Effects of total-body irradiation on bone marrow erythroid burst-forming units (BFU-E) and hemopoietic regeneration in dogs.

The acute and long-term effects of total-body X irradiation (TBI) on early erythroid progenitors, burst-forming units (BFU-E), in the bone marrow of beagles were studied for midline tissue doses of 1.6 and 2.4 Gy. After both radiation doses the initial reduction in the concentration of BFU-E was greater than that found for the granulocyte-macrophage progenitor cells (GM-CFC) and thus was in general agreement with the higher in vitro radiosensitivity of BFU-E compared to GM-CFC. After TBI with 1.6 Gy the GM-CFC and BFU-E returned to their normal levels within 2-4 weeks without showing long-term radiation effects. In contrast, after TBI with 2.4 Gy the concentrations of GM-CFC and BFU-E remained below the pretreatment levels up to 1 year after exposure. For a given midline tissue dose, the extent of the long-term effect of radiation on the BFU-E in a certain bone marrow site seems to be dependent on the local radiation dose in the respective bone marrow space. The minor radiation effects observed in the erythrocyte concentration in the peripheral blood, the hematocrit, and the hemoglobin concentration point to the enormous compensatory capacity of the more mature erythropoietic transit population to increase the proliferative capacity upon demand.

Haematological effects of rhGM-CSF in dogs exposed to total-body irradiation with a dose of 2.4 Gy.

It was the specific aim of this study to test the stimulatory effects of recombinant human GM-CSF (rhGM-CSF) on haemopoietic regeneration in dogs which had received total-body irradiation (TBI) with a dose of 2.4 Gy. In normal dogs rhGM-CSF given subcutaneously at 10 microgram/kg per day or 30 microgram/kg per day for 21 days caused strong but transient increases in the peripheral blood neutrophils. The monocyte counts also showed a transient rise during treatment in a dose-dependent fashion, whereas the lymphocyte counts increased only at the higher dose of rhGM-CSF and the platelet counts were transiently depressed during the course of the treatment. In the irradiated animals treatment with rhGM-CSF decreased the severity and shortened the duration of neutropenia but had no significant influence on monocyte or lymphocyte recovery. The granulocyte values showed a characteristic pattern of fluctuations with the first peak occurring at the same time (day 10 to day 13) when the abortive rise was observed in the untreated dogs. In contrast the GM-CFC in the peripheral blood remained depressed during the whole treatment course, similar to the untreated irradiated controls. These results indicate that treatment with GM-CSF can be an effective biological monotherapy for radiation-induced bone marrow failure, but that for higher radiation doses the number of GM-CSF responsive target cells will become a critical determinant of therapeutic efficacy.

In vitro studies on the radiosensitivity of multipotent hemopoietic progenitors in canine bone marrow.

The in vitro radiation response to 280-kV x-rays (does rate 72 cGy/min) of multipotent hemopoietic progenitor cells, mixed colony-forming units (CFU-mix), from canine bone marrow was assayed and compared to the radiation response characteristics of early erythroid progenitors, erythroid burst-forming units (BFU-E). To improve the colony-forming efficiency, the effect of various bone marrow cell separation techniques on colony formation of both progenitors was examined. The separation of bone marrow aspirates by discontinuous buoyant gradient centrifugation using the lymphocyte separation medium Lymphoprep with a density of 1.070 g/ml allowed the establishment of reproducible survival curves. The survival curves for both progenitors were strictly exponential, and CFU-mix were found to be more radiosensitive (D0 = 12 +/- 2 cGy) than BFU-E (D0 = 16 +/- 2 cGy).

Early indicators of response to accidental radiation exposure and the relevance for clinical management strategies.

In the development of clinical strategies to manage radiation accident casualties, the medical doctor in charge should be encouraged to use a "decision tree" to establish by a "sequential diagnosis procedure". This should be done within the first few days after exposure to determine whether or not a spontaneous recovery of hemopoietic function can be expected. With the assistance of a computer simulation model it appears possible to relate certain granulocyte response patterns to the extend and quality of damage caused in the hematopoietic stem cell pool. The determination of this damage is of great importance because it quantifies the strain inflicted upon the hemopoietic system by radiation exposure. It must be remembered that some stem cells are intact or are able to repair the damage completely. These stem cells serve as the ultimate source of hemopoietic recovery. The other stem cells that are left with the restricted hematopoietic potential are the source for the so called abortive recovery. On this basis it must be recognized that the day-to-day detailed analysis of documentation of blood cell changes for the first 5-10 days after exposure is of critical importance in order to be able to answer the question whether a spontaneous hemopoietic recovery can be expected or not. If the stem cell pool is sufficiently damaged (less than 6-8 in 10,000 stem cells) then one can expect a particular constellation of blood cells around day 5-7 characterized by severe granulocytopenia, severe lymphopenia and beginning thrombocytopenia. This blood cell response pattern is indicative of an irreversible stem cell damage. In this case, a transplantation of stem cells may well be life saving if done using the criteria developed for the treatment of severe aplastic anemia by bone marrow transplantation including an appropriate conditioning regimen for immune suppression.

Prediction of clinical outcome of radiation accident victims.

On the basis of the analysis of more than 350 individuals that were exposed to ionizing radiation in the course of more than 25 radiation accidents reported world wide since 1945, a biomathematical computer model was developed that simulates the pattern of granulocyte changes seen. It allows one to calculate the number of stem cells remaining intact to initiate recovery. It is shown that the major question to be asked is whether a spontaneous stem cell recovery can be expected or not. This question can be readily answered within 3-5 days after radiation exposure on the basis of the constellation of hematopoietic findings and devised stem cell pool size calculations.