Significance of Quantitation of Autoantibodies in the Eluate of Sensitized Red Cells in Warm Autoimmune Hemolytic Anemia

Abstract A statistical study of 128 cases of autoimmune hemolytic anemias, serologically followed up in the same laboratory, led to some conclusions on classification, prognosis and treatment. Five forms were distinguished: 1. Idiopathic autoimmune hemolytic anemia with warm autoantibodies (IAHA-wa) was the most frequent form (65 per cent of the cases). It was observed in all peroids of life. A slight predominance among females was noted. This form was characterized clinically by a generalized or conjunctival icterus and a moderate splenomegaly. Hematologically a macrocytic normochromic anemia was present and serologically warm incomplete autoagglutinins, often nonspecific, or sometimes specific for a group antigen, were detected. Hemolysins were not found. The average course was 13 months followed by recovery (54 per cent) and 16 months followed by death (46 per cent). These two groups of patients were compared extensively. No differences in the age, sex, blood group and severity of the initial anemia were noted. A low reticulocyte count, leukopenia and association with thrombocytopenic purpura were more frequent in fatal cases. Tile persistence of a positive indirect Coombs test was unfavorable. Those with free antibodies in the plasma were the most serious. Fifty-two per cent of fatal cases had a positive indirect Coombs test. Of those who recovered, 18.5 per cent had this serologic finding. Transfusions were usually done at the begining of the disease. The efficacy of corticosteroid hormones was confirmed; the percentage of recoveries has risen since this therapy has been used fully (37.5 to 70 per cent). Early or late splenectomy had no influence on final desensitization (long-term effect), but led in 58 per cent of the cases to good clinical results (immediate effect). The spleen destroys red cells coated with noncomplement-fixing antibodies, so that splenectomy leads to compensation for the anemia. One must also describe the acute autoimmune hemolytic anemia observed especially in children, in which warm hemolysins could be detected at the very early stage of the disease. Complement was diminished or absent and the serum often showed anticomplementary activity. Complete recovery was rapid. 2. Symptomatic autoimmune hemolytic anemia with warm autoantibodies (SAHA-wa) accompanied mostly malignant conditions of the lymphocytic or reticuloendothelial systems as well as more rarely disseminated lupus erythematosus (17.6 per cent of the cases). Except for the causal disease, these cases were not different from IAHA-wa and their prognosis depended on the prognosis of the causal disorder. 3. Idiopathic autoimmune hemolytic anemia with cold antibodies (IAHAca) was less frequent (7.7 per cent of the cases). Clinically it was characterized by the rarity of splenomegaly, the chance of cold paroxysmal hemoglobinuria (1 case out of 10) and of Raynaud’s syndrome (1 case out of 10), and serologically by the presence of a cold acid-hemolysin (7 cases out of 8) along with an increased titer of complete agglutinins. Complement was diminished or absent. A positive Coombs test was possibly due to complement fixation. The course of these forms seemed to be very chronic: Nine cases of the 10 of the series were in progress for an average of 26 months, without any apparent trend to densensitization. The action of hormone therapy was less striking than in the warm variety. Splenectomy was probably not effective (1 case), since the red cells sensitized by complement-fixing antibodies were mainly recovered by the liver. 4. Symptomatic autoimmune hemolytic anemia wiith cold antibodies (SAHA-ca) was divided into two distinct forms: (a) one symptomatic of a malignant condition of the blood of the same type as in SAHA-wa (7 per cent of cases). The serology was identical to that of IAHA-ca. The prognosis was determined by that of the causal disease; (b) one symptomatic of a virus or a presumed virus infection (3.9 per cent of cases). Here an acid-hemolysin usually accompanied a very high complete cold agglutinin titer. Complete recovery occurred rapidly. In all cases with cold antibodies exposure to cold had to be carefully avoided. In cases of hemolysins, washed red cells had to be used for transfusions.

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Expression of CD47, CD35, CD55 and CD59 on Red Cells from Patients with Warm Autoimmune Hemolytic Anemia.

Blood ◽

10.1182/blood.v108.11.3738.3738 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3738-3738

Author(s):

Melca O. Barros ◽

Mihoko Yamamoto ◽

Maria S. Figueiredo ◽

Elisa Y. Kimura ◽

Jose Orlando Bordin

Keyword(s):

Hemolytic Anemia ◽

Lupus Erythematosus ◽

Autoimmune Hemolytic Anemia ◽

Experimental Studies ◽

Red Cells ◽

High Dose ◽

Healthy Controls ◽

Life Threatening ◽

Positive Direct ◽

Cd59 Expression

Abstract Autoimmune hemolytic anemia (AIHA) is defined as an increased destruction of red cells (RBC) in the presence of anti-RBC autoantibodies. CD47 is an integrin-associated protein expressed on all cells including RBCs. Animal models show that CD47 deficiency contributes to accelerated development of AIHA, while CD35 (CR1- complement receptor 1), CD55 (decay accelerating factor), and CD59 (membrane inhibitor of reactive lysis) are complement inhibitory proteins. Using flow cytometry analysis, in this study we evaluated the expression of CD47, CD35, CD55, and CD59 on RBCs of patients with warm AIHA before any treatment had been initiated. The study population consisted of 12 patients with active AIHA [M:F = 6:6, median age: 32 yrs (3 – 73)], and 20 healthy controls [M:F = 9:11, median age: 36 yrs (25 – 71)]. Ten patients presented idiopathic AIHA while 2 subjects had secondary AIHA (systemic lupus erythematosus and non-Hodgkin’s lymphoma). At presentation the median Hb level was 6.6 mg/dL (range: 2.9 to 10 mg/dL), and the median absolute reticulocyte count was 324 × 109/L (range: 215 to 756 × 109/L). At the time of the analyses, all 12 patients had a positive direct antiglobulin test (DAT), 12 (100%) had IgG on their RBCs, 5 (41.7%) had IgG plus C3, and none had C3 alone. The strength of agglutination of all positive DATs showed a strong reaction. The RBC eluates prepared by a dichloromethane technique from the cell samples were positive in all 12 patients, but the retrieved autoantibodies were pan-reactive showing no specific reactivity. The mean fluorescence intensity (MFI) of the expression of CD47, CD35 and CD55 on RBCs of AIHA patients and healthy individuals were not statistically different (CD47 = 464.4 and 464.4; CD35 = 186.8 and 194.3; CD55 = 396.9 and 381.1, respectively). Four patients with life-threatening AIHA were treated with high dose of steroids and RBC transfusions, but 3 patients evolved to death. Two patients who died presented low CD55 expression on their RBCs at diagnosis. AIHA patients showed significant lower CD59 expression on RBCs than healthy controls (MFI = 512.3 ± 28.0 and 553.7 ± 36.6, P = .03). Although CD59 expression in patients that evolved to remission was not significantly different from healthy controls (MFI = 538.5 ± 14.4 and 553.7 ± 36.6), the expression of CD59 on RBCs of 3 AHAI patients who died were significantly lower than that seen on RBCs of healthy controls (MFI = 433.6 ± 69.6 and 553.7 ± 36.6, P = .0001). Although experimental studies have suggested that CD47 has a profound influence on the severity of AIHA in mice, our preliminary data on 12 patients with AIHA did not demonstrate difference on the expression of CD47 on RBCs of patients with warm AIHA or healthy indibiduals. On the other hand, complement regulatory proteins (CD35, CD55, and CD59) may play an important role in protecting RBC destruction through the activation of complement. Our results suggest that patients with life-threatening warm AIHA may present significant CD59 deficiency on their RBCs that may increase the susceptibility of cells to complement-mediated lysis resulting in severe clinical hemolysis.

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Leukocyte reduction of red cells when transfusing patients with autoimmune hemolytic anemia: a strategy to decrease the incidence of confounding transfusion reactions

Immunohematology ◽

10.21307/immunohematology-2019-752 ◽

2020 ◽

Vol 12 (2) ◽

pp. 84-86

Author(s):

Jeanne A. Lumadue ◽

Rosetta Sue Shirey ◽

Thomas S. Kickler ◽

Paul M. Ness

Keyword(s):

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Red Cells ◽

Transfusion Reactions

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Teardrop-shaped red cells in autoimmune hemolytic anemia

American Journal of Hematology ◽

10.1002/ajh.2830210410 ◽

1986 ◽

Vol 21 (4) ◽

pp. 415-418 ◽

Cited By ~ 6

Author(s):

Deborah L. Farolino ◽

Pradip K. Rustagi ◽

Mark S. Currie ◽

Thomas D. Doeblin ◽

Gerald L. Logue

Keyword(s):

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Red Cells

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Hemolytic Anemia with Cold Detectable IgG Antibodies

Blood ◽

10.1182/blood.v36.5.549.549 ◽

1970 ◽

Vol 36 (5) ◽

pp. 549-558 ◽

Cited By ~ 13

Author(s):

MALCOLM R. MACKENZIE ◽

NANCY C. CREEVY

Keyword(s):

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Red Cells ◽

Coombs Test ◽

Igg Antibodies ◽

Igg Antibody ◽

Configurational Change ◽

Direct Coombs Test ◽

Type 3

Abstract Erythrocytes obtained from patients who manifest autoimmune hemolytic anemia can be divided into at least three categories by the nature of their protein coats as determined by direct antiglobulin (Coombs) test: IgG alone, IgG and complement (C), C alone. IgG antibodies were detected by direct Coombs Test at 4°C but not at 37°C in patients of type 3 A.H.A. Experiments at 4, 10, 15, 20, 30 and 37°C demonstrated that the IgG antibody was not eluted from the red cells at 37°C but apparently underwent a configurational change above 10°C such that agglutination no longer occurred with the Coombs reagent. This change was reversible. The presence of cold detectable IgG antibodies provides a mechanism for C deposition on erythrocytes in some cases of A.H.A., ostensibly due to complement alone.

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Immuno-Electronmicroscopic Studies of Surface Antigens of Blood Elements

Blood ◽

10.1182/blood.v33.2.179.179 ◽

1969 ◽

Vol 33 (2) ◽

pp. 179-185

Author(s):

GIORGIO TONIETTI ◽

GIUSEPPE A. ANDRES ◽

LIDIA ACCINNI ◽

MARIA PURPURA ◽

KONRAD C. HSU

Keyword(s):

Blood Group ◽

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Similar Pattern ◽

Surface Antigens ◽

Red Cells ◽

Human Igg ◽

Normal Individuals ◽

Blood Elements

Abstract Tagging, by means of the immunoferritin technic, of autoantibody on the erythrocytes of a patient with autoimmune hemolytic anemia, and on cells from a panel of blood of normal individuals of blood group O which have been incubated either with the patient’s serum or eluate from his cells, is described. It was found that: 1) Ferritin-labeled antibody to human IgG was localized on the surface of the patient’s erythrocytes at fairly even intervals. 2) Ferritin-labeled antibody to human IgM, β1C, rat globulin or pure ferritin alone was not bound to the patient’s cells. 3) None of the ferritin-conjugates mentioned in 1) or 2) or pure ferritin was bound to red cells from normal individuals represented in the panel. 4) Only ferritin-conjugated antibody to human IgG was localized, in a similar pattern, on the surface of the normal red cells which had been incubated either with the patient’s serum or the eluate from his cells, whereas none of the conjugates in 2) or pure ferritin was bound to these treated cells.

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Autoimmune hemolytic anemia warm-antibody type (Warm AIHA) in an 8-year-old Balinese girl

Paediatrica Indonesiana ◽

10.14238/pi41.1.2001.64-8 ◽

2017 ◽

Vol 41 (1) ◽

pp. 64

Author(s):

Putu Tri Yasa ◽

Ida Bagus Mudita ◽

Hendra Santoso ◽

Sudaryat Suraatmadja

Keyword(s):

Complete Remission ◽

Hemolytic Anemia ◽

Clinical Features ◽

Autoimmune Hemolytic Anemia ◽

Oral Prednisone ◽

Type A ◽

Red Cells ◽

Coombs Test ◽

Laboratory Findings

A case of autoimmune hemolytic anemia warm antibody type A (warm AIHA) in an 8-year-old Balinese girl was reported. The diagnosis was established based on clinical features, laboratory findings including positive Coombs' test positive. The etiology was probably primary or Idiopathic. The child was transfused with packed red cells and treated with oral prednisone. The response of the treatment was good and she experienced complete remission. The prognosis in patients with idiopathic warm AIHA are unpredictable. The girl underwent further follow-up in the child hematologic division every two weeks.

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Autoantibodies in Acquired Hemolytic Anemia With Special Reference to the LW System

Blood ◽

10.1182/blood.v42.3.445.445 ◽

1973 ◽

Vol 42 (3) ◽

pp. 445-453 ◽

Cited By ~ 18

Author(s):

G. H. Vos ◽

L. D. Petz ◽

G. Garratty ◽

H. H. Fudenberg

Keyword(s):

Special Reference ◽

Hemolytic Anemia ◽

Autoimmune Hemolytic Anemia ◽

Guinea Pigs ◽

Red Cells ◽

Normal Cells ◽

Cross Absorption ◽

Rare Cells

Abstract Most autoantibodies in patients with warm antibody autoimmune hemolytic anemia (AIHA) have specificity within the Rh system. Using rare cells such as -D- and Rhnull cells, Weiner and Vos (1963) described specificity against normal cells (nl), partially deleted cells (pdl), and deleted cells (dl). Recently, autoantibodies which failed to react with Rhnull cells that were of anti-U specificity have been described. It was suggested that the "Rh related" autoantibodies that cannot be identified as specific Rh antibodies may be anti-U. In the present study we examined eluates from the red cells of eight patients with AIHA using a panel of extremely rare cells and cross-absorption and elution techniques. We demonstrated autoantibody specificities not definable without the rare cells and, further, defined heterogeneity of the LW antigen. Autoantibodies with U specificity occurred in three eluates only. It was always present with an antibody of another specificity. Six of the eluates contained anti-LW, two anti-nl, five anti-pdl, three anti-dl, and one anti-e. Absorption and elution studies using the rare Rh-positive LW-negative (Mrs. Bigelow) showed that anti-pdl may in fact represent anti-LW + LW1 and that Mrs. Bigelow may represent a weak variant of LW. Injection of her red cells into guinea pigs produced an anti-LW that reacted similarly to the antibody produced by injecting Rh-positive LW-positive cells. An analogy to the ABO is suggested that normal Rh-positive LW-positive cells represent LW1, Rh-negative LW-positive cells represent LW2, Mrs. Bigelow represents LW3 and Rhnull cells represent the only true LW-negative (lw).

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