Autoimmune Anemia Research Articles

Identification of autoantigens in autoimmune haemolytic anaemia by a non-radioisotope immunoprecipitation method.

In human autoimmune anaemia (AIHA), warm autoantibodies frequently appear to have serological specificity for the Rh complex, but to date, immunochemical techniques have failed to demonstrate that the antibodies react with Rh-associated polypeptides. We describe the immunoprecipitation of red blood cell (RBC) autoantigens, using biotin labelling and a luminescent detection method. In three cases of warm AIHA, a band of 32 kD and a diffuse zone of 38-51 kD or 40-51 kD were specifically precipitated by eluted RBC autoantibody. This pattern corresponds closely with that precipitated by two Rh-specific monoclonal antibodies, BRIC 207 and AB5. Antibody from the three eluates also showed serological specificity for the Rh complex in a haemagglutination assay against a panel of RBC with a range of Rh phenotypes, including rare -D-/-D- and Rh null cells. Eluted autoantibody from another warm AIHA patient immunoprecipitated a peptide of 67 kD that did not correspond in apparent molecular mass either with Rh-associated bands, or with major RBC membrane proteins or sialoglycoproteins (SGP). The haemagglutination assay showed that this eluate contained both Rh-specific and Rh-unrelated antibody. Warm autoantibody eluted from the RBC of a clinically normal, but direct antiglobulin test positive, blood donor was serologically unreactive with the Rh complex, and immunoprecipitated unknown peptides of 26, 29, 35, 48 and 51 kD, together with a band of 90 kD that comigrated with SGP alpha 2 (glycophorin A). In six further warm AIHA cases, no antigens were precipitated by autoantibody-containing RBC eluates. Overall, the results demonstrate that autoantibodies bind to Rh polypeptides in some, but not all, patients with warm AIHA, suggesting that the aetiology of the disease may vary.

Murine autoimmune hemolytic anemia resulting from Fc gamma receptor- mediated erythrophagocytosis: protection by erythropoietin but not by interleukin-3, and aggravation by granulocyte-macrophage colony- stimulating factor

We have evaluated the therapeutic activity of recombinant erythropoietin (rEpo), in comparison with recombinant interleukin-3 (rIL-3) and granulocyte-macrophage colony-stimulating factor (rGM-CSF), on a lethal form of acute anemia resulting from Fc gamma receptor-mediated erythrophagocytosis after a single injection (500 micrograms) of a monoclonal anti-mouse red blood cell (MRBC) autoantibody. Continuous perfusion of rEpo before the administration of anti-MRBC monoclonal antibody completely protected animals from death due to anemia with a rapid recovery, while no protection was obtained by rIL-3 perfusion. In contrast, rGM-CSF perfusion markedly accelerated the progression of anemia and the mortality rate. This was found to result from an enhancement of erythrophagocytosis by Kupffer cells and by polymorphonuclear leukocytes that massively infiltrated the livers. Even after the injection of a sublethal dose (100 micrograms) of anti-MRBC monoclonal antibody, rGM-CSF-perfused mice died of a severe form of acute anemia. Furthermore, we have shown that rEpo was able to treat efficiently a spontaneous form of autoimmune hemolytic anemia in a majority of anemic NZB mice, whereas rGM-CSF markedly aggravated anemia. This may be of clinical importance, because GM-CSF administration could exhibit an adverse effect in some autoimmune diseases that involve autoimmune anemia.

Murine Autoimmune Hemolytic Anemia Resulting From FCγReceptor-Mediated Erythrophagocytosis: Protection by Erythropoietin but not by Interleukin-3, and Aggravation by Granulocyte-Macrophage Colony-Stimulating Factor

Abstract We have evaluated the therapeutic activity of recombinant erythropoietin (rEpo), in comparison with recombinant interleukin-3 (rIL-3) and granulocyte-macrophage colony-stimulating factor (rGM-CSF), on a lethal form of acute anemia resulting from Fc gamma receptor- mediated erythrophagocytosis after a single injection (500 micrograms) of a monoclonal anti-mouse red blood cell (MRBC) autoantibody. Continuous perfusion of rEpo before the administration of anti-MRBC monoclonal antibody completely protected animals from death due to anemia with a rapid recovery, while no protection was obtained by rIL-3 perfusion. In contrast, rGM-CSF perfusion markedly accelerated the progression of anemia and the mortality rate. This was found to result from an enhancement of erythrophagocytosis by Kupffer cells and by polymorphonuclear leukocytes that massively infiltrated the livers. Even after the injection of a sublethal dose (100 micrograms) of anti- MRBC monoclonal antibody, rGM-CSF-perfused mice died of a severe form of acute anemia. Furthermore, we have shown that rEpo was able to treat efficiently a spontaneous form of autoimmune hemolytic anemia in a majority of anemic NZB mice, whereas rGM-CSF markedly aggravated anemia. This may be of clinical importance, because GM-CSF administration could exhibit an adverse effect in some autoimmune diseases that involve autoimmune anemia.

The occurrence of auto‐antibodies in patients with gastro‐duodenal lesions

The occurrence of auto-antibodies in patients with the autoimmune disease pernicious anaemia and in patients with active duodenal ulcers was investigated. In order to characterize antigenic structures, various cellular and subcellular fractions were prepared from pig gastric mucosa and from a homogenate of duodenal mucosa. By means of an enzyme-linked immunosorbent assay and immunoblotting, both the H+,K(+)-ATPase and pepsinogen/pepsin were shown to constitute the major antigens. All of the seven pernicious-anaemia sera that were tested contained auto-antibodies against both antigens, and the epitopes of the H+,K(+)-ATPase were shown to be localized on its cytoplasmic face. In 75% (18/24) of the sera from patients with duodenal ulcers, auto-antibodies were detected when using purified antigens. Six sera reacted with H+,K(+)-ATPase and twelve reacted with pepsinogen, one reacted with both antigens, and four sera reacted with the duodenal mucosal antigen. The occurrence of auto-antibodies indicates that there is a mucosal lesion and that immunological factors may be involved in the pathogenesis of the disease in some patients.

Incomplete warm hemolysins: II. Corresponding antigens and pathogenetic mechanisms in autoimmune hemolytic anemias induced by incomplete warm hemolysins

The pathogenesis of autoimmune hemolytic anemias induced by incomplete warm hemolysins (IWHs) is only poorly understood. Sera from eight patients with mostly severe IWH-related autoimmune anemias allowed us to address some key elements of the pathogenetic process. With the use of a modified enzyme-linked immunoassay we could show that (i) IWHs bound to native RBCs and (ii) IWHs initiated the activation of the complement cascade on native RBCs up to the level of C3. The interaction of IWHs with their corresponding antigens on red blood cell membranes, which were pretreated with different enzymes, showed that despite serological uniformity there are at least two different types of IWHs: (i) IWHs reacting with phospholipase susceptible antigens (six of the studied sera) and (ii) IWHs reacting with antigens which are hardly, if at all, phospholipase susceptible (two of the studied sera). Aside from Rh antibodies, most IWHs apparently belong to a second category of RBC antibodies whose corresponding antigens can be inactivated by phospholipase treatment of the cell membranes. IWHs showed no specificity for Rh antigens or the antigens of the I i -system .

Long-Term Immunoregulatory Effects of Therapy with Corticosteroids and Anti-Thymocyte Globulin

Corticosteroids and anti-thymocyte globulin (ATG) have been extensively used in the treatment of autoimmune diseases, aplastic anemia and organ graft rejection; nonetheless, the precise mechanisms of action of these agents are unknown. Studies of their long term immunoregulatory effects, particularly in humans, have been limited. We examined the long term effects of therapy with ATG given for 2-4 weeks and prednisone for 2 months in 4 patients with newly diagnosed insulin dependent diabetes (IDD). Three matched newly-diagnosed untreated IDD patients and 17 healthy volunteers served as controls. No differences in total lymphocyte count, percentage of B cells, percentage of total T cells (CD3), helper-inducer T cells (CD4) or cytotoxic-suppressor cells (CD8), lymphocyte blastogenesis assays, or pokeweed mitogen-induced IgG secretion in T & B cell co-cultures were detected before therapy. A transient lymphopenia following ATG administration was the only immunological defect found in the first month of therapy. At 2 months, however, patients treated with ATG and prednisone had diminished immunoregulatory T cell function demonstrated by production of only 28 +/- 3% IgG expected in T & B co-culture, compared to 205 +/- 35% for untreated IDD patients and 107 +/- 13% for normals (p less than 0.01). This diminished IgG production resulted from excessive suppressor function, since co-cultures of T cells from treated patients with T and B cells from normal volunteers suppressed the latter's IgG production by 76 +/- 9%. This enhanced suppressor activity persisted for 3-6 months following therapy. Other immunological functions were not statistically different from those present at the inception of the study. Thus, treatment with corticosteroids and ATG produces long-term enhanced suppressor activity, a finding which suggests that treatment with combination ATG and Prednisone is a rational form of immunomodulation in conditions associated with decreased suppressor function.

Splenic Enlargement and Hyperfunction as Indications for Splenectomy in Chronic Leukemia

The chronic leukemias are associated with significant morbidity from splenic enlargement and hyperfunction. Although some patients with chronic leukemia benefit from splenectomy, the indications for operation are unclear. To identify those patients who benefit most from splenectomy, nine patients with chronic lymphocytic leukemia (CLL) and eight patients with chronic granulocytic leukemia (CGL) who had splenectomy to palliate the symptoms of massive splenic bulk or to improve the hematologic sequelae of splenic hyperfunction were studied. Splenectomy for bulk symptoms provided good palliation of symptoms, but the duration of the benefit was limited by the stage of the disease. Five of eight patients with CGL with bulk symptoms died within 6 months of operation. Splenectomy for hyperfunction was limited to a short-term hematologic response. In three of four patients with CLL who were Coombs positive, the presence of autoantibodies correlated with a recurrent transfusion requirement within 3 months of splenectomy. Thus, the benefit of splenectomy for bulk symptoms must be weighed against the risk of surgery and the patient's limited life expectancy. The benefit of splenectomy for treatment of splenic hyperfunction depends on the stimulus to hyperfunction and may not be beneficial for patients with refractory autoimmune anemias.

寒冷凝集素性溶血性貧血を合併したマイコプラズマ肺炎の1例

寒冷凝集素性溶血性貧血を合併したマイコプラズマ肺炎の1例

CYTIDINE DEAMINASE DEFICIENCY AND IMMUNODEFICIENCY MORE THAN A COINCIDENCE ?: 155

A 9 month-old boy, the fourth and the only surviving child of consanguineous parents of tunisian origin, was investigated for severe immunodeficiency associated with autoimmune anemia and thrombocytopenia. Increased serum immunoglobulin levels contrasted with an absence of antibody response after sensitization and blood T lymphocytes increased in number were incapable of proliferating normally in the presence of mitogens, antigens or of OKT3 antibody. Defective secretion of IL2 and of IFNrby patient's leukocytes was observed whatever the inducer used. Proliferative response as well as IFNr secretion were restored by incubation in the presence of exogenous IL2. Enzyme study disclosed a profound deficiency in cytidine deaminase (CDA) in the patient's lymphocytes (10-21-23 pmol.min−1.10−6 cells, normal value 130 ± 30, n = 39). Low values were found in the father (20-21) and the mother (18-55). Cytidine (100 μM) did reverse the inhibitory effect of pyrazofurin on parents' lymphocytes PHA-induced proliferation but not that of patient's lymphocytes. During a 6-months survey lymphocyte CDA activity of the patient, his father and mother rose to 63, 140 and 130 pmol.min−1.10−6 cells, respectively. The fact that CDA deficiency has been transitory in the parents rules out an hereditary enzymatic defect. Nevertheless, our results suggest that CDA activity should be systematically determined in immunodeficient patients.

Methyldopa: Effects on the murine immune system

Mice were treated for 7 weeks with doses of methyldopa somewhat exceeding those given to man, and mixed immunotoxic effects were observed. Daily subcutaneous injections of 5 mg (in 0.1 ml) methyldopa or saline (0.1 ml) did not generally alter body weights, except on day 19, when the methyldopa-treated mice weighed significantly less. During treatment, all mice were immunized twice with sheep red blood cells (SRBC) and bled four times. Anti-SRBC titers were not affected by methyldopa treatment, but leukocyte counts were dramatically decreased, and hematocrits to a lesser degree. Although in methyldopa-treated mice spleen and kidneys were increased in size, liver, lung, heart, and thymus size was not affected. These results are discussed in the context of other studies on the mode of action of methyldopa in eliciting an autoimmune anemia in man treated therapeutically with this drug.

Tolerance and autoimmunity to erythroid differentiation (B-G) major histocompatibility complex alloantigens of the chicken.

Hematopoietic chimeras were produced at four different stages of ontogeny between two allogeneic strains of chickens. All chimeras produced by parabiosis at day 12 of embryogenesis and the majority (83%) of the ones produced at day 15 by intravenous injection of allogeneic stem cells remained healthy, chimeric, and specifically tolerant at both the humoral and cell-mediated level throughout a long examination period. Chimeras generated at day 17 of embryogenesis demonstrated specific unresponsiveness at the cell-mediated level but produced specific anti-donor alloantibodies directed against erythrocyte-associated major histocompatibility complex (MHC) (B-G) antigens. These chimeras and a minority (17%) of the chimeras generated at day 15 of embryogenesis developed severe antibody-mediated autoimmune hemolytic anemia after the 5th mo of age and succumbed to massive bursal lymphomas and metastases by the 10th mo of age. The immunological and pathological characteristics of these birds appear to reflect an autoimmune state rather than one of tolerance. Erythroid chimeras generated at day 21 of ontogenic development displayed normal levels of GVH reactivity. These birds were eventually able to eliminate the chimeric state and remained healthy until deliberately killed. These results show that there is a critical period in embryogenesis during which the induction of allogeneic erythrocytic chimerism leads to the development, in adult life, of severe autoimmune anemia, B cell lymphomas, and death. B-G MHC antigens are erythroid differentiation antigens of the chicken. Polymorphic determinants on B-G antigens appear to be important cross-reactive determinants (with environmental bacteria), against which a high background immunity exists. Evidence is presented that the immune response to B-G antigens is responsible for the development of autoimmunity and other pathological events that follow and that tolerance to class I MHC antigens (B-F antigens) shared by lymphocytes erythrocytes is maintained at the same time that B-G tolerance is broken.

Immune Pancytopenia

Circulating T-lymphocytes from a 13-year-old boy with autoimmune anaemia, severe neutropenia and thrombocytopenia inhibited autologous and normal homologous bone marrow myeloid colony formation in vitro. This inhibition was abolished when the patient's antithymocyte globulin and complement-treated T-lymphocytes were used. T-lymphocytes from normal individuals did not cause such an inhibition. The patient's lymphocytes showed no inhibitory effect on erythroid colony formation. Investigation of the patient's serum failed to disclose any leucoagglutinin, lymphocytotoxin or humoral factor against myeloid colony formation. These findings indicate that T-lymphocytes may play a role in the pathogenesis of neutropenia in immune pancytopenia.

Severe autoimmune anemia and thrombopenia in mixed connective tissue disease

Severe autoimmune anemia and thrombopenia in mixed connective tissue disease

IgM antibodies to red cells and autoimmune anemia in patients with malaria.

Rising titers of IgM antibodies to red blood cells were found in 56 patients following infection with Plasmodium falciparum parasites. These antibodies, detected by indirect immunofluorescence using fluorescein-conjugated monospecific antisera, were not present in the sera of 23 non-malaria patients with high IgM concentration, 9 patients with infectious, malignant, and autoimmune diseases, and 16 normal subjects. Rising titers of IgM erythrocyte antibodies were associated with an increase of mean IgM from 100 to 500 mg/100 ml and a decrease of mean C3 from 150 to 75 mg/lOO ml over 24 days. Mean concentrations of serum IgA, IgG, IgD, IgE, albumin, globulin, and total protein were not significantly different in patients and controls throughout the study. IgM erythrocyte antibody titers and serum IgM levels were highest, and serum C3 were lowest in the most anemic patients. Survival time of 51Cr-tagged normal red cells was reduced in 10 of 11 patients studied following the clearance of malarial parasitemia and was the shortest in the most anemic patients.

The susceptibility to autoxidation of human red cell lipids in health and disease.

Summary. The susceptibility of human red cells to autoxidation was measured (i) in various groups of normal subjects, (ii) in haemolytic states, and (iii) in non‐haemolytic disease. Many types of haemolytic disease—notably thalassaemia major and autoimmune haemlytic anaemia—were associated with a greatly and persistently increased susceptibility of the red cell lipids to autoxidation. In two patients with haemolytic disease this was the only biochemical abnormality found. Susceptibility to autoxidation is about three times higher in the newborn period than it is in adult life; and there is a comparatively wide scatter in old age.

Myasthenic syndromes in hypothyroidism. Electrophysiological study of neuromuscular transmission and muscle contraction in two patients.

Two hypothyroid patients, one with autoimmune thyroiditis and pernicious anemia and the other with idiopathic primary myxedema, were studied electrophysiologically. The former showed findings of the Eaton-Lambert myasthenic syndrome, and the latter showed findings of the desensitization block of neuromuscular transmission or the conduction block at the fine terminal filaments of the motor axon. The muscle contraction study revealed the posttetanic muscle contracture without electrical discharges in the former, and the posttetanic twitch depression and the decreased tetanic force in the latter.

The Effect of Irradiation and Infusion of Marrow and Spleen Cells on Autoimmune Anemia in NZB/B1 Mice

The Effect of Irradiation and Infusion of Marrow and Spleen Cells on Autoimmune Anemia in NZB/B1 Mice

The Hæmolytic Anæmias, Congenital and Acquired—Part 2. The Auto Immune Anæmias

The Hæmolytic Anæmias, Congenital and Acquired—Part 2. The Auto Immune Anæmias