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Ferrokinetics and erythropoiesis in man: the measurement of effective erythropoiesis, ineffective erythropoiesis and red cell lifespan using 59Fe.

Existing ferrokinetic methods do not provide a direct and quantitative measurement of effective and ineffective red cell production. A new method is described for calculating the daily uptake of iron by maturing red cells and the mean red cell lifespan. Ineffective erythropoiesis and non-erythroid iron turnover are also measured. The method involves standard laboratory techniques but the analysis requires access to a computer. The preliminary results suggest that it will be a clinically useful tool for the investigation of erythroid disorders.

Anemia↗

Ferrokinetic study of splenic erythropoiesis: relationships among clinical diagnosis, myelofibrosis, splenomegaly, and extramedullary erythropoiesis.

Splenic erythropoiesis was demonstrated by surface counting of 59Fe in 129 of 1,350 ferrokinetic studies performed over a 15 year period. These 129 studies were carried out in 108 patients, including 40 with chronic myelogenous leukemia (CML), 24 with agnogenic myeloid metaplasia (AMM), 18 with polycythemia vera (PV), six with a myelodysplastic syndrome, five with acute leukemia, three with prostate or breast carcinoma, two each with aplastic anemia or Hodgkin's disease, and one each with idiopathic thrombocythemia, multiple myeloma, chronic renal failure, or treated hypopituitarism. Splenomegaly was present in 83% of the studies and hepatomegaly in 72%. Grade II-III myelofibrosis was demonstrated in 62% of the cases. Hepatic erythropoiesis was present in 77% of the studies (only 38% in PV), and marrow erythropoiesis was undetectable in 33%. Total erythropoiesis was about twice normal (range 0.2 to 8 times normal) but was ineffective to varying degrees in 86% of the studies. Relationships between organomegaly, myelofibrosis, and extramedullary erythropoiesis, as well as differences among clinical disorders, are discussed. Differences observed between CML in chronic or blastic phase suggested that the erythroid cell line was involved in the proliferative process. It is concluded that splenic erythropoiesis 1) is encountered in a variety of clinical conditions; 2) is not necessarily associated with splenomegaly or myelofibrosis, even in the myeloproliferative disorders; 3) is part of a predominantly extramedullary (in the liver as well as in the spleen), expanded, and largely inefficient total erythropoiesis; and 4) can be evaluated in a semiquantitative manner by surface counting.

Erythropoiesis↗

A mathematical model of erythropoiesis in mice and rats. Part 2: Stimulated erythropoiesis.

A mathematical model of erythropoietic cell production and its regulation process has been proposed in a preceding paper. It is primarily based on the assumption that the number of cell divisions taking place in the CFU-E and erythropoietic precursor stages is regulated depending on the oxygen supply of the tissue. Quantitative dose-response relationships for in vivo erythropoiesis are suggested. Here, we demonstrate that this model adequately reproduces data obtained in situations of stimulated erythropoiesis in mice and rats. In detail, this implies a quantitative description of the following processes: (1) Changes in tissue oxygen tension (Pto2) following removal of red cells (bleeding, haemolytic anaemia) or increase in plasma volume (dilution anaemia) or decrease in atmospheric oxygen pressure (hypoxia). (2) Pto2 dependent erythropoietin (EPO) production. (3) Dose-response of EPO on erythropoietic amplification (up to two to four additional mitoses). (4) The changes of the marrow transit time. Model simulations are compared with experimental data for changes of erythropoiesis during hypoxia, EPO-injection, and different forms of anaemia. A satisfactory agreement suggests that the model adequately describes and correlates different direct and indirect ways to stimulate erythropoiesis. It quantifies the role and relative contribution of the haematocrit, haemoglobin concentration, atmospheric oxygen pressure, tissue oxygen pressure, and plasma volume as triggers in erythropoietic stimulation under various conditions. Furthermore, the model may allow to optimize the scheme of EPO-administration and to find the maximum increase of erythropoiesis for a given amount of erythropoietin.

Anemia↗

Impaired splenic erythropoiesis in phlebotomized mice injected with CL2MDP-liposome: an experimental model for studying the role of stromal macrophages in erythropoiesis.

Erythropoiesis occurs in the presence of erythropoietin (EPO) without macrophages in vitro. In hematopoietic tissues, however, erythroid cells associate closely with stromal macrophages, forming erythroblastic islands via interactions with adhesion molecules. To elucidate the role of macrophages in erythropoiesis, we selectively abrogated stromal macrophages of splenic red pulp of phlebotomized mice by injection with dichloromethylene diphosphonate encapsulated in multilamellar liposomes (CL2MDP-liposome). In the spleen, no erythropoietic activity occurred until 5 days after the treatment. Colony assay revealed that the erythropoiesis was suppressed at the level of CFU-E. The splenic erythropoietic activity gradually developed from day 6 after the treatment, when F4/80+ macrophages began to appear in the red pulp. EPO mRNA was expressed in kidney but not in liver or spleen of phlebotomized mice injected with CL2MDP-liposome, and the serum EPO concentration in these mice was higher than that in phlebotomized mice. These findings suggest that abrogation of stromal macrophages by injection with CL2MDP-liposome impairs the splenic microenvironment for erythropoiesis induced by hypoxic stress, and this may be an excellent experimental model for further characterization of the in vivo role of splenic macrophages in erythropoiesis.

Anemia↗

The ontogeny of erythropoiesis in the mouse detected by the erythroid colony-forming technique. I. Hepatic and maternal erythropoiesis.

Employing the erythroid colony-forming technique, it is shown that throughout hepatic erythropoiesis in the mouse, the CFU-E population remains sensitive to erythropoietin. Maximum stimulation was achieved during this period using an erythropoietin concentration of 0.075 units/ml. The peak in the CFU-E concentration occurs between the 11th and 12th day while absolute values show a maximum on the 14th day of gestation. These results are discussed in terms of changing cell populations, both of erythropoietic precursors and hepatocytes from which it is concluded that at no time during foetal erythropoiesis does the CFU-E population change or become unresponsive to erythropoietin. The BFU-E population follows closely that of the CFU-E, but declines about 24 h earlier on the 16th day of gestation. The effect of the foetus on the mother was also studied during the second half of pregnancy. During this period of natural perturbation both femoral and, in particular, splenic erythropoiesis are increased. However, during this time an erythropoietin concentration of 0.3 units/ml was required to maximally stimulate the CFU-E population derived from these tissues. The fact that both adult and foetal erythroid tissue maintain a rather constant requirement for erythropoietin for their growth in vitro, indicates that it is an intrinsic property of the cells. It is concluded that increased maternal erythropoiesis is due to an increased oxygen requirement causing hypoxia due to the growing foetus.

Animals↗

A mathematical model of erythropoiesis in mice and rats. Part 3: Suppressed erythropoiesis.

A mathematical model of erythropoietic cell production and its regulation process has been proposed in a preceding paper. It is primarily based on the assumption that the number of cell divisions taking place in the CFU-E and erythropoietic precursor stages can be regulated depending on the oxygen supply to the tissue. Here we provide evidence that this model adequately describes situations of suppressed erythropoiesis. In detail this implies a quantitative description of the following processes: (1) changes in tissue oxygen tension (Pto2) due to increase in red cell numbers (red cell transfusion, posthypoxia), decrease in plasma volume (dehydration) or increase in atmospheric oxygen pressure (hyperoxia), (2) Pto2 dependent reduction of erythropoietin (EPO) production, (3) dose-response of reduced EPO-levels on erythropoietic amplification (omission of three to five mitoses). Model simulations are compared to experimental data obtained from red cell transfusion, posthypoxia, hyperoxia and dehydration. A satisfactory agreement suggests that the model adequately describes and correlates different ways to suppress erythropoiesis. It quantifies the role and relative contribution of the haematocrit, haemoglobin concentration, atmospheric oxygen pressure, tissue oxygen pressure and plasma volume as triggers in erythropoietic suppression under various conditions. In conjunction with the preceding two papers it could be shown that one unique set of model parameters is sufficient to describe erythropoiesis in steady state, stimulation and suppression. Limitations of the model are discussed and experiments for a more detailed investigation of the feedback mechanisms are proposed.

Animals↗

Inhibition of Friend virus (FVP)-induced erythropoiesis by an erythropoiesis-inhibitory factor (EIF).

An erythropoietin-independent murine erythroleukemia (FVp) has been used to evaluate the effects of an erythropoiesis-inhibitory factor (EIF) isolated from human urine. The consequent inhibition of FVp-induced erythropoiesis suggests that EIF exerts its effect independently of ESF. The inhibitory effect of EIF on FVp-induced erythropoiesis may reveal a potential for the physiological control of some types of erythroleukemia.

Animals↗

[Erythropoiesis and osteogenesis (II. The mechanism of the suppression of erythropoiesis during the reparative process in bone tissue)].

The experiment on rats and mice has shown that the presence of a developing callus in the organism after fracture of the tubular bones inhibits erythropoiesis. Inhibition of erythropoiesis is due to delay of erythropoietin biosynthesis by serotonin. A problem on two-fold serotonin effect is under discussion: on the one hand, serotonin inhibits the erythropoietin formation and delays erythropoiesis, on the other hand, it intensifies the effect of ready erythropoietin on the developing bone marrow erythroid cells.

Anemia↗

Erythropoiesis-stimulating factor(s), erythropoiesis and erythrocyte 2,3-diphosphoglycerate in young rabbits with marked post-natal fall in haemoglobin.

The erythropoietic activity and erythrocyte 2,3-diphosphoglycerate (2,3-DPG) were studied during and after the nadir of the post-natal anaemia in normal, rapidly growing rabbits, from the 12th to the 35th day after birth. Whole blood haemoglobin (Hb) decreased from 9.3 g dl-1 on the 12th to 4.9 g dl-1 on the 25th day, while erythropoiesis-stimulating factor(s) (ESF) in plasma (determined by a cell culture assay) concomitantly rose from undetectable to high levels. In spite of marked rise in body weight, from 250 to 480 g, estimated haemoglobin mass (Hb mass) and reticulocyte mass production rate (Rt prod) remained essentially the same, about 1.8 g and 0.3 ml day-1. From the 25th to the 35th day, ESF decreased to a lower level, while Hb increased to 10.8 g dl-1 and Hb mass and Rt prod rose sharply, to 6.9 and 1.2 ml day-1. The 2,3-DPG rose markedly during the observation period, but showed a transient decline on the 29th day, simultaneously with the peak in reticulocyte counts (Rt) (24%) and release of young erythrocytes with low 2,3-DPG. The data indicate that the regions governing the erythropoietin production/release became increasingly sensitive to hypoxia during the observation period. The possibility also exists that the increase in ESF was due only in part to hypoxic stimulation. It could be related to the maturation of the animal in other ways, such as shift from extra-renal to renal erythropoietin production and the growth. The lack of response to increasing stimulation indicates that the erythropoiesis was restricted by the availability of iron and/or other factors necessary for erythrocyte and haemoglobin production.

2,3-Diphosphoglycerate↗

[Erythropoiesis and functional characteristics in bone marrow erythroblastic islets during stimulated adn inhibited erythropoiesis].

When erythropiesis is stimulated (acute blood loss) or inhibited (posttransfusion polycythemia), there are early changes in the cytochemical values of erythroblastic islets (EI): in the levels of acid and neutral glucoconjugates and in the activity of nonspecific esterase. A close correlation has been found between the erythropoiesis in EI and its functional characteristics. It is concluded that central macrophages play the key role in the modulation of EI erythropoiesis. It is suggested that EI macrophages are involved in the provision of bioenergetic and reparative processes in EI.

Animals↗

Investigations on erythropoiesis in newborn rats. Dependence of erythropoiesis in newborn rats on the intensity of the haemopoietic processes in the lactating mothers.

The intensity of haemopoietic processes was investigated in 7, 9, 11, 14 and 19-day-old suckling rats in relation to the intensity of these processes in their mothers. The rate of the haemopoietic processes in newborn rats was determined on the basis of 59Fe incorporation into the blood and haemopoietic organs. The activity of the erythropoietic system in lactating rat females was stimulated by haemorrhage and inhibited by erythrocyte transfusion. Anaemization of lactating rats by haemorrhage did not stimulate erythropoiesis in the suckling rats. Posttransfusion polycythaemia in the lactating mothers inhibited erythropoiesis in the suckling rats beginning with the 9th day of life. This phenomenon became more pronounced with the age of the rats.

Animals↗

Regulation of erythropoiesis in suckling rabbits with and without postnatal anemia: partial suppression of production/release of erythropoiesis stimulating factor(s) by iron supplements.

The postnatal anemia in rabbits is accompanied by a marked rise in the plasma erythropoiesis stimulating factor(s) (ESF). The purpose of this study was to establish whether the increase in plasma ESF is only related to the anemia, or whether other mechanisms also are involved. Two matched groups of rabbits were studied from the 15th to the 36th day after birth. One group received iron parenterally and had no postnatal fall in hemoglobin concentration (Hb), the other developed the usual anemia. In both groups plasma ESF was undetectable on the 15th day, and also on the 22nd day, despite a marked fall in Hb in the untreated group and rise in the iron-treated group. Thereafter plasma ESF showed a slight, continuous rise in the nonanemic rabbits, in contrast to a marked, transient rise with maximum on the 29th day in the untreated group. On the 36th day there was no difference between the groups. In the iron-treated group the reticulocyte production rate remained unchanged, while the Hb mass rose continuously. In the untreated animals there was an initial decline in reticulocyte production rate, while Hb mass showed a slight increase. From the 29th day, however, reticulocyte production rate rose to the same level as in the iron-treated group and Hb mass rose markedly. In conclusion, the rise in plasma ESF during the postnatal anemia in rabbits is only in part related to the low Hb. Hypoxia-independent mechanisms, probably related to the growth and maturation per se also are involved. The lack of erythropoietic response to the rise in plasma ESF is due to lack of available iron.

Anemia↗

[Cytological characteristics of erythropoiesis in thalassemia. I. Functional characteristics of the nucleated elements of erythropoiesis in the bone marrow].

In 24 patients affected with thalassaemia of various degrees of seriousness the functional condition of nuclear cells or red serie was investigated in the bone-marrow. The investigation was carried out by analyzing partial erythroblastogrammes, evaluating proliferative activity according to the 3H-thymidine marking index and determining ineffective erythropoiesis by means of nucleated PAS-positive erythroblasts. The findings reveal the degree of seriousness of the disease being directly dependent on the extent of functional disturbances in the cells of the erythropoietic system.

Bone Marrow Cells↗