - 8.75 Gy. The LD30/60,
LD50/60, LD70/60 and LD 90/60 [95% CI] were estimated at 6.88 [5.98, 7.31], 7.43 [6.92, 7.90], 7.98 [7.56,
8.79], and 8.77 [8.20, 10.36]. The slope of the DRR was 0.96 (±0.51) probits per linear dose. The respective
overall mean survival time (MST) for each cohort ranged from 15 - 20 d with a MST of decedents at all
doses of 16.7 d (range 11 - 23 d). No deaths were observed following d24 post TBI. There was a reduction
-1
-1
in absolute neutrophil count (ANC) to <500 µL 6 - 7 d after irradiation and to <100 µL within 8 – 13 d.
-1
The ANC for all animals fell to <100 µL (grade 4 neutropenia). The 7.25 Gy cohort, evaluated for its
-1
proximity to the LD50/60 of 7.43 Gy on the probit plot, had an average nadir of 3 µL on day 13. The ANC
-1
of survivors recovered to ≥100 µL between days 14 – 22. Only surviving animal’s attained recovery to
-1
ANC ≥500 µL (d 17– 24). The first day the PLT count decreased to <20,000 µL was similar between all
-1
-1
dose cohorts and ranged from days 10 – 13. The duration of thrombocytopenia (PLT <20,000 µL ) at 7.25
Gy was 8.6 d (range, 4 - 14 d). The DRR established at SNBL is consistent with the previously published
DRR in rhesus macaques administered medical management (Health Physics 103, 2012 pg.367). The
comparable LD50/60 and slope provides definitive validation of the radiation physics and medical
management administered in the study.
1
1
1
(PS6-41) X-ray irradiation of mice and blood. David J. Sandgren ; Sereyratana Som ; Cielita L. Page ;
1
1
1
1
1
Norbert Makori ; Steven M. Glaza ; Ronald G. Manning ; Thomas W. Beck ; Koichiro Fukuzaki ; and Ryoichi
Nagata , SNBL USA, Ltd., Everett, WA and Shin Nippon Biomedical Laboratories, Ltd., Kagoshima, Japan
1
2
2
Radiation research capabilities at SNBL USA, Ltd. have expanded to include two total-body
irradiation (TBI) mouse models and ex vivo blood irradiation. SNBL USA can support drug development
efforts around mitigation of and treating radiation injury specifically related to Acute Radiation Syndrome
(ARS) with mouse survival studies and mouse time course biosampling studies. We have successfully
established and herein present a 30 day dose response curve for TBI (0-14 Gy, X-Ray, 160 kV at 25 mA)
male C57BL/6 mice, including LD10 through LD90. During the study, it was essential to ensure objectivity
among staff, better animal welfare, and consistent endpoint recording. We accomplished this by
establishing an ARS severity scoring system based on animal appearance and behavior. Individual scores
for posture, coat, and behavior, as well as combined scores, increased as dose increased and time
progressed. Differences in total scores (out of 12 possible) compared to control mice from 7, 9, and 14 Gy
show up as early as 10, 3, and 2 days post-TBI, respectively. At day 5 post-TBI, 14 Gy mice mean total
scores were 6 counts higher than 8 Gy mean total scores. A daily plasma and biomarker collection post-
TBI is another established male C57BL/6 mouse model. The volume of blood and plasma was inversely
proportional to radiation dose as time progressed, correlating well with loss in body weights. Neutrophil
counts show a significant increase (R < 0.01) over control at 6 h post-TBI (4.3-, 5.6-, and 6.2-fold for 3, 8,
and 14 Gy, respectively) followed by the classic fall in concentration. Lymphocyte counts show a 4.2-, 4.7-
, and 4.1-fold decrease at 6 h for 3, 8, and 14 Gy vs control, respectively (R < 0.001). Both neutrophils and
lymphocytes show a dose dependant change through day 6. Select histopathological results on 2 d, 6 d,
30 d, or last day of survival post-TBI will also be presented. Part of SNBL USA’s full supportive care
paradigm in TBI nonhuman primate studies includes blood transfusions. On-site X-ray irradiation
346 | P a g e
