losartan-potassium and Anemia--Aplastic

Immunosuppressive therapy is the treatment for aplastic anemia patients ineligible for transplantation. The role of hematopoietic growth factors as adjunct to treatment in these patients is unclear. We conducted a systematic review and meta-analysis of randomized controlled trials comparing treatment with immunosuppressive therapy and hematopoietic growth factors to immunosuppressive therapy alone in patients with aplastic anemia. Two reviewers appraised the quality of trials and extracted data. For each trial, results were expressed as relative risks with 95% confidence intervals (CI) for dichotomous data. The addition of hematopoietic growth factors yielded no difference in overall mortality at 100 days, one year and five years [relative risks 1.33 (95% CI 0.56-3.18), relative risks 0.90 (95% CI 0.50-1.63) and relative risks 0.89 (95% CI 0.55-1.46), respectively]. There was no difference in overall hematologic response and in the occurrence of infections. HGF significantly decreased the risk for relapse, relative risks 0.45 (95% CI 0.30-0.68, 3 trials). Hematopoietic growth factors were not associated with higher occurrence of myelodysplastic syndrome and acute myeloid leukemia or paroxysmal nocturnal hemoglobinuria. The addition of hematopoietic growth factors does not affect mortality, response rate or infections occurrence. Therefore, it should not be recommended routinely as an adjunct to the immunosuppressive therapy for patients with aplastic anemia.

Topics: Anemia, Aplastic; Drug Therapy, Combination; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Humans; Immunosuppressive Agents; Randomized Controlled Trials as Topic; Survival Analysis; Survival Rate; Treatment Outcome

Association between androgens and erythropoiesis has been known for more than seven decades. Androgens stimulate hematopoietic system by various mechanisms. These include stimulation of erythropoietin release, increasing bone marrow activity and iron incorporation into the red cells. Before the discovery of recombinant erythropoietin (rhEpo), androgens were used in the treatment of anemia associated with renal disease, bone marrow suppression, and hypopituitarism. Anabolism is an additional advantage of androgen therapy. Furthermore, in light of recent reports regarding adverse effects of rhEpo, the role of androgen therapy in various types of anemias should be readdressed. Polycythemia remains a known side effect of androgen therapy. In this review, we will briefly discuss the initial animal and human studies which demonstrated the role of androgens in the treatment of anemia, their mechanism of action, a detailed account of the efficacy of androgens in the treatment of various anemias, the erythropoietic side effects of androgens and finally, the relationship between hematocrit levels and cardiovascular disease.

Topics: Aging; Androgens; Anemia; Anemia, Aplastic; Animals; Bone Marrow; Coronary Disease; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Humans; Iron; Kidney Failure, Chronic; Male

Haematological abnormalities are common in systemic lupus erythematosus (SLE) and may be manifested by anaemia of different pathogenesis. The objective of this article was to describe some data concerning autoimmune haemolytic anaemia, aplastic and megaloblastic ones accompanying SLE and also to present erythropoietin (EPO) function in the above mentioned diseases. In SLE many factors are produced which disturb the organism haematological balance both on the peripheral level and in the bone marrow. It is assumed that the autoantibodies produced in SLE are the main cause of anaemia. However it should be considered that quantitative changes in the number of erythrocytes observed in this disease are also caused by chronic inflammatory condition, which as the element of autoimmune disease impairs the endocrine function of the kidneys in EPO production. It influences bone marrow, iron metabolism and then haemopoiesis. Apart from humoral factors the role of mechanisms connected with immune cellular response is also considered.

Topics: Anemia, Aplastic; Anemia, Hemolytic, Autoimmune; Anemia, Megaloblastic; Erythropoietin; Humans; Immunity, Cellular; Kidney; Lupus Erythematosus, Systemic

Hematologic dysfunction, including thrombocytopenia, anemia, neutropenia, thromboses, and coagulopathy, occur commonly during critical illnesses. A major challenge is to identify drug-induced causes of hematologic dysfunction. Given the wide variety of drug-induced hematologic effects, clinicians always should consider any concomitant drugs in the differential diagnosis of acquired hematologic dysfunction. The most severe effects include drug-induced aplastic anemia, heparin-induced thrombocytopenia, and drug-induced thrombotic microangiopathy. Certain drugs are associated with multiple hematologic effects. For example, cisplatin can cause hemolytic uremia syndrome and erythropoietin deficiency, and quinine can precipitate immune-mediated thrombocytopenia, immune-mediated thrombocytopenia, and thrombotic microangiopathy.

Topics: Anemia, Aplastic; Anticoagulants; Erythropoietin; Hematologic Diseases; Heparin; Humans; Methyltransferases; Severity of Illness Index; Thrombocytopenia

Immunosuppressive therapy (IST) is essential to treat aplastic anemia. The pharmacological mechanism, therapeutic effect of main drugs and their application method, reasonable dosage, synergistic action are briefly reviewed in this article. These reviewed drugs include ATG/ALG, CsA, ALG/ATG + CsA, IIST (ALG + CsA) + HGFs, McAb-T, HD-MP and HD-IVIG. The purpose of this review was to direct to clinical therapy for patients with aplastic anemia.

Topics: Anemia, Aplastic; Antilymphocyte Serum; Cyclosporine; Drug Therapy, Combination; Erythropoietin; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunosuppressive Agents; Recombinant Proteins

Thirty per cent of cancer patients suffer from anemia before any treatment. This anemia is caused by haematopoiesis troubles related to cytokines production and by endogenous erythropoietin deficiency. Clinically, its symptoms, including fatigue, spoils patients'quality of life. Known as a prognostic factor for several cancers, anemia also lowers radiotherapy or chemotherapy efficiency by tumor hypoxia. Recombinant EPO restores normal haemoglobin level, quality of life and treatment efficiency.

Topics: Anemia, Aplastic; Cell Hypoxia; Erythropoietin; Fatigue; Female; Humans; Neoplasms; Oxygen Consumption; Quality of Life; Uterine Cervical Neoplasms

As the major regulator of erythropoiesis in man, erythropoietin inhibits the programmed cell death of committed erythroid precursors. In cancer patients, a relative erythropoietin deficiency is coupled with a decreased responsiveness to the substance mediated by the effects of inflammatory cytokines on the marrow and on ferrokinetics, leading to a high incidence of anemia. Two recombinant human erythropoietin (rhEPO) preparations--epoetin alfa (Epogen, Procrit) and epoetin beta (Marogen)--as well as a modified erythropoietic compound (darbepoetin alfa [Aranesp]) are in clinical use. Part 2 of this two-part series on hematopoietic agents reviews the use of these erythropoietic factors and their effect on the anemia that develops in cancer patients. Thrombopoietic factors and progenitor cell-mobilizing factors are also briefly addressed.

Topics: Anemia, Aplastic; Erythropoietin; Hematinics; Humans; Medical Oncology; Myelodysplastic Syndromes; Neoplasms; Practice Patterns, Physicians'; Quality of Life; Recombinant Proteins

Topics: Adult; Anemia; Anemia, Aplastic; Autoradiography; Bone Marrow; Bone Marrow Cells; Cell Cycle; Cytophotometry; DNA; Erythroblasts; Erythrocytes; Erythroid Precursor Cells; Erythropoiesis; Erythropoietin; Female; Gestational Age; Heme; Hemoglobins; Humans; Infant, Newborn; Iron; Male; Mitosis; Oxidation-Reduction; Polycythemia; Pregnancy; Reticulocytes; RNA

The use of hematopoietic growth factors has increased rapidly during the last decade. Among the growth factors available, erythropoietin (EPO) was the first growth factor to be used clinically. To date, EPO has shown activity in the treatment of the tumor-associated anemia and for correction of tumor hypoxia, however, when compared with transfusion of erythrocytes EPO treatment did not significantly prolong survival in cancer patients in any published study so far. Recently, novel extramedullary EPO receptors have been identified leading to a better understanding of the molecular mechanisms of action of EPO. Results from these experiments and from several clinical studies suggest that EPO treatment may be beneficial for patients with (chronic) infections (HIV, inflammatory bowel disease, septic episodes) and for treatment of the fatigue syndrome following cancer chemotherapy. In addition, EPO may also improve stem cell engraftment following high-dose chemotherapy and can increase survival rates of patients with aplastic anemia and myelodysplastic syndrome. Currently, new EPO derivatives, synthetic fusion proteins and gene therapeutic studies are under clinical investigation suggesting that the EPO-induced effects may be increased significantly by these agents in the future.

Topics: Anemia, Aplastic; Chronic Disease; Erythropoietin; Fatigue; Genetic Therapy; Hematopoietic Stem Cell Mobilization; Hematopoietic Stem Cell Transplantation; HIV Infections; Humans; Infections; Myelodysplastic Syndromes; Receptors, Erythropoietin; Recombinant Proteins

The use of hematopoietic growth factors, although well established for the management of chemotherapy-induced neutropenia, remains controversial for the treatment of aplastic anemia and inherited bone marrow failure syndromes. The most commonly used factors are granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, and erythropoietin. Newer growth factors such as stem cell factor, thrombopoietin, Flt3 ligand, and interleukins have shown promising results in the laboratory, and some have been used in clinical trials. This article reviews the clinical use of old and new hematopoietic growth factors in acquired and inherited bone marrow failure, and discusses emerging concerns about long term toxicity of these factors.

Topics: Anemia, Aplastic; Bone Marrow Diseases; Clinical Trials as Topic; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Humans; Stem Cell Factor

Topics: Anemia; Anemia, Aplastic; Erythropoietin; Humans; Multiple Myeloma; Myelodysplastic Syndromes

An 89-year-old man with a severe aplastic anaemia is described. The patient was proven to be cortico-steroid and cyclosporin resistant, but had a trilineage response during subsequent treatment with recombinant human erythropoietin.

Topics: Aged; Aged, 80 and over; Anemia, Aplastic; Blood Platelets; Erythropoietin; Hemoglobins; Humans; Leukocyte Count; Male

The anemia of malignancy is common and is related to several etiologic factors, a major one being relative erythropoietin (Epo) deficiency. Blood transfusions, the traditional therapy, provides a quick solution but is associated with complications. This was the rationale for recombinant human Epo (rHuEpo) in the treatment of anemia of cancer. Over the past few years, about 20 publications have reported the results of rHuEpo in the treatment of cancer-associated anemia in more than 850 patients with a variety of malignancies. In general, more than half of the patients responded with a significant increase in their hemoglobin level, a decrease in blood transfusion requirements, and an improved performance status and quality of life. About 4 weeks are required till the onset of effect. The hormone is well tolerated with minimal adverse effects and subcutaneous injections appear to be the preferred method of administration. Additional studies will hopefully answer several questions including optimal dosage and duration of treatment, Epo resistance, and the possibility of predicting the response.

Topics: Anemia, Aplastic; Blood Transfusion; Drug Resistance; Erythropoietin; Hemoglobins; Humans; Neoplasms; Quality of Life; Recombinant Proteins; Time Factors; Treatment Outcome

Topics: Anemia, Aplastic; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Leukemia; Myelodysplastic Syndromes

The human recombinant hematopoietic growth factors have attracted widespread interest because of their potential efficacy in a number of clinical settings. Recombinant human erythropoietin is now an established therapy to treat and prevent the anemia associated with chronic renal failure in children and adults. This agent also shows extraordinary promise to stimulate erythrocyte production and reduce transfusion requirements in premature infants. Granulocyte and granulocyte-macrophage colony-stimulating factors stimulate the production and function of myeloid cells and have provided major clinical benefit to children with congenital disorders of neutrophil production. The myeloid growth factors also show great promise in reducing the duration and severity of neutropenias associated with cytotoxic cancer treatment and in improving granulocyte production in patients with human immunodeficiency virus infections.

Topics: Anemia; Anemia, Aplastic; Antineoplastic Agents; Bone Marrow Transplantation; Child; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; HIV Infections; Humans; Infant, Newborn; Neutropenia; Recombinant Proteins; Stem Cell Factor

The hemopoietic growth factors are peptide hormones that are known to be responsible for the in vitro and in vivo proliferation of bone marrow progenitor cells into mature differentiated cells. These cytokines have had a major impact on the management of patients with cytopenias and have been extensively used as an adjunct to the management of patients with hematologic malignancies, with or without prior intensive chemotherapy. Other potential uses, being rigorously studied, include the potential mobilization of stem cells as well as recruitment phase-specific cells into the cell cycle, thus providing a more sensitive environment for targeting specific chemotherapeutic agents.

Topics: Acute Disease; Anemia; Anemia, Aplastic; Bone Marrow Transplantation; Colony-Stimulating Factors; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Humans; Leukemia; Myelodysplastic Syndromes; Neutropenia

Treatment of aplastic anemia may raise considerable problems in some patients. This report concerns a boy whose illness started at 11 years of age. At first admission laboratory data were: hemoglobin 7.5 g/l, and counts of leucocytes, neutrophils and platelets were 2.3, 0.6, and 8 x 10(9)/l, respectively. His bone marrow was hypoplastic with sparse erythropoiesis. The patient did not respond to traditional medical treatment. Serum contained a high concentration of erythropoietin but no antibodies against erythropoietin. The patient's serum did neither alone, nor supported with recombinant erythropoietin, stimulate erythropoiesis in a bioassay, suggesting that some factor(s) inhibiting erythropoietic activity was present. Based on this hypothesis, plasma exchange was performed. After 26 weeks of plasmapheresis the hematological parameters were normalized. We conclude that plasmapheresis might be an alternative in treatment of resistant aplastic anemia. Further diagnostic tools to identify patients who might benefit from such a treatment are required.

Topics: Anabolic Agents; Anemia, Aplastic; Antilymphocyte Serum; Blood Cell Count; Child; Cyclosporins; Erythropoietin; Hemoglobins; Humans; Immunotherapy; Male; Plasmapheresis; Prednisone; Remission, Spontaneous

Topics: Acquired Immunodeficiency Syndrome; Anemia, Aplastic; Colony-Stimulating Factors; Erythropoietin; Hematologic Diseases; Hematopoiesis; Humans; Myelodysplastic Syndromes; Recombinant Proteins

In summary, anemia developing in a patient with cancer can be due to several different factors. A relative failure of erythropoiesis, in conjunction with a modestly shortened erythrocyte survival, is the most likely explanation for the anemia and can occur in patients with or without bone marrow invasion. Several theories have been proposed to explain the mechanism of limited red cell production in cancer. Internal iron starvation and cancer toxic factors have been widely implicated. Immunoglobulin inhibitors of erythropoiesis occur in the rare entity, pure red cell aplasia, which is sometimes associated with thymomas. Autoimmune hemolytic anemia and microangiopathic hemolytic anemia can also occur in patients with solid cancers, pointing out the need for a complete evaluation of anemia in any patient with recent-onset anemia. Successful treatment and prognostic implications of anemia in cancer is dependent on proper diagnosis.

Topics: Anemia; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Megaloblastic; Anemia, Myelophthisic; Bone Marrow; Erythropoiesis; Erythropoietin; Humans; Intestinal Absorption; Iron; Mononuclear Phagocyte System; Neoplasms

In the mouse, control of the proliferation of erythroid precursors is complex and probably based on the sequential expression of membrane receptors for different regulator molecules during erythroid differentiation. Purified GM-CSF, G-CSF and multi-CSF are all able to initiate proliferation of the earlier erythroid precursors, the multi-CFC and BFU-E, and multi-CSF can stimulate the proliferation of erythroid precursors through all subsequent stages of differentiation. Erythropoietin acts as proliferative stimulus only for the terminal stages of erythropoiesis. There is as yet no evidence for the existence of a distinct BPA regulator with proliferative effects restricted to early erythroid precursors.

Topics: Anemia, Aplastic; Animals; Cell Division; Chemical Phenomena; Chemistry, Physical; Colony-Stimulating Factors; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Hematopoietic Stem Cells; Liver; Mice; Models, Biological; Pregnancy; Receptors, Cell Surface; Receptors, Colony-Stimulating Factor

Topics: Anemia, Aplastic; Bone Marrow Cells; Cell Division; Cells, Cultured; Colony-Stimulating Factors; Erythropoietin; Hematopoiesis; Humans; Immunoglobulin M; Immunosuppression Therapy; Interleukin-2; Lymphocytes; Phytohemagglutinins; T-Lymphocytes

Topics: Anemia, Aplastic; Animals; Cell Division; Colony-Stimulating Factors; Cyclic AMP; Erythrocytes; Erythrocytes, Abnormal; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Polycythemia Vera; Prostaglandins E

Topics: Anemia, Aplastic; Animals; Bone Marrow Cells; Cell Differentiation; Cell Division; Cells, Cultured; Clone Cells; Colony-Forming Units Assay; Erythrocytes; Erythropoiesis; Erythropoietin; Hematologic Diseases; Hemoglobins; Humans; Immunosuppression Therapy; Mice; Myeloproliferative Disorders; Syndrome; T-Lymphocytes; Time Factors

Pure red cell aplasia is a selective aplasia of the marrow erythroid cells. Unlike aplastic anemia, the marrow has a normal cellularity and the patients generally have normal leukocyte and platelet blood counts. The congenital form of the disease occurs in the firlst 1 1/2 years of life and is often responsive to corticosteroids. The acquired form may be secondary to infections, drugs, chemicals, or hemolytic anemia (aplastic crisis). In these cases it is often acute and self-limited with cessation of the infection or drug ingestion. It may also be secondary to systemic lupus erythematosus, rheumatoid arthritis, acute severe renal failure, severe nutritional deficiency, or diverse neoplasms, and may remit with treatment of the primary condition. When a thymoma is present, it should be resected since a remission is produced in 29 per cent of these patients. The remaining patients have an acquired primary form of the disease that tends to be chronic and in some cases may have an immune pathogenesis. A cytotoxic immunoglobulin inhibitor of the marrow erythroid cells or erythropoietin has been described and these patients may respond to prednisone and/or to cytotoxic immunosuppressive drugs such as cyclophosphamide and 6-mercaptopurine. Pure red cell aplasia appears to be more common than the literature has revealed and has stimulated much investigation into an immune pathogenesis for marrow failure.

Topics: Acute Kidney Injury; Anemia, Aplastic; Antilymphocyte Serum; Arthritis, Rheumatoid; Blood Cell Count; Blood Transfusion; Cyclophosphamide; Deficiency Diseases; Erythropoietin; Humans; Immune System Diseases; Infections; Lupus Erythematosus, Systemic; Mercaptopurine; Prednisone; Remission, Spontaneous; Splenectomy; Thymoma; Thymus Neoplasms

There have been numerous new contributions to the knowledge of foetal haemoglobin over the last few years. It is, therefore, timely to review them together. They throw light on the arrangement on the chromosome of non-alpha chain genes, and on the condition generally known as Hereditary Persistence of Foetal Haemoglobin (HPFH) and have contributed to other aspects of human ontogeny and physiology.

Topics: Adult; Amino Acid Sequence; Anemia, Aplastic; Anemia, Sickle Cell; Child; Erythrocytes; Erythropoietin; Female; Fetal Blood; Fetal Hemoglobin; Genes; Genetic Linkage; Genetic Variation; Genetics, Population; Hemoglobinopathies; Humans; Infant; Infant, Newborn; Leukemia; Oxygen; Pregnancy; Protein Denaturation; Suppression, Genetic; Thalassemia

Topics: Anemia; Anemia, Aplastic; Antibodies, Anti-Idiotypic; Antigen-Antibody Complex; Autoimmune Diseases; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Chromium Radioisotopes; Complement System Proteins; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Heme; Humans; Immunoglobulin G; Immunosuppressive Agents; Iron; Iron Radioisotopes; Middle Aged

Topics: Anabolic Agents; Androgens; Anemia, Aplastic; Bone Marrow Transplantation; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Heme; Hepatitis; Humans; Iron; Leukemia; Transplantation, Homologous; Tuberculosis, Miliary

Topics: Age Factors; Aged; Androgens; Anemia, Aplastic; DNA; Erythropoietin; Female; Hematopoiesis; Humans; Male; Methenolone; Oxymetholone; Prognosis; Remission, Spontaneous; RNA; Testosterone

Topics: Adult; Anemia, Aplastic; Animals; Antibodies, Antinuclear; Antigen-Antibody Complex; Bone Marrow; Bone Marrow Cells; Child, Preschool; Cyclophosphamide; Erythrocytes, Abnormal; Erythropoiesis; Erythropoietin; Heme; Humans; Immunoglobulin G; Infant; Mercaptopurine; Prednisone; Stimulation, Chemical; Thymectomy; Thymoma

Topics: Adrenal Cortex Hormones; Alpha-Globulins; Androgens; Anemia, Aplastic; Animals; Base Sequence; Blood Proteins; Cell Division; Chemical Phenomena; Chemistry; Dogs; Drug Synergism; Endocrine Glands; Erythropoiesis; Erythropoietin; Glycoproteins; Hematopoiesis; Humans; Kidney; Leukocytes; Leukocytosis; Leukopenia; Mice; Polycythemia; Polycythemia Vera; Radiation Effects; Rats; Thrombopoietin; Thyroid Hormones

Topics: Anemia; Anemia, Aplastic; Animals; Blood Transfusion; Bone Marrow; Bone Marrow Cells; Cell Differentiation; Chronic Disease; Erythropoiesis; Erythropoietin; Humans; Immune Sera; Kidney; Kidney Diseases; Lipids; Mercaptopurine; Polycythemia Vera

Topics: Adrenal Cortex Hormones; Adrenocorticotropic Hormone; Anabolic Agents; Anemia, Aplastic; Bone Marrow; Bone Marrow Transplantation; Capillaries; Deficiency Diseases; Erythropoietin; Hepatitis; Humans; Lectins; Splenectomy; Testosterone

Topics: Adrenocorticotropic Hormone; Anabolic Agents; Anemia, Aplastic; Erythropoietin; Gonadal Steroid Hormones; Hormones; Humans; Thyroxine

This study was designed to investigate four different immunosuppressive therapy (IST) regimens as treatment of acquired severe aplastic anemia (SAA).. 142 consecutive SAA patients were randomized to receive one of the following IST regimens: equine anti-human thymocyte immunoglobulin (E-ATG) alone (IST regimen I); E-ATG and cyclosporine A (CSA) (IST regimen II); E-ATG, CSA plus recombinant human granulocyte-macrophage colony-stimulating factor (rhuGM-CSF) and rhu erythropoietin (rhuEPO) (IST regimen III); or rabbit ATG (ATG-F), CSA, rhuGM-CSF, and rhuEPO (IST regimen IV). No repeated courses of E-ATG or ATG-F were given for nonresponders. All patients also received stanozolol or testosteron propionate.. The overall response rate to IST regimen I was 58%. The response to IST regimen II (79%) was significantly higher (p = 0.04), more rapid and complete than after IST regimen I. The response rate to IST regimen IV (53%) was significantly lower than that of IST regimen III (73%, p = 0.039). The additional use of growth factors did not reduce early deaths and did not accelerate hematopoietic recovery after IST. Of the 142 patients enrolled in this trial, 92 (65%) are alive at a median follow-up time of 102 months (range, 54-166 months). The 5-year actuarial survival for IST regimens I, II, III, and IV was 58%, 81%, 80%, and 66%, respectively.. The combination of E-ATG and CSA remains the best combination for the treatment of SAA patients, producing a survival advantage at 5 years. The addition of growth factors did not improve these results. Rabbit ATG-F appeared less effective than E-ATG.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Child; Child, Preschool; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunosuppressive Agents; Male; Middle Aged; Multivariate Analysis; Prospective Studies; Recombinant Proteins; Survival Analysis

Allogeneic hematopoietic stem cell transplantation (HSCT) is associated with prolonged anemia caused by defective erythropoietin (Epo) production. We enrolled 34 recipients of an allogeneic HSCT in three consecutive trials to determine the optimal utilization of recombinant human erythropoietin (rhEpo) therapy in this setting.. In the first trial (n = 7), rhEpo 1400 U/kg/week was given from day 1 until a hemoglobin (Hb) level of 10 g/dL was achieved, for a maximum of 60 days. In the second trial, rhEpo 500 U/kg/week was given to achieve Hb levels of 13 to 14 g/dL in 13 anemic patients with fatigue 56 to 1440 days after transplant. In the third trial, rhEpo was scheduled to start on day 35 in 14 patients at a dose of 500 U/kg/week with the aim of achieving Hb levels of 13 to 14 g/dL.. In trial 1, erythroid recovery to 1% reticulocytes and red blood cell transfusion independence were faster, but the number of transfusions was not reduced compared to 10 controls. Responses were brisk in trial 2, with transfusion independence achieved after a median of 1 week in 12 of 13 patients, and 2-g Hb increments or Hb values of 11, 12, and 13 g/dL after 6, 7, 10, and 10 weeks, respectively. Transfusions were significantly reduced in the first month of rhEpo therapy. In trial 3, transfusion independence was obtained after a median of 1 week in 13 of 14 patients, and 2-g Hb increments or Hb values of 11, 12, and 13 g/dL after 3, 4, 6, and 8 weeks, respectively. Transfusions rates were considerably reduced compared to the previous month in the same patients or compared to controls undergoing peripheral blood or marrow transplant without rhEpo.. Anemia after allogeneic HSCT is exquisitely sensitive to rhEpo. The benefit is minimal when it is given early post-transplant, as used in all trials to date. However, the rate of major response is greater than 90% when rhEpo is started after day 35. These data provide the basis on which to conduct a prospective, randomized, placebo-controlled trial of rhEpo therapy after allogeneic HSCT.

Topics: Adolescent; Adult; Anemia, Aplastic; Bone Marrow Transplantation; Child; Cyclosporine; Drug Administration Schedule; Erythropoietin; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Immunosuppressive Agents; Leukemia; Lymphoma, Non-Hodgkin; Male; Middle Aged; Myelodysplastic Syndromes; Recombinant Proteins; Thrombocytosis; Transplantation, Homologous

Interleukin-11 (IL-11) is a thrombopoietic cytokine that attenuates postchemotherapy thrombocytopenia at doses of 50 microg/kg/d subcutaneously. Very little is known about the activity of IL-11 in patients with bone marrow failure states.. Our preliminary experience with IL-11 at doses of 50 microg/kg/d suggested that patients with bone marrow failure developed significant peripheral and pulmonary edema after the prolonged dosing necessary for treating these conditions. We, therefore, initiated a study of low-dose IL-11 (starting dose, 10 microg/kg/d).. Sixteen patients were assessable for response. Six patients had diploid cytogenetics; the others had a variety of chromosomal abnormalities. Six (38%) of 16 patients showed a platelet response to IL-11, and two had a multilineage response (to IL-11 alone, n = 1; to IL-11 plus G-CSF and erythropoietin, n = 1). The median increase in peak platelet counts was 95 x 10(9)/L above baseline in the responders (range, increase of 55 x 10(9)/L to 130 x 10(9)/L above baseline). Responders included five of 11 patients with myelodysplasia and one of four patients with aplastic anemia. Response durations were 12, 13, 14+, 25, 30, and 30+ weeks. Side effects of IL-11 were mild (peripheral edema, n = 7; conjunctival injection, n = 7; myalgia, n = 1; all grade 1). Seven patients had no side effects.. Our pilot study suggests that administration of low-dose IL-11 (10 microg/kg/d) can raise platelet counts without significant toxicity in selected thrombocytopenic patients with bone marrow failure.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Bone Marrow Cells; Child; Child, Preschool; Drug Administration Schedule; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-11; Karyotyping; Male; Middle Aged; Myelodysplastic Syndromes; Pilot Projects; Platelet Count; Thrombocytopenia

To evaluate the therapeutic potential of hematopoietic growth factors (HGFs) during immunosuppressive treatment (IST) of severe aplastic anemia (SAA), 38 patients with newly diagnosed SAA received IST alone (group I), or IST plus recombinant human erythropoietin and granulocyte-macrophage colony-stimulating factor (rhEPO + rhGM-CSF) (group II). Eleven patients in each group received antilymphocyte globulin (ALG) for IST, and eight patients in each group received cyclosporine (CSA). Complete remission rates at one year were 26% and 74% for group I and group II patients, respectively. The ALG-treated subgroup showed the greatest differences between treatments. Compared with patients receiving ALG alone, patients treated with ALG plus HGFs had significantly better one-year survival (100% vs. 54.5%, P < 0.05), complete remission rates (91% vs. 36%, P < 0.05), more rapid and complete hematologic recovery, greater reductions in transfusion requirements, and lower infection rates. The data suggest a potential role for rhEPO + rhGM-CSF therapy in SAA patients receiving IST.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Blood Cell Count; Blood Transfusion; Bone Marrow Cells; Cell Count; Child; Drug Therapy, Combination; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunosuppressive Agents; Infections; Male; Middle Aged; Recombinant Proteins; Remission Induction; Survival Rate; Time Factors; Treatment Outcome

A multicenter randomized controlled study was undertaken in order to determine whether epoetin beta (EPO) ameliorates the anemia in aplastic anemia (AA) patients treated with granulocyte colony-stimulating factor (G-CSF). Enrolled patients were randomized into 3 groups: group C receiving G-CSF alone as the control; group L receiving G-CSF and 200 IU/kg of EPO; group H receiving G-CSF and 400 IU/kg of EPO. Throughout the study, the dose and the administration interval of G-CSF were adjusted to maintain neutrophil counts between 1000 and 5000 microliters EPO was administered subcutaneously for 12 wk as the first step in treatment and when favorable effects were observed over this period, treatment was continued for another 12 wk as the second step in treatment. Significant erythroid responses were defined as increases in untransfused hemoglobin values > 1.0 g/dl or > 50% decreases in RBC transfusion requirements over the treatment period. Of 131 patients enrolled, 88 patients allocated to groups L and H were evaluated for toxicity to EPO and 110 were evaluated for erythroid responses. Four of the 31 patients (12.9%) in group C, 6 of the 41 patients (14.6%) of group L, and 14 of the 38 patients (36.8%) of group H showed erythroid responses in the first step in treatment. The erythroid responses of group H were significantly higher than those of the other 2 groups (p < 0.05). The significant effects of EPO were due to erythroid responses in non-severe AA. Responding patients were significantly different from non-responders with regard to disease severity, hemoglobin concentration, reticulocyte count, serum endogenous erythropoietin levels and serum transferrin receptors; non-severe AA patients were more likely to respond to EPO, and responding patients had lower serum EPO and higher hemoglobin concentration, reticulocyte count and serum transferrin receptors than non-responders. The response rate increased in the second step in treatment, suggesting that long-term treatment improved the efficacy of EPO. No serious side-effects were observed. From these results, we conclude that EPO given in combination with G-CSF is a safe and effective alternative for the treatment of anemia of a subset of AA patients.

Topics: Adult; Anemia, Aplastic; Biomarkers; Blood Transfusion; Drug Therapy, Combination; Erythropoiesis; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Hemoglobins; Humans; Male; Receptors, Transferrin; Recombinant Proteins; Reticulocyte Count; Time Factors

A therapeutic trial of interleukin-3 (IL-3) was carried out in four patients with aplastic anemia refractory to the prior therapies. Daily subcutaneous doses of 2.5, 5.0 or 7.5 micrograms/kg was given for 7 or 14 days. In a patient who had co- and immediate boost-administration of granulocyte colony-stimulating factor (G-CSF) and/or erythropoietin (Epo) and another who had sequential administration of G-CSF and Epo two weeks after IL-3, definite hematological response was obtained during the course after IL-3. In one patient, moderate to severe side effects consisting of facial edema, conjunctival bleeding, chills and fever, were observed after two days' administration of IL-3. Co- or sequential administration of other hemopoietic factor(s) may be essential in IL-3 therapy for aplastic anemia.

Topics: Adult; Aged; Anemia, Aplastic; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-3; Male; Middle Aged; Recombinant Proteins

Serum erythropoietin (EPO) concentrations were measured by radioimmunoassay in 46 patients with idiopathic aplastic anemia. The concentrations of serum EPO in aplastic anemia were shown to correlate inversely both with hemoglobin concentration and % bone marrow hematopoietic tissue. Serial determination of EPO levels showed a progressive decline in proportion to improvement of anemia. The results suggest that serial determination of serum EPO is useful in monitoring the hematopoiesis of aplastic anemia.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Biomarkers; Bone Marrow Cells; Erythropoietin; Female; Hematopoiesis; Hemoglobins; Humans; Male; Middle Aged; Monitoring, Physiologic; Radioimmunoassay

Ten patients with aplastic anemia (AA) and seven patients with refractory anemia (RA) were treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and erythropoietin (rhEpo) in combination. rhG-CSF (5-20 micrograms/kg) and rhEpo (120-720 U/kg) were administered by s.c. injection three times a week for at least six months, and the administration was continued as maintenance therapy for as long as possible when hematological responses were observed. Six (60%) of the ten AA patients and four (58%) of the seven RA patients showed multilineage responses. Of these responders, six patients achieved trilineage recovery. While all of the responders were dependent on red blood cell transfusions and eight of them required platelet transfusions before treatment, they now no longer need transfusions of either red blood cells or platelets. A median treatment duration of 9 (range 1 to 28) months was required to achieve multilineage recovery. The responders showed an ability to maintain the multilineage recovery for 9+ to 47+ months and to tolerate long-term treatment. These results indicate that the long-term treatment with rhG-CSF and rhEpo may benefit a substantial percentage of patients with AA and RA and provide an optional therapy for these patients.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Anemia, Refractory; Drug Therapy, Combination; Erythrocyte Transfusion; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Male; Middle Aged; Platelet Transfusion; Recombinant Proteins; Time Factors

The safety and efficacy of recombinant human erythropoietin (epoetin alpha) were investigated in adult aplastic anemia patients whose hemoglobin (Hb) concentration was less than 10g/dl. Epoetin alpha was given subcutaneously every day at a dose of 3,000IU/body for two weeks, and the dosage was increased to 6,000IU, 12,000IU and 24,000IU every two weeks when the increment of Hb was insufficient. In cases in whom Hb concentration increased by more than 1g/dl or whose transfusion requirements reduced to less than 50%, treatment was judged to be effective. The whole rate of efficacy was 34.5% (10/29). Response to epoetin alpha treatment was better in patients whose symptoms were relatively mild. Mild cases responded to the treatment with 6,000IU/body/day, although a dosage of 24,000IU/body/day was required in moderate or severe cases. Neither serious adverse effect nor abnormal laboratory findings were observed. These results suggest that high dose subcutaneous epoetin alpha treatment is effective for the aplastic anemia in terms of increasing Hb concentration and reducing blood transfusions.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Blood Transfusion; Bone Marrow Examination; Erythropoietin; Female; Hemoglobins; Humans; Male; Middle Aged; Recombinant Proteins

The anemia of patients with familial amyloidotic polyneuropathy (FAP) was evaluated. Anemia was seen in 32 (91%) of the 35 FAP patients, more often with progression of the disease. The incidence of macrocytic hypochromic anemia was the most common type (40%). In 14 autopsied and 2 biopsied cases, no amyloid deposition was detected in the bone marrow. Thirteen (81%) of the 16 FAP patients showed hypoplastic bone marrow. Bone marrow aspiration of 2 patients revealed a decreased ratio of erythrocytic/myelocytic cells. The plasma levels of vitamin B12 and folate were within normal ranges. Neither oral nor intravenous administration of iron had any effect on the anemia of FAP patients. Intravenous erythropoietin elevated blood hemoglobin levels and blood pressure in 2 patients. Orthostatic hypotension, one of the most common symptoms of FAP, was unexpectedly improved. Secondary hypoplastic anemia is common in FAP, but treatment of anemia in this disease using erythropoietin is promising.

Topics: Adult; Aged; Amyloid Neuropathies; Anemia, Aplastic; Biopsy, Needle; Bone Marrow; Erythropoietin; Female; Humans; Male; Middle Aged; Recombinant Proteins

Topics: Adult; Aged; Aged, 80 and over; Anemia; Anemia, Aplastic; Blood Transfusion; Erythrocyte Indices; Erythroid Precursor Cells; Erythropoietin; Female; Humans; Male; Middle Aged; Myelodysplastic Syndromes; Recombinant Proteins

Topics: Adolescent; Adult; Anemia, Aplastic; Child; Child, Preschool; Erythropoietin; Humans; Infant; Recombinant Proteins

Recent advances in genomics have enabled the successful identification of a number of rare pathogenic mutations. Uncovering these mutations is essential as the first step towards devising a cure for the often debilitating and life-limiting diseases arising from them. For many of these mutations targeted agents do not yet exist. Here, we describe the case of a patient who has a novel pathogenic mutation in the erythropoietin (. The patient aged 5 months, was started on recombinant erythropoietin, at a standard dose of 500 units (50 U/kg) and subsequently 800 units three time weekly and her blood counts were monitored over 4 years.. Patient-customized therapies can be highly effective in the treatment of rare genetic disorders and for many of these disorders effective treatment may already exist in the clinical domain, as described for the patient in this report.. This work was supported by the New York Stem Cell Foundation (V.G.S.), a gift from the Lodish Family to Boston Children's Hospital (V.G.S.), and National Institutes of Health Grants R01 DK103794 and R01 HL146500 (V.G.S.).

Topics: Anemia, Aplastic; Child; Epoetin Alfa; Erythropoietin; Female; Hemoglobins; Humans; Mutation; Recombinant Proteins; United States

Aplastic anemia (AA) is characterized as hypoplasia of bone marrow hematopoietic cells and hematopenia of peripheral blood cells. Though the supplement of exogenous erythropoietin (EPO) has been clinically approved for AA treatment, the side-effects hinder its further application. Here a robust treatment for AA induced by chemotherapy drugs is explored using gadofullerene nanoparticles (GFNPs).

Topics: Anemia, Aplastic; Animals; Antineoplastic Agents, Alkylating; Bone Marrow Cells; Busulfan; Cell Differentiation; Cyclophosphamide; Disease Models, Animal; Erythropoiesis; Erythropoietin; Female; Fullerenes; Hematopoietic Stem Cells; Mice; Mice, Inbred ICR; Nanoparticles

The patients with severe aplastic anemia (SAA) usually rely on red cell transfusion which lead to secondary iron overload. Transforming growth differentiation factor-15 (GDF-15) plays an important role in erythropoiesis and iron regulation. In this study, we investigated the level of GDF-15 and other indexes of iron metabolism in SAA patients to explore the correlation with GDF-15 and iron overload in SAA.. The levels of serum GDF-15, hepcidin (Hepc), and erythropoietin (EPO) were determined by ELISA. The levels of serum iron (SI), ferritin, TIBC, and transferrin saturation (TS) were measured by an auto analyzer. Iron staining of bone marrow cells was used for testing extracellular and intracellular iron.. The GDF-15 level in the experimental group was higher than that of the case-control group and normal control group (all p < 0.05). The Hepc level in the experimental group and case-control group were both higher than that of healthy controls (all p < 0.05). The Hepc level was significantly lower in the experimental group patients who had excessive GDF-15 (r = -0.766, p = 0.000). There was a positive correlation between the level of GDF15 and EPO in the experimental group (r = 0.68, p < 0.000). The level of GDF15 in SAA patients was positively correlated with SI levels (r = 0.537, p = 0.008), TS levels (r = 0.466, p = 0.025), and sideroblasts (%) (r = 0.463, p = 0.026). Moreover, there was a positive correlation between GDF-15 level and blood transfusion-dependent time (r = 0.739, p = 0.000).. Our data indicated that GDF-15 plays an important role in iron metabolism in SAA. GDF-15 might be a novel target for SAA therapy.

Topics: Adult; Anemia, Aplastic; Biomarkers; Blood Transfusion; Bone Marrow; Bone Marrow Cells; Case-Control Studies; Erythropoietin; Female; Growth Differentiation Factor 15; Hepcidins; Humans; Iron; Male; Middle Aged; Prognosis; Severity of Illness Index; Young Adult

To evaluate the efficacy and safety of Shengxue mixture combined with intraosseous blood infusion for treatment of aplastic anemia patients.. From 2011 to 2015, Institute of blood diseases of Shaanxi Medical University admitted 53 patients with aplastic anemia. The patients were treated with shengxue mixture 200 ml, orally, twice a day. Stanozolol tablets, Adult 2 mg, three times a day, mycophenolate mofetil 1.0 g, twice a day. Intraosseous infusion of the following medicine were administered in patients: recombinant human EPO 10000 U, recombinant human G-CSF 450 µg, recombinant human IL-11 4.5 mg, dexmethasone 20 mg, once a week, a total of four times. One month later, the blood cell counts and bone marrow biopsy were performed. Consolidation treatment continued for 3 to 6 months after discharge, and therapeutic effect was observed and followed-up for more than a year.. After one month of treatment, 40 patients were basically cured (75.47%), 8 patients were remitted(15.09%), Hemoglobin level, white blood cell count and platelet count were significantly improved after treatment (P<0.01). The overall response rate was 90.57%(48 patients). Patients with bone marrow hyperplasia was 46 (86.79%), versus 9(16.98%) before treatment. There was a difference (P<0.05). After 3 to 6 months of treatment, 40 patients were cured (75.47%); 8 patients were remitted(15.09%); 3 patients were obviously improved(5.66%); 2 patients were ineffective(3.77%). The overall response rate was 96.23%(51 cases). No obvious side effects were observed. No patients were relapsed after one year.. Shengxue mixture combined with Intraosseous infusion is a fast, efficient, safe method for the treatment of aplastic anemia.

Topics: Anemia, Aplastic; Drugs, Chinese Herbal; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Infusions, Intraosseous

Erythroid hypoplasia (EH) is a rare complication associated with recombinant human erythropoietin (rHuEPO) therapies, due to development of anti-rHuEPO antibodies; however, the underlying mechanisms remain poorly clarified. Our aim was to manage a rat model of antibody-mediated EH induced by rHuEPO and study the impact on iron metabolism and erythropoiesis. Wistar rats treated during 9 weeks with a high rHuEPO dose (200 IU) developed EH, as shown by anemia, reduced erythroblasts, reticulocytopenia, and plasmatic anti-rHuEPO antibodies. Serum iron was increased and associated with mRNA overexpression of hepatic hepcidin and other iron regulatory mediators and downregulation of matriptase-2; overexpression of divalent metal transporter 1 and ferroportin was observed in duodenum and liver. Decreased EPO expression was observed in kidney and liver, while EPO receptor was overexpressed in liver. Endogenous EPO levels were normal, suggesting that anti-rHuEPO antibodies blunted EPO function. Our results suggest that anti-rHuEPO antibodies inhibit erythropoiesis causing anemia. This leads to a serum iron increase, which seems to stimulate hepcidin expression despite no evidence of inflammation, thus suggesting iron as the key modulator of hepcidin synthesis. These findings might contribute to improving new therapeutic strategies against rHuEPO resistance and/or development of antibody-mediated EH in patients under rHuEPO therapy.

Topics: Anemia, Aplastic; Animals; Antibodies; Erythropoiesis; Erythropoietin; Gene Expression Regulation; Hepcidins; Humans; Iron; Membrane Proteins; Rats; Rats, Wistar; Recombinant Proteins; Serine Endopeptidases

to estimate the regulation of erythropoiesis and the coagulation system in patients with suppressed hematopoiesis in a mountain hospital (3200 m above sea level).. The investigation included 12 patients with aplastic anemia (AA) and 10 with idiopathic thrombocytopenic purpura (ITP). Blood was received at a Bishkek hospital, then on days 20 and 40 of stay in the mountains. The authors studied erythropoietin (EPO) by enzyme immunoassay (Protein Contour kit, Russia), serum ferritin (SF) by immunoradioassay (Immunotech kit, Czech Republic), hypoxia-inducible factor-1alpha (HIF-1alpha), homocysteine (HC), hepcidin, endothelin (ET), and thrombomodulin (TM) by sandwich enzyme immunoassay, by applying monospecific antisera and monoclonal antibodies against relevant antigens (IDG Int Inc, USA).. On staying in the mountains, there was a gradual increase in the content of hemoglobin in patients with AA and ITP. On day 40, in keeping with higher hemoglobin (Hb) levels, both groups showed a decrease in HIF-1alpha concentrations to the normal values (from 8.2 to 4.5 pg/ml). Due to the anemic syndrome, baseline EPO was increased by 5-7 times in the patients from both groups. On days 20-40, the content of EPO showed a 1.3-2.5-fold increase. In AA, HC was almost 3 times greater than the normal values; in ITP, it was 1.5-fold increased. On day 20 and during the patients'stay in the mountains, the level of HC remained in the normal range in both groups.. Hypoxic hypoxia positively affects a number of hematological parameters, by normalizing erythropoiesis (Hb, EPO, and HIF-1alpha), iron metabolism (SF), and the coagulation system (HC, ET, and TM).

Topics: Adolescent; Adult; Altitude; Anemia, Aplastic; Biomarkers; Climatotherapy; Erythropoiesis; Erythropoietin; Ferritins; Follow-Up Studies; Hematopoiesis, Extramedullary; Hemoglobins; Homocysteine; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoenzyme Techniques; Kyrgyzstan; Middle Aged; Purpura, Thrombocytopenic, Idiopathic; Radioimmunoassay; Thrombomodulin; Treatment Outcome; Young Adult

The aim of the study is to report results of erythropoietin treatment for late hyporegenerative anemia in the hemolytic disease of the newborn (HDN). Reports previously published concern only a few cases, with controversial results.. Case series report concerning 50 neonates with HDN due to Rh, ABO or KpA antigens, aged more than 7 days. Erythropoietin treatment started when hematocrit dropped to levels requiring transfusion, with an inappropriate reticulocyte response (Reticulocyte Production Index <1).. At start of treatment mean age was 24.3 +/- 12.0 days (range 8-65 days), hematocrit 24.1 +/- 2.8% (range 18-30%), and Reticulocyte Production Index 0.34 +/- 0.25 (range 0.05-0.98). Hematocrit and Reticulocyte Production Index showed significant increases after 7 and 14 days of treatment (p <0.001). No difference was observed either between infants with Rh-HDN and ABO-HDN or between Rh-HDN patients with or without intrauterine transfusions. Seven infants (14%) required one packed RBC transfusion during erythropoietin therapy, 2 of them within 72 hours from starting treatment. The percentage of transfused infants showed no difference either between ABO-HDN and Rh-HDN or between Rh-HDN with and without intrauterine transfusions. Moderate, short-lasting neutropenia, not associated to infections, was observed in 11 patients. No other adverse effect was observed.. The administration of erythropoietin appears to be a safe and useful therapy. Its efficacy should be confirmed by randomized studies.

Topics: Anemia, Aplastic; Erythroblastosis, Fetal; Erythropoietin; Humans; Infant; Infant, Newborn; Recombinant Proteins

The objective of this study was to investigate the expressions of hematopoietic cytokines EPO, SCF and GM-CSF in bone marrow cells of patients with chronic aplastic anemia (CAA) and their significance. The mRNA expressions of EPO, SCF and GM-CSF from 35 CAA patients and 10 healthy individuals were detected by semi-quantitative RT-PCR. The results showed that the levels of EPO, SCF and GM-CSF in CAA patients were obviously lower than those in the healthy individuals (p < 0.01). It is concluded that the immunity disorder mediated by hematopoietic cytokines is one of the pathogenesis in the CAA patients, it provided theoretical basis for guiding clinical treatment.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Bone Marrow; Child; Chronic Disease; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Male; Middle Aged; RNA, Messenger; Stem Cell Factor; Young Adult

The aim of this study was to investigate the curative effects of amifostine (AMF) combined with recombinant human beta-erythropoietin (rhbeta-EPO) on patients with pure erythroid aplasia (PEA). Two patients with PEA were treated with amifostine and rhbeta-EPO. The therapeutic regimen was adopted with AMF 0.4 g/day given by intravenous injection for 5 days first, then after a break of 2 days it went on for 3 weeks consecutively, that was considered as one treatment cycle. The rhbeta-EPO 6 000 U was used by subcutaneous injection for 3 times per week. The results showed that the red cell count, hemoglobin and reticulocyte count of two patients obviously increased after treatment. The erythroid ratio in bone marrow increased. Bone marrow biopsy showed that the erythroid proliferation improved. Intervals of red cell transfusions (RCT) in the two patients who live by red cell transfusion were prolonged after AMF treatments, and the amounts of each RCT was decreased obviously. The main side effect of amifostine was discomfort of digestive system, but was tolerated by all patients. In conclusion, amifostine plus rhbeta-EPO may be a new, effective and safety method especially for the elder PEA patients. The long-term curative effects and mechanism of amifostine still need further evaluation.

Topics: Aged, 80 and over; Amifostine; Anemia, Aplastic; Drug Therapy, Combination; Erythropoietin; Humans; Male; Recombinant Proteins; Treatment Outcome

Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by intravascular hemolysis leading to anemia and other clinical manifestations. Transfusions are often required to support hemoglobin at tolerable levels. A PNH patient with aplastic anemia was treated with the complement inhibitor eculizumab, followed by concurrent treatment with recombinant human erythropoietin (rHuEpo). Eculizumab alone reduced hemolysis, increased PNH red blood cell (RBC) mass, and decreased transfusions. Addition of rHuEpo during eculizumab therapy, enhanced erythropoiesis, further increased PNH RBC mass and hemoglobin levels, and rendered the patient transfusion independent for more than two years. These data show that driving erythropoiesis during eculizumab treatment provided further benefit to a patient with PNH and underlying bone marrow failure.

Topics: Anemia, Aplastic; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Blood Transfusion; Complement C5; Drug Synergism; Erythrocyte Indices; Erythropoiesis; Erythropoietin; Hemoglobins; Hemoglobinuria, Paroxysmal; Hemolysis; Humans; Male; Middle Aged; Recombinant Proteins; Treatment Outcome

Topics: Algorithms; Anemia, Aplastic; Bone Marrow Transplantation; Cyclosporine; Erythropoietin; Humans; Red-Cell Aplasia, Pure; Severity of Illness Index

Topics: Acquired Immunodeficiency Syndrome; Anemia, Aplastic; Autoantibodies; Erythropoietin; Heart Transplantation; HIV-1; Humans

Topics: Anemia, Aplastic; Erythropoiesis; Erythropoietin; Humans; Neoplasms; Recombinant Proteins; Red-Cell Aplasia, Pure

A line of transgenic mice overexpressing erythropoietin was created. These mice retained their capacity to reduce their gastro-intestinal absorption of iron and to regulate the changes in their iron metabolism and they could serve as a model for the in vivo study of iron homeostasis and erythropoiesis. NEW INDICATIONS FOR RECOMBINANT HUMAN ERYTHROPOIETIN: After chronic terminal kidney failure, the treatment of chronic dialysed kidney failure patients and patients treated with azathioprine or patients having undergone surgery and requiring transfusion, other indications have been proposed. Such as anaemia in children following inadequate production of endogenous erythropoietin and/or direct inhibition of the erythroid cell line in the bone marrow or anaemia during pregnancy and, since the Sixties, anaemia during cancer. TO ASSESS THE PHYSIOPATHOLOGY OF ANEMIA:In anaemic patients suffering from a malignant blood disease, it would be useful to calculate the relationship between the predicted and observed rates of erythropoietin as well as the transferin serum receptors.

Topics: Adult; Anemia, Aplastic; Animals; Child; Erythropoiesis; Erythropoietin; Female; Humans; Male; Mice; Mice, Transgenic; Neoplasms; Pregnancy; Receptors, Transferrin; Recombinant Proteins; Red-Cell Aplasia, Pure; Treatment Outcome

IMPROVED QUALITY OF LIFE WITH EPOETIN BETA: In a study against a placebo, there was evidence that the quality of life scores were significantly improved in patients treated with epoetin beta, whether they exhibited a solid tumour or a malignant lymphoma. The same was noted in children with cancer exhibiting severe neoplasia and treated with chemotherapy. The efficacy and tolerance to treatment were equivalent, whatever the administration regimen. IN PATIENTS SUFFERING FROM MYELODYSPLASTIC SYNDROMES: A particular entity among malignant blood diseases, myelodysplastic syndromes are at the origin of anaemia against which repeated transfusions and growth factors are proposed with varying results and disadvantages, and against which erythropoietin may be moderately effective (a mean of 25% in non-selected cohorts of patients). DEPENDING ON THE PROTOCOLS OF ERYTHROPOIETIN ADMINISTRATION IN CASES OF MYELODYSPLASTIC SYNDROMES: Recombinant human erythropoietin, irrespective of its concentration in myelodysplastic cell culture, does not appear capable of restoring normal erythropoiesis. The influence of prolonged treatment is not admitted by all. The effects of the addition of growth factors (notably G-CSF) are obvious but some are controversial because of the costs and the prolonged duration of such treatments.

Topics: Adult; Anemia, Aplastic; Blood Transfusion; Child; Erythropoiesis; Erythropoietin; Hemoglobinometry; Humans; Myelodysplastic Syndromes; Neoplasms; Quality of Life; Randomized Controlled Trials as Topic; Recombinant Proteins; Red-Cell Aplasia, Pure; Treatment Outcome

PURE RED CELL APLASIA: Designated by the acronym PRCA or the term erythroblastopenia, pure red cell aplasia is characterised by severe anaemia with reticulocytopenia. It may occur in acute form induced by infectious agents, following drug toxicity or transplantation of allogeneic haematopoietic cells, associated with autoimmune haemolytic anaemia. The chronic form is rarely constitutional but can be acquired and is usually associated with blood or idiopathic diseases. IMMUNOLOGICAL INHIBITION OF ERYTHROPOIESIS: Among the mechanisms responsible for PRCA is immunological erythropoiesis inhibition. This may be of lymphocyte T cell origin or due to the presence of antibodies in the patient's serum. Although observations of PRCA with presence of neutralising antierythropoietin antibodies in patient's serum have multiplied over the past 5 years, they still remain extremely rare. From a therapeutic point of view, they require withdrawal of epoetin and often the administration of immunosuppressors and transfusion for symptomatic treatment. GROWTH FACTORS: The role of growth factors in restoring aplastic anaemia appears to be only partial, at random and temporary.

Topics: Acute Disease; Anemia, Aplastic; Autoantibodies; Chronic Disease; Erythropoietin; Humans; Immunosuppressive Agents; Recombinant Proteins; Red-Cell Aplasia, Pure; Risk Factors

THE DEBATE: Although some believe that rHu-EPO should not be widely used in malignant affections, others think that because of the varied impact of these anaemia, its wider use should be recommended. FOR A TARGET USE: Various observations (influence of the degree of extension of the myelomas to the skeleton and response to specific treatment in the case of myeloma, whatever the haemoglobin concentration, degree of prevention of rHu-EPO chemo-induced anaemia) are in favour of its use in selected patients. FOR A WIDER USE: The benefits of treatment with rHu-EPO are not limited to the symptomatology of anaemia but extend to its potential complications in the most fragile patients. Other than the risks of infection, the heavy costs of transfusions must also be taken into account.

Topics: Anemia, Aplastic; Blood Transfusion; Cost-Benefit Analysis; Erythropoietin; Hemoglobinometry; Humans; Multiple Myeloma; Neoplasms; Prognosis; Recombinant Proteins; Red-Cell Aplasia, Pure

Topics: Anemia, Aplastic; Blood Transfusion; Erythropoietin; Humans; Neoplasms; Quality of Life; Recombinant Proteins; Red-Cell Aplasia, Pure

Previously, we and others have shown that fetal liver infusion (FLI) leads to autologous hematopoietic improvement in 40-54% of patients with aplastic anemia. However, whether this recovery was spontaneous or the effect of the infused liver cells was not clear. To dissect the role of FLI in autologous hematopoietic recovery, the colony-supporting potential of fetal liver-conditioned medium (FLCM) was evaluated in bone marrow (BM) cells of normal adult and aplastic anemia patients. In both cases, each sample of FLCM supported the growth of colony-forming cells in semi solid culture medium. The FLCM was assayed for the presence of four principal colony-stimulating cytokines, namely stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and erythropoietin (Epo). While GM-CSF, IL-3, and Epo were present in insignificant amounts or were altogether absent, 50-635 pg/ml of SCF was found in 8 of the 13 FLCM samples tested. Preliminary results of bioneutralization assay indicated the possible role of SCF, secreted by the FL cells, in colony-supporting activity of aplastic anemia and normal BM cells. Overall, our in vitro study implicates the paracrine role of infused FL cells in regenerating autologous hematopoiesis in aplastic anemia patients.

Topics: Anemia, Aplastic; Antibodies, Monoclonal; Bone Marrow Cells; Cells, Cultured; Colony-Forming Units Assay; Colony-Stimulating Factors; Culture Media, Conditioned; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Female; Fetus; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Stem Cells; Hepatocytes; Humans; Interleukin-3; Pregnancy; Stem Cell Factor

Recombiant human erythropoietin (epoetin) has greatly contributed to improvement of the anemia of chronic renal failure patients on hemodialysis. However, the reduced erythropoietic effect to epoetin and its high cost have induced lots of supplementary treatments. Therefore, we performed a prospective study to evaluate the effects of adjuvant low-dose androgen therapy in patients using a lower-dose of epoetin than the commonly recommended dose on anemia and the nutritional parameters.. 17 patients of hemoglobin (Hgb) less than 9 g/dL even after being treated with 1,000 U epoetin subcutaneously (s.c.) 3 times per week on a stable status for more than 6 months, who were on hemodialysis at our institution were examined. They were injected with the same dose of epoetin s.c. and nandrolone decanoate 100 mg intramuscularly (i.m.) weekly for another 6 months. Blood test was performed every month before therapy for 6 months and after therapy for 6 months and the mean values were reviewed for comparison.. Hgb (7.75 +/- 0.9 vs 8.99 +/- 1.39 g/dL, p < 0.01) and hematocrit (Hct) (23.68 +/- 2.85 vs 26.66 +/- 3.91%, p < 0.01) were apparently changed before and after adjuvant therapy. Hgb and Hct, weekly dose of epoetin were not statistically different in 9 male patients before and after adjuvant therapy. The weekly dose of epoetin was not statistically different in 8 female patients, but Hgb and Hct (8.02 +/- 0.6 vs 9.72 +/- 1.31 g/dL, 24.54 +/- 1.7 vs 28.74 +/- 3.06%, p < 0.01) were statistically different before and after adjuvant therapy. In comparison between male and female groups, weekly doses of epoetin and nandrolone decanoate were significantly greater in the female group than the male group (epoetin: 50.66 +/- 6.23 vs 61.18 +/- 8.76 U/kg/week, nandrolone decanoate: 1.69 +/- 0.2 vs 2.04 +/- 0.29 mg/kg/week, p < 0.05).. Our data show that the adjuvant androgen therapy is effective for the anemia of hemodialysis patients who did not recover from anemia even after being continuously treated with low-dose epoetin.

Topics: Adult; Anabolic Agents; Anemia, Aplastic; Chemotherapy, Adjuvant; Erythropoiesis; Erythropoietin; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; Nandrolone; Nutritional Status; Prospective Studies; Recombinant Proteins; Renal Dialysis

In our previous study, approximately 60% of aplastic anemia (AA) and refractory anemia (RA) patients treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human erythropoietin (rhEpo) showed a multilineage response. In this study, we analyzed the long-term follow-up of the multilineage responders (multi-R). In the follow-up analysis of 11 multi-R (6 AA and 5 RA), 10 patients (5 AA and 5 RA) were evaluable. The range of time from the start of treatment to the final contact was 50 to 125 months. Analysis of survival times revealed a significant difference between multi-R and non-multi-R among AA patients given this treatment (P = .016). One AA and 1 RA patient among the multi-R developed acute leukemia. Of 7 living multi-R, 3 AA and 2 RA patients did not need transfusion at final contact. Four of them maintained the target hemoglobin concentration of more than 11 g/dL for quality-of-life benefit. The findings suggested that this result is an important advantage of this treatment.

Topics: Acute Disease; Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Refractory; Blood Transfusion; Erythropoietin; Female; Follow-Up Studies; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Male; Middle Aged; Survival Analysis

A 9-year-old girl who had hepatitis-associated aplastic anemia was treated intermittently with methylprednisolone pulse therapy and growth factors (granulocyte-colony stimulating factor (G-CSF), recombinant human erythropoietin (rhEpo) and cyclosporin A (CyA) for over two years. At this time, there was hematological improvement, but chromosome analysis revealed monosomy 7. After six months, there was progression to myelodysplastic syndrome (MDS) (stage in refractory anemia of excess blasts (RAEB)) with monosomy 7, monosomy 6, marker chromosome and with hematological deterioration. She received bone marrow (1.57 x 10(5) cells kg(-1) (patient body weight)) plus cord blood cell (0.3 x 10(7) cells kg(-1) (patient body weight)) transplantation from her brother, 2 years and 7 months after the diagnosis of hepatitis-associated aplastic anemia. Engraftment was achieved after two weeks, and acute graft-versus-host disease occurred in a mild form after four weeks. Hematological remission has been continuous for 20 months after bone marrow transplantation. Transformation of hepatitis-associated aplastic anemia to MDS with the monosomy 7, monosomy 6 and marker chromosome in this patient was considered to have been related to the administration of high doses of immunosuppressive drugs plus growth factors.

Topics: Anemia, Aplastic; Bone Marrow Transplantation; Child; Chromosomes, Human, Pair 6; Chromosomes, Human, Pair 7; Cord Blood Stem Cell Transplantation; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Growth Substances; Hepatitis; Humans; Immunosuppressive Agents; Monosomy; Myelodysplastic Syndromes; Recombinant Proteins; Transplantation, Homologous; Treatment Outcome

Topics: Aged; Anemia, Aplastic; Erythropoietin; Humans; Kidney Failure, Chronic; Male; Recombinant Proteins; Renal Dialysis

An 83-year-old woman received a diagnosis of moderate aplastic anemia in November 1990. Immunosuppressive therapy consisting of anti-lymphocyte globulin combined with high-dose corticosteroids was effective until pancytopenia developed in August 1993. The patient was hospitalized again and recurrence of aplastic anemia was diagnosed on the basis of hematologic findings, including RBC 129 x 10(4)/microliter, Hb 5.5 g/dl, Ret 23,200/microliter, WBC 2,200/microliter with 27% neutrophils, platelets 2.2 x 10(4)/microliter, and hypoplastic bone marrow. Recombinant human granulocyte-colony stimulating factor (G-CSF) of 125 micrograms/day combined with recombinant human erythropoietin (EPO) of 6,000 U/day were started in November 1993. The doses of G-CSF and EPO were increased to 250 micrograms/day and 12,000 U/day, respectively. We stopped combination therapy in March 1995, after trilineage hematopoietic cell recovery had been achieved. Complete recovery in peripheral blood was sustained for more than 2 years despite the termination of G-CSF and EPO therapy. Combination therapy with G-CSF and EPO may be safe and effective for elderly patients with aplastic anemia when the choice of therapy is limited.

Topics: Aged; Aged, 80 and over; Anemia, Aplastic; Drug Therapy, Combination; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Recombinant Proteins; Remission Induction; Time Factors; Treatment Outcome

We report the case of a 41-year-old black man with acquired immunodeficiency syndrome who developed a severe chronic anemia due to parvovirus infection. Bone marrow biopsy revealed erythroid aplasia. The infectious nature of the anemia was not recognized, and the patient was treated with erythropoietin. The patient's reticulocyte response was inadequate, however, and he remained anemic. A second bone marrow biopsy showed erythroid hyperplasia and prominent intranuclear parvovirus inclusions within erythroid progenitors. Erythropoietin was discontinued and was followed by a course of intravenous immunoglobulin, which resulted in rapid correction of anemia. To our knowledge, this is the first reported case of fulminant human parvovirus infection exacerbated by erythropoietin administration and documented by sequential bone marrow histologic examination. This case illustrates the critical importance of considering parvovirus in the etiology of chronic anemia with erythroid aplasia in immunocompromised patients.

Topics: Adult; AIDS-Related Opportunistic Infections; Anemia, Aplastic; Antibodies, Viral; Erythrocytes; Erythropoietin; Fatal Outcome; Humans; Hyperplasia; Immunoglobulins, Intravenous; Male; Parvoviridae Infections; Parvovirus B19, Human; Recombinant Proteins; Virus Activation

Topics: Adult; Anemia, Aplastic; Antilymphocyte Serum; Bone Marrow Transplantation; Erythropoietin; Hemoglobinuria, Paroxysmal; Humans; Male; Myelodysplastic Syndromes; Splenectomy; Transplantation Conditioning

The relationship between aplastic anemia and viral hepatitis is well recognized, and such patients usually have a high mortality. We successfully treated a case of aplastic anemia following living-related orthotopic liver transplantation (LROLT) for non-A, non-B, non-C hepatitis. A 2-yr-old boy with fulminant hepatic failure from non-A, non-B, non-C hepatitis received LROLT. Before transplantation, he had pancytopenia which was probably hepatitis associated, and viral suppression was suspected after bone marrow (BM) biopsy. After the transplantation, he developed progressive pancytopenia and a diagnosis of aplastic anemia was made via BM biopsy. With immunosuppressant agents (cyclosporine, methylprednisolone), cytokine therapy (granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), recombinant human erythropoietin (rhEPO)) was effectual and the patient recovered from pancytopenia. He was discharged from the hospital 57 d after the liver transplantation and remains well 1 yr after LROLT. Combined cytokine therapy with high doses of G-CSF, M-CSF and rhEPO appeared to be effective in the treatment of aplastic anemia following liver transplantation for non-A, non-B, non-C hepatitis. Since M-CSF activates macrophages, it may have contributed to the graft rejection. Careful consideration should be given to the use of high-dose M-CSF in liver transplant patients.

Topics: Anemia, Aplastic; Bone Marrow; Child, Preschool; Drug Therapy, Combination; Erythropoietin; Granulocyte Colony-Stimulating Factor; Hepatitis, Viral, Human; Humans; Immunosuppressive Agents; Liver Transplantation; Living Donors; Macrophage Colony-Stimulating Factor; Male; Recombinant Proteins

We have recently shown that in patients with aplastic anemia (AA) recovering following immunosuppressive therapy, the persistent reduction in the bone marrow clonogenic potential is unrelated to suppressive effects of the myeloid stroma and intrinsic to the hematopoietic progenitors. We examined the mechanisms of this defect by determining the response of the aplastic CD34+ clonogenic precursors to proliferative signals induced by hematopoietic growth factors and comparing their results with those of a control population. Light density bone marrow mononuclear cells were lymphocyte and monocyte depleted and enhanced for the CD34+ progenitors by immunomagnetic selection. Selected progenitors were then cultured in the mixed colony assay with incremental concentrations of combinations containing erythropoietin (Epo), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3) and c-kit ligand. Bone marrow from aplastic patients had significantly fewer light density cells displaying the CD34 antigen (mean 0.65%, SD 0.35 vs. 1.62%, SD 1.4; p=0.002). Dose response studies on aplastic CD34+ cells demonstrated that at low concentrations of Epo, IL-3 and GM-CSF, clonogenic growth was significantly impaired but achieved normal values at concentrations giving plateau growth in control cultures. However, for all colony types, responses to effective concentrations of c-kit ligand corresponded with those of controls. These data suggest abnormalities at the receptor or signal transduction levels.

Topics: Adolescent; Adult; Anemia, Aplastic; Antigens, CD34; Cohort Studies; Colony-Forming Units Assay; Dose-Response Relationship, Drug; Erythropoietin; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Hematopoietic Stem Cells; Humans; Immunophenotyping; Immunosuppressive Agents; Interleukin-3; Middle Aged; Multivariate Analysis; Pilot Projects; Stem Cell Factor

Immunoreactive serum erythropoietin (EPO) was measured in anemic and non-anemic patients with acquired non-severe aplastic anemia (AA; n = 22) and myelodysplastic syndromes (MDS; n = 31) receiving or not androgens to examine the effect of androgen therapy and anemia on EPO levels in these disorders. Soluble transferrin receptor (TfR) and absolute reticulocyte count (ARC) were also assayed in order to evaluate erythropoietic activity. AA and MDS patients were stratified for anemia and androgen treatment as follows: 12 untreated anemic patients; 17 anemic patients during androgen therapy; 14 non-anemic patients without any treatment (> 1 year); and 10 non-anemic patients on androgen therapy. Although EPO levels in non-anemic patients were significantly higher than in healthy controls (n = 29) no statistically significant differences in Hb and EPO values were found between non-anemic patients receiving or not androgen therapy. In the linear regression analysis between Hb and log EPO concentration, no statistically significant differences in the slopes between untreated and androgen-treated anemic groups nor between both groups and patients with iron deficiency anemia (n = 23) were observed. However, the y intercept (log EPO) of regression line was significantly higher in androgen-treated anemic patients than in the androgen therapy-free anemic group. Serum TfR levels were higher in treated than in untreated anemic patients, whereas ARC was not different between both groups. These data seemingly indicate that (1) androgens at pharmacological doses do not increase serum EPO levels in non-anemic AA and MDS patients, and (2) in patients with AA and MDS, androgen-driven EPO stimulation is appreciably enhanced by anemia.

Topics: Adult; Androgens; Anemia, Aplastic; Erythropoietin; Humans; Myelodysplastic Syndromes

Serum erythropoietin (sEpo) concentration is primarily related to the rate of renal production and, under the stimulus of hypoxia, increases exponentially as hemoglobin (Hb) decreases. Additional factors, however, appear to influence sEpo, and in this work, we performed studies to evaluate the role of the red blood cell precursor mass. We first compared the relationship of sEpo with Hb in patients with low versus high erythroid activity. The first group included 27 patients with erythroid aplasia or hypoplasia having serum transferrin receptor (sTfR) levels < 3 mg/L (erythroid activity < 0.6 times normal), while the second one included 28 patients with beta-thalassemia intermedia having sTfR levels > 10 mg/L (erythroid activity > 2 times normal). There was no difference between the two groups with respect to Hb (8.3 +/- 1.6 v 8.0 +/- 1.3 g/dL, P > .05), but sEpo levels were notably higher in patients with low erythroid activity (1,601 +/- 1,542 v 235 +/- 143 mU/mL, P < . 001). In fact, multivariate analysis of variance (ANOVA) showed that, at any given Hb level, sEpo was higher in patients with low erythroid activity (P < .0001). Twenty patients undergoing allogeneic or autologous bone marrow transplantation (BMT) were then investigated. A marked increase in sEpo was seen in all cases at the time of marrow aplasia, disproportionately high when compared with the small decrease in Hb level. Sequential studies were also performed in five patients with iron deficiency anemia undergoing intravenous (IV) iron therapy. Within 24 to 72 hours after starting iron treatment, marked decreases in sEpo (up to one log magnitude) were found before any change in Hb level. Similar observations were made in patients with megaloblastic anemia and in a case of pure red blood cell aplasia. These findings point to an inverse relationship between red blood cell precursor mass and sEpo: at any given Hb level, the higher the number of red blood cell precursors, the lower the sEpo concentration. The most likely explanation for this is that sEpo levels are regulated not only by the rate of renal production, but also by the rate of utilization by erythroid cells.

Topics: Anemia; Anemia, Aplastic; Anemia, Hypochromic; Anemia, Megaloblastic; Antineoplastic Combined Chemotherapy Protocols; beta-Thalassemia; Bone Marrow Transplantation; Erythrocyte Indices; Erythroid Precursor Cells; Erythropoiesis; Erythropoietin; Feedback; Folic Acid; Hodgkin Disease; Humans; Iron; Kidney; Receptors, Transferrin; Transplantation Conditioning; Vitamin B 12

In an attempt to evaluate the role of thrombopoietin (TPO) in the pathobiology of aplastic anaemia (AA), we have examined TPO levels in sera from 54 AA patients and 119 healthy controls. A total of 92 samples were collected from AA patients: 43 samples were harvested at diagnosis, 23 samples in the cytopenic period after treatment, and 26 samples when patients were in partial (n=10) or complete remission (n=16) following immunosuppressive treatment. TPO serum levels were assessed by a sandwich-antibody ELISA that utilized a polyclonal rabbit antiserum for both capture and signal. Serum samples from normal donors revealed a mean TPO level of 95.3 +/- 54.0 pg/ml (standard deviation). Mean TPO levels in AA sera collected at diagnosis and before onset of treatment were 2728 +/- 1074 pg/ml (P<0.001 compared to normal controls: mean platelet count at that time: 27x10(9)/l). TPO serum levels of AA patients in partial or complete remission after immunosuppressive treatment were significantly lower than TPO levels at diagnosis (P<0.001). However, despite normal platelet counts (mean 167x10(9)/l), TPO levels remained significantly elevated in complete remission (mean TPO 1009 +/- 590 pg/ml, P<0.001 compared to normal controls). There was a significant inverse correlation between serum TPO levels and platelet counts in AA patients who were not transfused for at least 2 weeks prior to sample collection (coefficient of correlation (r) = -0.70, P<0.0001). In summary, TPO levels were highly elevated in sera of patients with AA. Thus there is no evidence to suggest an impaired TPO response contributing to thrombocytopenia in AA. Thrombopoietin did not return to normal levels in remission, indicating a persisting haemopoietic defect in remission of AA. We hypothesize that elevated levels of TPO may be required to maintain normal or near normal platelet counts in remission of AA.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Female; Humans; Immunosuppression Therapy; Male; Middle Aged; Platelet Count; Thrombopoietin

Topics: Adolescent; Anemia, Aplastic; Delayed-Action Preparations; Erythropoietin; Ferrous Compounds; Glomerulonephritis; Humans; Kidney Failure, Chronic; Male; Parvoviridae Infections; Parvovirus B19, Human; Renal Dialysis; Uremia

Topics: Anemia, Aplastic; Bone Marrow; Erythropoietin; Follow-Up Studies; Hemoglobins; Humans; Recombinant Proteins; Treatment Outcome

We report the successful treatment of pancytopenia with G-CSF and high dose erythropoietin (Epo) in an elderly patient diagnosed with aplastic anemia (AA). Furthermore this effect is dose dependent for Epo in vivo. Detection of apoptosis by gel electrophoresis shows that high dose Epo protects bone marrow mononuclear cells from spontaneous apoptosis in vitro. These findings may explain some of the mechanisms of aplastic anemia.

Topics: Aged; Aged, 80 and over; Anemia, Aplastic; Cells, Cultured; Dose-Response Relationship, Drug; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Male; Remission Induction

Iron overload is a serious complication of many forms of anaemia, arising in part from mechanisms associated with compensatory increases in erythropoiesis. To investigate other mechanisms by which anaemia itself may perturb iron metabolism, without the confounding effects of compensatory erythropoiesis, we studied transgenic mice with a partially disabling insertion in the erythropoietin gene, which manifested as incomplete erythropoietin deficiency. Mice were studied aged 7-8 weeks. Haemoglobin concentrations were 6.6 +/- 0.8 g/dl in mice homozygous for the modified erythropoietin gene, 12.9 +/- 2.2 g/dl in heterozygous mice and 14.1 +/- 1.0 g/dl in controls. Homozygous mice showed significant hepatic iron loading (2-fold increase in liver non-haem iron, compared with heterozygous mice and normal controls, with iron staining principally in the periportal hepatocytes). Absorption studies using 59Fe showed increased uptake from the lumen of an in vivo isolated duodenal segment in homozygous mice, although at this point in time overall transfer of radioiron to the circulation and other tissues (mucosal transfer) was not different from controls. These observations demonstrate that anaemia can lead to hepatic iron loading even in the absence of increased erythropoiesis, and are consistent with the possibility that anaemic hypoxia can enhance mucosal iron uptake by the duodenal enterocyte.

Topics: Absorption; Anemia, Aplastic; Animals; Erythropoietin; Ferritins; Heterozygote; Homozygote; Iron; Liver; Mice; Mice, Transgenic

A 63-year-old woman was found to have thrombocythemia and was referred to our hospital for further evaluation in September 1990. Peripheral blood showed platelet 170.0 x 10(4)/microliter, WBC 14,900/microliter and Hb 9.8 g/dl. Bone marrow was hypercellular with increased megakaryocytes and normal karyotype. She was diagnosed as essential thrombocythemia (ET), and treated with 2 mg of busulfan daily for 3 months until her platelet count decreased to 33.1 x 10(4)/microliter. Busulfan was given again for 40 days (a total of 80 mg) in another hospital when the platelet count increased to 71.1 x 10(4)/microliter in September 1991. In December 1991, she was admitted to our hospital because of pancytopenia. Examination of blood revealed platelet 0.4 x 10(4)/microliter, WBC 1,800/microliter and Hb 7.0 g/dl with hypocellular marrow. A diagnosis of busulfan-induced severe bone marrow aplasia was made. We administered metenolone acetate 15 mg and G-CSF 300 micrograms daily. Blood transfusions were given frequently. However, no effect was observed during her hospitalization. After discharge, G-CSF 600 micrograms and erythropoietin 24,000 units were continued twice a week in combination with metanolone acetate. Pancytopenia gradually began to improve as of June 1992, and then trilineage recovery was achieved in March 1994 with platelet 13.3 x 10(4)/microliter, WBC 5,500/microliter and Hb 12.1 g/dl. The platelet count has been within the normal range for more than 2 years after recovery.

Topics: Anemia, Aplastic; Busulfan; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Methenolone; Middle Aged; Remission Induction; Thrombocythemia, Essential

Aplastic anaemia is both frequent and difficult to manage in patients with dyskeratosis congenita (DC). We recently treated a 23-year-old male for a year with granulocyte colony-stimulating factor (G-CSF) and erythropoietin (Ep), with an excellent neutrophil response, and a transient effect on haemoglobin levels. G-CSF alone or combined with other cytokines may provide at least a partial effect in pancytopenic patients with DC.

Topics: Adult; Anemia, Aplastic; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Hyperpigmentation; Male; Nail Diseases; Syndrome; X Chromosome

Topics: Aged; Anemia, Aplastic; Blood Cell Count; Erythropoietin; Humans; Male

A cytogenetically normal man with severe aplastic anemia was treated with granulocyte colonystimulating factor (G-CSF), erythropoietin (EPO), cyclosporin A, anti-thymocyte globulin, and interleukin-6 (IL-6), which resulted in a gradual improvement in his neutrophil count and hemoglobin level. After 2 years of the therapy, monosomy 7 was detected during cytogenetic analysis of his bone marrow, which evolved during a period of 5 months into acute myeloblastic leukemia. An in vitro proliferation assay of cytokine responses showed that leukemic blasts were sensitive only to G-CSF, and not to EPO or IL-6. Although allogeneic bone marrow transplantation from an HLA-matched unrelated donor was carried out in the non-remission stage, the patient died of systemic fungal infection on day 25, without any evidence of hematological engraftment. As long-term use of cytokines and immunomo-suppressants in patients with severe aplastic anemia may induce or hasten the onset of a malignant transformation, careful attention must be paid to clonal evolution. Due to the poor prognosis of secondary myelodysplasia and leukemia, allogeneic bone marrow transplantation for such patients must be carried out early in the course of the disease.

Topics: Adult; Anemia, Aplastic; Antilymphocyte Serum; Bone Marrow Transplantation; Chromosomes, Human, Pair 7; Cyclosporine; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-6; Leukemia, Myeloid, Acute; Male; Monosomy

Topics: Acquired Immunodeficiency Syndrome; Anemia, Aplastic; Antineoplastic Agents; Erythropoietin; Hematopoietic Cell Growth Factors; Humans; Leukemia, Myeloid, Acute; Myelodysplastic Syndromes; Neutropenia; Transplantation

A 67-year-old female was admitted with fatigue. Peripheral blood examination showed severe pancytopenia. Bone marrow biopsy revealed hypoplastic marrow. She was diagnosed as having aplastic anemia. Steroid pulse therapy was not effective. After treatment with erythropoietin (EPO) and granulocyte colony-stimulating factor (G-CSF), blasts which were positive for CD13, CD33, CD34 and HLA-DR and negative for myeloperoxidase appeared in the peripheral blood. At this time, bone marrow biopsy revealed myelofibrosis with increased blasts. Chromosome analysis showed 46XX, add (1) (p36), add (1) (q44), -2, -5, del (7) (q11), -12, +3mar. She died of pneumonia despite chemotherapy with etoposide. Administration of EPO and G-CSF may have led to the rapid development of leukemia and myelofibrosis.

Topics: Aged; Anemia, Aplastic; Erythropoietin; Fatal Outcome; Female; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid, Acute; Primary Myelofibrosis

Rejection after allogeneic BMT for aplastic anemia is a complication with a high risk of mortality. We describe a patient who, following a second episode of rejection after a second BMT entered a third durable remission subsequent to treatment with ALG, donor lymphocyte infusions, GM-CSF, and erythropoietin. Therapy was well tolerated. At 5 years after rejection treatment, his hematopoiesis is of complete donor origin as determined by analyses of short tandem repeats. Thus, donor lymphocyte infusions can be considered as a therapy option for marrow rejection after allogeneic BMT for aplastic anemia.

Topics: Adult; Anemia, Aplastic; Antilymphocyte Serum; Bone Marrow Transplantation; Cyclosporine; Erythropoietin; Graft Rejection; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunosuppression Therapy; Immunosuppressive Agents; Lymphocyte Transfusion; Male; Microsatellite Repeats; Polymerase Chain Reaction; Remission Induction; Transplantation, Homologous

The plasma erythropoietin (Epo) concentration was measured by radioimmunoassay in 75 patients with acquired aplastic anaemia. Overall there was an inverse relationship between the concentration of plasma Epo and the degree of anaemia. Plasma Epo concentrations were lower in patients who were sampled soon after diagnosis as opposed to those studied at later times. Although a decrease in the plasma Epo concentration was noted in all erythroid responders following immunosuppressive (IS) therapy or bone marrow transplantation (BMT), it was lower in patients undergoing BMT than in those receiving IS therapy for any given degree of anaemia.

Topics: Adolescent; Adult; Age Factors; Anemia, Aplastic; Biomarkers; Bone Marrow Transplantation; Child; Child, Preschool; Erythropoietin; Female; Hematocrit; Humans; Immunosuppressive Agents; Infant; Male; Middle Aged; Radioimmunoassay; Regression Analysis; Time Factors

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) and erythropoietin (rhEPO) were used to treat patients with aplastic anemia (AA). In terms of effects on erythrocyte recovery, the combined use of rhG-CSF and rhEPO showed a favorable response in 6 of 14 (42.9%) patients with moderate AA following 10 weeks treatment and in 3 of 14 (21.4%) patients thereafter. However, the response was poor in patients with severe AA (3/13). A favorable response in severe AA was observed in 1 of 13 (7.7%) patients following 10 weeks treatment and in 2 of 13 (15.4%) patients thereafter. The overall effect on erythrocytes was observed in 44.4% patients. A dose of 400 micrograms/m2 G-CSF was sufficient to cause an increase in neutrophil count and 100 IU/kg rhEPO appeared to be sufficient to cause an increase in erythrocyte count. In 6 of 27 (22.2%) patients, a trilineage response was observed. Interestingly, a delayed and long-lasting effect was obtained in 5 of 27 (18.5%) patients. These results suggest that rhG-CSF can synergize with rhEPO in erythrocyte response, especially in patients with moderate AA.

Topics: Adult; Anemia, Aplastic; Drug Therapy, Combination; Erythrocyte Count; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Leukocyte Count; Male; Middle Aged; Neutrophils; Recombinant Proteins

Correction of anaemia with recombinant human erythropoietin (rHu-EPO) improves the responsiveness of thyroidal and gonadal axes to exogenous TRH and GnRH in chronic haemodialysis patients, but the mechanisms remain to be fully elucidated. In order to assess the influences of endogenous erythropoietin on the hypothalamo-hypophyseal thyroidal and gonadal axes, we studied the response of polycythaemic and anaemic patients, in comparison to normal controls, after the administration of exogenous TRH and GnRH.. Exogenous hypothalamic factors, 500 micrograms TRH and 100 micrograms GnRH, were administered as a bolus and blood samples were obtained over a 3-hour period at 30, 60, 90, 120 and 180 minutes.. Five male polycythaemic patients (low EPO), three male anaemic patients (high EPO) and six normal age and sex matched controls were studied.. Blood samples were centrifuged immediately and the serum was stored at -20 degrees C until assayed for total T4, free T4, free T3, TSH, prolactin, growth hormone (TRH test), and FSH, LH, testosterone (GnRH test). Haematological parameters and biochemical profiles were also measured.. After TRH administration, both patient groups showed a normal TSH response; however, their free T4 and free T3 secretion was blunted compared to controls. Normal basal PRL levels increased in an exaggerated fashion, whereas, when compared to chronic renal failure patients on chronic haemodialysis, we did not see a paradoxical GH response or a basal GH increase in these 5 patients. GnRH administration in the study groups elicited a normalization in the LH response without an increase in testosterone levels; however, an exaggerated FSH response was found in the polycythaemic patients (low EPO).. Thus by investigating the role of low endogenous EPO levels in non-anaemic and anaemic patients with high EPO levels, our study suggests that the underlying chronic disease state may be the major contributing factor in the regulation of the hypothalamo-hypophyseal thyroid and gonadal axes, rather than the EPO levels.

Topics: Aged; Anemia, Aplastic; Erythropoietin; Follicle Stimulating Hormone; Gonadotropin-Releasing Hormone; Growth Hormone; Humans; Luteinizing Hormone; Male; Middle Aged; Pituitary Hormones, Anterior; Polycythemia Vera; Prolactin; Testosterone; Thyroid Hormones; Thyrotropin; Thyrotropin-Releasing Hormone

A 19-year-old man was diagnosed as having severe aplastic anemia and received high-dose methylprednisolone treatment without hematological response. A second course of high-dose mPSL treatment together with granulocyte colony-stimulating factor (G-CSF) plus erythropoietin (EPO) was then started and resulted in trilineage blood cell response. Ten months after the combination therapy thrombocytopenia developed and cytogenetic analysis showed 45,XX,-7, indicating an evolution to myelodysplastic syndrome (MDS) associated with monosomy 7.G-CSF and EPO treatment together with immunosuppression may be an effective therapy in SAA patients, but such a therapy may increase the risk of evolution to MDS.

Topics: Adult; Anemia, Aplastic; Chromosomes, Human, Pair 7; Drug Therapy, Combination; Erythropoietin; Glucocorticoids; Granulocyte Colony-Stimulating Factor; Humans; Male; Methylprednisolone; Monosomy; Myelodysplastic Syndromes

Two patients with severe aplastic anemia received combination therapy consisting of granulocyte colony-stimulating factor (G-CSF) and erythropoietin (EPO) by subcutaneous injection. During the first 8 weeks, we administered only G-CSF, but subsequently we administered EPO with G-CSF. Administration of G-CSF caused rapid increase in neutrophil counts in each case. In the first case, a 23-year-old woman, the first sign of improvement of anemia appeared in the 12th week but 50 more weeks elapsed before improvement of platelet count. In the second case, a 59-year-old woman, marked increase of reticulocytes appeared in the 32nd week, and the RBC count become normal in the 46th week. Minor improvement of platelet count was obtained in the 40th week. After 53 weeks of treatment, we stopped administration of G-CSF and EPO, and all of peripheral blood cell counts decreased. Therefore G-CSF and EPO were given to her again, and she showed the same response as first administration. It showed that she was dependent upon G-CSF and EPO. Finally she obtained normalization of all blood cell counts. In both cases, no serious side effects were observed. Combination therapy consisting of G-CSF and EPO may be beneficial for patients with aplastic anemia.

Topics: Adult; Anemia, Aplastic; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Middle Aged; Remission Induction

Topics: Anemia, Aplastic; Anemia, Sickle Cell; Erythropoietin; Female; gamma-Globulins; Humans; Infant

A 6-year-old girl with post-hepatitic severe aplastic anaemia was referred to our hospital. Haematological examination showed a haemoglobin level of 5.2 g/dl, platelet count of 8,000/microliters, and white blood cell count of 130/microliters with 17% neutrophils. She was treated with recombinant human granulocyte-colony stimulating factor (15 micrograms/kg/day i.v.) and cyclosporin A (6 mg/kg/day p.o.). The absolute neutrophil count gradually increased, but Hb and platelets were not improved. The intravenous administration of recombinant human erythropoietin (100 U/kg three times a week) was started, and the reticulocyte count reached 20,000/microliters on day 12. The platelets increased to 81,000/microliters after 16 months of combined administration of recombinant human granulocyte-colony stimulating factor, recombinant human erythropoietin and cyclosporin A. After 20 months of combined administration, the haematological results were: Hb, 13.1 g/dl; platelets 80,000/microliters; WBC, 9500/microliters with 40% neutrophils. After recombinant human granulocyte-colony stimulating factor treatment, the myeloid elements of the bone marrow and the number of granulocyte-macrophage colony forming units increased. Bone marrow erythropoiesis and erythroid colonies also increased after recombinant human erythropoietin administration. The clinical course suggested a beneficial effect of haemopoietic growth factors and cyclosporin A in post-hepatitic aplastic anaemia.

Topics: Anemia, Aplastic; Bone Marrow; Child; Cyclosporine; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Hematopoietic Stem Cells; Hemoglobins; Humans; Leukocyte Count; Platelet Count; Recombinant Proteins; Reticulocyte Count; Time Factors

Serum erythropoietin (EPO) and soluble transferrin receptor levels were serially measured in 74 patients with aplastic anaemia (AA). As control groups we investigated healthy controls (n = 24) and patients with iron-deficiency (n = 23) or haemolytic anaemia (n = 16). There was a significant negative correlation of log EPO on haematocrit both in AA patients and in the anaemic control group. However, for the same degree of anaemia, log EPO levels in AA were significantly higher than in iron-deficiency or haemolytic anaemia. EPO levels at diagnosis did not correlate with severity of aplastic anaemia, nor did they predict outcome after immunosuppression. During immunosuppressive treatment of AA with anti-thymocyte globulin and cyclosporine A, EPO levels were significantly lower compared with pre-treatment values without a corresponding change in haematocrit. This impaired EPO response to anaemia during immunosuppression might affect recovery of erythropoiesis. In AA patients, EPO levels declined with haemopoietic recovery. However, compared with normal controls, EPO levels in remission patients were still higher with respect to their haematocrit. Results of this study argue against the model of a simple feedback regulation of EPO via hypoxic anaemia. Our data support the hypothesis that cytokines and the erythropoietic progenitor pool are involved in the regulation of EPO production. The results illustrate that serial measurements of EPO along with therapeutic interventions are necessary to identify patients who might benefit from treatment with exogenous recombinant human EPO.

Topics: Adolescent; Adult; Age Factors; Aged; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Iron-Deficiency; Biomarkers; Erythropoietin; Female; Follow-Up Studies; Hematocrit; Humans; Immunosuppression Therapy; Male; Middle Aged; Receptors, Transferrin; Sex Factors; Treatment Outcome

A 3-year-old girl was admitted with a one-month history of a tendency to bleed to Jikei Kashiwa hospital in May, 1992. She developed pancytopenia as follows; hemoglobin: 8.6 g/dl, red blood cell: 316 x 10(4)/microliters, reticulocyte: 9,480/microliters, white blood cell: 2,500/microliters (neutrophil: 400/microliters) and platelet count: 2.7 x 10(4)/microliters. Her bone marrow was hypoplastic, but was so dysplastic in 3 cell-lines as to be diagnosed as hypoplastic refractory anemia. After two courses of methylprednisolone pulse therapy followed by oral prednisolone therapy which were not effective and were supplemented by blood transfusions, the treatment of 20mg/day oral Cepharanthin, a biscoclaurine alkaloid, and intravenous recombinant human erythropoietin (rhEPO) twice a week at dose of 6,000 U/week was initiated in January, 1993. About 3 months later she showed a steady rise in hemoglobin concentration (from 4.1 to 11.9 g/dl) and platelet count (from 4,000 to 39,000/microliters). Although the rhEPO was tapered and ceased in September, 1993, her hemoglobin concentration has ranged from 11.0 to 11.9 g/dl and her platelet count from 30,000 to 40,000/microliters by giving her Cepharanthin and low dose prednisolone.

Topics: Alkaloids; Anemia, Aplastic; Anemia, Refractory; Anti-Inflammatory Agents, Non-Steroidal; Benzylisoquinolines; Child, Preschool; Drug Synergism; Drug Therapy, Combination; Erythropoietin; Female; Humans; Recombinant Proteins

Topics: Anemia, Aplastic; Erythropoietin; Hepatitis, Viral, Human; Humans; Male; Middle Aged; Recombinant Proteins

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Arthritis, Rheumatoid; Biomarkers; Cyclosporine; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Hematocrit; Humans; Middle Aged

Physiological platelet synthesis is thought to require the humoral activities of meg-CSF and thrombopoietin, which respectively promote proliferation and maturation of megakaryocytic cells. A meg-CSF/thrombopoietin-like protein that is present in plasma of irradiated pigs has been purified and cloned. This protein binds to and activates the c-mpl protein, a member of the cytokine receptor superfamily. The isolated Mpl ligand shares homology with erythropoietin and stimulates both megakaryocytopoiesis and thrombopoiesis.

Topics: Amino Acid Sequence; Anemia, Aplastic; Animals; Base Sequence; Blood Platelets; Cell Differentiation; Cell Division; Cell Line; Cloning, Molecular; Erythropoietin; Humans; Ligands; Megakaryocytes; Mice; Molecular Sequence Data; Receptors, Immunologic; Recombinant Proteins; Sequence Homology, Amino Acid; Swine; Thrombopoietin; Tissue Distribution

Aplastic anaemia is characterized by multilineage bone marrow failure resulting in pancytopenia. We have successfully treated a young woman with severe aplastic anaemia (SAA) who was resistant to antilymphocyte globulin (ALG) and corticosteroids, with a combination therapy consisting of erythropoietin, cyclosporin A and granulocyte-colony stimulating factor (G-CSF). The patient received erythropoietin and CSA for a period of 10 months without success before G-CSF treatment was started. After 6 weeks of G-CSF therapy she responded with a sustained trilineage recovery. This suggests that immunosuppression together with haemopoietic growth factors may be an effective treatment in patients with SAA who are ALG resistant and cannot be treated by BMT.

Topics: Adolescent; Alkaline Phosphatase; Anemia, Aplastic; Antilymphocyte Serum; Cyclosporine; Drug Resistance; Drug Therapy, Combination; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Prednisolone

We examined the combined effects of stem cell factor (SCF), or interleukin-3 (IL-3) with erythropoietin on the development of haemopoietic progenitors in 19 patients with aplastic anaemia (AA) and eight normal controls by using an in vitro clonal assay. SCF significantly enhanced the growth of total erythroid colonies (erythroid bursts, mixed colonies) in 11 patients and all normal controls, whereas IL-3 did so in only three patients. The number of SCF- or IL-3-dependent erythroid colonies was substantially lower in AA patients than in the controls. Comparison of the capacity of SCF and IL-3 to increase total erythroid colony growth indicated that half of the AA patients responded more strongly to SCF than the normal controls, while few patients responded in such a manner to IL-3. These findings suggest that SCF in vivo will have a more dramatic effect than IL-3 in improving anaemia in patients with AA.

Topics: Adolescent; Adult; Anemia, Aplastic; Cell Division; Child; Child, Preschool; Colony-Forming Units Assay; Dose-Response Relationship, Drug; Erythroid Precursor Cells; Erythropoietin; Female; Hematopoietic Cell Growth Factors; Humans; Interleukin-3; Male; Stem Cell Factor

The white-spotting (Ws) locus of rats represents a 12-base deletion of the c-kit receptor tyrosine kinase. Homozygous Ws/Ws rats are deficient in melanocytes, mast cells, and erythrocytes. Although mice possessing two mutant alleles at the c-kit (W) locus, such as mice of W/Wv genotype, show severe anemia even in adult age, the anemia of Ws/Ws rats remarkably ameliorated with age. We investigated the mechanism of the age-dependent amelioration. Bone marrow cells of Ws/Ws rats did not form macroscopic colonies in the spleen of irradiated rats, and the concentration of burst-forming unit-erythroid in the marrow of Ws/Ws rats was comparable with that of +/+ rats. Therefore, the increase in morphologically identifiable erythroid precursors in the marrow of Ws/Ws rats was attributed to the increased concentration of colony-forming unit-erythroid (CFU-E). Furthermore, the increase in CFU-E appeared to result from the increased concentration of erythropoietin (EPO). Because injections of relatively low doses of EPO cured the slight anemia that remained in adult Ws/Ws rats, CFU-E and/or its immediate precursors of Ws/Ws rats appeared to be more sensitive to EPO than those of W/Wv mice, in which a huge dose of EPO was necessary to cure the anemia.

Topics: Age Factors; Anemia, Aplastic; Animals; Erythropoietin; Gene Deletion; Genotype; Hematocrit; Mice; Mice, Inbred C57BL; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-kit; Rats; Rats, Mutant Strains; Receptor Protein-Tyrosine Kinases; Receptors, Colony-Stimulating Factor; Recombinant Proteins

Topics: Anemia, Aplastic; Child; Erythropoietin; Humans; Male; Recombinant Proteins; Splenic Infarction

An immunologic crossreactant of erythropoietin seemed to develop and persist in serum samples from a patient during treatment and remission of idiopathic aplastic anemia. It had a steeper slope to radioimmunoassay log-dose response lines and a larger molecular size than erythropoietin. On fractionation of serum, the apparent crossreactant was bound by staphylococcal Protein A at pH 7.5 and recovered by elution from it at pH 3.0. Adsorption of serum from the patient, and from one of two similarly affected children, with rabbit IgG linked to agarose appeared to remove completely the apparent crossreactant. These treated sera gave radioimmunoassay log-dose response lines essentially parallel to that given by the International Reference Preparation (IRP) for erythropoietin and estimates of immunoreactive erythropoietin appropriate to the normal hemoglobin concentrations. The apparent crossreactant of erythropoietin is thus accounted for by heterophilic antibodies to rabbit IgG. These developed in the patient following treatment with rabbit antilymphocyte globulin but seem to have arisen spontaneously in the children. Thus iatrogenic and idiopathic antibodies to rabbit IgG interfered in a radioimmunoassay for erythropoietin in serum through their ability to react with the radioimmunoassay anti-erythropoietin antiserum raised in rabbits.

Topics: Adult; Anemia, Aplastic; Animals; Antibodies, Anti-Idiotypic; Antibodies, Heterophile; Erythropoietin; False Positive Reactions; Humans; Immunoglobulin G; Immunosorbent Techniques; Male; Quality Control; Rabbits; Radioimmunoassay

Because GM-CSF possesses burst-promoting activity (BPA) and megakaryocyte colony-stimulating activity (Meg-CSF) as well as stimulating activity on granulocyte-macrophage progenitors, and erythropoietin (Epo) has thrombopoietin-like activity, the combination therapy of GM-CSF and Epo seems to be more effective for stimulating erythropoiesis and thrombocytopoiesis in patients with pancytopenia. For this reason, the combination therapy of recombinant human GM-CSF (rhGM-CSF) and rhEpo was performed in two patients with refractory anemia (RA) and aplastic anemia (AA). Epo-unresponsive anemia was remarkably improved by adding rhGM-CSF to Epo and the effect lasted for 1 1/2 months in a patient with RA, but severe anemia occurred again immediately after the discontinuation of Epo. The neutralizing antibodies against GM-CSF were not demonstrated at the phase when anemia re-progressed in this patient. In a patient with AA, anemia and thrombocytopenia, which were refractory to previous administration of rhGM-CSF, responded to the combined administration of GM-CSF and Epo. Although the effects were maintained for 3 1/2 months, the anemia and thrombocytopenia became worse again after the administration of rhGM-CSF was changed from daily to every other day. These findings suggest the usefulness of combination therapy of GM-CSF and Epo for patients with pancytopenia.

Topics: Adult; Anemia, Aplastic; Anemia, Refractory; Drug Therapy, Combination; Erythropoietin; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Male; Recombinant Proteins

The regulation of megakaryocytopoiesis and platelet production has not yet been clearly elucidated. Several cytokines have been shown to be capable of producing megakaryocyte colonies from bone marrow [i.e. Interleukin (IL)-3, granulocyte-macrophage (GM)-colony-stimulating factor (CSF), erythropoietin (Epo)]. In addition, other activities have been reported to stimulate megakaryocyte precursors, yet a megakaryocyte-CSF (Meg-CSF) has not been purified to homogeneity and IL-3, GM-CSF and/or Epo often contaminate purification attempts which could account for the activities. A Meg-CSF has been isolated from the urine of patients with aplastic anaemia and purified by sequential ultrafiltration, cation exchange, G-50 chromatography, preparative PAGE, chromatofocusing and cation exchange HPLC. The activity of this material is 2-4 x 10(4) CFU-Meg/mg as measured in a murine marrow, serum-containing assay. This activity also stimulates CFU-Meg in the absence of adherent accessory cells and in serum-free cultures, indicative of the direct stimulation on CFU-Meg. Immunoassays, colony forming assays, and proliferation assays demonstrate that purified Meg-CSF has no GM-CSF, IL-3, M-CSF, G-CSF or IL-1 alpha, -3, -6, -9 and -11. In confirmation of these results, neutralizing antibody to IL-6 also did not abrogate Meg-CSF activity. Therefore the previously-reported megakaryocyte colony-stimulating activity in purified aplastic anaemia patient urine is due to a unique cytokine: Meg-CSF.

Topics: Anemia, Aplastic; Animals; Colony-Forming Units Assay; Colony-Stimulating Factors; Erythropoietin; GPI-Linked Proteins; Humans; Interleukins; Megakaryocytes; Membrane Glycoproteins; Mesothelin; Mice; Proteins

Blood erythropoietin (EPO) concentration was measured by radioimmunoassay in 513 patients with various diseases. Untreated polycythemia vera showed lower EPO concentration than normal. Aplastic anemia (AA) revealed the highest EPO level among all anemic diseases in relation to hematocrit value. EPO level of AA patients who underwent bone marrow transplantation was as low as normal subjects even when the anemia has not fully recovered. Paroxysmal nocturnal hemoglobinuria (PNH) showed unusually high EPO concentration among hemolytic anemias. In normal subjects, blood EPO concentration showed a diurnal rhythm that was higher at night than during the daytime. These findings suggest the diagnostic usefulness of measurement of EPO in blood diseases.

Topics: Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Bone Marrow Transplantation; Circadian Rhythm; Erythropoietin; Female; Hemoglobinuria, Paroxysmal; Humans; Male; Middle Aged; Polycythemia Vera; Radioimmunoassay; Reference Values

Less is known about the mucin-type sugar chains attached to human erythropoietin as compared with N-linked sugar chains which structures and function have been well studied. In this study, we purified urinary human erythropoietin from three independent groups of aplastic anemia patients, and analyzed the structures of mucin-type sugar chains as well as that obtained from recombinant human erythropoietin produced by Chinese hamster ovary cells. Unlike the N-linked sugar chains, the mucin-type sugar chains are totally different between the urinary and the recombinant erythropoietins. All of the three independent samples of urinary erythropoietin contained oligosaccharides with the structures of +/- Neu5Ac alpha 2-->6GalNAc, while recombinant human erythropoietin contained those with the structures of Neu5Ac alpha 2-->3Gal beta 1-->3(Neu5Ac alpha 2-->6)GalNAc, Neu5Ac alpha 2-->3Gal beta 1-->3GalNAc, and Gal beta 1-->3(Neu5Ac alpha 2-->6)GalNAc.

Topics: Anemia, Aplastic; Animals; Carbohydrate Sequence; CHO Cells; Cricetinae; Erythropoietin; Humans; Molecular Sequence Data; Mucins; Oligosaccharides; Recombinant Proteins; Urine

Recombinant human erythropoietin (rhEpo) was administered to 14 patients with myelodysplastic syndrome (MDS) and seven patients with aplastic anemia (AA). In 19 patients, doses of 6000 units were given intravenously three times a week (t.i.w.) with the dose being doubled up to 24,000 units every 8 weeks until a response was obtained. RhEpo was given subcutaneously in two patients. Seven patients, four with MDS and three with AA, showed a significant response with an increase of hemoglobin concentration during therapy. The response occurred at doses of 12,000 units in five and 24,000 units in two patients. Responding patients with both MDS and AA had a relatively low serum Epo (s-Epo) level prior to Epo therapy. MDS responders had either refractory anemia (RA) or RA with ring sideroblasts (RARS), while two of the Epo responders in AA had a severe form of the disease. However, since some of the Epo responders had a high initial s-Epo concentration, a high s-Epo level does not preclude the use of rhEpo. Serial determination of s-Epo levels showed a progressive decline in six of the seven responders even when they were on rhEpo therapy, while the s-Epo levels remained elevated or further increased with time in most nonresponders. RhEpo was well tolerated by all patients. The results suggest that rhEpo is a safe and effective treatment for a certain proportion of patients with MDS and AA. Moreover, serial determination of s-Epo during therapy may be useful in monitoring and predicting the therapeutic effect of rhEpo.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Refractory; Anemia, Refractory, with Excess of Blasts; Erythropoietin; Female; Humans; Male; Middle Aged; Prognosis; Recombinant Proteins

A 32 year-old female admitted to our hospital with pancytopenia. The hematological data on admission were: RBC: 247 x 10(4)/microliters, Hb: 8.8 g/dl, Plts: 13,000/microliters, WBC: 2,500/microliters. Bone marrow aspirate and biopsied specimen showed marked hypocellularity without infiltration of abnormal cells. A diagnosis of aplastic anemia was made. Neither high-dose methyl-prednisolone pulse therapy nor anti-lymphocyte globulin were effective. With combination of oxymetholone (30 mg/day), recombinant erythropoietin (rHuEpo; 12,000 U/day, three times a week) and recombinant granulocyte-colony simulating factor (rHuG-CSF; 33 micrograms/day) for 3 months, remarkable improvements of hematological data were obtained. Her hemoglobin level reached 11.4 g/dl, and platelets count 49,000/microliters. However, 4 weeks after the withdrawal of erythropoietin and G-CSF administrations, her platelet count fell to 12,000/microliters. It was suggested that combination therapy with erythropoietin and G-CSF were effective for aplastic anemia.

Topics: Adult; Anemia, Aplastic; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Recombinant Proteins

Topics: Adolescent; Adult; Anemia, Aplastic; Erythrocyte Count; Erythropoietin; Granulocyte Colony-Stimulating Factor; Humans; Leukocyte Count; Male; Platelet Count; Recombinant Proteins; Reticulocytes

Aplastic anemia (AA) is a rare human bone marrow disorder of unknown etiology manifested by a strongly impaired growth of hematopoietic precursors. In this study, we examined the ability of recombinant human stem cell factor (SCF) to stimulate proliferation in vitro of bone marrow cells from 15 AA patients. All patients had been previously treated with antilymphocyte globulin (ALG). SCF, in combination with erythropoietin (Epo), interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF), increased the number of hematopoietic colonies formed in a semisolid medium by AA marrows. Maximal colony numbers reached 30% of the numbers observed with normal bone marrow cells. Proliferation of AA cells cultured in a liquid medium containing SCF together with Epo, IL-3, GM-CSF, and G-CSF approached 70% of the control level, as measured by 3H-thymidine incorporation. The effect of the combination of SCF with the other growth factors was more than 10 times stronger than that of the growth factors alone. The most marked effect of SCF was on the generation of erythroid colonies by precursor cells. The results demonstrate synergism between CSF and other hematopoietic growth factors, resulting in the most efficient stimulation of the in vitro growth of AA bone marrow cells described to date. Use of SCF, either alone or in combination with other factors, may be of potential value in treatment of AA.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Bone Marrow; Cell Division; Cells, Cultured; Child; Colony-Forming Units Assay; DNA; Erythroid Precursor Cells; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Hematopoietic Stem Cells; Humans; Interleukin-3; Male; Middle Aged; Recombinant Proteins; Stem Cell Factor

Effects of recombinant human erythropoietin (rhEpo) and the combination of recombinant human interleukin-3 (rhIL-3) or recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) with rhEpo on erythroid colony formation were examined in vitro in 13 patients with aplastic anemia and 16 with myelodysplastic syndromes (MDS). The methylcellulose cultures of marrow cells from normals and the patients yielded no erythroid colonies in the absence of rhEpo. In normals, CFU-E and BFU-E colony formation was significantly increased by adding either rhIL-3 or rhGM-CSF with rhEpo, compared with rhEpo alone, and rhIL-3 was more potent than rhGM-CSF to form colony-forming units and burst-forming units of erythroid (CFU-E) (BFU-E) colonies. By adding rhIL-3 with rhEpo, CFU-E colony formation was increased in half of patients with RA, compared with rhEpo alone, and by rhGM-CSF, in one third. Approximately one third or one fourth of the patients with MDS showed increased BFU-E colonies when rhIL-3 or rhGM-CSF were added to rhEpo. Cultures containing rhIL-3 or rhGM-CSF with rhEpo yielded larger numbers of BFU-E colonies in half of the patients with nonsevere aplastic anemia than those containing rhEpo alone. These observations suggest that the combination of these growth factors, especially rhIL-3 with rhEpo, is applicable to the treatment of anemia in some patients with aplastic anemia and MDS.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Child; Child, Preschool; Colony-Forming Units Assay; Drug Therapy, Combination; Erythropoiesis; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Substances; Humans; In Vitro Techniques; Interleukin-3; Male; Middle Aged; Myelodysplastic Syndromes; Prospective Studies; Recombinant Proteins

In patients with idiopathic aplastic anaemia (n = 34) and Fanconi's anaemia (n = 8), sampled once or on several occasions, serum erythropoietin (Epo) increased with increasing severity of anaemia with apparently similar rates of increase in each group. However, after adjustment for Hb, log Epo values for the Fanconi's anaemics tended to be greater than those for the idiopathic aplastic anaemics (P < 0.01). Erythropoietin concentrations in serum samples from patients with Fanconi's and idiopathic aplastic anaemias tended to be greater than in samples from patients with anaemias from protein energy malnutrition, myelodysplasia and iron deficiency. The results suggest that there is no deficiency of erythropoietin in Fanconi's and idiopathic aplastic anaemias and that if exogenous erythropoietin is of any benefit it would need to be administered in doses large enough to induce a significant increase in log Epo. Results of the study illustrate the need to take account of the assumptions which underlie interpretation of the statistical analysis. Use of erythropoietin values in place of log Epo gives misleading conclusions demonstrable as invalid as the conditions for normality of distribution of the data and homogeneity of variances were not satisfied.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Blood Component Transfusion; Child; Child, Preschool; Erythropoietin; Fanconi Anemia; Female; Hemoglobins; Humans; Male; Middle Aged; Oxymetholone; Sex Factors

To assess the effects of decreased erythrocyte production on the levels of serum erythropoietin in children, we measured simultaneous hemoglobin concentrations and erythropoietin in 18 children with iron deficiency anemia, 17 children with transient erythroblastopenia of childhood (TEC), and 7 children with aplastic anemia. In all but two patients (one with TEC; one with aplastic anemia), erythropoietin was measured at diagnosis, before institution of specific therapy for the anemia. There was a statistically significant inverse linear correlation between log10 erythropoietin and hemoglobin values for all patient groups (r = 0.904 to 0.912; p less than 0.005). A comparison of the slopes of the regressions for each patient group by analysis of covariance revealed a significantly steeper slope for the iron deficiency group (-0.553) versus the TEC (-0.287) and aplastic anemia (-0.256) groups (p = 0.0001). The difference in erythropoietin levels appeared greatest in patients whose presenting hemoglobin level was greater than 5 gm/dl. Decline in serum erythropoietin levels was more precipitous in the less severely anemic patients with iron deficiency anemia than in the patients with TEC or aplastic anemia. These data reveal quantitative and qualitative differences in the relationship between serum erythropoietin and hemoglobin levels when children with severe iron deficiency anemia versus those with TEC or aplastic anemia are considered, even though all three conditions are characterized by hypoproliferation of erythrocytes.

Topics: Analysis of Variance; Anemia, Aplastic; Anemia, Hypochromic; Child; Child, Preschool; Erythropoietin; Hemoglobins; Humans; Infant; Red-Cell Aplasia, Pure

Two children affected by severe aplastic anaemia (SAA) underwent allogeneic bone marrow transplantation (BMT) using partially matched family donors. In both cases there was a successful engraftment of donor haemopoietic stem cells. However, after an initial erythropoietic recovery, 5 months following BMT both children became severely anaemic. Although multiple factors were responsible for anaemia, in both cases there was a markedly impaired erythropoietin response to anaemia, as indicated by the inappropriately low levels of serum erythropoietin (EPO). Treatment with recombinant human erythropoietin (rHuEPO) induced a sustained erythropoietic response with complete correction of anaemia. This pilot study suggests that rHuEPO can be effective in correcting long-lasting anaemia after marrow transplantation, characterized by inadequate erythropoietin production.

Topics: Anemia; Anemia, Aplastic; Bone Marrow Transplantation; Child, Preschool; Erythropoietin; Female; Hemoglobins; Humans; Pilot Projects; Recombinant Proteins

We investigated the interactions between human erythropoietin (hEpo) and serum factor(s) on murine megakaryocyte (MK) colony formation. Serum-free cultures supported the growth of a large number of murine MK colonies in the presence of murine interleukin-3 (mIL-3). The addition of fetal calf serum (FCS) to mIL-3-containing cultures resulted in only a minimal increase in the number of murine MK colonies. In contrast, hEpo alone had no murine MK colony-stimulating activities in serum-free cultures. hEpo required the presence of FCS, murine serum, or human serum in cultures to promote murine MK colony growth and synergized with these sera to stimulate murine MK colony formation. Furthermore, sera from patients with aplastic anemia showed higher synergistic activities with hEpo than sera from hematologically normal persons (normal human serum). When normal human serum was fractionated by gel-filtration chromatography, two peaks with the synergistic activity were observed in the eluent. However, serum did not show any synergistic effects with hEpo on the growth of murine GM colonies or murine colony-forming unit-erythroid-derived colonies. Although human serum synergized with hEpo to stimulate murine MK colony formation, human cytokines such as IL-3, IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-CSF (G-CSF) failed to induce murine MK colony formation in Epo-containing cultures. In cultures containing human IL-1 alpha + human IL-6 + hEpo as well as in cultures containing hEpo, human IL-3 and human GM-CSF failed to show stimulatory effects on murine MK colony formation. Moreover, the synergistic activity of human serum with hEpo could not be neutralized by antibodies such as antihuman IL-1 alpha, antihuman IL-3, antihuman IL-4, antihuman IL-6, antihuman G-CSF, and antihuman GM-CSF. Our data show that serum contains a growth factor(s) that synergizes with Epo to stimulate the proliferation and differentiation of MK precursors, and strongly suggest that this factor(s) is an unique growth factor(s) that is distinct from IL-1 alpha, IL-3, IL-4, IL-6, G-CSF, and GM-CSF.

Topics: Anemia, Aplastic; Animals; Erythropoietin; Female; Growth Substances; Hematopoiesis; Humans; In Vitro Techniques; Interleukin-3; Megakaryocytes; Mice; Recombinant Proteins

PURPOSE AND RATIONALE: There has been no previously published experience with granulocyte-macrophage colony-stimulating factor (GM-CSF) at doses less than 15 micrograms/m2/d in patients with aplastic anemia, and most observations have been made at doses of 100 to 500 micrograms/m2/d (2.5 to 12.5 micrograms/kg/d). The benefits of using considerably lower doses, if effective, should include a decrease in cost and in side effects. We have therefore used very low doses of GM-CSF to treat a group of patients with aplastic anemia. Additionally, since severe anemia is often a problem in these patients, we recently started administering erythropoietin along with the GM-CSF. Herein we report the results of very-low-dose GM-CSF therapy in patients with aplastic anemia and our preliminary findings in those individuals who received combination therapy.. We administered recombinant human GM-CSF subcutaneously at doses of 5 to 20 micrograms/m2/d ("very-low-dose GM-CSF") to 12 patients with aplastic anemia. In addition, a 13th patient received erythropoietin together with the GM-CSF regimen, and three of the 12 individuals who initially received 1 or more months of GM-CSF alone were later also given erythropoietin (4,000 U/d subcutaneously).. In five of 12 patients (42%) treated with very-low-dose GM-CSF, an increase in neutrophil counts (2.0- to 6.7-fold) was noted, and one of these subjects attained a bilineage response (neutrophil counts, 0.3 to 1.75 x 10(9)/L; platelet counts, 8 to 169 x 10(9)/L). Moreover, a sixth patient showed a rise in platelet counts (19 to 80 x 10(9)/L) without a concomitant increase in neutrophils. Constitutional side effects were minimal. Combining erythropoietin and very-low-dose GM-CSF produced a bilineage response (neutrophils, 1.0 to 3.0 x 10(9)/L; hemoglobin, 7.4 to 9.4 g/dL) in the one patient who received erythropoietin together with the GM-CSF from the time that GM-CSF was initiated. In one of the other patients who were given combination therapy, the addition of erythropoietin appeared to enhance the response; this patient demonstrated a neutrophil response to GM-CSF alone and a trilineage response (neutrophils, 0.8 to 3.75 x 10(9)/L; hemoglobin, 7.0 to 13.1 g/dL; and platelets, 10 to 34 x 10(9)/L) to the combination. No toxicity was associated with the addition of erythropoietin.. Our observations suggest that (1) very low doses of GM-CSF (5 to 20 micrograms/m2/d subcutaneously) may be used initially in neutropenic patients with aplastic anemia, and the dose subsequently increased only in patients who do not respond; and (2) the administration of erythropoietin together with GM-CSF is well tolerated, can augment responsiveness in some patients, and deserves further study.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Bacterial Infections; Bone Marrow; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hemoglobins; Humans; Injections, Subcutaneous; Leukocyte Count; Lymphocytes; Male; Middle Aged; Neutropenia; Neutrophils; Platelet Count; Recombinant Proteins; Thrombocytopenia; Time Factors

The serum concentration of erythropoietin in 79 cases with various blood diseases, uremia, chronic obstructive pulmonary disease etc was determined. At comparable degrees of anemia, patients with myelodysplastic syndrome and aplastic anemia had the highest levels of erythropoietin in our study. The high level of erythropoietin titer in patients with aplastic anemia should be taken as the nom for renal synthesis and release of this hormone. The erythropoietin level in patients with uremic anemia was lower than the level in patients with anemia of other causes but still higher than that of the normal controls. Patients suffering from polycystic kidney disease with or without uremia had a high level of erythropoietin due to local hypoxia of remnant kidney tissue resulting from the pressure of cystic formation. Different methods are used to determine the erythropoietin level, which varies with the stage and etiology of the diseases. There are other stimulating or inhibitory factors of erythropoiesis when the assay is processed. Transfusion and administration of certain drugs also influence the growth of erythroid cells, thus the serum titers of erythropoietin differed markedly between patients at comparable hemoglobin concentration.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Child; Erythropoietin; Female; Hemoglobins; Humans; Lung Diseases, Obstructive; Male; Middle Aged; Uremia

Serial serum erythropoietin levels were measured in 10 consecutive patients undergoing allogeneic bone marrow transplantation. Observed erythropoietin levels are compared with those predicted from a large control population of anaemic patients not receiving chemotherapy. There was an initial acute rise in serum erythropoietin, peaking between days 1 and 4 after marrow transfusion, which was unrelated to changes in haemoglobin concentration. Patients maintained serum erythropoietin concentrations at around twice the predicted level for the first 2 weeks following transplantation, with a gradual fall into the expected range by wk 3. Erythropoietin levels did not change with episodes of bacterial infection or acute graft-versus-host disease. A patient with severe aplastic anaemia had initial successful engraftment with normalisation of erythropoietin levels, but showed a marked and amplified rise in erythropoietin 2 wk before falling peripheral blood counts indicated failure of the bone marrow graft.

Topics: Adolescent; Adult; Anemia, Aplastic; Bone Marrow; Bone Marrow Transplantation; Child; Cyclosporins; Erythropoietin; Female; Hemoglobins; Humans; Infant; Leukemia; Male; Middle Aged; Radiography; Radioimmunoassay

A 69-year-old female was admitted for pancytopenia. The hematological examination showed leukocytes 1,800/microliters, hemoglobin 5.3 g/dl and platelets 9.6 x 10(4)/microliters. A bone marrow aspiration revealed hypoplasia, but no abnormal cells. Serum erythropoietin titer was 5,100 mU/ml. Diagnosis of aplastic anemia was made. She received 400 ml of blood transfusion twice, and was then treated with recombinant human erythropoietin (rHuEPO) (12,000 U/day) three times a week for eight weeks. Hemoglobin level gradually increased to the level of 12.0 g/dl. This case suggests that there are some cases of aplastic anemia which can respond to treatment with rHuEPO.

Topics: Aged; Anemia, Aplastic; Drug Administration Schedule; Erythropoietin; Female; Humans; Recombinant Proteins

Burst-promoting activity (BPA) was measured in the sera from 31 children with aplastic anaemia (AA). BPA levels were elevated in most of the children with AA (65.2%), the mean value (137.7 +/- 18.4%) being significantly higher than that in normal children (69.6 +/- 9.4%), in children in the recovery period and in children with non-aplastic anaemia. There was a negative relationship between the BPA level in children with AA and the peripheral haemoglobin concentration. The BPA level was higher in those whose duration of illness was shorter than 1 year. In three cases of AA caused by chloramphenicol and benzene hexachloride and one case of congenital pure red cell AA, the BPA level was not elevated. Eleven patients received fetal liver cell suspensions intravenously (FLI). After FLI the BPA level in their sera was significantly reduced. According to these results, it appears that the elevation of BPA level is a special phenomenon of AA. The measurement of BPA in serum is helpful for differentiation between AA and other kinds of anaemia. The elevation of the BPA level in serum is a biological compensation for the haematopoietic disorder, and the measurement of BPA in the serum of patients with AA may be helpful in evaluating the haematopoietic condition.

Topics: Adolescent; Anemia, Aplastic; Child; Child, Preschool; Chloramphenicol; Dose-Response Relationship, Drug; Erythropoietin; Female; Hematopoiesis; Hemoglobins; Hexachlorocyclohexane; Humans; Immunosuppressive Agents; Injections, Intravenous; Interleukin-3; Liver; Male; Phytohemagglutinins

Human urinary erythropoietin has been purified to homogeneity. The seven-step procedure yielded a preparation with a potency of 225,000 U/mg protein. SDS-polyacrylamide gel electrophoretic analysis of the purified hormone revealed a single protein band with a molecular weight of about 35,000 that migrated with the biological activity. As to its stability, the purified hormone retained its activity in the presence of 0.001% Tween 20.

Topics: Anemia, Aplastic; Animals; Biological Assay; Chromatography, High Pressure Liquid; Drug Stability; Electrophoresis, Polyacrylamide Gel; Erythropoietin; Humans; Hydrogen-Ion Concentration; Molecular Weight; Pancytopenia; Rats; Rats, Inbred Strains

Eight patients with myelodysplastic syndromes (MDS) and four patients with aplastic anemia (AA) were treated with recombinant erythropoietin (rEpo) to investigate its effect on the anemia of these patients. rEpo was administered by i.v. injection three times a week for at least four weeks. The doses were 3,000, 6,000, or 12,000 U/day. Despite an elevated "endogenous" Epo level, a greater than 1.5 g/dl increase in hemoglobin (Hb) concentration was observed in one patient with refractory anemia (RA), one patient with refractory anemia with excess of blasts (RAEB), and one patient with AA. A greater than 50% decrease in red cell transfusion requirement was observed in one patient with RA and one patient with AA. One RA patient and one AA patient have received rEpo as maintenance therapy for more than 64 and 100 weeks, respectively. They no longer need red cell transfusions and have had a normal Hb concentration and normal ferrokinetics. No side effect was seen. These results indicate that rEpo may benefit some patients with MDS and AA who are dependent on red cell transfusions while further studies will be necessary to elucidate the mechanism by which rEpo stimulates erythropoiesis and improves anemia in patients with these diseases.

Topics: Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Erythropoietin; Female; Humans; Iron; Male; Middle Aged; Myelodysplastic Syndromes; Recombinant Proteins

Congenital hypoplastic (Diamond-Blackfan) anemia is a rare macrocytic anemia, generally presenting during infancy or childhood. The condition usually occurs sporadically or in a pattern consistent with autosomal recessive inheritance, although autosomal dominant transmission has been proposed in some kindreds. We report the largest known kindred of congenital hypoplastic anemia, with at least 7 affected individuals over 3 generations, and propose that studies of this kindred may be useful for identifying the mechanism by which their genetic abnormality results in congenital hypoplastic anemia. Erythropoietic investigations on relatives show no inhibitors of erythropoiesis in serum, T-lymphocytes, or macrophages. Their erythroid progenitor cells (CFU-E and BFU-E) were generally quantitatively normal, and were capable of rapid proliferation, as judged by cell-cycle shortening. However, their erythroid progenitors displayed a relative insensitivity to recombinant erythropoietin, and produced relatively few normoblasts per erythroid progenitor cell. We propose that these and subsequent studies may be helpful in selecting candidate genes responsible for the molecular defect in this kindred.

Topics: Adolescent; Adult; Anemia, Aplastic; Anemia, Macrocytic; Child, Preschool; Erythroid Precursor Cells; Erythropoiesis; Erythropoietin; Female; Humans; Male; Pedigree; Recombinant Proteins

Physicochemical and biological properties of recombinant human erythropoietin (rhEPO) were compared with human urinary erythropoietin (uEPO). uEPO and rhEPO were purified to apparent homogeneity from the urine of patients with aplastic anemia and from the conditioned medium of Chinese hamster ovary (CHO) cells transfected with a cDNA clone for human EPO, respectively. The microheterogeneous nature of both factors, observed on isoelectric focusing, is derived from the difference of the number of terminal sialic acid residues bound to the carbohydrate chains of the EPO molecule. The primary structure of rhEPO, consisting of 165 amino acid residues, was determined, and the C-terminal arginine predicted from the cDNA sequence was confirmed to be missing, as described previously (Recny et al. (1987) J. Biol. Chem. 262, 17156). Three N-glycosylation and one O-glycosylation sites of both factors were determined as Asn24, Asn38, and Asn83 and Ser126, respectively. Two disulfide linkages are located between Cys7 and Cys161, and between Cys29 and Cys33, in both EPOs. Hematogenic potencies of rhEPO and uEPO compared in normal and in partially nephrectomized rats were approximately the same. Both factors also stimulated the colony formation of CFU-E, BFU-E, and CFU-Meg in a dose-dependent manner. From these results, it is concluded that rhEPO produced in CHO cells transfected with cDNA clone for human EPO is indistinguishable from uEPO physicochemically and biologically, and is valuable for further research and for clinical use.

Topics: Amino Acid Sequence; Amino Acids; Anemia, Aplastic; Animals; Cells, Cultured; Chemical Phenomena; Chemistry, Physical; Cloning, Molecular; Disulfides; Electrophoresis, Polyacrylamide Gel; Erythropoietin; Hematopoiesis; Humans; Iron; Iron Radioisotopes; Mice; Molecular Sequence Data; Peptide Fragments; Rats; Recombinant Proteins; Sulfhydryl Compounds

The question as to whether the serum concentration of erythropoietin is relatively high for the degree of anemia in patients with erythrocytic hypoplasia has regained interest, since recombinant human-like erythropoietin has become available as a drug for replacement therapy. We have compared the concentration of serum immunoreactive erythropoietin in nonrenal anemic patients with erythrocytic hypoplasia (22 cases) or active erythropoiesis (82 cases). In both groups a negative correlation was determined between the blood hemoglobin concentration and the logarithm of the erythropoietin concentration. However, the two regression lines were not identical, and the serum erythropoietin concentration was significantly higher for the degree of anemia in the patients with erythrocytic hypoplasia. Additional measurements in four patients suffering from acute leukemia with marrow failure showed that the erythropoietin concentration decreased towards the values observed in anemic patients with active erythropoiesis when the erythron recovered in the early phase of complete remission. These data support the idea that, independent of the O2 offer, the proliferating erythrocytic progenitors by negative feedback lower the blood level of erythropoietin.

Topics: Adult; Anemia, Aplastic; Bone Marrow Cells; Erythropoiesis; Erythropoietin; Female; Hemoglobins; Humans; Leukemia; Male; Radioimmunoassay; Regression Analysis

Responsiveness of bone marrow erythropoietic stem cells (CFU-E and BFU-E) to recombinant human erythropoietin (rh-Ep) was examined in vitro in 23 patients with aplastic anemia and 14 with myelodysplastic syndrome (MDS) to investigate the clinical use of rh-Ep for these diseases. Bone marrow mononuclear cells were cultured by methylcellulose methods for CFU-E and BFU-E assays. In normals, the CFU-E numbers reached a plateau of increase at Ep doses of almost 2-5 units, and no further increase was observed with the addition of larger Ep doses. In aplastic anemia, the responses of CFU-E to Ep were relatively good in nonsevere type and generally poor in severe type. However, the CFU-E numbers increased with increasing doses of Ep in some of the patients with aplastic anemia. Among the patients with MDS, the responses of CFU-E to Ep were relatively good in primary acquired refractory anemia (PARA) and primary acquired sideroblastic anemia. On the other hand, the responses of CFU-E to Ep were poor in refractory anemia with an excess of blasts (RAEB) and RAEB in transformation among the MDS patients. BFU-E responses to Ep were poor in severe aplastic anemia, RAEB, and RAEB-T. However, there are Ep responsive patients in some of aplastic anemia and PARA. High titers of rh-Ep were suggested to be effective clinically in some patients with aplastic anemia and those with PARA.

Topics: Anemia, Aplastic; Anemia, Refractory; Anemia, Refractory, with Excess of Blasts; Anemia, Sideroblastic; Bone Marrow; Cells, Cultured; Dose-Response Relationship, Drug; Erythroid Precursor Cells; Erythropoietin; Humans; Myelodysplastic Syndromes; Recombinant Proteins; Reference Values

In eleven patients with uraemia on intermittent haemodialysis treatment, recombinant human erythropoietin (rHuEpo) was used at a dosage schedule of 100 IU/kg bodyweight thrice weekly. Erythrokinetic studies (blood volume, RBC survival and iron kinetics) were performed in nine cases before and after 6 months of treatment. The remaining two patients had only RBC and plasma volume determinations before and after treatment. Although total blood volume remained unchanged, RBC volume was increased in all cases. Red cell loss was not modified, and quantitative improvement of RBC production was noted in all cases. No qualitative defect of erythroid maturation or release was observed in the treated patients. In conclusion, rHuEpo treatment improves the anaemia of haemodialysis patients by normalising circulating RBC volume only through an increase in red cell production.

Topics: Adolescent; Adult; Anemia, Aplastic; Erythrocyte Aging; Erythrocyte Indices; Erythropoiesis; Erythropoietin; Female; Hematocrit; Humans; Kidney Failure, Chronic; Male; Middle Aged; Recombinant Proteins; Renal Dialysis

Topics: Anemia, Aplastic; Congresses as Topic; Erythropoietin; Europe; Genetic Engineering; Humans; Kidney Failure, Chronic; United States

Topics: Anemia, Aplastic; Child, Preschool; Erythropoietin; Fanconi Anemia; Fanconi Syndrome; Humans; Recombinant Proteins

Using an enzyme linked immunosolvent assay kit supplied by Toyobo Co., erythropoietin (EPO) concentration in plasma was measured. Normal 100 samples showed a logarithmic distribution in EPO concentrations and normal range was between 5.4-32.5 mU/ml (mean +/- 2 SD). Coefficient variations (C.V.) of 3 samples continuously assayed were 3.8%, 6.5%, and 9.1% and C.V. of 3 samples assayed day by day were 3.6%, 7.2%, and 11.5%. Dilution test revealed that 3 samples diluted to 75%, 50%, and 25% were on each line which go through zero point. After the addition of 5.0, 15.0, and 35.0 mU/ml of EPO to the 3 samples, assay of EPO revealed 95.5 +/- 6.3% (mean +/- SD) of recovery. Assay of EPO in plasma from 19 patients with aplastic anemia revealed that all samples were higher than normal range and that 4 samples from the patients with severe aplastic anemia (less than 8.0 g/dl of hemoglobin) showed higher than 3,000 mU/ml. Furthermore, 67% (14/21) of samples from patients with leukemia showed higher than normal range in EPO concentration. EPO concentration in plasma from 17 patients with chronic renal failure were within normal range although the patients showed anemia.

Topics: Adult; Anemia, Aplastic; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; Reagent Kits, Diagnostic

To investigate the role of erythropoietin in aplastic anemia, the effects of high titers of recombinant human erythropoietin (rh-Ep) on CFU-E and BFU-E in patients with aplastic anemia were studied in vitro. Colony assays were performed by methylcellulose culture methods added with 1 to 500 units of rh-Ep. In normal bone marrow, the maximum CFU-E colony formation was observed at 2 to 5 units of rh-Ep, and BFU-E at 2 to 10 units. Colonies did not increase by addition of higher titer of rh-Ep to the cultures. In aplastic anemia, the numbers of CFU-E and BFU-E were low at 2 units of rh-Ep in culture system. In most patients with aplastic anemia studied, erythroid colonies were increased in accordance with the increase of rh-Ep added to cultures. These results suggest that the administration of high titers of rh-Ep in vivo may be useful for the improvement of anemia in aplastic anemia.

Topics: Adult; Aged; Anemia, Aplastic; Erythrocytes; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Male; Middle Aged; Recombinant Proteins

To determine whether the erythropoietin (epo) insensitivity of erythroid progenitor differentiation in congenital pure red cell aplasia or Diamond-Blackfan anaemia is intrinsic to the progenitor itself or is due to defective accessory cell function or active suppression, progenitors from normals and two patients (one steroid resistant and one spontaneously remitting), separated from all known accessory cells using sequential negative selection techniques (adherence, E-rosetting, and direct and indirect immune-panning), were studied. Initially, we evaluated three patients with DBA using unfractionated bone marrow mononuclear cells. Progenitors from two steroid non-responsive patients showed insensitivity to crude epo (c-epo) while one steroid responsive patient demonstrated normal in vitro sensitivity to c-epo. When recombinant epo (r-epo) was used in place of c-epo, the two steroid non-responders continued to demonstrate in vitro progenitor epo insensitivity. However, sensitivity of progenitors from the steroid responder, which was normal in the presence of c-epo, became abnormal when recombinant epo (r-epo) was substituted. Thus, using unfractionated bone marrow, the abnormal response to epo of progenitors from some patients with DBA appears to be obscured by stimulating factors termed erythroid burst-promoting activity (BPA) which are present in c-epo. Using fractionated highly enriched progenitors, from normals and a steroid responsive patient a final 3-10-fold enrichment of progenitors was achieved, but no such enrichment was seen when marrow from a steroid resistant patient was cultured. The epo sensitivities of normal and of patient erythroid progenitors were similar. However, at sub-optimal epo concentrations in both patients CFU-E responsiveness to crude BPA was abnormal compared to the three controls. We conclude from these studies that in DBA: (a) the failure of erythropoiesis is due to an intrinsic progenitor defect; (b) this defect involves progenitor insensitivity to factors in addition to erythropoietin: and (c) there exists a spectrum of disease reflected in the degree of the in vitro abnormality observed.

Topics: Adolescent; Anemia, Aplastic; Bone Marrow; Cell Separation; Cells, Cultured; Child; Child, Preschool; Dose-Response Relationship, Drug; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Infant; Interleukin-3; Recombinant Proteins

Several murine monoclonal antibodies (MAbs) specific for human erythropoietin (HuEpo) were produced by hybridomas obtained from the fusion of murine myeloma cells, P3X63-Ag.8-653, with the splenocytes of mice immunized with recombinant human Epo (rHuEpo). Based on epitope analysis by a competitive binding assay, these MAbs could be classified into at least three groups: (1) 1E10, (2) 1H7, (3) 2D6, 3D6 and 3D8. In a sandwich enzyme-linked immunosorbent assay (ELISA), using these MAbs as the solid-phase antibodies, MAb-bound HuEpo was detected with rabbit anti-HuEpo sera. Some combinations of two different classes of MAbs, such as 1H7 and 3D8, were found to capture much more HuEpo than each MAb used individually. Urinary HuEpo (U-HuEpo) was highly purified from the urine of patients with severe aplastic anemia with about 50% final recovery using an immunoaffinity column on which a mixture of 1H7 and 3D8 was immobilized. The purified U-HuEpo had a specific activity of 77,340 U/mg in a radioimmunoassay (RIA) and of 76,673 U/mg using an in vivo bioassay.

Topics: Anemia, Aplastic; Animals; Antibodies, Monoclonal; Antibody Specificity; Binding Sites, Antibody; Binding, Competitive; Cricetinae; Cricetulus; Epitopes; Erythropoietin; Humans; Immunosorbent Techniques; Male; Mice; Mice, Inbred BALB C; Recombinant Proteins

To clarify the control mechanism of production of erythropoietic growth factors in anemic states, we compared erythropoietin (Epo) and burst-promoting activity (BPA) in patients with aplastic anemia and iron deficiency anemia, using in vitro erythroid progenitor assays. Although serum levels of Epo activity increased in the presence of anemia, the rise was more marked in patients with aplastic anemia. BPA was high only in the sera of aplastic anemia patients. Serum levels of BPA of patients with aplastic anemia negatively correlated with hemoglobin concentrations, while those of patients with iron deficiency anemia did not correlate. In 2 patients with aplastic anemia who responded well to androgen therapy, serum levels of Epo activity and BPA decreased after the hemopoiesis had recovered. These results suggest that serum levels of BPA do not rise in response to anemia only. The elevated BPA levels in sera in cases of aplastic anemia are probably related to a reduction in the number of hemopoietic stem cells. Moreover, we observed that BPA in bone-marrow-conditioned medium (BMCM) from patients with severe aplastic anemia increased more than in the BMCM from patients with severe iron deficiency anemia. Therefore, our findings suggest that the enhanced BPA production depends on a decrease in hemopoietic precursors rather than the anemic state.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Hypochromic; Bone Marrow; Culture Media; Erythropoietin; Female; Hematopoiesis; Humans; Lymphokines; Male; Middle Aged; Tissue Inhibitor of Metalloproteinases

Topics: Anemia, Aplastic; Erythropoietin; Humans; Kidney Diseases; Recombinant Proteins

We investigated the in vivo effects of a crude extract from the urine of aplastic anemia patients (AA urinary extract) on erythroid precursor cells in the femoral bone marrow and spleens of normal adult mice. A single intraperitoneal injection of AA urinary extract induced a significant increase in the number of splenic erythroid burst-forming units (BFU-e) and erythroid colony-forming units (CFU-e) within 24 h after injection. We then injected pure recombinant erythropoietin (Epo) equivalent to the amount present in the urinary extract. This addition increased the number of splenic CFU-e by almost the same degree as the amount induced by the AA urinary extract 24 h after injection, but failed to elicit any change in the number of splenic BFU-e. In other studies, mice were injected with the same amount of lipopolysaccharide (LPS) and/or pure Epo as that present in the AA urinary extract. Experiments with Limulus amebocyte lysate-adsorbed (endotoxin-depleted) or nonadsorbed (endotoxin-containing) AA urinary extracts showed that endotoxin contamination interfered with the increase in numbers of marrow CFU-e and enhanced the increase in splenic CFU-e numbers induced by pure Epo or Epo activity in the AA urinary extract. The number of splenic BFU-e, however, was not affected by administration of LPS and/or Epo or by adsorbed endotoxin. These data suggest that AA urinary extract contains a stimulating activity for mouse splenic BFU-e, and that this activity is not attributable to the Epo activity or endotoxin contamination within the urinary extract.

Topics: Adsorption; Anemia, Aplastic; Animals; Colony-Forming Units Assay; Endotoxins; Erythrocytes; Erythropoietin; Hematopoiesis; Hematopoietic Stem Cells; Horseshoe Crabs; Lipopolysaccharides; Male; Mice; Mice, Inbred Strains; Recombinant Proteins

Radioimmunoassays for erythropoietin are limited so far to a few specialized laboratories and this requires transport and storage of samples. We therefore tested the stability of immunoreactive erythropoietin in serum and plasma samples obtained from a uremic and a nonuremic anemic patient. No significant change in the concentration of immunoreactive erythropoietin was found in either serum or plasma samples for up to 14 days of storage. This type of stability was observed no matter whether the samples were stored at room temperature, 4 degrees C, or -20 degrees C. There was no difference between the estimates of erythropoietin in serum and heparinized plasma. Validity of the radioimmunoassay used in this study was demonstrated by parallelism of dilution curves of test specimens and the 2nd International Reference Preparation for erythropoietin and by a close correlation between the immunoreactivity and the bioactivity of the hormone, as assessed in the same samples by the exhypoxic polycythemic mouse bioassay. In conclusion the data obtained clearly indicate that the necessity of storage and transport of clinical samples does not limit the practicability of the radioimmunoassay for erythropoietin.

Topics: Anemia, Aplastic; Blood Preservation; Erythropoietin; Freezing; Humans; Kidney Failure, Chronic; Polycystic Kidney Diseases; Radioimmunoassay; Reference Standards; Renal Dialysis

Erythropoietin titers when related to the hematocrit percentage and measured by bioassay in 33 normal volunteers and in 61 patients with anemias not complicated by renal or chronic disease were found to overlap with titers measured by radioimmunoassay in 20 normals and 28 patients with similar anemias. Erythropoietin titers measured by radioimmunoassay in 34 patients with rheumatoid arthritis, 25 patients with sickle cell anemia (58 separate samples), and 28 patients with erythroid hypoplasia caused by hematologic malignancies were compared with those in the control group of patients with uncomplicated anemias and found not to differ significantly from titers in this group. Erythropoietin titers measured by bioassay in 12 patients with aplastic anemia also fell within the range of those in the control group. Consequently, erythropoietin titers in these anemias appear to be determined primarily by the degree of anemia and not by any specific effect of these illnesses on the production of erythropoietin.

Topics: Anemia; Anemia, Aplastic; Anemia, Sickle Cell; Arthritis, Rheumatoid; Biological Assay; Erythropoietin; Female; Hematologic Diseases; Humans; Male; Radioimmunoassay; Red-Cell Aplasia, Pure

Recent studies suggest that megakaryocytopoiesis is governed by a dual-level regulatory process, with megakaryocyte colony-stimulating factor (Meg-CSF) primarily influencing proliferation of the committed precursors and thrombopoietin required for megakaryocyte ploidy amplification and for maturation. The authors have examined different sources of Meg-CSF in a microagar culture system with a view to their capacity to enhance megakaryocyte colony formation directly or via an indirect T-lymphocyte- or monocyte-mediated effect. The comparative influences of phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM), erythropoietin (Epo), sera of patients with severe aplastic anemia, and direct PHA addition to the culture were evaluated for their capacity to enhance megakaryocytic colony formation as well as for the maturation rate of megakaryocytes (Mk) grown in our microagar culture system. Each treatment by itself enhanced colony formation from unseparated low-density cells. Removal of T-lymphocytes and monocytes from the bone marrow sample caused a cessation of the enhancing effect of direct PHA addition to cultures stimulated with Epo, but did not influence the enhancing activities of severe aplastic anemia serum (SAA), PHA-LCM, and Epo. The results show that SAA serum, Epo, and PHA-LCM induced Mk colony formation directly and therefore may act via a common mechanism. Differences, however, were observed concerning their colony-stimulating potency and their influence on the Mk maturation rate.

Topics: Anemia, Aplastic; Bone Marrow Cells; Cell Division; Cell Survival; Colony-Stimulating Factors; Culture Media; Erythropoietin; Humans; Leukocytes; Megakaryocytes; Monocytes; Phytohemagglutinins; Stem Cells; T-Lymphocytes

Highly purified and cloned preparations of interleukin-1 (IL-1) were found to antagonize the capacity of erythropoietin (Epo) to stimulate the proliferation of mouse spleen and bone marrow erythroid precursor cells (EPC) in culture. Cloned murine IL-1 and purified and cloned human IL-1 alpha and IL-1 beta were approximately equipotent in this assay. IL-1 inhibited the proliferation response of EPC even when added as long as 17 h after Epo, suggesting that IL-1 does not affect binding of Epo to receptors or biochemical events following shortly thereafter. Indomethacin did not influence the inhibitory effect of IL-1 on Epo-induced proliferation, and PGE2 had no demonstrable effect on the process. Tumor-necrosis factor-alpha and interferons beta 1, and gamma did not affect Epo-induced proliferation. It is suggested that IL-1 mediated antagonism of the effects of Epo on erythroid precursors is a factor in the pathogenesis of many types of hypoplastic anaemia, including those associated with infections, rheumatoid arthritis and systemic lupus erythematosus, giant-cell arteritis, graft-versus-host disease and disorders associated with lymphocyte-mediated suppression of erythropoiesis.

Topics: Anemia; Anemia, Aplastic; Animals; Bone Marrow; Cell Division; Cells, Cultured; Erythroblasts; Erythropoietin; Female; Growth Inhibitors; Hematopoietic Stem Cells; Interleukin-1; Mice; Spleen; Tumor Necrosis Factor-alpha

It is apparent that multiple cellular stages and biologic processes can be identified during megakaryocytopoiesis that are potentially subject to control by hematopoietic growth factors and marrow accessory cell populations. Two classes of megakaryocyte progenitor cells, the colony forming unit-megakaryocyte (CFU-MK) and the burst forming unit-megakaryocyte (BFU-MK), have now been detected in normal human bone marrow cells. The BFU-MK by virtue of the greater cellular content of its resultant colonies and the delayed time of appearance of these colonies appears to be a more primitive progenitor cell with a greater proliferative potential than the CFU-MK. A number of hematopoietic growth factors including megakaryocyte colony stimulating factor, (MK-CSF), recombinant erythropoietin (EPO) and granulocyte macrophage colony stimulating factor (GM-CSF) are each capable of increasing cloning efficiency of human megakaryocyte progenitor cells. It is presently unknown whether these factors act directly on the CFU-MK or whether they stimulate marrow accessory cells to elaborate growth factors that influence CFU-MK proliferation. In order to answer this question, the effect of these growth factors on the cloning efficiency of a human megakaryocytic cell line, EST-IU, was examined. Each of these factors was capable of increasing leukemia cell colony formation. One can conclude from these studies that MK-CSF, EPO, and GM-CSF act directly on cells of the megakaryocytic lineage. The physiologic significance of the lineage nonspecific effects of EPO and GM-CSF on megakaryocytopoiesis is yet to be determined. On the basis of these observations, a model of human megakaryocytopoiesis was suggested. Several factors appear able to influence multiple steps in megakaryocytic development, whereas others influence only specific stages or cellular events occurring during megakaryocytopoiesis.

Topics: Anemia, Aplastic; Blood Platelets; Bone Marrow Cells; Cell Differentiation; Cell Line; Colony-Stimulating Factors; Erythropoietin; Growth Substances; Hematopoiesis; Hematopoietic Stem Cells; Humans; Interleukin-3; Megakaryocytes; Tumor Cells, Cultured; Tumor Stem Cell Assay

Erythropoietin (Epo) titers in various hematological states were determined by a radioimmunoassay. Epo titers in patients with uremic anemia and iron deficiency anemia were inversely correlated with their respective hemoglobin concentrations. Epo titers in patients with uremic anemia were significantly lower than those in patients with iron deficiency anemia with comparable hemoglobin concentrations.

Topics: Anemia; Anemia, Aplastic; Anemia, Hypochromic; Erythropoietin; Humans; Kidney Failure, Chronic; Polycythemia Vera; Renal Dialysis; Uremia

We produced an antiserum by immunizing rabbits with purified human megakaryocyte colony stimulating factor (Meg-CSF). With the use of an anti-Meg-CSF IgG fraction (AM-IgG), we detected immunoreactive Meg-CSF both in human aplastic anemia serum (AAS) and normal serum. Based on our immunological and biological analyses, Meg-CSF appeared to be antigenically as well as functionally distinct from human urinary erythropoietin (EPO) and thrombopoietic stimulating factor. The AM-IgG fraction was able to suppress the ability of both aplastic anemia serum and purified Meg-CSF to promote megakaryocyte colony formation. In addition, the supernatant formed after immune precipitation of the AAS with AM-IgG no longer possessed Meg-CSF-like activity. The AM-IgG did not suppress the ability of EPO, phytohemagglutinin-stimulated leukocyte conditioned medium (PHA-LCM), or PHA-LCM + EPO to promote erythroid, granulocyte-macrophage, or mixed colony formation, respectively. The use of this antibody has further defined the dependency of human megakaryocytopoiesis on Meg-CSF.

Topics: Anemia, Aplastic; Colony-Forming Units Assay; Colony-Stimulating Factors; Electrophoresis, Polyacrylamide Gel; Erythropoietin; Hematopoiesis; Humans; Immune Sera; Immunodiffusion; Immunoelectrophoresis; Immunoglobulin G; Megakaryocytes; Phytohemagglutinins

Human urinary neuraminidase, an enzyme that releases sialic acid from hematopoietic factors found in urinary preparations, was partially characterized, and a method was developed to derive these hematopoietic factors free of enzyme activity. Neuraminidase in urinary preparations from healthy humans and aplastic anemic (AA) patients had optimal activity at pH 5.3 and hydrolyzed both alpha 2----3 and alpha 2----6 type ketosidic linkages of N-acetyl-neuramin lactose and alpha 1-acid glycoprotein. When subjected to Sephacryl S-300 gel filtration, urinary neuraminidase showed a single peak of activity with an apparent molecular weight of 380,000 daltons, even under denaturing conditions (6 M guanidine hydrochloride). Furthermore, among a variety of compounds tested, no potent inhibitor of the enzyme was found. Heat treatment of AA urinary preparations eliminated about 80% of neuraminidase activity, while successive two-step ethanol precipitation eliminated residual enzyme. Erythropoietin, megakaryocyte colony-stimulating factor (CSF) and granulocyte/macrophage phage CSF activities were retained after these treatments.

Topics: Anemia, Aplastic; Chemical Precipitation; Erythropoietin; Hot Temperature; Humans; Hydrogen-Ion Concentration; Interleukin-3; Kinetics; Megakaryocytes; Molecular Weight; Neuraminidase

Untreated human serum is known to be toxic to in vitro assays for erythropoietin, including the mouse spleen cell assay system (MSCA). This phenomenon had previously been shown to be mediated by complement-dependent IgM heteroantibodies and can be overcome by heating the serum at 56 degrees C for 30 minutes. Using the MSCA, we have found that the toxic effect of serum could also be removed by treatment with a precipitating antibody against the C3c component of complement. The effects of the two methods of complement inactivation on the measurement of stimulatory activity in serum have been compared. For normal serum, the results after heat inactivation and antibody treatment were similar. In contrast, serum from a patient with aplastic anemia gave a result equivalent to 327 mU erythropoietin/ml after heat treatment, but after antibody treatment equivalent to 1,520 mU erythropoietin/ml. Gel permeation chromatography of unheated, heated, and antibody-treated sera showed that heating markedly reduced the activity of the erythropoietin peak. Seventy percent of the activity of partially purified urinary erythropoietin was lost during heating in the presence of normal serum. In addition, heating caused the appearance of high molecular weight compounds that are stimulatory in the MSCA. The level of this activity appeared to be directly related to the stimulatory activity of the unheated serum.

Topics: Anemia, Aplastic; Animals; Biological Assay; Blood Physiological Phenomena; Complement System Proteins; Erythropoiesis; Erythropoietin; Hot Temperature; Humans; Mice; Spleen

I have examined Ferrokinetics and the activity of serum erythropoietin (by Frieds method) for determining the in vivo erythropoiesis, additionally the presence of "effective" or "ineffective" erythropoiesis in patients with hypoplastic anemia. The number of reticulocytes and the reticulocyte production index also were analysed concerning erythropoiesis. Based on above-mentioned findings, the prognosis of these patients was analysed. Furthermore, from the relationship between the morphological examination of the erythroid cells in the bone marrow and the time course of the activity of serum erythropoietin, the change of erythropoiesis was investigated. From these results, a higher level of serum erythropoietin was observed in patients with more severe type in erythropoiesis, whereas the prognosis in these patients was much poorer in relation to an elevated value of serum erythropoietin. Furthermore, it might be suggested that a high level of erythropoietin which is enough to help the erythroid progenitor cells to differentiate and maturate to normal erythroblasts was able to shorter the maturation time of the erythroid cells and also effectively abolished "ineffective erythropoiesis" inducing the proper erythropoiesis.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Erythropoiesis; Erythropoietin; Female; Humans; Iron; Kinetics; Male; Middle Aged; Prognosis

Serum erythropoietic activity was determined in 32 patients with beta thalassemia major and intermedia. Quantitation was performed by an in vitro bioassay using rabbit erythroid precursor cells (CFU-E) either by colony assay or by 3H-thymidine uptake. 20 polytransfused beta-thalassemic major patients had erythropoietic activity (mean 89.3 +/- 36 milliunits/ml) which was not significantly different (p greater than 0.2) from normal individuals (51.3 +/- 32 milliunits/ml). 12 untransfused patients with beta thalassemia intermedia were found to have comparable serum erythropoietic activity (p greater than 0.01). These levels were much lower than those found in patients with aplastic anemia who had a comparable degree of anemia. We have shown that the low EPO activity in thalassemic patients was not due to experimental conditions (excess of ferritin, low transferrin) nor to specific inhibitors appearing in this disease. No correlation was found between the erythropoietic activity and sex or other clinical parameters of the patients such as severity of the anemia, splenectomy, iron chelation or transfusion therapy. 4 young thalassemic children (1-2 yr of age) studied had high erythropoietic activity ranging from 661 to 5793 milliunits/ml--significantly different from normal children of the same age. It is suggested, therefore, that a decrease in serum erythropoietin levels develops during the course of the disease.

Topics: Anemia, Aplastic; Blood Transfusion; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Female; Ferritins; Hot Temperature; Humans; Iron; Male; Thalassemia

To explore the etiology of congenital hypoplastic or Diamond-Blackfan anemia (DBA) we investigated in vitro erythropoiesis in nine patients. Of the nine, seven were clinically responsive to prednisone. Four were infants evaluated at the time of diagnosis. Six were never or were only minimally transfused. Those for whom prednisone had been prescribed had discontinued the drug a minimum of five months prior to study. The bone marrows of these nine patients were compared with those of hematologically normal individuals and with those of four patients with transient erythroblastopenia of childhood (TEC) whose erythroid aplasia was as severe as that of the patients with DBA. Using the plasma clot semisolid culture technique to enumerate erythroid progenitors and to evaluate the growth characteristics of the colonies to which they give rise, we concluded that at the onset of DBA: (a) erythroid progenitor frequency does not correlate with the degree of anemia and erythroblastopenia; (b) erythroid progenitor differentiation may in some cases be abnormally insensitive to crude preparations of erythropoietin; and (c) progenitor erythropoietin insensitivity in vitro does not necessarily indicate prednisone insensitivity in vivo. Thus, DBA does not appear to be solely the result of deficient formation of erythroid progenitors but is, in addition, a disorder that is due to defective progenitor differentiation in vivo.

Topics: Adult; Anemia, Aplastic; Cell Differentiation; Child, Preschool; Colony-Forming Units Assay; Erythroblasts; Erythrocytes; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Infant; Time Factors

Burst-promoting activity (BPA) in the sera of patients with various types of anemia and polycythemia was compared with that of normal subjects by an in vitro method using mouse bone marrow cells. The control culture contained normal human AB serum instead of sample materials. Results were expressed as a percentage of burst numbers in control cultures. Serum erythropoietin (Epo) levels were determined by a radioimmunoassay. Serum BPA in patients with aplastic anemia (155.4 +/- 56.7%, mean +/- SD) was significantly higher than that in normal subjects (112.1 +/- 29.1%, Wilcoxon's rank sum test, P less than 0.05). However, serum BPA in patients with uremic anemia (122.2 +/- 26.5%), polycythemia vera (101.9 +/- 19.5%) and stress polycythemia (115.5 +/- 25.6%) was not significantly different from normal subjects. There was a correlation between serum BPA and Epo titers in patients with aplastic anemia and paroxysmal nocturnal hemoglobinuria (r = 0.81, t test, P less than 0.001).

Topics: Anemia; Anemia, Aplastic; Animals; Erythropoiesis; Erythropoietin; Female; Hematopoiesis; Humans; Mice; Pokeweed Mitogens; Polycythemia; Spleen

A two-stage cell culture assay specific for human erythroid burst-promoting activity (BPA) is described. Human peripheral blood mononuclear cells were cultured in suspension with or without a BPA test sample for two days, then transferred to methylcellulose medium with added erythropoietin (EPO) and incubated for ten more days, and finally BFU-E-derived colonies were scored. An increase in number of colonies due to the presence of BPA was observed that was proportional to the concentration of BPA in the test sample. This response was linear with respect to number of cells plated between 2 and 5 X 10(5)/ml. The system was standardized with a partially purified human urinary BPA preparation. Dose responses to urinary protein preparations, plasma, and serum were parallel. The assay system was found to be nonresponsive to highly purified EPO and to bacterial endotoxin. It was determined that BPA action was confined to the suspension culture stage of the assay, while EPO presence was an absolute requirement during methylcellulose culture. In the two-stage assay optimal amounts of BPA caused up to 358% increases of BFU-E-derived colonies; the same amounts of BPA added to conventional methylcellulose cultures caused only up to 54% increases over the number of colonies obtained with EPO alone. Plasma and serum BPA levels of hematologically normal and abnormal individuals showed no correlation with EPO levels and hemoglobin (Hb) concentrations. This seems to rule out the possibility that BPA elaboration is regulated by oxygen availability or the amount of EPO circulating in an organism.

Topics: Anemia, Aplastic; Animals; Cell Count; Cells, Cultured; Colony-Forming Units Assay; Colony-Stimulating Factors; Dose-Response Relationship, Drug; Endotoxins; Erythropoiesis; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Mice; Polycythemia Vera

Methylcellulose culture assay was used to detect committed haemopoietic stem cells, CFU-C and CFU-E, in aplastic anaemia patients with autologous haemopoietic reconstitution. Severe diminution of CFU-C was found in all the patients studied and the absence of a dose-response to colony stimulating factor (CSF) was demonstrated. A reduced number of CFU-E and lower erythropoietin (Ep) sensitivity of those progenitors was detected as well. Autologous serum added to the bone marrow cultures of these patients enhanced the growth of CFU-C but inhibited CFU-E growth. According to the results presented, some residual damage at the stem cell level is suggested.

Topics: Adult; Androgens; Anemia, Aplastic; Bone Marrow Cells; Cells, Cultured; Colony-Forming Units Assay; Colony-Stimulating Factors; Erythropoiesis; Erythropoietin; Female; Hematopoiesis; Hematopoietic Stem Cells; Humans; Male; Middle Aged

The erythropoietin response to anaemia was compared in 30 children with haemolytic anaemia and in 5 children with Fanconi's hypoproliferative anaemia. Serum erythropoietin was measured by radioimmunoassay. In children with haemolytic anaemia the serum erythropoietin concentration increased exponentially with decreasing haematocrit values (r = 0.74; p less than 0.001). Serum erythropoietin levels also correlated with reticulocyte counts (r = 0.62; p less than 0.001). Children with Fanconi's hypoproliferative anaemia had considerably higher serum erythropoietin levels than children with haemolysis for the same degree of anaemia. These data indicate that erythropoietin production in Fanconi's anaemia may be dependent on other factors in addition to the degree of anaemia and relative hypoxaemia.

Topics: Adolescent; Anemia, Aplastic; Anemia, Sickle Cell; Cell Count; Child; Child, Preschool; Erythropoietin; Fanconi Anemia; Female; Hematocrit; Humans; Male; Radioimmunoassay; Reticulocytes; Sickle Cell Trait

Topics: Anemia, Aplastic; Anemia, Hypochromic; Animals; Erythropoietin; Humans; Kidney Failure, Chronic; Male; Mice; Polycythemia; Rats; Renal Dialysis

Thrombocytopoietin seems to be only partially responsible for the regulation of platelet production. We determined precisely the differences between a second thrombocytopoiesis-stimulating factor (TSF2), and thrombocytopoietin (TSF1) and erythropoietin (Epo). Fractionation was carried out, first on a DEAE-cellulose phosphate column and then on a Sephadex G-75 column. Thrombocytopoietic activity in the various fractions was assessed using 75Se-methionine platelet incorporation into normal recipients. Epo concentrations were determined using a radioimmunoassay. We showed that the apparent molecular weight of TSF2 is 14,000 daltons. It differs from TSF1 (48,000 daltons) and from Epo (39,000 daltons). For doses of 8-12 mU Epo/rat, found in whole serum injected, no effect on thrombocytopoiesis was found. On the contrary, a significant effect (p less than 0.01) was found when the same quantities of Epo present in the Sephadex G-75 F4' fraction were injected (2-10 mU/rat). TSF2 can be separated from TSF1 and Epo, using biochemical techniques.

Topics: Anemia, Aplastic; Animals; Chromatography, DEAE-Cellulose; Chromatography, Gel; Erythropoietin; Glycoproteins; Hydroxyurea; Molecular Weight; Rats; Rats, Inbred Strains; Thrombopoietin

We have analysed the contribution to megakaryocyte colony formation in methylcellulose made by human plasma, serum, media conditioned by phytohemagglutinin (PHA) stimulated leukocytes (PHA-LCM), erythropoietin (EPO) preparations, and platelets. The culture system was used as a bioassay for megakaryocyte colony stimulating activity (Meg-CSA) in plasma samples of patients with perturbed megakaryocytopoiesis. Preparations of heparinized platelet-poor plasma yielded the most consistent results. Platelet-poor plasma of normal subjects will at best facilitate the occasional growth of small megakaryocyte colonies. Colony frequency and size are reproducibly enhanced in the presence of PHA-LCM as a source of exogenous Meg-CSA. Commercially available EPO preparations may vary in their content of activities that influence megakaryocyte colony formation. Addition of these preparations to cultures that contain plasma and PHA-LCM usually does not enhance colony formation. In contrast to platelet-poor plasma, platelet rich plasma and serum are less supportive of megakaryocyte colony growth. It is suggested that this loss of activity may be related to the release of inhibitors by activated platelets or alternatively caused by absorption of activities by platelets. Plasma samples from patients with megakaryocytopoietic dysfunction may contain components that promote colony formation without addition of PHA-LCM or EPO. This phenomenon is consistently observed for patients with severe aplastic anemia and bone marrow transplant recipients after completion of their ablative preparative regimen.

Topics: Anemia, Aplastic; Biological Assay; Blood; Blood Platelets; Colony-Forming Units Assay; Erythropoietin; Hematopoietic Stem Cells; Humans; Lymphocyte Activation; Megakaryocytes; Phytohemagglutinins

The activity capable of promoting the growth of human erythroid burst-forming cells (BFU-E) in culture was measured in the sera from 39 patients with aplastic anemia (AA) and compared with similar activity in patients with various other hematologic disorders and 31 normal subjects. Burst-promoting activity (BPA) was determined by its ability to support erythroid burst growth from adherent cell-depleted normal human marrow cells. The results were expressed as the percentage of burst growth supported by test serum compared with cultures established in the presence of 20% test serum and 2.5% phytohemagglutinin-stimulated lymphocyte conditioned medium. The mean BPA level in normal serum was 18.5% (1.5 +/- SEM) and was not significantly different from BPA levels in patients with various forms of nonhypoplastic anemia or polycythemia (10.2% +/- 1.2%). In contrast, 15 of the 39 patients with AA had elevated BPA levels, ranging from 40.0% to 106.0%. These elevated levels did not correlate with serum erythropoietin or hematocrit values, white blood cell count, platelet count, time from diagnosis, or the presence or numbers of BFU-E in circulation. The BPA was shown not to be T cell growth factor (interleukin-2), and the effect was not blocked by the addition of cyclosporine to culture, consistent with a direct effect of this activity on BFU-E. When the 39 patients with AA were treated with antithymocyte globulin, 20 obtained a complete or partial remission. BPA levels determined from sera obtained before treatment did not correlate with response or duration of survival but did correlate with granulocyte-macrophage colony-stimulating activity (GM-CSA).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Anemia, Aplastic; Bone Marrow Cells; Child; Child, Preschool; Cyclosporins; Erythropoietin; Female; Hematocrit; Hematopoietic Stem Cells; Humans; Leukocyte Count; Male; Middle Aged; Monocytes; Platelet Count; Reference Values

Patients with various types of anaemia, but with comparable haemoglobin levels, show a wide range of serum erythropoietic activity. We have developed a method for the fractionation of serum by HPLC followed by bioassay of the individual fractions, using the mouse spleen cell microassay. Up to three distinct peaks of erythropoietic activity corresponding to molecular weights (MW) greater than 300,000, 250,000-300,000 and 40,000 have been found in serum from both normal and anaemic subjects. The erythropoietic profiles of the sera examined differ markedly in anaemias of different aetiology. The chemical nature and the physiological significance of the stimulators remain to be investigated.

Topics: Adolescent; Aged; Anemia; Anemia, Aplastic; Anemia, Megaloblastic; Animals; Biological Assay; Chromatography, High Pressure Liquid; Erythropoietin; Female; Hemoglobins; Humans; Leukemia; Male; Mice; Mice, Inbred C57BL; Middle Aged; Spleen

Highly purified erythropoietin was not available in quantities needed to carry out planned investigations and, therefore, the use of high performance liquid chromatography was explored. This technique permits the separation of proteins with high efficiency and resolution. Three types of chromatography were used. Size exclusion or gel permeation, reversed phase, and ion exchange columns were utilized with different solvent systems. The chromatographic fractions were assayed either by an exhypoxic polycythemic mouse assay or by the fetal liver cell assay. In addition, selected fractions were tested for their capability to stimulate CFU-E and BFU-E colony formation in methyl cellulose. The results of the techniques of size exclusion and ion exchange chromatography were found to be rapid and reproducible. Although reversed phase chromatography gave excellent resolution, the results were somewhat variable. Using different chromatographic combinations, erythropoietin with a specific bioactivity in the range of 50,000 u/mg protein was isolated. Although the erythropoietin gene has now been cloned and the hormone purified by utilizing monoclonal antibodies, high performance liquid chromatography may be useful in the removal of unwanted contaminants, as a tool in chemical characterization, and as a possible method for the hormone's identification and measurement in clinical laboratories.

Topics: Anemia, Aplastic; Animals; Biological Assay; Chromatography, Gel; Chromatography, High Pressure Liquid; Erythropoietin; Fetus; Humans; Liver; Mice; Polycythemia

Human erythropoietin was isolated from urine of aplastic anemic patients in a high yield with a simple purification procedure using an immunoadsorbent column of monoclonal antibodies and a Sephadex G-100 column. About 6 mg of erythropoietin was isolated from 700 liters of urine and the specific activity was estimated to be 81,600 units/mg of protein with an in vivo 59Fe incorporation assay method, using starved rats. Activity measurement of the extracts from sliced gels after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the Western blotting technique revealed heterogeneity of the isolated erythropoietin, which is probably caused by variable amounts of carbohydrates attached to the polypeptide chain. Thirty amino acids in the NH2-terminal portion of the isolated hormone were sequenced.

Topics: Anemia, Aplastic; Antibodies, Monoclonal; Chromatography, Affinity; Erythropoietin; Humans; Molecular Weight

An in vitro method for the determination of erythropoietin (EPO) titres in human serum samples was developed to detect low levels of EPO titres in various haematological states. Fetal mouse liver cells (FMLC) were incubated in methylcellulose-containing media, and radioiron incorporation into haem was estimated in cultured FMLC. We could determine the concentrations of EPO from around 20 to 600 mU per ml of a very small amount of serum samples from patients and normal adults. Using this method, EPO titres of normal adults (54 +/- 31 mU/ml), chronic renal failure patients (29 +/- 23 mU/ml), aplastic anaemia patients (545 +/- 74 mU/ml), and polycythaemia vera patients (16 +/- 7 mU/ml) were determined.

Topics: Anemia, Aplastic; Animals; Cells, Cultured; Culture Media; Erythropoietin; Humans; Kidney Failure, Chronic; Methods; Mice; Mice, Inbred ICR; Polycythemia Vera

Erythropoietin concentrations were increased significantly (p less than 0.025) in nine cats with natural feline leukemia virus infection and associated erythroid aplasia compared to six clinically normal cats. Adult cats experimentally inoculated with the Kawakami-Theilen isolate of feline leukemia virus developed a progressive simultaneous increase in erythropoietin activity and decrease in packed cell volume. These findings indicate that erythroid aplasia associated with feline leukemia virus infection is not caused by a failure in erythropoietin production.

Topics: Anemia, Aplastic; Animals; Cat Diseases; Cats; Erythropoietin; Female; Hematocrit; Leukemia; Leukemia Virus, Feline; Male; Methylprednisolone; Methylprednisolone Acetate; Specific Pathogen-Free Organisms

We have studied the growth of erythroid bursts in methylcellulose cultures from bone marrow (7 cases) and peripheral blood cells (17 cases) of patients with primary acquired refractory anemias. In most cases, erythroid burst-forming units (BFU-E) were either absent or present in lower than normal numbers, and these levels remained low to absent in sequential studies. In 2 patients, circulating BFU-E were initially higher than normal, but subsequently declined to normal levels. 1 patient had low-normal numbers of circulating BFU-E initially, but these declined to zero in a later study. No clinical or routine hematological features permitted distinction between patients with initially high BFU-E and those with initially low or absent BFU-E. These findings parallel those reported for granulocyte-macrophage progenitors in refractory anemias, and support the concept that erythroid progenitor cells in these disorders are influenced by the disordered hematopoiesis.

Topics: Adult; Aged; Anemia, Aplastic; Blood Cells; Bone Marrow Cells; Cells, Cultured; Child; Child, Preschool; Colony-Forming Units Assay; Culture Media; Erythrocyte Count; Erythropoiesis; Erythropoietin; Female; Follow-Up Studies; Humans; Male; Middle Aged

A 70-year-old woman developed typical clinical symptoms of pure red cell anemia (PRCA) following a history of rheumatoid arthritis (RA). The patient's bone marrow erythropoietic progenitors cells were cloned in a micro agar culture system several times over a period of 11 months, revealing a diminished frequency of bone marrow erythroblasts paralleled by a markedly reduced number of CFU-e and BFU-e in vitro. No inhibitory activity in the patient's IgG fraction could be detected either by preincubation with IgG and/or rabbit complement, or in the continuous presence of IgG. Depletion of T lymphocytes from the patient's bone marrow cells led to an improved in vitro erythroid proliferation. Cytostatic therapy with cyclophosphamide clinically induced a marked increase in the bone marrow erythroblast and reticulocyte number, correlated in vitro by normalization of CFU-e levels and increase in the number of BFU-e. Nevertheless, BFU-e values never attained normal levels, which could be attributed to a reduced stem cell pool resulting from previous therapy with cyclophosphamide and/or antirheumatic drugs. Two independent factors, a reduced pool of committed stem cells as well as an autoimmune cell-mediated suppression, may both contribute to the pathomechanism of the disease in this patient.

Topics: Aged; Anemia, Aplastic; Bone Marrow Cells; Clone Cells; Colony-Forming Units Assay; Cyclophosphamide; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Immunoglobulin G; Lymphocyte Depletion

Using procedures that were effective in the purification of human urinary erythropoietin (Epo), we attempted initial purification of megakaryocyte colony-stimulating factors (CSF) in urinary extracts from patients with aplastic anemia (AA) and idiopathic thrombocytopenic purpura (ITP). Comparison of colony stimulation by purified human Epo and crude urinary extracts revealed: (1) that the pure Epo augments megakaryocyte colony formation in culture and (2) MEG-CSF activity is also present in materials other than Epo in the crude urinary extracts from the two types of patients. Similar to purification of Epo, ethanol precipitation and sulfopropyl-Sephadex chromatography provided twofold and threefold increases in the specific activity of MEG-CSF, respectively. In contrast to Epo, however, significant inactivation of MEG-CSF activity was seen with phenol treatment. The elution profile of MEG-CSF seen on hydroxylapatite chromatography of urinary extracts was different from that of Epo. These data provided a basis for initial steps for purification of MEG-CSF and support the notion that MEG-CSF is distinct from Epo.

Topics: Aminosalicylic Acids; Anemia, Aplastic; Cells, Cultured; Chromatography; Colony-Stimulating Factors; Erythropoietin; Ethanol; Humans; Hydroxyapatites; Megakaryocytes; Purpura, Thrombocytopenic

3 different methods, (1) assays of CFU-Cs and CFU-Es, (2) responsiveness of CFU-Cs and CFU-Es to low doses of CSF and ESF, respectively and (3) effects of ALG on CFU-C colony formation in vitro, were used to evaluate the quantitative and qualitative defects of stem cells in 28 patients with aplastic anaemia. Some patients with aplastic anaemia who had attained complete remission several years previously, exhibited severely depressed in vitro CFU-C colony formation. This suggests that the defect persists for a long time after clinical complete remission. Residual marrow CFU-Cs and CFU-Es did not have defective responses to the humoral stimulating factors, CSF and ESF. No patient showed a rise of colony number after treatment with ALG in vitro.

Topics: Anemia, Aplastic; Colony-Forming Units Assay; Colony-Stimulating Factors; Dose-Response Relationship, Drug; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Male; Middle Aged; Pancytopenia

Topics: Anemia, Aplastic; Animals; Chromatography, Affinity; Erythropoietin; Glycoproteins; Hematopoiesis; Humans; Neuraminidase; Rats

To study the action of corticosteroids on erythroid precursors, (burst forming unit-erythroid and colony forming unit-erythroid) in Diamond-Blackfan Syndrome (DBS), marrow from a newly diagnosed untreated infant was studied in vitro with prednisone and dexamethasone. This patient subsequently proved to be steroid responsive. Colony numbers increased linearly in an erythropoietin (EPO) dose response study. There was marked enhancement of colony numbers at all EPO doses after adding either prednisone (10(-6) M) or dexamethasone (10(-9) M) to the cultures. The data indicate that corticosteroids augment erythropoiesis at both early (BFU-E) and late (CFU-E) stages of development in DBS. In contrast, marrow from a second infant with DBS, clinically steroid resistant, failed to respond to steroids in vitro.

Topics: Anemia, Aplastic; Colony-Forming Units Assay; Dexamethasone; Dose-Response Relationship, Drug; Drug Resistance; Erythropoiesis; Erythropoietin; Humans; In Vitro Techniques; Infant; Infant, Newborn; Prednisone; Syndrome

Topics: Anemia, Aplastic; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Middle Aged

A radioimmunoassay (RIA) method for erythropoietin (Epo) was developed and validated against the polycythaemic mouse assay. The correlation was good, with a r = 0.94. Several other criteria of specificity were also filled by the RIA, which had a lower detection limit of 5 microU/ml. The mean serum-Epo level in 6 patients with secondary polycythaemia, 50.2 +/- 26.2 microU/ml, was significantly higher than in a group of 11 normal subjects, 28.7 +/- 7.2 microU/ml (P less than 0.0002). However, the Epo level in 31 polycythaemia vera (PV) patients, M = 21.9 +/- 6.6 microU/ml, was not significantly different from normal (P = 0.006). Since previous studies with bioassay of heat-treated and concentrated plasma samples have shown a decreased serum-Epo level in PV, Epo levels were measured before and after heat treatment and concentration of samples from normals and polycythaemics. It was found that the levels of immunoreactive material increased after heat treatment and 40 times concentration in samples from normals and patients with secondary polycythaemias, but decreased in PV. We conclude that the Epo levels in serum in the low range measured by our and previous RIA:s probably are not true Epo levels but are partly due to an unspecific serum effect, that was removed by heat treatment.

Topics: Anemia, Aplastic; Animals; Antibody Affinity; Antibody Specificity; Erythropoietin; Humans; Immune Sera; Mice; Polycythemia Vera; Radioimmunoassay

A chromatofocusing column was used to separate burst-promoting activity (BPA) and erythropoietin (Ep) activity in sera from patients with aplastic anemia and, interestingly a fraction containing a considerable level of BPA without Ep activity was found at around pH 5.0. Another fraction at lower pH contained both activities. This is the first report of the existence of a fraction with BPA but without Ep activity in sera from aplasia patients. In trying to measure BPA in sera and urine, a new assay procedure was used involving counting the number of erythroid bursts in normal human marrow cell cultures initiated with test samples to which 1-2 units of standard Ep were added 5-6 days later. Separation of BPA from Ep activity in patients sera was also attempted by Sephadex G-100 but this was unsuccessful due to the presence of inhibitors to burst formation. Also, in a urine separation, BPA and Ep activities were eluted in the same fraction. Sephadex G-100 chromatography gave results which seemed to indicate separation of BPA from Ep activity, however, this may have been due to the presence of inhibitor(s). This idea was supported by the fact that the peaks of both activities coincided when a urine preparation containing no inhibitors was tested.

Topics: Anemia, Aplastic; Animals; Bone Marrow Cells; Cell Line; Chromatography, Gel; Culture Media; Erythropoiesis; Erythropoietin; Female; Humans; Isoelectric Focusing; Leukemia, Experimental; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Molecular Weight; Pregnancy

Topics: Alkalies; Anemia, Aplastic; Erythropoietin; Female; Fetal Hemoglobin; Hemoglobins; Humans; Male; Protein Denaturation

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Bone Marrow; Child; Child, Preschool; Erythropoietin; Humans; Infant; Iron; Leukemia; Middle Aged

Bioassays for human erythropoietin are cumbersome, time-consuming, and insensitive. The purification of human erythropoietin (EP) had provided small quantities of highly bioactive EP (approximately 70,000 U/mg) required for the development of an EP radioimmunoassay (RIA). The RIA for EP described in this investigation, can detect 5 mU/ml of EP in the assay tube; the serum concentration of EP in normal individuals ranged from less than 18 to 81 mU/ml with a mean value of 29 mU/ml. In contrast, nine patients with severe aplastic anemia had markedly elevated serum EP concentrations with a mean value of 3,487 mU/ml, range 984--6,434 mU/ml. In this RIA, patients with Polycythemia vera had consistently undetectable EP concentrations, less than 18 mU/ml. Eleven patients with chronic renal failure had a higher mean serum EP concentration (40.5 mU/ml) than normal individuals, but the range (less than 18-115 mU/ml) overlapped that of normals. By immunologic and gel chromatographic criteria, EP measured in serum was similar to standard urinary EP. The EP immunoassay that we have developed has sufficient sensitivity and specificity not only to quantitate the elevated serum EP levels found in aplastic anemia but also to discriminate decreased from normal serum concentrations of EP in most circumstances. This simple, reliable RIA has provided the necessary framework upon which to increase our understanding of the importance of EP in hematopoiesis.

Topics: Adult; Aged; Anemia, Aplastic; Blood Transfusion; Chromatography, Gel; Erythrocyte Transfusion; Erythropoietin; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; Polycythemia Vera; Radioimmunoassay

The results from a biological test for erythropoietin (using the rate of iron absorption in polycythemic mice) and a commercially-available immunological test (haemagglutination-inhibition test) were compared. Of 19 batches of the immunological test which were investigated, 7 batches were completely inactive and a further 3 batches reacted only with the test serum supplied with the test. There was a poor correlation between the results from the biological and the immunological measurements, both on patients with high and those with low serum erythropoietin levels. The difficulty of the immunological erythropoietin test is that pure erythropoietin is not sufficiently available. The immunological test investigated here does not use pure erythropoietin. Aside from this, pathophysiological considerations would lead one to expect basic differences between the results from immunological and biological tests.

Topics: Adrenal Gland Neoplasms; Anemia, Aplastic; Erythropoietin; Hemagglutination Inhibition Tests; Humans; Kidney Failure, Chronic; Leukemia; Pheochromocytoma; Polycythemia Vera

Growth of human erythroid and erythroid-granulocytic colonies was investigated in a fibrinogen clot culture system. CFU-E, BFU-E, CFU-C and the mixed colonies were grown in cultures in medium conditioned by phytohaemagglutinin-stimulated human mononuclear cells, with small amounts of aplastic anaemia serum, but no exogeneous erythropoietin (Epo). The level of Epo in the culture was 0.015 U. The mixed colonies constituted 8-22% of the total colonies grown. The results indicate that high dose Epo is not required to grow BFU-E and the mixed colonies in cultures containing erythroid potentiating factor(s), and the frequency of bipotent haemopoietic stem cells in human bone marrow or peripheral blood may be higher than that reported previously.

Topics: Adult; Anemia, Aplastic; Cells, Cultured; Child; Colony-Forming Units Assay; Erythrocytes; Erythropoietin; Fibrinogen; Granulocytes; Hematopoietic Stem Cells; Humans; Infant, Newborn; Time Factors

A number of studies have demonstrated that certain immunocompetent cells play a role in the regulation of normal erythropoiesis. These regulatory cells (monocytes--macrophages, lymphocytes) modulate almost every phase of the erythropoietic process, and along with erythropoietin represent the major controlling force in erythropoiesis. Evidence indicate that pathological alterations of these cell-mediated activities can lead to clinical disturbances of red cell production such as is seen in patients with Diamond-Blackfan syndrome and aplastic anemia.

Topics: Anemia, Aplastic; Animals; Bone Marrow; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; HLA Antigens; Lymphocytes; Mice; Monocytes; T-Lymphocytes, Regulatory

Published reports of the levels of erythropoietin (Ep) in normal human serum have been reviewed. Current evidence suggests that the level of Ep in normal human serum is 30 +/- 10 mU Ep/ml. When serum was concentrated before assay somewhat lower values were obtained and the mouse fetal liver cell assay gave slightly higher values. Three new potential Ep assay techniques are presented. These are: 1) The use of synthetic microbeads as indicators in an agglutination assay, 2) the use of wheat germ lectin in affinoelectrophoresis techniques, and 3) an immunoradiometric assay as an alternative to radioimmunoassays. The results indicate that the use of all three assays should be explored further.

Topics: Anemia, Aplastic; Animals; Chromatography, Affinity; Erythropoietin; Hemagglutination Inhibition Tests; Hemagglutination Tests; Humans; Immunoelectrophoresis, Two-Dimensional; Lectins; Mice; Microspheres; Rabbits; Radioimmunoassay; Wheat Germ Agglutinins

In order to study the changes in erythroid precursor cells in primary acquired and secondary sideroblastic anemia, bone marrow cells from 4 patients with primary acquired sideroblastic anemia (PASA) and 3 patients with refractory anemia with excess of myeloblasts (RAEM) or erythroleukemia associated with an excess of ringed sideroblasts were cultured for erythroid colony-forming units (CFU-E). The number of CFU-E was markedly decreased in all 7 cases, and erythroid colonies formed consisted exclusively of normal-appearing erythroblasts, while ringed sideroblasts were observed in scattered single erythroblasts or in small aggregates of erythroblasts in primary as well as in secondary sideroblastic anemia. These findings may indicate the presence of 2 populations of erythroid progenitor cells in the bone marrow of patients with primary acquired and secondary sideroblastic anemia. A slight to moderate decrease in granulocyte-macrophage colony-forming units (GM-CFU) was observed in 3 cases of PASA. The decrease in GM-CFU, however, was marked in sideroblastic anemia associated with RAEM or erythroleukemia.

Topics: Aged; Anemia, Aplastic; Anemia, Sideroblastic; Colony-Forming Units Assay; Erythroblasts; Erythropoiesis; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Iron; Leukemia, Erythroblastic, Acute; Male; Middle Aged

The capability of chronic beta 2-adrenoceptor activation to effectively stimulate erythrocyte production in vivo was investigated in mice which had been treated with the hematopoiesis inhibiting agent busulfan. A relatively low dose (5 mg/kg i.p.) of busulfan produced moderate depression of erythropoiesis 10 days after a single injection, as determined by 59Fe-incorporation into erythrocytes. Administration of albuterol (1 mg/kg s.c. twice daily), a selective beta 2-adrenergic agonist, significantly enhanced erythropoiesis for 5--10 days after the injection of busulfan. In a long-term study with albuterol at the same dose a significant increase in hematocrit values as well as in the circulating erythrocyte mass was found in busulfan (5 mg/kg i.p. weekly) treated mice when compared to saline-busulfan treated control mice. Simultaneous injections of the beta-adrenergic blocking agent propanolol (4 mg/kg i.p.) diminished the effect of albuterol on erythropoiesis. Albuterol at a lower dose (0.1 mg/kg) had no significant effect on erythrocyte mass. In view of recent findings, which have shown that the proliferation of the pluripotent hemopoietic stem cell pool is blocked by busulfan, it is concluded that the main site of beta 2-adrenergic action on erythropoiesis is on the erythroid committed stem cell pool. In addition, enhanced release of erythropoietin from the kidney following the application of albuterol may contribute to beta 2-adrenergic stimulation of erythropoiesis.

Topics: Albuterol; Anemia, Aplastic; Animals; Busulfan; Erythrocyte Indices; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Iron; Kidney; Mice

Topics: Adult; Anemia, Aplastic; Bone Marrow; Erythropoietin; Female; Heme; Humans; In Vitro Techniques; Pregnancy; Pregnancy Complications, Hematologic

Erythropoietin measurements were performed in 2 patients with refractory hypoplastic anaemia who required regular blood transfusion. Erythropoietin levels varied inversely with the haemoglobin level consistent with the accepted role of erythropoietin in reflecting the adequacy of tissue oxygenation. A particularly step rise in erythropoietin occurred when the haemoglobin concentration fell below 8.0 g/dl. This coincided with the deterioration in clinical symptoms necessitating imminent admission for transfusion. Change from a regime involving a 5--6 unit transfusion every 6 weeks to that involving a regular 2 unit per 2 weeks transfusion resulted in a lower and more stable pattern of erythropoietin levels and was also associated with a considerable improvement in general well being.

Topics: Aged; Anemia, Aplastic; Blood Transfusion; Erythropoietin; Female; Hemoglobins; Humans; Middle Aged; Time Factors

Urinary proteins from patients with iron deficiency anemia and acquired aplastic anemia were fractionated by chromatography on QAE-Sephadex A-50 and Sephadex G-25. Fractions containing erythroid burst promoting activity (code named regulatory protein RP) were separated from erythropoietin. Mouse bone marrow cells were preincubated for one day in suspension culture, in the presence or absence of RP, transferred to a methylcellulose based system and incubated for six more days with erythropoietin (EPO). It was found that the presence of RP in the preincubation medium had a 2 to 4 fold enhancing effect on subsequent erythroid burst colony formation. However, when RP was added to methylcellulose based cultures simultaneously with EPO, the erythroid burst response was reduced or abolished. Addition of RP to marrow cell suspension cultures increased the number of self replicating, pluripotent (erythroid/granuloid, E/G ratio = 3) spleen colony forming units (CFU-S) found at the end of 2 days incubation 3-5 fold over their number in control cultures incubated without the factor. In marked contrast to this, addition of EPO to the cultures caused an increased persistence of CFU-S with a predominantly erythroid commitment (E/G ratio = 19) and a low self replicating ability, as measured by retransplantation of spleen cells into secondary recipients. These observations are compatible with the presence in RP of a factor, or factors, capable of maintaining the size of certain early precursor cell compartments.

Topics: Anemia, Aplastic; Anemia, Hypochromic; Animals; Cells, Cultured; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Humans; Male; Mice; Mice, Inbred C3H; Mice, Inbred DBA; Proteinuria; Spleen

In the last decade a large increase of our basic understanding concerning erythropoietin and the regulation of erythropoiesis has led to improved methods for the cell culture of human bone marrow cells. These culture methods in turn have been applied to bone marrow failures with a remarkable increase in our knowledge of the pathogenesis of some of these conditions, particularly the aplasias. The pathogenesis of pure red cell aplasia has been elucidated, and 60% of these patients have been shown to respond to cytotoxic, immunosuppressive treatment. Bone marrow transplantation has proved to be very helpful in the treatment of aplastic anemia and has provided impetus for increased knowledge concerning the pathogenesis of the aplasia. Some of these patients may have suppression of marrow hematopoiesis by the marrow T-cells and can be successfully treated with antilymphocyte globulin or high-dose prednisolone. The future looks bright for further clinical advances concerning the bone marrow failures, but more must be learned about the pathogenesis of these anemias if improved methods of treatment are to be developed.

Topics: Aged; Anemia, Aplastic; Anemia, Sideroblastic; Animals; Bone Marrow; Cyclophosphamide; Erythropoiesis; Erythropoietin; Female; Humans; Pancytopenia; Prednisone; Primary Myelofibrosis

The development since 1966 of a technology for growing stem cells in vitro has provided new insights into the controls of blood cell production. Hematopoietic hormones have been purified and important cellular interactions in hematopoiesis have been defined.

Topics: Anemia, Aplastic; Cell Communication; Erythrocytes, Abnormal; Erythropoietin; Forecasting; Granulocytes; Growth; Hematopoiesis; Hematopoietic Stem Cells; Leukemia, Myeloid, Acute; Models, Biological; Polycythemia; Preleukemia; Syndrome; Technology

Nine glycoprotein-protein fractions prepared of erythropoietin-active crude urinary protein and separated by gel filtration on Sephadex G-50, ion exchange chromatography on Sephadex DEAE A-50, gel filtration on Sephacryl S-200 and Sepharose 6B were investigated by analytical isotachophoresis. 7 to 38 components could be separated of every chromatographic fraction investigated using chloride as leading and glycinate as terminating electrolyte. Improvement of resolution is possible by variation of buffer system conditions.

Topics: Anemia, Aplastic; Chromatography, Gel; Chromatography, Ion Exchange; Electrophoresis; Erythropoietin; Glycoproteins; Humans; Proteins

The level of erythropoietin (Ep) was measured in sera and urine from aplastic anemia patients. Increased levels of Ep were demonstrated in sera from all 25 patients studied. An elevated level of Ep was found in the urine of 17 of 23 patients in whom the urine was tested. No correlation between blood hemoglobin and Ep level was observed. A higher serum Ep level was noted in patients with aplastic anemia than in patients with sideropenic anemia of the same severity. To explain the discrepancy diminished Ep consumption in bone marrow of aplastic anemia patients is discussed.

Topics: Anemia, Aplastic; Anemia, Hypochromic; Animals; Erythropoietin; Female; Hemoglobins; Humans; Mice

A case of adult pure red cell aplasia (PRCA) with a serum IgG inhibitor to erythropoiesis and an IgG lambda M component is presented. The study of lymphocyte populations revealed a slight but definite decrease of E and EA rosettes, with dissociation between E rosettes and PHA blastic transformation of blood lymphocytes and increase of membrane IgM-bearing lymphocytes. The relationship between PRCA and paraproteinemia is discussed: it is suggested that the serum M component may derive from an immunological unbalance between T and B lymphocytes. Since a survey of the literature reveals 5 similar cases, it is suggested that paraproteinemia may be the hallmark of a particular variety of chronic PRCA 'type 1'.

Topics: Aged; Anemia, Aplastic; Autoimmune Diseases; Erythropoietin; Female; Fluorescent Antibody Technique; Humans; Immunoglobulin G; Lymphocytes; Paraproteinemias; Receptors, Antigen, B-Cell; Rosette Formation

Topics: Anemia, Aplastic; Cell Differentiation; Cell Division; Cells, Cultured; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; T-Lymphocytes

Topics: Anemia, Aplastic; B-Lymphocytes; Bone Marrow; Cells, Cultured; Cytotoxicity, Immunologic; Erythropoiesis; Erythropoietin; Hemoglobins; Humans; Lymphocyte Activation; Lymphocytes; T-Lymphocytes

To explore the etiology of congenital hypoplastic anemia (CHA) or the Diamond-Blackfan anemia, erythropoietin responsive committed erythroid precursors were enumerated by the plasma clot method. These included blood and marrow erythroid burst-forming units (BFU-E) and marrow erythroid colony-forming units (CFU-E). The peripheral blood nucleated cells of 11 patients and the marrow cells of seven of these patients were examined. Studies were repeated in several patients during relapse and after induction of remission. BFU-E were undetectable in the marrow and blood of all but one relapsed patient, and the numbers of marrow CFU-E were depressed in all relapsed patients. Blood BFU-E remained low in all of the patients in remission. No evidence was obtained for suppression of normal CFU-E or BFU-E by CHA lymphocytes. Erythropoietin dose-response curves performed in two patients revealed a 10-fold increase in erythropoietin requirement for marrow CFU-E colony growth. This marked unresponsiveness to erythropoietin was strikingly improved by steroid therapy in one patient. We suggest that CHA is the result of a qualitative and/or quantitative deficiency of BFU-E. If BFU-E are produced, they must be relatively unresponsive to erythropoietin. The abnormal BFU-E give rise to erythropoietin unresponsive CFU-E and, thence, to proerythroblasts that are, in turn, trapped in that early stage of development because of their poor erythropoietic response. Hence, red cell production is deficient. Steroids appear to improve the erythropoietin response of CHA erythroid precursors.

Topics: Adolescent; Adult; Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Child; Child, Preschool; Erythrocytes; Erythropoietin; Female; Humans; Infant

Topics: ABO Blood-Group System; Adolescent; Adult; Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Cell Count; Cell Differentiation; Cells, Cultured; Child; Erythropoietin; Female; Humans; Liver Diseases; Liver Function Tests; Male; Thymidine; Time Factors

Human urinary erythropoietic-active crude protein was isotachophoretic fractionated in a LKB Uniphor apparatus equipped with a plastic column using polyacrylamide as a stabilizing medium, phosphat ions as leading and glycinat ions as terminating electrolytes and Ampholine carrier ampholytes (pH: 5--7) as spacers. The exhypoxic polycythemic mouse method to assay erythropoietic activity showed that the majority of the hormon was sharply resolved. Under these conditions preparative isotachophoresis demonstrated a higher purification factor compared to other biochemical preparation-methods.

Topics: Anemia, Aplastic; Anemia, Hypochromic; Electrophoresis; Erythropoietin; Humans

A teenage boy with mixed-cellularity Hodgkin disease presented with severe anemia secondary to pure red cell aplasia of marrow without evidence of lymphomatous infiltration or hemolysis. In vitro studies of the patient's serum demonstrated an inhibitor of erythropoietin activity which appeared to be an IgG but which did not directly bind erythropoietin. The patient's anemia resolved and the inhibitor disappeared following chemotherapy for Hodgkin disease. Presumably, the inhibitor was directed at a very early stage of red blood cell production. This phenomenon may be related to other autoimmune manifestations occasionally seen in patients with lymphomas. The case is presented to bring attention to the unusual occurrence of red cell aplasia in Hodgkin disease. Several hypotheses concerning significance and etiology of the anemia are detailed.

Topics: Adolescent; Anemia, Aplastic; Binding Sites, Antibody; Bone Marrow; Erythropoiesis; Erythropoietin; Hemoglobins; Hodgkin Disease; Humans; Male

A summary of the historical developments associated with congenital anemias are discussed. The clinical status, the peripheral blood and bone marrow picture of these patients are reviewed. The effectiveness of treatment and prognosis for these patients is evaluated.

Topics: Adrenal Cortex Hormones; Adult; Anemia; Anemia, Aplastic; Blood Transfusion; Bone Marrow; Erythropoietin; Hemosiderosis; Humans; Syndrome

A 29-year-old Chinese male developed severe aregenerative anemia. The bone marrow was diffusely hypercellular with increased marrow reticulin and a persistent failure of erythroid differentiation beyond the pronormoblast stage. Although he did not manifest classic features of systemic lupus erythematosus, multiple serologic studies were in accord with this diagnosis. The patient's defect in erythropoiesis was studied by an in vitro technique for the growth of erythroid colonies. Despite the severe erythroid hypoplasia, the patient's marrow yielded abundant large erythroid colonies. Serum erythropoietin activity was high as judged by use of this in vitro assay. Although the patient's native serum did not affect colony formation, a separated IgG fraction was markedly inhibitory to colony growth. This suggests that the erythroid hypoplasia may have resulted from a unique autoantibody. The patient's hematologic abnormalities completely reversed following treatment with corticosteroids.

Topics: Adrenal Cortex Hormones; Adult; Anemia, Aplastic; Bone Marrow; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Male; Reticulin

Human erythropoietin, derived from urine of patients with aplastic anemia, has been purified to apparent homogeneity. The seven-step procedure, which included ion exchange chromatography, ethanol precipitation, gel filtration, and adsorption chromatography, yielded a preparation with a potency of 70,400 units/mg of protein in 21% yield. This represents a purification factor of 930. The purified hormone has a single electrophoretic component in polyacrylamide gels at pH 9, in the presence of sodium dodecylsulfate at pH 7, and in the presence of Triton X-100 at pH 6. Two fractions of the same potency and molecular size, by sodium dodecyl sulfate gel electrophoresis, but differing slightly in mobility at pH 9, were obtained at the last step of fractionation. The nature of the difference between these two components is not yet understood.

Topics: Anemia, Aplastic; Electrophoresis, Polyacrylamide Gel; Erythropoietin; Humans; Hydrogen-Ion Concentration; Molecular Weight

The responsivenes to erythropoietin of cultured bone marrow cells, obtained from 7 patients with refractory anemia with hyperplastic marrow, was studied. 5 of these patients' marrows also exhibited sideroblasti changes. Heme synthesis in cultured bone marow cells was either responsive to stimulation by erythropoietin, or completely refractory. The sensitivity of the bone marrow cells to the hormone was not related to either the clinical or laboratory findings.

Topics: Aged; Anemia, Aplastic; Anemia, Sideroblastic; Cells, Cultured; Erythropoiesis; Erythropoietin; Female; Heme; Humans; Male; Middle Aged

The effect of erythroid aplasia on the clearance rate of exogenous erythropoietin from the circulation has been studied in irradiated (200, 400 and 800 r) and cyclophosphamide (50 mg/kg) treated rats. After irradiation, the T1/2 was increased from 1.5 h in controls to 2.3 h. In spite of similar aplasia, the T1/2 observed after cyclophosphamide administration was identical to the T1/2 in controls. These results suggest that the action of X-rays on the catabolism of erythropoietin is independent of erythroid aplasia and could be related to an extramedullary effect of irradiation.

Topics: Anemia, Aplastic; Animals; Bone Marrow; Bone Marrow Cells; Cyclophosphamide; Erythropoietin; Half-Life; Male; Radiation Effects; Rats

Topics: Adolescent; Aged; Anabolic Agents; Anemia, Aplastic; Animals; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Male; Mice

The concentration and erythropoietin dependence of erythropoietic progenitor cells (CFU-E) were examined in 13 patients with aplastic anaemia at different stages of their disease. The CFU-E incidence was shown to be quantitatively diminished in aplastic anaemia but tended to recover to normal values if the disease recovered. In addition the CFU-E showed a qualitatively different response to stimulation by erythropoietin, being resistant to low concentrations but responsive to concentrations greater than 0.2 U/ml whereas there was a linear response in the controls up to 0.5 U/ml.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Bone Marrow; Clone Cells; Dose-Response Relationship, Drug; Erythropoiesis; Erythropoietin; Female; Humans; In Vitro Techniques; Male; Middle Aged; Stimulation, Chemical

The results of the erytropoietin level determination in serum and urine of patients with congenital anemias are presented and compared to the results obtained in children with acute aplastic anemias. Three patients with congenital hypoplastic anemia Diamond-Blackfan, two with Fanconi's anemia, one with congenital pancythopenia with hyperplastic marrow and five patients with acute aplastic anemia were studied. The increased serum erythropoietin level was found in every patient whose blood hemoglobin was less than 12g%. Erythropoietin was detected in nonconcentrated urine when serum erythropoietin level was higher than 0,5 units/ml. The statistically significant negative correlation between the serum erythropoietin level and blood hemoglobin concentration was found. In two patients suffering from congenital anemias, in whome the significantly increased erythropoietin level (about 1.0 units/ml) was detected, increased ammount of hemoglobin F in peripheral blood as well as increased MCV--signs of so called "stress" erythropoiesis-were noted. The results presented, together with the results obtained by other authors, indicate that congenital anemias studied here are not due to the disturbance in erythropoietin production.

Topics: Adolescent; Adult; Anemia, Aplastic; Child; Child, Preschool; Erythrocytes, Abnormal; Erythropoietin; Fanconi Anemia; Female; Humans; Infant; Male; Syndrome

Marver & Gurney ((1968)Ann.N.Y.Acad.Sci. 149, 570-575) reported that the erythropoietin excretion in two anaemic patients increased 17 and 3.5-fold, respectively, during the production of alkaline urine by the administration of NaHCO3. We have studied the excretion of erythropoietin in 6 persons, following the daily administration of 3.5 g of NH4Cl, which resulted in an average urine pH of 5.9; and following the administration of hexapotassium hexasodium pentacitrate, which increased the urine pH TO 7.5. In the alkaline urine, erythropoietin excretion showed an average, but non-significant, two-fold increase. The increase was significant in only two experimental persons.

Topics: Adult; Aged; Ammonium Chloride; Anemia, Aplastic; Citrates; Erythropoietin; Female; Humans; Hydrogen-Ion Concentration; Kidney Diseases; Male

The combination of aplastic anaemia and thymona was observed in a 49-year old woman. Concomitant with severe normochromic anaemia and reticulocytopenia was a cell-rich bone marrow smear in which erythropoietic precursors were almost absent. Immunological phenomena could not be demonstrated. The erythropoietin level was maximally increased. Pathological anatomy typically revealed a predominantly spindle-cell thymonal with only minor capsular invasion.

Topics: Anemia, Aplastic; Bone Marrow Examination; Erythrocyte Count; Erythropoietin; Female; Humans; Middle Aged; Reticulocytes; Thymoma; Thymus Neoplasms

An erythropoietic inhibitor was detected in the serum of a patient with refractory anemia. Using an in vitro heme synthesis method, the patient's serum produced tenfold inhibition of erythropoietin-stimulated radioactive iron (Fe59) incorporation into heme of normal human marrow at 72 hours, as compared with AB serum. In a separate experiment the patient's serum produced threefold inhibition, whereas immunoglobulin G (IgG) prepared from the same serum sample produced 12-fold inhibition. To identify the site of action of the inhibitor, serum was tested in a cell culture system whereby human marrow cells, grown in a plasma clot, respond to exogenous erythropoietin with the appearance of nucleated erythroid colonies. Each colony arises from a committed erythroid progenitor. The patient's serum produced a two- or tenfold reduction in the number of colonies from normal human marrow. The effect was also demonstrated on autologous marrow obtained when the patient was in "partial clinical remission". Serum samples obtained at various times during the course of the patient's illness all demonstrated a suppressive effect on colony growth. All serums were heat-inactivated, and total hemolytic complement could not be detected in either culture system. It is concluded that the anemia is due to an inhibitor, probably of IgG class, that acts on the erythroid progenitor cell. The absence of heat-labile complement components in the culture systems suggests that the mechanism is not due to immune cytolysis.

Topics: ABO Blood-Group System; Anemia, Aplastic; Cells, Cultured; Complement System Proteins; Depression, Chemical; Erythropoiesis; Erythropoietin; Female; Heme; Humans; Immunoglobulin G; Middle Aged

Peripheral blood lymphocytes from six patients with congenital hypoplastic anemia suppressed erythroid cell formation by normal human bone marrow cells in response to erythropoietin in vitro. The results suggest that the anemia in these children has an immunologic basis.

Topics: Adrenal Cortex Hormones; Anemia, Aplastic; Animals; Bone Marrow Cells; Cell Differentiation; Cricetinae; Erythrocytes, Abnormal; Erythropoiesis; Erythropoietin; Lymphocytes

Topics: Adult; Anemia, Aplastic; Erythropoiesis; Erythropoietin; Female; Humans; Oxymetholone

Four children with congenital hypoplastic anemia (Diamond-Blackfan syndrome) and 30 control children with normal erythropoiesis were studied by a cell culture method in which human marrow, grown in a plasma clot, responds to added erythropoietin (EPO) with the appearance of discrete colonies of nucleated erythroid cells. The colonies arise from EPO-responsive stem cells and are not related to the number of morphologically identifiable marrow erythroids plated. Results of studies on control marrow indicated that without EPO there was little or no colony formation. Increasing EPO doses or nucleated marrow cells per culture resulted in a linear increase in colony numbers. The optimal EPO concentration of 2.5 U/ml yielded a mean of 158 +/- 79 colonies/1 x 10(5) nucleated cells on day 7 of incubation. Even in the absence of recognizable erythroids, marrows of all four patients with anemia grew erythroid colonies. Two patients on no therapy had decreased colony numbers. The other two, on prednisone, had normal numbers. Sera from patients did not inhibit colony formation from either autologous or control marrow. In contrast, serum from an adult with acquired pure red cell aplasia produced striking inhibition of colony growth. It appears that the red cell failure in this disorder is not due to an absence of erythroid stem cells, and a serum inhibitor to erythropoiesis as seen in the acquired disease is unlikely.

Topics: Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Cell Differentiation; Child, Preschool; Culture Techniques; Erythrocytes, Abnormal; Erythropoiesis; Erythropoietin; Humans; Infant; Infant, Newborn; Syndrome

Topics: Adenylyl Cyclases; Androgens; Anemia, Aplastic; Erythropoiesis; Erythropoietin; Heme; Humans; Kidney; RNA; Testosterone

Three cases of adult pure red cell aplasia (PRCA) ARE REPORTED. All patients proved refractory to various combinations of androgens and corticosteroids. The first case, harboring a thymoma, showed a complete clinical remission following cyclophosphamide therapy. The second and third responded similarly to either a combined cyclophosphamide + antilymphocyte globulin (ALG) treatment or to ALG administration preceded by a small dosage of cyclophosphamide, which had proved ineffective when administered alone. Serum IgG inhibitors to erythropoiesis were demonstrated in all cases by means of in vivo and/or in vitro techniques. The inhibitor(s), although directed against the erythroid marrow in both the first and third patients (PRCA type A), apparently functioned as an antibody to circulating erythropoientin (Ep) in the second case (PRCA type B). The inhibitor(s) was always absent in postremission samples. Additionally, experimental models for both types of human PRCA were established in normal rodents. The present studies support the contention that adult PRCA is an autoimmune disease. The therapeutic role of cytotoxic-immunodepressive agents in PRCA patients is confirmed. It is emphasized that ALG may represent an additional therapeutic tool in cases resistant to cyclophosphamide and/or steroids. In addition, cyclophosphamide proved effective in a patient harboring a thymoma not amenable to surgery. Finally, it is postulated that IgG serum autoantibodies, directed against either an early erythroid precursor (PRCA type A) or, more rarely, circulating Ep (PRCA type B), play a major role in the pathogenesis of the disease.

Topics: Aged; Anemia, Aplastic; Antibodies, Antinuclear; Antilymphocyte Serum; Autoantibodies; Blood Transfusion; Bone Marrow Examination; Cyclophosphamide; Dexamethasone; Diabetic Coma; Erythrocytes; Erythropoietin; Female; Heart Failure; Hematocrit; Hepatitis; Humans; Immunoglobulin G; Iron; Male; Middle Aged; Radiography, Thoracic; Sarcoma, Kaposi; Serum Globulins; Skin Manifestations; Thymoma

The serum from five children with congenital pure red cell aplasia demonstrated no erythropoietic inhibitory effect either in vivo in animals or in vitro. The significance of immunosuppressive therapy in this disease is discussed.

Topics: Anemia, Aplastic; Animals; Antibodies; Blood Proteins; Bone Marrow; Bone Marrow Cells; Erythropoiesis; Erythropoietin; Female; Goats; Heme; Humans; Infant; Iron Radioisotopes; Male; Mice; Neutralization Tests; Rabbits; Rats

Two experimental models for pure red cell aplasia (PRCA) were established. In the first one, administration of PRCA serum IgG in normal mice induced a sustained inhibitory effect on erythropoiesis, a progressive decline of the hematocrit values and an inverse rise of erythropoietin (Ep) levels in serum. Thus, the physiopathological pattern of PRCA type I (or A) was established, In the second model a rabbit producing anti-Ep crossreacting with endogenous Ep was subjected to a booster injection of Ep. The rise of the immune response was associated with decrease of Gct values and disappearance of erythroid precursors from marrow smears, and its subsequent decline with reticulocytosis and regression of the anemia, thus reproducing the physiopathological pattern of PRCA type II (or B).

Topics: Anemia, Aplastic; Animals; Autoantibodies; Autoimmune Diseases; Chronic Disease; Disease Models, Animal; Erythropoietin; Immune Sera; Immunoglobulin G; Mice; Rabbits

An inhibitor of erythropoiesis was found in the urine of patients with aplastic anemia. The inhibitor was concentrated with kaolin and separated from erythropoietin by subsequent fractionation with alcohol and gel filtration on Sephadex G-100 and DEAE-Sephadex A-50. The inhibitor blocked the effect of erythropoietin (standard C) when injected into mice with hypoxic polycythemia 3 H before or along with exogenous erythropoietin; partial neurtralization of standard C also was observed on incubation with the inhibitor.

Topics: Anemia, Aplastic; Chromatography, Gel; Erythropoiesis; Erythropoietin; Humans; Proteins

The immunochemical assay (HAI) for erythropoietin is accurate enough to detect small quantities and sensitive enough to demonstrate consistently and quantitatively the presence of erythropoietin in serum and urine. It will be the purpose of this report to describe in detail the procedure for the assay, the usefulness and value of erythropoietin testing in the clinical laboratory. The relationship of erythropoietin levels in serum and urine in the classification of diseases of the red blood cells are discussed. There is presently no other method available for the measurement of erythropoietin that permits the detection of extremely small differences, sensitive enough to demonstrate consistently and quantitatively the presence of small amounts of erythropoietin.

Topics: Anemia, Aplastic; Blood Transfusion; Erythropoietin; Hemagglutination Inhibition Tests; Hematocrit; Humans; Polycythemia; Polycythemia Vera; Urologic Diseases

An 18-yr-old female with chronic active hepatitis developed a severe anemia due to a lack of red cell production. Her bone marrow showed many large proerythroblasts but an almost complete lack of more mature erythroblasts. Incubation of the marrow cells in a normal medium with erythropoietin concentrate led to increased erythropoiesis as indicated by the development of mature erythroblasts as well as a ninefold increase in hemoglobin synthesis. The patient's plasma was cytotoxic for erythroblasts. Following splenectomy, a remission of the disease occurred. This study indicates that in some cases the anemia associated with abundant marrow proerythroblasts and the absence of mature erythroblasts has the same pathogenesis as pure red cell aplasia and that splenectomy may be beneficial when there is a lack of response to immunosuppressive drugs.

Topics: Adolescent; Anemia, Aplastic; Autoantibodies; Bone Marrow Cells; Bone Marrow Examination; Cells, Cultured; Cyclophosphamide; Cytotoxicity Tests, Immunologic; Erythroblasts; Erythrocytes; Erythropoietin; Female; Hemoglobins; Hemosiderosis; Hepatitis B; Humans; Immunosuppression Therapy; Iron; Iron Radioisotopes; Karyotyping; Liver Function Tests; Prednisone; Splenectomy; Transfusion Reaction

Topics: Anemia, Aplastic; Erythropoiesis; Erythropoietin; Humans; Polycythemia

Topics: Anemia, Aplastic; Anemia, Hypochromic; Erythropoietin; Humans; Polycythemia Vera

The response of bone marrow to erythropoietin (EPO) from five children with the Diamond-Blackfan syndrome, also known as congenital hypoplastic anaemia (CHA), was tested in tissue culture by measurement of haem synthesis. Studies of 13 control marrows indicated that the maximum EPO effect occurred at approximately 70 h incubation using an EPO concentration of 0.2-0.3 units/ml and a nucleated cell concentration of 5 x 10(6) per culture. Under these conditions, haem synthesis was 121% greater in EPO-stimulated than in unstimulated cultures. Patients with CHA with anaemia and diminished marrow erythroids had reduced or absent haem synthesis. In one patient, haem production became normal after a spontaneous remission, and was not inhibited by autologous plasma drawn at the time of diagnosis. Plasma from three patients did not show inhibitory activity when cultured with control marrow. In contrast, plasma from an adult with acquired pure red cell aplasia produced striking inhibition of haem synthesis when cultured with control marrow. We conclude that, in comparison to some cases of the adult acquired condition, CHA is not due to inhibitors or antibodies. When present, erythroid precursors in children with CHA are capable of responding normally to EPO with increased haem synthesis.

Topics: Anemia, Aplastic; Bone Marrow; Child, Preschool; Dose-Response Relationship, Drug; Erythrocytes, Abnormal; Erythropoietin; Female; Heme; Humans; Male; Syndrome

Topics: Aged; Anabolic Agents; Anemia, Aplastic; Erythropoietin; Hematopoietic Stem Cells; Humans; Male; Nephrotic Syndrome

Topics: Anemia, Aplastic; Animals; Antibodies; Bone Marrow; Bone Marrow Cells; Bone Marrow Examination; Erythropoiesis; Erythropoietin; Female; Fetus; gamma-Globulins; Goats; Humans; Immune Sera; Immunity, Maternally-Acquired; Iron; Iron Radioisotopes; Liver; Mice; Polycythemia; Rabbits; Reticulocytes; Sheep; Spleen

Topics: Anemia, Aplastic; Animals; Antibodies; Autoradiography; Cell Differentiation; Cell Division; Cells, Cultured; Centrifugation; Dialysis; DNA; Erythrocytes, Abnormal; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Microscopy, Electron; Rabbits; Staining and Labeling; Stimulation, Chemical; Syndrome; Thymidine; Tritium

Topics: Anemia, Aplastic; Anemia, Hemolytic; Animals; Antibodies; Antibody Formation; Chromatography; Erythropoietin; Humans; Immune Sera; Immunodiffusion; Rabbits; Radioimmunoassay; Rats

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Blood Cell Count; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Erythrocytes, Abnormal; Erythropoietin; Female; Heme; Humans; Iron; Iron Radioisotopes; Male; Middle Aged; Stimulation, Chemical

Topics: Adenosine Triphosphate; Anemia, Aplastic; Anemia, Sideroblastic; Aspartate Aminotransferases; Blood Transfusion; Carbon Dioxide; Diphosphoglyceric Acids; Electrophoresis, Starch Gel; Erythrocyte Count; Erythrocytes; Erythropoietin; Fructose-Bisphosphate Aldolase; Glutathione; Glyceraldehyde-3-Phosphate Dehydrogenases; Glycolysis; Hematocrit; Hemoglobins; Hexokinase; Humans; Hydrogen-Ion Concentration; Lactates; Methemoglobin; Oxygen Consumption; Phosphofructokinase-1; Phosphorus; Potassium; Reticulocytes; Sodium; Thalassemia

Topics: Anemia, Aplastic; Animals; Chromatography, DEAE-Cellulose; Chromatography, Gel; Erythropoiesis; Erythropoietin; Humans; Injections, Subcutaneous; Mice; Polycythemia; Time Factors

Topics: Agranulocytosis; Anemia, Aplastic; Anemia, Macrocytic; Bone Marrow Examination; Bone Neoplasms; Breast Neoplasms; Dihydrotestosterone; Erythropoiesis; Erythropoietin; Female; Hepatitis; Humans; Iron; Iron Radioisotopes; Male; Oxymetholone; Prednisolone; Sepsis; Transfusion Reaction

Topics: Anemia, Aplastic; Anemia, Sideroblastic; Animals; Biological Assay; Circadian Rhythm; Computers; Dialysis; Erythropoietin; Evaluation Studies as Topic; Female; Freeze Drying; Hemagglutination Tests; Hemoglobinuria, Paroxysmal; Humans; Iron Radioisotopes; Male; Methods; Mice; Mice, Inbred Strains; Microchemistry; Oxygen Consumption; Thalassemia; Ultrafiltration

The marrow cells of a patient with pure red cell aplasia markedly increased their rate of heme synthesis when they were freed from the host environment and were incubated in vitro. When the red cell aplasia was treated with cyclophosphamide and prednisone, marrow cell incorporation of (59)Fe into heme in vitro increased several weeks before a reticulocytosis was apparent, and was the earliest effect noted. The plasma gammaG-globulins of this patient inhibited heme synthesis by normal marrow cells or the patient's own marrow cells obtained after remission of the disease. Since the inhibition of heme synthesis could be the result of damage to erythroblasts, the patient's posttreatment marrow cells or normal marrow cells were labeled with (59)Fe and were then incubated with the patient's pretreatment, treatment, and posttreatment gammaG-globulins as well as normal gammaG-globulins. At the end of this incubation the supernatant and cells were separated and counted. Heme was extracted and also was counted. Treatment of the cells with the patient's pretreatment gammaG-globulins resulted in a release of 40% of the radioactive heme from the cells. This represented the loss of radioactive hemoglobin and was an index of erythroblast cytotoxicity. A progressive disappearance of the cytotoxic factor in the gammaG-globulins occurred in the 3 wk period preceding the onset of reticulocytes in the patient's blood. Posttreatment and normal gammaG-globulins did not produce this effect and increased injury of red cells and lymphocytes was not produced by the patient's pretreatment gammaG-globulins. These studies demonstrate a method for measuring erythroblast cytoxicity and show that red cell aplasia is associated with gammaG-globulins that specifically damage erythroblasts. Whether interference with new erythroblast development also occurs and contributes to the inhibition of heme synthesis has not yet been ascertained.

Topics: Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Cyclophosphamide; Erythrocytes, Abnormal; Erythropoietin; gamma-Globulins; Heme; Hemoglobins; Humans; Iron Isotopes; Male; Middle Aged; Prednisone

Topics: Acetates; Anemia, Aplastic; Blood Transfusion; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Chromatography, DEAE-Cellulose; Chromium Isotopes; Complement Inactivator Proteins; Complement System Proteins; Cytotoxicity Tests, Immunologic; Erythrocytes; Erythropoietin; Freezing; gamma-Globulins; Glycols; Hot Temperature; Humans; Iron Isotopes; Isotope Labeling; Methods; Suramin; Zymosan

Topics: Anemia, Aplastic; Animals; Antibodies; Antibodies, Antinuclear; Biological Assay; Cyclophosphamide; Erythrocytes; Erythropoietin; Female; Humans; Male; Mice; Middle Aged; Muscle, Smooth; Prednisone

Topics: Anemia, Aplastic; Blood Coagulation; Blood Transfusion; Erythropoiesis; Erythropoietin; Female; Fluoxymesterone; Humans; Iron; Iron Isotopes; Kidney; Kidney Failure, Chronic; Liver; Liver Function Tests; Long-Term Care; Male; Prothrombin Time; Warfarin

Topics: Anemia, Aplastic; Erythropoietin; Humans; Methods

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Sickle Cell; Bone Marrow Diseases; Creatinine; Erythropoietin; Female; Hematocrit; Hemolysis; Humans; Iron; Leukemia; Lymphoma; Male; Middle Aged; Multiple Myeloma; Sex Factors

Topics: Adult; Anemia, Aplastic; Blood Cell Count; Blood Cells; Chronic Disease; Erythropoiesis; Erythropoietin; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; Renal Dialysis; Reticulocytes; Uremia

Topics: Anemia, Aplastic; Erythropoiesis; Erythropoietin; Humans

Topics: Anemia, Aplastic; Erythropoietin; Humans; Male; Middle Aged; Thymectomy; Thymoma; Thymus Neoplasms

Topics: Adult; Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Erythropoietin; Female; Humans; Methenolone

Topics: Acute Disease; Anemia, Aplastic; Child; Erythropoietin; Hemoglobins; Humans; Leukemia

Topics: Anemia, Aplastic; Animals; Erythrocytes; Erythropoiesis; Erythropoietin; Humans; Kinetics; Mice; Mitosis; Rabbits

Topics: Anemia, Aplastic; Animals; Antibodies; Antilymphocyte Serum; Bone Marrow Cells; Cells, Cultured; Chromatography, DEAE-Cellulose; Cyclophosphamide; Cytotoxicity Tests, Immunologic; Erythropoiesis; Erythropoietin; Female; Fluorescent Antibody Technique; gamma-Globulins; Heme; Horses; Humans; Immunoelectrophoresis; Immunoglobulin G; Immunosuppression Therapy; Iron Isotopes; Leukocyte Count; Middle Aged; Sheep; Thymus Gland; Ultracentrifugation

Topics: Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Erythropoiesis; Erythropoietin; Humans; Leukemia, Myeloid

Topics: Anemia, Aplastic; Blood Volume; Bone Marrow; Bone Marrow Cells; Child; Chromium Isotopes; Craniopharyngioma; Dwarfism, Pituitary; Erythropoiesis; Erythropoietin; Growth Hormone; Humans; Hypophysectomy; Hypopituitarism; Iron; Kinetics; Leukocyte Count; Lymphocytes; Male; Radioimmunoassay; Testosterone; Transferrin

Topics: Adult; Anemia, Aplastic; Ceruloplasmin; Erythropoietin; Female; Humans; Male; Middle Aged

Topics: Anemia; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Hypochromic; Anemia, Macrocytic; Bilirubin; Erythropoietin; Humans; Iron; Neoplasms

Topics: Adult; Aged; Alkaline Phosphatase; Anemia, Aplastic; Aspartate Aminotransferases; Bilirubin; Blood Transfusion; Bone Marrow Diseases; Chromium Isotopes; Erythropoiesis; Erythropoietin; Female; Hematocrit; Humans; Iron; Iron Isotopes; Lymphoma; Male; Middle Aged; Multiple Myeloma; Oxymetholone; Primary Myelofibrosis

Topics: Anemia, Aplastic; Animals; Blood Cell Count; Busulfan; Cell Differentiation; Erythropoiesis; Erythropoietin; Female; Hematopoiesis; Hematopoietic Stem Cells; Iron Isotopes; Leukocytes; Mice; Radiation Chimera; Spleen; Thymidine; Tritium

Topics: Anemia; Anemia, Aplastic; Animals; Blood; Blood Cells; Busulfan; Cell Differentiation; Cell Division; Chromatography, Gel; Erythropoiesis; Erythropoietin; Female; Humans; Hyperplasia; Iron; Iron Isotopes; Mice; Neuraminidase; Phenylhydrazines; Polycythemia; Sheep

Topics: Adult; Anemia, Aplastic; Animals; Biological Assay; Blood Transfusion; Child; Chromatography; Chromatography, Gel; Erythropoiesis; Erythropoietin; Female; Growth Inhibitors; Humans; Infant; Iron Isotopes; Male; Mice; Molecular Weight; Polycythemia; Pregnancy; Spectrophotometry; Urine

Topics: Anemia, Aplastic; Animals; Black People; Blood Proteins; Chromatography; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Humans; Immunodiffusion; Immunoelectrophoresis; Immunoglobulin G; Iron Isotopes; Mice; Polycythemia; Rodent Diseases; Thymectomy; Thymoma

Topics: Anemia; Anemia, Aplastic; Anemia, Sickle Cell; Animals; Biological Assay; Breast Neoplasms; Cachexia; Erythrocytes; Erythropoiesis; Erythropoietin; Iron; Iron Isotopes; Leukemia; Lymphoma; Male; Mice; Plasma; Polycythemia; Prostatic Neoplasms

Topics: Anemia, Aplastic; Erythropoiesis; Erythropoietin; Humans

Topics: Anemia; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Hypochromic; Anemia, Macrocytic; Diagnosis, Differential; Erythrocytes; Erythropoietin; Half-Life; Hemorrhage; Humans; Iron; Neoplasms; Vitamin B 12 Deficiency

Topics: Adult; Aged; Anemia; Anemia, Aplastic; Anemia, Hypochromic; Anemia, Macrocytic; Animals; Bone Marrow; Bone Marrow Diseases; Daunorubicin; Erythropoietin; Female; Hemoglobins; Humans; Male; Middle Aged; Phenylhydrazines; Rabbits; Radiation Effects

Topics: Anemia; Anemia, Aplastic; Animals; Erythropoiesis; Erythropoietin; Humans; Iron Isotopes; Kidney Failure, Chronic; Mice; Polycythemia; Proteins; Proteinuria; Spectrophotometry

Topics: Adrenal Cortex Hormones; Anemia, Aplastic; Erythropoietin; Humans; Lectins; Pancreatectomy

Topics: Anemia, Aplastic; Anemia, Hemolytic; Animals; Bone Marrow; Erythrocytes; Erythropoietin; Female; Humans; Male; Mice; Phenylhydrazines; Rabbits

The quantitative relationship between red cell volume, erythropoietin level, and erythropoiesis was evaluated in 43 human beings. Results in normal man were compared with studies in patients with anemia from bone marrow failure and with polycythemia vera. The maximum erythropoietin excretion after bleeding normal men was similar to the basal levels found in patients with chronic anemia of similar magnitude. Although erythropoietin values were low in patients with polycythemia vera, bleeding evoked a normal response. In patients anemic from bone marrow failure, basal levels were elevated, and phlebotomy resulted in an increase consistent with the new level of anemia. These observations indicate that erythropoietin level is affected primarily by the degree of anemia and is not influenced by the duration of anemia. In normal subjects, a fivefold increase in urinary erythropoietin was associated with a doubling of erythropoiesis. Despite similar degrees of erythropoietin production, anemic patients with evidence of bone marrow in the lower extremities had greater red cell production. In patients with polycythemia vera, red cell production was inappropriately elevated with regard to the urinary erythropoietin excretion. Bone marrow maturation time was not shortened in patients anemic from bone marrow failure to the same degree as in bled, normal volunteers. In addition to an adequate level of erythropoietin production, normal bone marrow function is necessary for maximal shortening of maturation time.

Topics: Adult; Aged; Anemia, Aplastic; Bloodletting; Bone Marrow; Erythrocyte Aging; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Hematocrit; Humans; Iron; Iron Isotopes; Leg; Male; Middle Aged; Polycythemia Vera

Topics: Anemia, Aplastic; Anemia, Hemolytic; Bone Marrow; Erythrocyte Count; Erythropoiesis; Erythropoietin; Humans; Reticulocytes

Topics: Anemia, Aplastic; Animals; Ascites; Blood Transfusion; Chronic Disease; Erythropoietin; Female; Glomerulonephritis; Humans; Hydronephrosis; Immune Sera; Iron Isotopes; Kidney Diseases; Kidney Failure, Chronic; Male; Mice; Phenacetin; Pyelonephritis; Renal Dialysis; Uremia

Topics: Anemia, Aplastic; Bone Marrow Cells; Cell Nucleus; Cellulose; Chromatography; Culture Techniques; Cyclophosphamide; Erythrocytes; Erythropoietin; Female; Fluorescent Antibody Technique; Humans; Immunoelectrophoresis; Immunoglobulin G; Immunosuppressive Agents; Iron Isotopes; Middle Aged; Transferrin

Topics: Anemia, Aplastic; Erythropoietin; Female; Humans; Kidney; Male

Topics: Adult; Aged; Anemia, Aplastic; Erythropoietin; Female; Humans; Kidney; Kidney Failure, Chronic; Male; Middle Aged; Uremia

Topics: Acetates; Anemia, Aplastic; Animals; Blood Transfusion; Bone Marrow; Bone Marrow Cells; Cell Differentiation; Cell Nucleus; Erythrocytes; Erythropoietin; Female; Hematopoiesis; Histones; Mice; Polycythemia; RNA; Spleen; Stimulation, Chemical; Time Factors; Tritium; Uridine

Topics: Anemia, Aplastic; Animals; Atmospheric Pressure; Biological Assay; Blood Cells; Bone Marrow; Bone Marrow Cells; Dactinomycin; Erythropoiesis; Erythropoietin; Hematocrit; Iron Isotopes; Methods; Mice; Nitrogen Mustard Compounds; Oxygen; Phenylhydrazines; Polycythemia; Rats; Starvation

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Sideroblastic; Bone Marrow Diseases; Erythropoietin; Female; Humans; Iron; Iron Isotopes; Kidney Failure, Chronic; Male; Middle Aged; Myeloproliferative Disorders; Oxymetholone

Topics: Acute Disease; Anemia, Aplastic; Animals; Biological Assay; Bone Marrow; Bone Marrow Cells; Bone Marrow Diseases; Erythropoiesis; Erythropoietin; gamma-Globulins; Hodgkin Disease; Humans; Iron Isotopes; Leukemia; Mediastinal Neoplasms; Mice; Osteosarcoma; Thymoma

Topics: Amniotic Fluid; Anemia, Aplastic; Animals; Biological Assay; Erythroblastosis, Fetal; Erythrocytes; Erythropoietin; Female; Fetal Diseases; Hemoglobinometry; Humans; Hypoxia; Infant, Newborn; Iron; Iron Isotopes; Maternal-Fetal Exchange; Mice; Pre-Eclampsia; Pregnancy; Pregnancy in Diabetics; Uremia

Topics: Adolescent; Anemia; Anemia, Aplastic; Anemia, Hemolytic; Anemia, Hypochromic; Anemia, Sickle Cell; Animals; Biological Assay; Blood Cell Count; Bone Marrow Examination; Child; Child, Preschool; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Hemoglobinometry; Humans; Iron; Iron Isotopes; Male; Rats; Reticulocytes; Thalassemia

Topics: Anemia; Anemia, Aplastic; Anemia, Hemolytic; Argentina; Blood Cell Count; Erythropoietin; Hemoglobinometry; Hookworm Infections; Humans; Reticulocytes; Sampling Studies

Topics: Acid-Base Equilibrium; Adolescent; Anemia; Anemia, Aplastic; Animals; Bicarbonates; Biological Assay; Erythrocytes; Erythropoietin; Humans; Hydrogen-Ion Concentration; Iron; Iron Isotopes; Kidney; Kidney Function Tests; Male; Mice; Middle Aged; Peptide Hydrolases; Primary Myelofibrosis

Topics: Adult; Aged; Anemia, Aplastic; Animals; Bone Marrow Diseases; Erythropoiesis; Erythropoietin; Female; Hematologic Diseases; Humans; Leukemia; Male; Middle Aged; Rats

Topics: Adult; Aged; Anemia, Aplastic; Animals; Chloramphenicol; Erythropoietin; Female; Hematopoiesis; Humans; Male; Mice; Middle Aged; Prognosis

Topics: Adult; Anemia, Aplastic; Animals; Bone Marrow; Erythropoietin; Female; Humans; In Vitro Techniques; Male; Methods; Middle Aged; Rabbits

Topics: Adult; Anemia, Aplastic; Erythropoietin; Female; Fluoxymesterone; Hemoglobinuria, Paroxysmal; Humans; Male; Methods; Middle Aged; Multiple Myeloma; Primary Myelofibrosis; Sex Factors; Time Factors

Topics: Adolescent; Adult; Anemia, Aplastic; Bilirubin; Blood Platelets; Body Weight; Bone Marrow Examination; Erythrocyte Count; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Hematocrit; Hemoglobinometry; Humans; Iron; Leukocyte Count; Methandrostenolone; Neutrophils; Prednisone; Reticulocytes

Topics: Anemia, Aplastic; Anemia, Hemolytic; Anemia, Hypochromic; Animals; Erythropoietin; Hodgkin Disease; Humans; Infant; Infant, Newborn; Leukemia; Lymphatic System; Mice; Mononuclear Phagocyte System; Polycythemia; Umbilical Cord

Topics: Adult; Aged; Anemia, Aplastic; Chronic Disease; Erythropoietin; Female; Humans; Male; Middle Aged; Prednisolone

Topics: Adolescent; Anemia, Aplastic; Anemia, Hemolytic, Autoimmune; Anemia, Hypochromic; Child; Child, Preschool; Erythroblastosis, Fetal; Erythropoietin; Female; Humans; Infant; Infant, Newborn; Kidney Diseases; Male; Pregnancy; Wilms Tumor

Topics: Anemia, Aplastic; Animals; Chromatography; Erythropoietin; Hematocrit; Humans; In Vitro Techniques; Mice; Rabbits

Topics: Adult; Anemia, Aplastic; Animals; Erythropoietin; Humans; In Vitro Techniques; Male; Rats; Urine

Topics: Anemia; Anemia, Aplastic; Anemia, Pernicious; Epoetin Alfa; Erythropoietin; Humans; Polycythemia Vera; Saliva; Splenectomy

Topics: Anemia; Anemia, Aplastic; Blood Transfusion; Epoetin Alfa; Erythropoiesis; Erythropoietin; Humans; Leukemia; Leukemia, Lymphoid; Metabolism

Topics: Anemia; Anemia, Aplastic; Animals; Biological Assay; Epoetin Alfa; Erythropoietin; Haplorhini; Hypoxia; Injections, Intraperitoneal; Iron Isotopes; Kidney; Mice; Nephrectomy; Pharmacology; Physiology, Comparative; Polycythemia; Rabbits; Radiometry; Research; Urine

Topics: Anemia; Anemia, Aplastic; Cerebellar Neoplasms; Epoetin Alfa; Erythropoietin; Hemangiosarcoma; Kidney; Kidney Diseases, Cystic; Radiation Effects; Radiation, Ionizing

Topics: Anemia; Anemia, Aplastic; Blood Chemical Analysis; Body Fluids; Epoetin Alfa; Erythropoietin; Humans; Leukemia; Urine

Topics: Anemia; Anemia, Aplastic; Aorta; Cell Division; Epoetin Alfa; Erythropoietin; Humans; Leukemia; Metabolism

Topics: Anemia; Anemia, Aplastic; Bone Marrow; Epoetin Alfa; Erythrocytes; Erythropoietin; Humans