Hematopoietic changes during fractional large-field irradiation of dogs with a subsequent bone marrow autograft

After the course of fractional large field irradiation of large volumes of bone marrow is completed (a total dose of 54 Cy), autotransplantation from the nonirradiated area of bone marrow contributes to a quicker restoration of blood cells amount injured at early stages after irradiation. The complete restoration of haemopoiesis is observed in a year, and in animals with automyelotransplantation--after a half of the year and remains at the same level for 3 years.     

Recovery course in mouse spleen and bone marrow after continuous irradiation.

The paper deals with the recovery process of some parameters in the spleen and bone marrow till day 60 after continuous irradiation with a daily dose of 476.5 mGy (50 R), 957 mGy (100 R) and 4785 mGy (500 R) up to the total accumulated dose of 9570 mGy (1000 R). The recovery process in the spleen and bone marrow are relatively significant and completed till day 28 or 60 respectively after irradiation.     

Mutagenic effect of cyclophosphan on bone marrow cells of irradiated rats]

The rate of chromosome aberrations in bone marrow cells of male rats was investigated in 24 hours after the cyclophosphan intraperitoneal injection (25 mg/kg). Cyclophosphan was given to rats exposed earlier (15 days, 1, 3, 4, 6 or 9 months before) to X- and gamma-irradiation (400 rads). It was found that preliminary irradiation led to the increase in the mutagenic effect of cyclophosphan as compared to that obtained for intact rats. This effect was demonstrated during 4 months after acute X-irradiation at a dose rate of 70 rads/min and during 1 month after chronic gamma-irradiation at a dose rate of 100 rads/day. Later the effect was shown to disappear in both cases. Chronic irradiation was found to be less efficient in the stimulation of chromosome damages caused by chemical mutagens. The increase of the mutagenic effect of cyclophosphan resulted in the increase of both the number of cells carrying chromosome breaks and the severity of a damage per cell. Different ways of the irradiation effect on the mutagenic action of chemicals are discussed.     

Bone marrow autotransplantation as a factor modifying the process of hematopoietic tissue recovery in irradiated guinea pigs

Guinea pigs were subjected to myeloexfusion 30-90 min before gamma-irradiation with a superlethal dose of 8 Gy. Immediately after irradiation, the bone marrow (4 X 10(7) cells) was autotransplanted. The automyelotransplantation provided a pronounced stimulation of haemopoiesis and substantially shortened the time of its regeneration. The myeloexfusion in some of the exposed control animals had a beneficial effect in their resistance and blood system recovery.     

B lymphocyte differentiation in lethally irradiated and reconstituted mice. I. The effect of Strontium-89 induced bone marrow aplasia on the recovery of the B cell compartment in the spleen.

The influence of ⁸⁹Sr-treatment on the recovery of the B cell compartment in lethally irradiated, fetal liver reconstituted mice was studied by means of membrane fluorescence. ⁸⁹Sr is a bone-seeking radio-isotope which causes in a dose of 3 μCi ⁸⁹Sr/g body weight a depletion of all nucleated cells, including immunoglobulin-bearing (B) cells, of the bone marrow.Treatment of irradiated and fetal liver reconstituted mice with 3 μCi ⁸⁹Sr/g body weight immediately and at 17 days after irradiation and reconstitution prevented recovery of the nucleated cell population, including B cells, in the bone marrow. In the spleen of such mice both nucleated cells and B cells reappeared at day 7 and 14 respectively. The B cell population in the spleen did not recover up to normal values during the experimental period of 45 days. It is concluded that B cell differentiation in lethally irradiated, fetal liver reconstituted mice can take place outside the bone marrow. The efficiency of this extra-medullary differentiation is discussed. The conclusion was drawn that mice with a ⁸⁹Sr-induced bone marrow aplasia are able to generate B lymphocytes. Consequently the bone marrow microenvironment seems not to be obligate to the differentiation of B lymphocytes. The peripheral lymphoid organs of such mice were found to be unable to compensate completely for the absence of B lymphocyte production in the bone marrow.     

Phospholipid metabolism in lymphoid cells at long-term periods following sublethal gamma-irradiation of rats

Dynamics of phospholipid metabolism in rat thymocytes and bone marrow cells was studied 1-6 months after fractionated irradiation. The rate of total and individual lipid synthesis was shown to increase in the exposed cells. The rate of lipid synthesis increased 1 and 2 months after irradiation and was normalized 3 and 6 months after irradiation.     

Allogeneic bone marrow transplantation for high risk non-Hodgkin's lymphoma during first complete remission

Allogeneic bone marrow transplantation from histocompatible sibling donors was performed in six patients with extranodal involvement of high grade lymphoma during first complete remission. Five patients had lymphoblastic lymphoma and one had diffuse undifferentiated lymphoma. The cytoreductive/immunosuppressive regimen consisted of total body irradiation and high dose cyclophosphamide. Four patients are alive in complete remission at 8 months, 14 months, 21 months and 47 months post transplantation. One patient who relapsed 7 months after his initial transplantation underwent a second transplantation but another relapse 17 months later led to his death. One patient died of chronic graft-versus-host disease and at autopsy there was no evidence of lymphoma. These data demonstrate that allogeneic bone marrow transplantation can produce durable remissions in patients with high grade lymphoma who present with bone marrow, central nervous system and/or skin involvement.     

THE EFFECT OF PARTIAL BODY IRRADIATION ON HAEMOPOIETIC STEM CELL MIGRATION

Data obtained after various types of partial body irradiation support the concept of a small rapidly exchangeable pool of CFUs, which seems to be exhausted rapidly after irradiation. The depletion of this pool is the most plausible explanation for the decrease in stem cell migration observed 3 hr after exposure in C₃H mice. After partial body irradiation the size of the rapidly mobilizable pool is reduced in proportion to the areas of bone marrow irradiated. When only one marrow area is shielded, the recovery of this pool does not occur during the first 24 hr after exposure.     

Treatment of Niemann-Pick disease type B by allogeneic bone marrow transplantation.

Allogenic bone marrow transplantation was carried out on a 3 year old girl with Niemann-Pick disease type B. Successful engraftment was achieved, and nine months after the procedure there was definite clearing of the sphingomyelin from the liver and pronounced clearing from the bone marrow. Any patient with Niemann-Pick disease type B complicated by early or severe hepatic impairment should be considered for bone marrow transplantation.     

The time-course of micronucleated polychromatic erythrocytes in mouse bone marrow and peripheral blood

The time-course of micronucleated polychromatic erythrocytes (MPCE) in mouse bone marrow and peripheral blood, induced by an acute 0.1 Gy dose of X-rays, was determined using flow cytometric analysis, which made frequent sampling possible and allowed use of a dose low enough not to affect erythroid cell proliferation. The frequency of MPCE (fMPCE) began to increase in the bone marrow at 10 h after irradiation and reached a maximum at 28 h after irradiation. In the peripheral blood fMPCE began to increase at 20 h after irradiation and peaked at about 40 h after irradiation. The time-course found is discussed on the basis of data on the differentiation of erythroid cells. The results indicate that the micronuclei registered in polychromatic erythrocytes may originate from lesions induced not only during the last cell cycle but also during earlier ones. After an acute dose of 1.0 Gy of X-rays the maximum fMPCE was delayed both in bone marrow and peripheral blood reflecting an effect on the cell cycle progression of erythroblasts.     

Transferrin binding of bone marrow cells and metabolic activity of erythrocytes after 5 Gy irradiation

The effect of total body irradiation (5 Gy) on functional mouse erythroid lineage has been studied. The transferrin binding capacity by bone marrow cells and the activity of glycolytic regulatory enzymes and intracellular levels of 2,3 bisphosphoglycerate in peripheral blood erythrocytes have been determined. Results obtained along one year post-irradiation period suggest a complete recovery in the erythroid cell lineage with respect to the biological endpoints investigated.     

Magnetic resonance (MR) evaluation of bone marrow in vertebral bodies.

Magnetic Resonance (MR) imaging was used to examine the hematopoietic bone marrow in the vertebral bodies of eight healthy subjects, and of 35 cancer patients who had been previously treated with radiation therapy. MR was instrumental in distinguishing viable hematopoietic tissue (red marrow) from adipose tissue (yellow marrow), whose presence reflected the extent of radiation-induced bone marrow injury. Different water content in proliferating hematopoietic tissue and adipose tissue enabled clear distinction of the two components even inside the same vertebral body. Three patterns of bone marrow viability were observed in irradiated patients: 1. Patients undergoing therapy at the time of MR study, and patients who had received low-intermediate dose several years before MR examination showed no alteration as compared with healthy controls (i.e. homogeneous presence of red marrow). 2. Patients who had received low-intermediate dose few years before MR, showed either partial re-colonization of yellow marrow or almost complete ablation of active red marrow with rare areas of re-colonization. 3. Patients who had received high dose, showed complete depletion of red marrow (fatty substitution) independently of the length of time elapsed since radiation therapy. Therefore, bone marrow recovery after radiation therapy was associate with two variables: received dose and length of time allowed for re-colonization by surviving hematopoietic tissue. In conclusion, our results provide evidence that MR can be purposively used to study composition and distribution of normal bone marrow, and to asses the extent of radiation-induced bone marrow injury; to monitor bone marrow recovery (or the lack of it); and in the general follow-up of treated cancer patients.     

Early and late effects in the bone marrow of mice following 2 Gy (6 MeV) neutron irradiation

Summary Following 5 Gy gamma irradiation, residual damage in bone marrow persisted up to one year and was ascribed to genetic defects in hemopoietic stem cells (von Wangenheim et al. 1986). To see whether high LET radiation is more efficient in inducing late effects, mice were whole-body irradiated with a single dose of 2 Gy neutrons ( = 6 MeV) and femoral cellularity, CFU-S number, proliferation ability of bone marrow cells (PF) and the compartment ratio (CR), i.e. the splenic 125-iodo-deoxyuridine incorporation per transfused CFU-S were measured up to one year after the radiation insult. Within 12 weeks, femoral cellularity, PF and CR recovered to control or near-control level, whereas CFU-S numbers remained significantly below control. No further recovery was observed. On the contrary, PF and CR deteriorated again after 12 and 26 weeks, respectively. CFU-S per femur tended to decrease as well. Thus it is demonstrated that a single dose of 2 Gy 6 MeV neutrons causes significant injury in function (PF) and structure (CFU-S numbers, CR) of bone marrow which persisted up to one year. While this residual injury can be attributed to genetic defects in hemopoietic stem cells, its increasing expression is probably due to late evolving damage in microenvironmental cells. The RBE of 6 MeV neutrons for the introduction of late effects in the bone marrow is in the range of 3.     

Autologous transplantation of a bone marrow graft manipulated by chemoseparation to eliminate residual tumor cells

An autologous bone marrow transplantation (ABMT) was performed in a 45-year-old male patient with AML in therapy-resistant first relapse including CNS-disease. The marrow graft was harvested 18 months before, at the beginning of first remission, and was subjected to an in-vitro incubation with 4-hydroperoxycyclophosphamide (4-HC) prior to cryopreservation to eliminate residual clonogenic tumor cells. After myeloablative therapy with high-dose Cyclo-phosphamide (CY), total body irradiation (TBI) and intrathecal application of Methotrexate (MTX), the manipulated marrow graft was reinfused. The myelopoietic reconstitution started already 6 days after ABMT and was completed 40 days thereafter. There is evidence for a complete remission in marrow and spinal fluid observed over a period of 4 months.     

Conservation of bone marrow CFUc in different phases of the cell cycle during cryopreservation

The effect of low temperature (-196 degrees C) preservation on the recovery of colon-forming units (CFUs) of bone marrow at different phases of the cell cycle before cryopreservation is dealt with. The intact bone marrow "enriched" with CFUs in S phase of the cell cycle and the bone marrow without colony-forming units in S phase were exposed to cryopreservation. After cryopreservation of the bone marrow enriched with CFUs in S phase and th bone marrow without colony-forming units in S phase the number of CFUs decreases by the same value as in the cryopreserved bone marrow obtained from intact mice.     

Evaluation of cell regeneration of bone marrow after fractional irradiation of the mouse in toto]

We have studied the recovery for mice bone marrow cells after fractionated irradiation of the whole body. The additional dose (Dr) to obtain a given biological effect if the irradiation is split in two equal subfractions (2 Di) separated by a short interval of time (i) is 40 rad per day when the interval of time between the two irradiations is lengthened of one day.     

Short- and long-term effects of whole-body irradiation with fission neutrons or X rays on the thymus in CBA mice

Young adult (6 weeks old) female CBA mice were exposed to whole-body irradiation with either 2.5-Gy fast fission neutrons of 1 MeV mean energy or 6.0-Gy 300 kVp X rays at centerline dose rates of 0.1 and 0.3 Gy/min, respectively. The weight of spleen and animal and the weight, cellularity, and histological structure of the thymus were studied at different times after irradiation. Thymic recovery after whole-body irradiation showed a biphasic pattern with minima at 5 and 21 days after irradiation and peaks of regeneration at Days 14 and 42 after X irradiation or at Days 14 and 70 after neutron irradiation. After the second phase of recovery, a marked decrease in relative thymus weight and cellularity was observed, which lasted up to at least 250 days after irradiation. Splenic recovery showed a monophasic pattern with an overshoot on Day 21 after irradiation. After neutron irradiation a late decrease in relative spleen and animal weight was observed. The observed late effects on thymus and spleen weight and thymus cellularity are discussed in terms of a persistent defect in the bone marrow.     

Study of mechanisms of anti-irradiation effects of interleukin-1beta in long-term bone marrow cultures

The influence of betaleukin (human recombinant interleukin-1 beta) on the processes of postirradiation recovery of haemopoietic precursors (GM-CFC) and the level of granulocyte-macrophag colony-stimulating factor (GM-CSF) were studied in long-term bone marrow cultures after gamma-irradiation with a dose 2 Gy. Then the betaleukin action on the contents of GM-CFC and induction of GM-CSF in the non irradiated cultures was studied. It was shown that betaleukin increased the induction of GM-CSF and raised the contents of GM-CFC in long-term bone marrow cultures, and the maximal increase of a GM-CSF level and GM-CFC amount was marked in 20 hours after introduction. At an irradiation of long-term bone marrow cultures in conditions of betaleukin introduction 20 hours prior to influence of radiation the smaller degree of damage and faster recovery of GM-CFC was observed. The data in this report suggest that one of the mechanisms of antiirradiation action of betaleukin apparently is connected to the action of the preparation on hematopoietic microenvironment cellular elements, that causes the release of a colony-stimulating factor and stimulation of recovery of haemopoietic precursors.     

Transfer of immunity by transfer of bone marrow cells: T-cell dependency.

Thymectomized, lethally irradiated mice reconstituted with normal bone marrow cells succumbed when challenged ip with rat Yoshida ascites sarcoma (YAS) cells 40 days after irradiation and reconstitution. In contrast, thymectomized irradiated mice reconstituted with bone marrow cells from YAS-immune donors rejected the subsequent tumor challenge. Pretreatment of the bone marrow cells from immune donors with anti-Thy 1.2 antiserum and complement completely abolished the transfer of anti-YAS resistance.Bone marrow cells from donors thymectomized 2 months before immunization enabled almost all recipients to reject YAS, but bone marrow cells from donors thymectomized 8 months before immunization protected only 50% of the recipients. Further analysis showed that mice thymectomized 8 months before immunization failed to generate anti-YAS antibody response, whereas the antibody response of mice thymectomized 2 months before immunization did not differ from that of non-thymectomized age-matched control mice. The data suggest that the immune reaction of mice against xenogeneic YAS requires long-lived T₂ lymphocytes.     

The stimulating effect of preliminary irradiation of the hematopoietic stroma with small doses on the content of hematopoietic stem cells in the bone marrow in vivo and in a long-term culture system

Preirradiation of mouse recipients with a dose of 1-2 Gy 24 and 48 h before lethal irradiation (8 Gy) made CFUs content of femur increase upon transplantation of bone marrow from exposed and intact donors. The same was with the long-term bone marrow culture: preirradiation of a stromal sublayer increased the number of CFUs in the transplanted bone marrow preirradiated with 6 Gy radiation. Retransplantation of bone marrow to irradiated donors after 5 day cultivation, a sublayer being activated, increased the number of CFUs in the femur in comparison with donors which were injected with the bone marrow from the culture without activation of the sublayer by low-level radiation.