The effect of internal exposure to 90Sr on hematopoietic stem cells in CBA mice

After acute intake of 90Sr the changes of d-9 CFUs number in mice (CBA) bone marrow, spleen and peripheral blood were investigated. The obtained results indicated similar quantitative changes in bone marrow and spleen CFUs on exposure to the 90Sr when radiation doses did not cause the decrease in life-time (1.11 kBq/g). Sarcomogeneous doses of 90Sr (29.6 kBq/g) resulted in drastic changes of hemopoietic system: spleen haematopoiesis activation and suppression of bone marrow functions. On the first day after 90Sr injection (29.6 kBq/g) the increase in number of peripheral blood CFUs (circulating pool) was observed.     

Compensation mechanisms in hemopoietic stem cell pool (CFUs) under the conditions of experimental chronic gamma-irradiation

The kinetics, proliferation and differentiation potentials of hemopoietic stem cells (CFUs) of bone marrow and spleen were investigated in CBA-line mice in the early period (1-30 days) of chronic gamma-irradiation at a dose rate of 0.16 Gy/day to attain a cumulative dose of 4.8 Gy. The results of the experimental study showed the prevalent maintenance of productivity of granulocytic and erythrocytic hemopoietic cell series within the range of reference values, persistent inhibition of the megakaryocytic series (in terms of all hemopoiesis parameters of interest), more marked suppression of the population of polypotential CFUs in the bone marrow as compared with that in the spleen. The obtained results indicated that the mechanisms of hemopoiesis compensation at stem cell pool level were as follows: the increase in proliferation potency of erythrocytic and in polypotential precursors, the rise in the proportion of granulocytic precursors in the real differentiation potential of CFUs, and the processes of repopulation manifested with different intensity in all stem cell populations under study. For maintenance of the necessary productivity of CFUs in each of hemopoietic cell series, consecutively or simultaneously, several compensatory-adaptive mechanisms are started, which allows the avoidance of a sharp competition between hemopoietic cell series under the conditions of stem cell pool depopulation, and preservation of the hemopoiesis as a whole.     

Kinetic models of hemopoietic stem cell populations

Summary Recent techniques enabled a series of quantitative studies to be made with various aspects of the stem cell functions. Results from several laboratories appear to agree on certain main points: the existence of a pluripotential stem cell in the small rodent, capable of forming visible colonies of hemopoietic foci in the spleen and the existence of some undifferentiated but committed precursor cells from which the differentiating and maturing cell populations originate. There is evidence that the primary stem cell is in a low turnover state in the normal animal, although on demand it is capable of fast and prolonged proliferative activity. The committed undifferentiated precursor cells differ greatly, depending on which cell line they represent. Some of these are in high turnover state in the normal animal (e.g., erythropoietin-responsive cells), others do not appear to be capable of proliferation (e.g., focus forming cells). Perturbation of the steady states by irradiation of the animal, or by treatment with cytotoxic drugs results in recovery patterns which yield valuable kinetic information.     

DNA immunization against tissue-restricted antigens enhances tumor immunity after allogeneic hemopoietic stem cell transplantation

Malignant relapse remains a major problem for recipients of allogeneic hemopoietic stem cell transplantation (HSCT). We hypothesized that immunization of allogeneic HSCT recipients against tissue-restricted Ags using DNA vaccines would decrease the risk of relapse without enhancing graft-vs-host disease (GVHD). Using the mouse B16 melanoma model, we found that post-HSCT DNA immunization against a single tumor Ag induces tumor rejection that is significantly greater than HSCT alone in a T cell-depleted MHC-matched minor Ag-mismatched allogeneic HSCT model (LP --> B6). In treatment models, post-HSCT DNA immunization provides significantly greater overall survival than the vaccine alone. Donor leukocyte infusion further enhances tumor-free survival, including in treatment models. There was no GVHD in HSCT recipients treated with DNA vaccination and donor leukocyte infusion. Further analysis demonstrated that these effects are dependent on CD8+ T cells of donor origin that recognize multiple epitopes. These results demonstrate that DNA immunization against tissue-restricted Ags after allogeneic T cell-depleted HSCT can induce potent antitumor effects without causing GVHD.     

Isolation of hemopoietic pluripotent stem cells from spleens of thiamphenicol-pretreated mice

The present report describes the bulk isolation of pluripotent stem cells (PSC) (assayed as day-9 CFU-S, colony-forming-units-spleen). As starting material, spleens, highly enriched with PSC, were used from mice that were bled and treated with thiamphenicol (TAP). In subjecting the spleen cells to a two-stage centrifugal elutriation procedure and a subsequent Percoll gradient centrifugation stage a 30-fold enrichment in the CFU-S concentration was achieved. The splenic PSC seeded with a characteristic low efficiency in the spleens of irradiated mice (f = 2%). Correcting the colony number for this, we obtained a cell mixture consisting of 88% PSC, contaminated with 4% committed precursor cells and about 10% ganuloid cells.     

Reconstruction of 90Sr intake for breast-fed infants in the Techa riverside settlements

The Techa River (Southern Urals, Russia) was contaminated as a result of radioactive releases by the Mayak plutonium production facility during 1949-1956. The persons born after the onset of the contamination have been identified as the "Techa River Offspring Cohort" (TROC). The TROC has the potential to provide direct data on health effects in progeny that resulted from exposure of a general parent population to chronic radiation. The purpose of the present investigation is the estimation of (90)Sr intake from breast milk and river water in the period from birth to 6 months of life, necessary for an infant dose calculation. The investigation is based on all available data concerning radioactive contamination due to global fallouts and Mayak releases in the Southern Urals where extensive radiometric and radiochemical investigations of human tissues and environmental samples were conducted during the second half of the twentieth century. The strontium transfer factor from mother's daily diet to breast milk was estimated as 0.05 (0.01-0.13) d L(-1). Based on this transfer factor and data on (90)Sr water contamination, the average total (90)Sr intake for an infant born in the middle Techa River region was found to be equal to 60-80 kBq in 1950-1951. For the same period, calculations of (90)Sr intake using ICRP models gave values of 70-100 kBq. From 1952 onwards, the differences in intakes calculated using the two approaches increased, reaching a factor of 2-3 in 1953. The Techa River data provide the basis for improving and adapting the ICRP models for application to Techa River-specific population.     

NUP98-HOXA9 expression in hemopoietic stem cells induces chronic and acute myeloid leukemias in mice

Here we describe hemopoietic chimeras serving as a mouse model for NUP98-HOXA9-induced leukemia, which reproduced several of the phenotypes observed in human disease. Mice transplanted with bone marrow cells expressing NUP98-HOXA9 through retroviral transduction acquire a myeloproliferative disease (MPD) and eventually succumb to acute myeloid leukemia (AML). The NUP98 portion of the fusion protein was shown to be responsible for transforming a clinically silent pre-leukemic phase observed for Hoxa9 into a chronic, stem cell-derived MPD. The co-expression of NUP98-HOXA9 and Meis1 accelerated the transformation of MPD to AML, identifying a genetic interaction previously observed for Hoxa9 and Meis1. Our findings demonstrate the presence of overlapping yet distinct molecular mechanisms for MPD versus AML, illustrating the complexity of leukemic transformation.     

Effect of a neutrophilia-inducing tumor on hemopoietic stem cells in mice

The influence of neutrophilic stimulation on hemopoietic stem cells was studied in mice with tumor-induced neutrophilia. Transfusions of marrow cells from normal and neutrophilic tumor-bearing mice into lethally irradiated normal and tumor-bearing mice were performed. The number and the erythroid:granuloid (E:G) ratio of day 7 colonies in the recipient spleens and bones as well as the size of spleen colonies of recipient animals were determined. The E:G ratio of spleen and bone marrow colonies between normal and tumor-bearing mouse recipients and the number of spleen colonies did not differ significantly in either experiment. However, spleen colonies which developed in tumor-bearing irradiated mice were significantly larger than those which developed in normal recipients in both experiments. These studies indicated that while the line of differentiation taken by hemopoietic stem cells was not affected by the neutrophilic influence of the tumor, the tumor-bearing host environment appeared to enhance proliferation of transfused stem cells and/or their descendants. The stimulators of granulocytopoiesis in this model of neutrophilia appear to act on a population of progenitor cells more mature than the stem cells capable of forming 7-day colonies in the spleen and bone marrow of irradiated recipient mice.     

Studies on the erythropoietic cell system in CBA mice after Rauscher virus infection.

Erythropoiesis in CBA mice was studied in Rauscher leukemia virus infected mice using the incorporation of 59Fe into spleen, bone marrow and peripheral blood. Beginning at day 4 an increased uptake into the spleen and a decrease in the bone marrow and the peripheral blood was observed. The increased uptake by the spleen was also found in plethoric mice. The erythropoietin responsive compartment was also enlarged in the spleen of these mice. The The dose-response-curve for erythropoietin was altered 4 days after infection, there was a higher background level of 59Fe incorporation and the response to low doses was better in infected animals. The reticulocytopenia which is usually seen in these mice, was overcome by administration of high doses of erythropoietin. It is concluded that the Rauscher virus acts in a similar manner to erythropoietin, but the erythropoiesis induced is ineffective since the cells do not mature. This maturation deficiency is influenced by administration of exogenous erythropoietin.     

The cell cycle of spermatogonial colony forming stem cells in the CBA mouse after neutron irradiation

In the CBA mouse testis about 10% of the stem cell population is highly resistant to neutron irradiation (D0, 0.75 Gy). Following a dose of 1.50 Gy these cells rapidly increase their sensitivity towards a second neutron dose and progress fairly synchronously through their first post-irradiation cell cycle. From experiments in which neutron irradiation was combined with hydroxyurea it appeared that in this cycle the S-phase is less radiosensitive (D0, 0.43 Gy) than the other phases of the cell cycle (D0, 0.25 Gy). From experiments in which hydroxyurea was injected twice after irradiation the speed of inflow of cells in S and the duration of S and the cell cycle could be calculated. Between 32 and 36 hr after irradiation cells start to enter the S-phase at a speed of 30% of the population every 12 hr. At 60 hr 50% of the population has already passed the S-phase while 30% is still in S. The data point to a cell cycle time of about 36 hr, while the S-phase lasts 12 hr at the most.