Diagnosis and gene analysis of congenital hemolytic anemia

Congenital hemolytic anemia is relatively uncommon and tends to be left undiagnosed because unusual special tests outside health insurance adaptation are needed for the diagnosis. To diagnose properly these patients, we instituted a comprehensive examination system for hemolytic anemia in Fukuoka University Hospital in 1990. In addition to basic laboratory tests, investigation of hemoglobin, SDS-PAGE analysis of red blood cell membrane proteins, a complete panel of red cell enzyme measurements and gene analysis were included. The red cell enzyme measurements was facilitated by the automation on the COBAS MIRA instrument (NIPPON ROCHE). By such a comprehensive examination, we have identified and confirmed 382 cases of congenital hemolytic anemia which otherwise would have escaped detection during the period October 1990 to September 2005 in our institution. SDS-PAGE procedure contributed to the diagnosis of 24 cases of band 3 deficiency, 13 cases of band 4.2 deficiency and 13 cases of band 4.1 deficiency. Hemoglobin studies uncovered 20 cases of abnormal hemoglobins, 126 cases of beta thalassemia and 7 cases of alpha thalassemia. Red cell enzyme measurements were also contributory to the diagnosis of 29 cases of hemolytic anemia in which enzyme levels were reduced.     

Clinicopathologic features of young and old sphha/sphha mice. Mutants with congenital hemolytic anemia.

A colony of mice with congenital hemolytic anemia, sphha/sphha, were evaluated over a 3-year period. Prominent findings included decreased survivability, reticulocytosis, increased peripheral blood leukocytes, extramedullary hematopoiesis in liver and spleen, lymphoid hyperplasia and membranoproliferative glomerulonephritis. Older (12 to 21 months) anemic animals had elevated serum levels of IgG1 and IgA. There was deposition of C3, IgG, IgM, and IgA in renal glomeruli of both control and anemic mice, but deposition of IgM and IgA was more prominent and widely distributed in anemic animals and correlated with mesangial expansion and the presence of electron dense deposits in the mesangium and in glomerular capillary walls. Prominent renal tubular hemosiderosis was noted in young and old anemic mice. The relation between the hemolytic anemia and glomerular disease is unclear but these mice may be an animal model useful for exploration of changes attendant with chronic hemolysis and evaluation of renal disease that accompanies hemolytic anemia.     

Mechanisms of congenital erythrocyte enzyme deficiencies associated with hemolytic anemia]

The search for a mechanism for red cell enzyme deficiency associated with congenital hemolytic anemia, requires one to determine the kinetic and thermodynamic properties of the enzyme reaction and study the physico-chemical and immunological characteristics of the protein which supports enzyme activity. The technique of iso-electric focalisation and the use of specific anti-enzyme antibodies, is the reason for recent progress in the understanding of the mechanism of these deficiencies. Examples of application of these techniques are given in relation to glucose-6-dehydrogenase, pyruvate kinase, glucose phosphate isomerase, phosphofructokinase and phosphoglycerate kinase of deficiencies showing the multiplicity of the molecular mechanisms.     

Experimental model of autoimmune hemolytic anemia induced in mice with levodopa

This paper describes an experimental model of autoimmune hemolytic anemia induced with multiple injections of levodopa. The serum antibodies were detected by indirect enzyme-linked immunosorbent assay (ELISA) and autoantibodies bound to syngeneic erythrocytes were detected by direct ELISA. A strain of mice injected with levodopa, at doses which are approximately equivalent to those used in human therapy, developed early and late cycles of IgM and IgG anti-mouse red blood cell (MRBC) autoantibody responses. These responses were followed by binding of IgM and IgG autoantibodies to the syngeneic erythrocytes and by two phases of anemia. Different strains of mice exhibited different sensitivities to Levodopa treatment, suggesting that genetic factors affect the anti-MRBC autoantibody response to levodopa. Indirect ELISA revealed that mice subjected to both X-irradiation (250 rad) and levodopa treatment had higher levels of IgG anti-MRBC autoantibodies than mice subjected to levodopa treatment alone, suggesting that a radiosensitive suppressor mechanism controls the autoantibody response. The autoantibodies induced with levodopa exhibited an unrestricted specificity and retained significant activity after absorption of MRBC. Serum autoantibodies were also detected by indirect ELISA in the sera of normal mice. This "natural" autoimmunity was mouse strain dependent and was possibly further influenced by environmental factors. The various implications of this model of autoimmune hemolytic anemia are discussed.     

Lethal congenital non-spherocytic,non-immune hemolytic anemia with genital and other anomalies in two brothers

Non-consanguineous healthy parents had 2 boys with severe, non-spherocytic, nonimmune hemolytic anemia, abnormalities of their external genitalia, flat occiput, dimpled earlobes, deep plantar creases, and increased space between their first and second toes. The birth of these children was separated by a spontaneous abortion at 3 months and delivery of a normal girl. We propose that these boys have a heretofore undescribed autosomal or X-linked recessive syndrome. © 1995 Wiley-Liss, Inc.     

Cold autoimmune hemolytic anemia with auto-anti-AI specificity: 51chromium survival studies

A 65-year-old woman was found to have severe autoimmune hemolytic anemia. The patient was group A1, Rho(D) positive. The direct antiglobulin test was strongly positive with anti-C3 and negative with anti- IgG. The serum contained two distinct IgM antibodies, auto-anti-I and auto-anti-AI. Both were reactive at 22 degrees C. However, the anti-AI also was reactive in saline and in albumin at 37 degrees C. An eluate revealed anti-AI and a weak anti-I. Sequential 51Chromium survival studies were done with group OI and AI red cells. The group OI red cells survived normally (97% at 24 hours) while the group A1I red cells were removed in a "two-component" pattern characteristic of IgM complement-fixing antibodies (62% survival at one hour, 49% at 24 hours). Based on these observations, the patient was subsequently transfused without incidence with six group O units of washed red cells prior to splenectomy. Although auto-anti-AI has been previously reported, this is the first case to demonstrate the use of 51Cr survival studies to determine its clinical significance.     

Autoimmune hemolytic anemia

Autoimmune hemolytic anemia (AHA) is characterized by the production of Coombs' antibodies, which are responsible for the destruction of red blood cells (RBCs). Analysis of both monoclonal anti-RBC autoantibodies derived from autoimmune New Zealand black mice and transgenic mice expressing a pathogenic IgM anti-RBC autoantibody has considerably improved our understanding of the B-cell responses involved in AHA, although our knowledge of T-cell immunity in AHA is still limited. The identification of the major T-cell epitope in the context of MHC class II molecules would be of paramount importance in helping to elucidate the cellular and molecular basis central to the development of AHA.     

Autoimmune hemolytic anemia

Autoimmune hemolytic anemias are a heterogenous group of disorders as evidenced by the multiple forms and classifications they have been commonly subjected to: acute or chronic, idiopathic or symptomatic, clinically latent or expressed, due to warm or cold autoantibodies. This work attempts to give an overview and synthesis of recent achievements in the knowledge of these syndromes within clinical, immunohematologic and pathophysiologic fields. In contrast, little progress has been made in the therapeutic measures the efficacy of which depends on the same variables as more than a decade ago.     

Immune hemolytic anemia associated with probenecid

Upon hospital admission a patient was found to have severe anemia and a strongly positive direct antiglobulin test (DAT). The patient was taking probenecid periodically for gout. An antibody was detected in the patient's serum that only reacted with red blood cells (RBCs) when probenecid was added. Eluates from the patient's RBCs, with and without the presence of drug, were nonreactive. Upon the discontinuation of probenecid, the patient's hemoglobin level improved steadily. We believe this to be the first reported case of immune hemolytic anemia associated with probenecid.