Page 369 - 2014 Printable Abstract Book
P. 369
(PS7-29) Investigation of time-consuming lymphocyte proliferation as a limiting factor in dicentric
analysis for individual biodosimetry. Christina Beinke, Dr.; Andreas Lamkowski, Dr.; Matthias Port, PD
Dr.; and Michael Abend, Prof. Dr. Bundeswehr Institute of Radiobiology affiliated to the University of Ulm,
Munich, Germany
Dicentric analysis (DA) is the most validated biodosimetry method and still serves as the gold
standard for individual radiation dose assessment. The two most limiting factors of DA are the time
consuming lymphocyte proliferation and the maximum sample capacity of a service lab. We started to
investigate the lymphocyte stimulation in order to identify targets suitable to be used for a time saving
modulation. As since 50 years phythemagglutinin (PHA-M) is used to produce mitotic lymphocytes, but
only few publications analyzing different mitogens exist, we examined the effectiveness of certain known
mitogens by comparing the mitotic indices (MI). Applied mitogens: 5 lectins (pokeweed [PW],
concanavalin A [CNA], LMA (maackia amurensis), LTV (tritium vulgaris) PHA-L and PHA-M (phaseolus
vulgaris) as well as LPS (E. coli) and streptolysin-O (SLO, S. pyogenes). We chose 3 concentrations for each
drug based on product specifications and literature search. Human peripheral whole blood supplemented
with mitogen and BrdU was cultivated in RPMI/FCS and colcemid was used to arrest cells in metaphase.
After automated cell fixation, slide preparation and FpG staining the MI were determined by manual
microscopy. Incubation of lymphocyte cultures for 48 h resulted on average in a 4-11-fold higher MI for
PHA-M stimulated cells (MI=6.7% at 26 µl/ml) over PHA-L (MI=1.7% at 10 µg/ml), CNA (MI=1.0% at 50
µg/ml) and PW (MI=0.6% at 10 µg/ml only). No metaphases were found for the other concentrations or
mitogens. Extending the incubation time to 72 h increased the average MI up to 27.0% (at 26 µl/ml) for
PHA-M, 12.5% and 4.9% (at 100 µg/ml and 50 µg/ml, respectively) for SLO, 5.5% (at 50 µg/ml) for CNA,
5.3% (at 10 µg/ml) for PHA-L and 3.9% (at 10 µg/ml) for PW. Finally, we used these selected lectines
combined with 3 different concentrations of SLO or LPS, respectively. Neither SLO nor LPS combined with
selected lectines increased the MI over the MI with the lectine alone. In summary, the widely used PHA-
M concentration for lymphocyte stimulation is 4-11 times more effective than administration of other
lectines and a combination of lectines with SLO or LPS.
1
2
(PS7-30) Novel radiation mitigators and anticancer drugs. Robert H. Schiestl, PhD ; Yelena Rivina ; and
1
1
Michael Davoren , UCLA School of Medicine and Public Health, Los Angeles, CA and Stanford, Palo Alto,
2
CA
The possibility of a radiation disaster from a nuclear detonation or accident has existed for over
50 years and spawned much of the basic research in radiobiology in the 1950-60s. The recent Fukushima
accident was yet another reminder that there remains a dire need to develop novel therapies against
radiation-induced toxicities. Here we report on the development of two novel radiation countermeasure
therapies: Yel001 and Yel002. These small, biologically active, drug-like molecules were uncovered in the
DEL high throughput assay reducing radiation-induced cyto- and geno-toxicity in yeast. Radiation-
modulating activity was further confirmed in yeast plate-based DEL Assay: addition of either Yel001 or
Yel002 to irradiated cultures reduced cell death and genomic instability. Further, Yel compounds increases
survival to 75% in vivo following an LD100/30 dose of ionizing radiation (IR) with the first therapeutic
injection administered 24 hours post exposure followed by injections at 48,72,96, and 120 hours.
Additionally, treatment with Yel001 and Yel002 compounds reduces radiation-induced leukemia from
90% to to 50% and 40% respectively. Of note, treatment with either Yel001 or Yel002 reduced
367 | P a g e
analysis for individual biodosimetry. Christina Beinke, Dr.; Andreas Lamkowski, Dr.; Matthias Port, PD
Dr.; and Michael Abend, Prof. Dr. Bundeswehr Institute of Radiobiology affiliated to the University of Ulm,
Munich, Germany
Dicentric analysis (DA) is the most validated biodosimetry method and still serves as the gold
standard for individual radiation dose assessment. The two most limiting factors of DA are the time
consuming lymphocyte proliferation and the maximum sample capacity of a service lab. We started to
investigate the lymphocyte stimulation in order to identify targets suitable to be used for a time saving
modulation. As since 50 years phythemagglutinin (PHA-M) is used to produce mitotic lymphocytes, but
only few publications analyzing different mitogens exist, we examined the effectiveness of certain known
mitogens by comparing the mitotic indices (MI). Applied mitogens: 5 lectins (pokeweed [PW],
concanavalin A [CNA], LMA (maackia amurensis), LTV (tritium vulgaris) PHA-L and PHA-M (phaseolus
vulgaris) as well as LPS (E. coli) and streptolysin-O (SLO, S. pyogenes). We chose 3 concentrations for each
drug based on product specifications and literature search. Human peripheral whole blood supplemented
with mitogen and BrdU was cultivated in RPMI/FCS and colcemid was used to arrest cells in metaphase.
After automated cell fixation, slide preparation and FpG staining the MI were determined by manual
microscopy. Incubation of lymphocyte cultures for 48 h resulted on average in a 4-11-fold higher MI for
PHA-M stimulated cells (MI=6.7% at 26 µl/ml) over PHA-L (MI=1.7% at 10 µg/ml), CNA (MI=1.0% at 50
µg/ml) and PW (MI=0.6% at 10 µg/ml only). No metaphases were found for the other concentrations or
mitogens. Extending the incubation time to 72 h increased the average MI up to 27.0% (at 26 µl/ml) for
PHA-M, 12.5% and 4.9% (at 100 µg/ml and 50 µg/ml, respectively) for SLO, 5.5% (at 50 µg/ml) for CNA,
5.3% (at 10 µg/ml) for PHA-L and 3.9% (at 10 µg/ml) for PW. Finally, we used these selected lectines
combined with 3 different concentrations of SLO or LPS, respectively. Neither SLO nor LPS combined with
selected lectines increased the MI over the MI with the lectine alone. In summary, the widely used PHA-
M concentration for lymphocyte stimulation is 4-11 times more effective than administration of other
lectines and a combination of lectines with SLO or LPS.
1
2
(PS7-30) Novel radiation mitigators and anticancer drugs. Robert H. Schiestl, PhD ; Yelena Rivina ; and
1
1
Michael Davoren , UCLA School of Medicine and Public Health, Los Angeles, CA and Stanford, Palo Alto,
2
CA
The possibility of a radiation disaster from a nuclear detonation or accident has existed for over
50 years and spawned much of the basic research in radiobiology in the 1950-60s. The recent Fukushima
accident was yet another reminder that there remains a dire need to develop novel therapies against
radiation-induced toxicities. Here we report on the development of two novel radiation countermeasure
therapies: Yel001 and Yel002. These small, biologically active, drug-like molecules were uncovered in the
DEL high throughput assay reducing radiation-induced cyto- and geno-toxicity in yeast. Radiation-
modulating activity was further confirmed in yeast plate-based DEL Assay: addition of either Yel001 or
Yel002 to irradiated cultures reduced cell death and genomic instability. Further, Yel compounds increases
survival to 75% in vivo following an LD100/30 dose of ionizing radiation (IR) with the first therapeutic
injection administered 24 hours post exposure followed by injections at 48,72,96, and 120 hours.
Additionally, treatment with Yel001 and Yel002 compounds reduces radiation-induced leukemia from
90% to to 50% and 40% respectively. Of note, treatment with either Yel001 or Yel002 reduced
367 | P a g e
