The human lung is among the most sensitive and critical tissues of concern in localized and systemic radiation exposures, and it is a subject of active preclinical research for evaluating mitigating therapies within the radiation countermeasures program. Our previous study comparing C57BL/6, CBA and C57L mice after whole-thorax irradiation pointed to the problems of late pleural effusions that prevented the full development of lung injury in C57BL/6 mice and suggested that the CBA and C57L strains are more favorable for modeling lung injury in humans (Jackson et al., Radiat. Res. 173, 10–20, 2010). We extended these comparisons to include three other mouse strains (BALB/c, C57BR/J and A/J mice) irradiated with 10, 12.5 or 15 Gy. Most of these mice were unable to survive the first 6 months and presented with a mixture of lung injury and pleural effusions as determined from gross pathology, histology and micro-CT. The independent and varying development of compressive pleural effusions of ill-defined etiology represents a concern for these strains in that they may not satisfy the preclinical requirements for approval of medical countermeasures (e.g. radiation mitigators) for human use. Thus, among the various different mouse strains studied so far for these pathologies, only three (CBA, C3H and C57L) appear to be desirable in exhibiting an early wave of pulmonary dysfunction attributed exclusively to radiation pneumonitis and for further assessment of radioprotective and mitigating therapies. C57L mice are particularly relevant in that they show significant lung damage at lower radiation doses that are closer to what is predicted for humans.

The mouse has been used extensively to model radiation injury to the lung, a major dose-limiting organ for radiotherapy. Substantial differences in the timing and sensitivity of this tissue between mouse strains have been reported, with some strains, including C57BL/6, being designated as “fibrosis-prone”. Pleural effusions have also been reported to be a prominent problem in many mouse strains, but it remains unclear how this affects the lung function and survival of the standard C57BL/6 mouse. The purpose of this investigation was to re-evaluate this strain in comparison with C57L and CBA mice after whole-thorax irradiation at doses ranging from 10 to 15 Gy. Breathing rate measurements, micro-computerized tomography, lung tissue weight, pleural fluid weight and histopathology showed that the most prominent features were an early phase of pneumonitis (C57L and CBA) followed by a late incidence of massive pleural effusions (CBA and C57BL/6). A remarkable difference was seen between the C57 strains: The C57L mice were exquisitely sensitive to early pneumonitis at 3 to 4 months while C57BL/6 mice showed a delayed response, with most mice presenting with large accumulations of pleural fluid at 6 to 9 months. These results therefore caution against the routine use of C57BL/6 mice in radiation lung experiments because pleural effusions are rarely observed in patients as a consequence of radiotherapy. Future experiments designed to investigate genetic determinants of radiation lung damage should focus on the high sensitivity of the C57L strain (in comparison with CBA or C3H mice) and the possibility that they are more susceptible to pulmonary fibrosis as well as pneumonitis.

The purpose of preclinical murine model development is to establish that the pathophysiological outcome of the rodent model of radiation-induced lung injury is sufficiently representative of the anticipated pulmonary response in the human population. This objective is based on concerns that the C57BL/6J strain may not be the most appropriate preclinical model of lethal radiation lung injury in humans. In this study, the authors assessed this issue by evaluating the relationship between morbidity (pulmonary function, histopathologic damage) and mortality among three strains of mice: C57BL/6J, CBA/J, and C57L/J. These different strains display variations in latency and phenotypic expression of radiation-induced lung damage. By comparing the response of each strain to the human pulmonary response, an appropriate animal model(s) of human radiation-induced pulmonary injury was established. Observations in the C57L/J and CBA/J murine models can be extrapolated to the human lung for evaluation of the mechanisms of action of radiation as well as future efficacy testing and approving agents that fall under the “Animal Rule” of the U.S. Food and Drug Administration (FDA) (21 CFR Parts 314 and 601).

Many studies in biomedical research and various allied fields, in which cells or laboratory animals are exposed to radiation, rely on adequate radiation dose standardization for reproducibility and comparability of biological data. Due to increasing concerns regarding international terrorism, the use of radioactive isotopes has recently been met with enhanced security measures. Thus, a growing number of researchers have considered transferring their studies from gamma-ray to kilovoltage X-ray irradiators. Current commercially-available X-ray biological irradiators produce radiation beams with reasonable field geometry and overall dose-homogeneity; however, they operate over a wide range of different energies, both between different models and for a specific unit as well. As a result, the contribution from Compton scattering and the photoelectric effect also varies widely between different irradiators and different beam qualities. The photoelectric effect significantly predominates at the relatively low X-ray energies in which these irradiators operate. Consequently, a higher dose is delivered to bony tissues and the adjacent hematopoietic cells of the bone marrow. The increase in average radiation absorbed dose to the bone marrow compartment of the mouse can be as high as 30%, causing higher hematological sensitivity of animals when exposed to kilovoltage X rays. Adjusting the radiation dose to simply provide biological equivalency is complicated due to steep dose gradients within the marrow tissue and the qualitatively different outcomes depending on the spatial location of critical stem and progenitor populations in relationship to bone. These concerns may be practically addressed by efforts to implement X rays of the highest possible beam energy and penetration and increased awareness that radiation damage to hematopoietic cells will not be identical to data obtained from standard 137Cs gamma rays.

Pulmonary tissue is sensitive and often treatment-limiting in patients exposed to total-body irradiation (TBI) in preparation for hematopoietic stem cell transplantation. Many rodent strains, however, exhibit a relatively high resistance to radiation lung damage that often requires extra radiation doses to be delivered locally to the thorax to generate significant levels of pulmonary injury. The present study compared the effects of TBI and bone marrow transplantation (BMT) on two mouse strains that are known to differ in lung radiosensitivity after whole-thorax irradiation, namely the relatively resistant CBA mice and the sensitive C57L mice. Evaluation by survival, microcomputerized tomography (micro-CT), lung tissue weights and histopathology showed that the C57L mice responded with severe lethal radiation pneumonitis at 4 months after 12.5 Gy while CBA mice showed only minimal sublethal damage at this dose. C57L mice receiving 10 Gy TBI also had focal fibrotic lesions in the lungs out to 8 months. The manifestation of both pneumonitis and focal fibrosis in the lungs of C57L mice at relatively low radiation doses points to the merits of using this strain in further studies aimed at exploring and ameliorating the high susceptibility of the lung as encountered in clinical TBI.

To ensure reliability and reproducibility of radiobiological data, it is necessary to standardize dosimetry practices across all research institutions. The photoelectric effect predominates over other interactions at low energy and in high atomic number materials such as bone, which can lead to increased dose deposition in soft tissue adjacent to mineral bone due to secondary radiation particles. This may produce radiation effects that deviate from higher energy photon irradiation that best model exposure from clinical radiotherapy or nuclear incidences. Past theoretical considerations have indicated that this process should affect radiation exposure of neighboring bone marrow (BM) and account for reported differences in relative biological effectiveness (RBE) for hematopoietic failure in rodents. The studies described herein definitively estimate spatial dose distribution and biological effectiveness within the BM compartment for ¹³⁷Cs gamma rays and 320 kVp X rays at two levels of filtration: 1 and 4 mm Cu half-value layer (HVL). In these studies, we performed: 1. Monte Carlo simulations on a 5 μm resolution model of mouse vertebrae and femur derived from micro-CT images; 2. In vitro biological experiments irradiating BM cells plated directly on the surface of a bone-equivalent material (BEM); and 3. An in vivo study on BM cell survival in irradiated live mice. Simulation results showed that the relative dose increased in proximity to bone at the lower radiation energies and produced averaged values of relative dose over the entire BM volume within imaged trabecular bone of 1.17, 1.08 and 1.01 for beam qualities of 1 mm Cu HVL, 4 mm Cu HVL and ¹³⁷Cs, respectively. In accordance with Monte Carlo simulations, in vitro irradiation of BM cells located on BEM and in vivo whole-body irradiation at a prescribed dose to soft tissue of 6 Gy produced relative cell killing of hematopoietic progenitors (CFU-C) that significantly increased for the 1 mm Cu HVL X rays compared to radiation exposures of higher photon energies. Thus, we propose that X rays of the highest possible kVp and filtration be used to investigate radiation effects on the hematopoietic system, as this will allow for better comparisons with high-energy photon exposures applied in radiotherapy or as anticipated in a nuclear event.