Fatal outcome in a patient with autoimmune hemolytic anemia associated with an IgM bithermic anti-ITP

BACKGROUND: Several cold autoantibodies (usually IgG) with IT specificity have been reported previously, as have autoantibodies with joint I and P blood group specificities (IP1, ITP1, iP1, IP). A fatal outcome associated with an IgM cold autoantibody of ITP specificity is reported. CASE REPORT: A 54-year-old man suffered from progressively severe cold autoimmune hemolytic anemia for 9 months. Hemoglobin concentration ranged from 6 to 7 g per dL (60-70 g/L) and reticulocytes from 3 to 5 percent (0.030-0.050). The direct antiglobulin test was weakly positive for IgM and strongly positive for C3d. The serum contained a cold agglutinin that reacted strongest with cord i red cells (RBCs) > adult I RBCs > adult i RBCs, which is consistent with IT specificity. The Donath-Landsteiner test was positive; the reaction was neutralized by globoside. The serum reacted weakly or was negative with RBCs from five group p blood donors, which suggests anti-ITP specificity. Dithiothreitol treatment of the serum abolished the cold agglutinin reactivity, which suggests that the anti-IT was IgM. The patient received > 40 RBC transfusions and failed to respond to oral steroids, oral cytoxan, high-dose pulse intravenous steroids, and plasma exchange at room temperature and at 35 degrees C. He died of sepsis following an unsuccessful trial of chlorambucil. Autopsy revealed unsuspected disseminated non-Hodgkin's lymphoma. CONCLUSION: Serologic studies are consistent with our patient's having a single IgM cold autoantibody with IT and P specificities (anti-ITP) and requiring both specificities on the same RBC to permit maximal antibody expression.     

A dual antiglobulin test for the detection of weak or nonagglutinating immunoglobulin M warm autoantibodies

BACKGROUND: Immunoglobulin (Ig)M warm autoantibodies (AABs) usually cause severe autoimmune hemolytic anemia (AIHA) and, in some cases, red blood cell (RBC)‐bound IgM cannot be detected. We describe a simple dual antiglobulin test (DDAT) for diagnosing such cases. STUDY DESIGN AND METHODS: A patient with erroneously suspected cold agglutinin syndrome was investigated. The direct antiglobulin test (DAT) was performed using standard techniques and dual (two stages) antiglobulin reagents (IgG rabbit anti‐human IgM, IgG goat anti‐rabbit IgG). RESULTS: A cold agglutinin syndrome was diagnosed initially, as the patient's serum was reactive with RBCs at a temperature of 28°C or less, and the DAT was strongly positive with anti‐C₃d. Six months later, the patient was reexamined at this hospital due to progressive hemolysis. His RBCs were found to be coated with IgM warm AABs that only became detectable using a DDAT, and his serum contained only a weak cold agglutinin. The hemolysis remained refractory to treatment with prednisolone and also prednisolone plus azathioprine, but gradually improved after treatment with prednisolone plus cyclophosphamide. CONCLUSION: Weak or nonagglutinating RBC‐bound IgM warm antibodies can be identified by the presented DDAT.     

A severe transfusion reaction associated with anti-EnaTS in a patient with an abnormal alpha-like red cell sialoglycoprotein

A 71-year-old woman (Ped) received 3 units of red cells (RBCs), compatible by the indirect antiglobulin test but strongly (4+) incompatible by direct agglutination at 37 degrees C. The next day, her plasma hemoglobin was 1252 mg percent and the direct antiglobulin test (DAT) was weakly positive (IgG and C3). Less than 5 percent of the transfused cells could be detected 48 hours posttransfusion. Her clinical condition deteriorated and renal failure developed. The patient died of pulmonary embolism. Her serum contained a strong (4+) IgM agglutinin and a weakly reactive (microscopically positive) IgG antibody, with anti-EnaTS specificity. EnaFS and EnaTS antigens were severely depressed or absent from the patient's RBCs; the ficin-resistant Ena antigen (EnaFR) appeared to be present. Pretransfusion RBC sialic acid level was 53 +/- 2 percent of normal. The patient's RBC membranes were shown to contain sialoglycoproteins beta and delta by sodium dodecyl sulphate polyacrylamide gel electrophoresis with periodic acid Schiff's base staining, with weak staining of components in the regions corresponding to alpha, alpha 2 and alpha delta. The nature of these components was not identified, but their presence suggested that the patient's RBCs expressed a previously undescribed sialoglycoprotein alpha variant.     

Hemolytic anemia associated with injection of fluorescein

A 49-year-old woman presented with a hemoglobin level of 9.5 g per dL (95 g/L), reticulocyte count of 6.7 percent (0.067), and hemoglobinuria. The next day, the hemoglobin had dropped to 5.8 g per dL (58 g/L), and total bilirubin was 8.8 mg per dL (150 mumol/L). The serum reacted 2+ with all red cells (RBCs). The direct antiglobulin test (DAT) was 3+ with anti-IgG and 1+ with anti-C3, but eluates prepared by two different methods did not react with untreated RBCs. The eluate reacted 2+ with amoxicillin-coated RBCs; amoxicillin had been listed in the patient's record as a previous medication. The patient denied recent ingestion of amoxicillin. Further investigation documented the injection of a dye, fluorescein sodium (AK-FLUOR-25%), for a ophthalmologic fluorescein angiographic study 2 days before admission. RBCs coated with AK-FLUOR reacted with the eluate. Controls consisting of normal serum, an eluate prepared from DAT-negative RBCs, and a serum known to contain anti-penicillin did not react with AK- FLUOR-coated RBCs. Nine days later, the DAT was negative and the serum did not react with untreated RBCs. In the presence of AK-FLUOR (1-in- 125) or amoxicillin (1 mg/mL), the serum reacted 2+ in the antiglobulin test. Antibodies to AK-FLUOR and amoxicillin appeared to react by two mechanisms, which is similar to results in recent reports of other drugs associated with hemolytic anemia. AK-FLUOR has not previously been reported to be associated with hemolytic anemia.     

Invalidation of antiglobulin tests by a high thermal amplitude cryoglobulin

A multiply transfused patient was referred for evaluation of a transfusion reaction. The direct and indirect antiglobulin tests (DAT, IAT) for alloantibody were negative. However, IgG-coated control cells failed to agglutinate in the negative reactions, casting doubt on their validity. At 4 degrees C, the patient's serum exhibited a large cryoprecipitate (2.9 mg/mL), made up predominantly of an IgG kappa paraprotein and having trace amounts of IgM and C3. Clear serum separated at 37 degrees C became cloudy within 10 minutes at room temperature (RT); within 4 hours, approximately 60 percent of the total precipitable cryoprotein had precipitated. Red cells (RBCs) incubated in fresh serum that had cooled to RT or RBCs obtained from RT or refrigerated samples contained cryoprecipitate that sedimented with the RBCs during washing with RT saline. On resuspension, enough IgG cryoglobulin redissolved to neutralize completely the commercial anti-IgG reagents. If the patient's samples were maintained at 37 degrees C, cryoprecipitate did not form, and RBCs washed four times at 37 degrees C gave valid DAT and IAT reactions. The removal of all cryoprecipitate from the patient's serum by centrifugation after overnight incubation at 4 degrees C also made possible valid antibody screening and compatibility tests.     

直接抗人球蛋白试验阳性的类型鉴别及临床意义

目的　通过直接抗人球蛋白试验 (DAT)阳性的类型鉴别 ,有助于临床对自身免疫性溶血性贫血 (AIHA)的正确诊断 ;对血库交叉配血不合的处理有指导意义。方法　DAT阳性标本分AIHA组和非AIHA组 ,进行免疫分型 ,然后把IgG或IgG C3的标本进行放散试验 ,利用放散液与标准抗体筛查细胞进行间接抗人球蛋白试验 (IAT)。结果　AIHA组患者的放散液均同标准抗体筛查细胞反应 ,非AIHA组除了 1例SLE患者外均为阴性反应。结论　DAT阳性患者可以分为两类 :自身抗体引起和免疫复合物引起     

Hypergammaglobulinemia can be associated with a positive direct antiglobulin test, a nonreactive eluate, and no evidence of hemolysis

To determine the cause of a positive direct antiglobulin test (DAT), blood banks routinely perform serologic tests on eluates prepared from DAT-positive red cells. Negative eluates traditionally have been suspected to be associated with drug reactions. This report confirms that the most frequent cause of a positive DAT and a nonreactive eluate is hypergammaglobulinemia. The results of 74 patient samples with positive DATs were analyzed retrospectively. Eluates prepared from the red cells of 54 patients (72.9%) reacted; eluates from 20 patients (27.1%) did not react. This latter group had identical serologic and clinical findings, suggesting that they made up a homogeneous group. In particular, the patients had a positive DAT, a negative indirect antiglobulin test, and a negative eluate; an increased serum concentration of IgG; and no evidence of hemolysis. In a subsequent study, DATs were performed prospectively on red cells from 44 consecutive patients with elevated serum IgG levels. The serum IgG concentration was highest in the three patients whose red cells had a positive DAT. The DAT also became positive in two patients treated with high-dose intravenous gammaglobulin (IV IgG). These studies indicate that a negative eluate from red cells with a positive DAT, a common serologic finding, is often caused by hypergammaglobulinemia. The authors postulate that IgG binds nonspecifically to the red cells because of the hypergammaglobulinemia.     

Severe immune-mediated hemolytic anemia secondary to treatment with cefotetan

Severe acute hemolytic anemia developed in a woman following treatment with multiple antibiotics for possible postpartum uterine infection. On admission, the hemoglobin was 5 g per dL (50 g/L), the reticulocytes were 35 percent (0.350), the direct antiglobulin test was strongly positive for IgG and C3d (mixed fields), and the indirect antiglobulin test was negative. Serologic studies revealed antibody to cefotetan that reacted by both the immune complex and the drug adsorption mechanisms. Before the diagnosis of cefotetan-related immune hemolysis was made, all medications had been discontinued, and the patient received 4 units of red cells and a short course of adrenocorticosteroids. Recovery was prompt and complete.     

Severe delayed hemolytic transfusion reaction secondary to anti-Ata

BACKGROUND: Anti-At(a) is a rare red cell (RBC) alloantibody found in the black population. It has been described as causing one case of mild hemolytic disease of the newborn, but its ability to cause hemolytic transfusion reactions is uncertain. CASE REPORT: The patient was a 60-year-old black female with a history of three uneventful pregnancies but no transfusions. On admission, her direct and indirect antiglobulin tests were negative, total bilirubin was 0.5 mg per dL, and lactate dehydrogenase was 224 IU per L. She received nine units of compatible RBCs in the perioperative period of a hemicolectomy. Her hemoglobin rose appropriately and stabilized at 12.6 g per dL by the 6th postoperative day. By Day 10 after surgery her hemoglobin had dropped to 6.8 g per dL, and her total bilirubin and lactate dehydrogenase had risen to 1.4 mg per dL and 783 IU per L, respectively. The direct and indirect antiglobulin tests were now newly positive with strengths of 3+. A warm hemolytic autoantibody was suspected. She was transfused two units of incompatible RBCs for a rapidly falling hemoglobin and symptomatic anemia. On Day 11, the total bilirubin rose to 3.5 mg per dL, and the lactate dehydrogenase was 1154 IU per L with a hemoglobin of 7.6 g per dL. Corticosteroids were begun. Studies of serum and an acid eluate revealed anti-At(a), but no other RBC antibodies. The patient stabilized, and further transfusion was avoided. CONCLUSION: Although anti-At(a) was previously described as being of uncertain clinical significance, this patient demonstrated the ability of the antibody to cause a severe delayed hemolytic transfusion reaction.     

Waldenstrom's macroglobulinemia with an IgM paraprotein that is both a cold agglutinin and a cryoglobulin and has a suppressive effect on progenitor cell growth

BACKGROUND: A patient with Waldenstrom's macroglobulinemia was admitted to the hospital with fever, leg pain, and dyspnea. The patient had gas gangrene of the left leg that required above-the-knee amputation. Plasmapheresis was instituted to treat hyperviscosity. STUDY DESIGN AND METHODS: The patient's serum contained an IgM-kappa paraprotein, a cryoglobulin, and a cold agglutinin. The serum was studied. RESULTS: The patient's red cells typed as A1, Rh-positive. The direct antiglobulin test was negative. The serum contained a cold agglutinin with anti-Pr cold agglutinin specificity (titer 4096). Maximal thermal range was 30 degrees C. Following dithiothreitol treatment, the cold agglutinin activity disappeared. The serum IgM concentration in the tested sample was 62.3 g per L. The cold agglutinin titer in the supernatant after removal of the cryoglobulin was 256, and the IgM level was 0.31 g per L. Redissolving the cryoglobulin in a equivalent volume of saline resulted in a cold agglutinin titer of 4096 and an IgM level of 68.4 g per L. These results indicate that the cryoglobulin and the cold agglutinin are the same paraprotein. Serum protein electrophoresis using agarose gel and immunofixation of the serum revealed an IgM-kappa monoclonal band. Progenitor cell assays were performed by adding the patient's serum at final concentrations of 0, 1, 5 and 10 percent (vol/vol) to patient's and normal donor's peripheral blood mononuclear cells. Inhibition of burst-forming units- erythroid and colony-forming units-granulocyte/macrophage by the patient's serum was demonstrated. Appropriate controls and the use of the serum of another patient with Waldenstrom's macroglobulinemia did not suppress progenitor cell growth. The patient's serum inhibited colony formation in a dose-response fashion. CONCLUSION: Reports of cryoprecipitable cold agglutinins are rare. This case is unusual because the IgM-kappa paraprotein was also a cold agglutinin with anti- Pr specificity and erythroid and granulocyte-macrophage progenitor cell- suppressive properties.     

Hemolytic transfusion reaction due to anti-Tca

BACKGROUND: Anti-Tc(a) detects a high-incidence antigen in the Cromer blood group system. Cromer system antibodies have not usually been associated with hemolytic transfusion reactions or hemolytic disease of the newborn. CASE REPORT: Anti-Tc(a) (initially identified in the patient's serum in 1982) was not detected when she was admitted to the hospital with upper gastrointestinal. bleeding. Three units of red cells were administered. The patient was discharged, but was readmitted to the hospital after her hemoglobin fell to 7.1 g per dL. Antibody detection tests remained negative and three additional units were transfused. Over the next 7 days, her hemoglobin steadily fell to 5.5 g per dL. The level of lactate dehydrogenase rose to 1257, the plasma hemoglobin rose to >16 mg per dL, and the haptoglobin decreased to <6 mg per dL. Five days after transfusion, her direct antiglobulin test was weakly reactive with complement-specific antiglobulin reagents. Eluates were nonreactive. Anti-Tc(a) was detected in her serum; no other antibodies were detected. Differential typing failed to detect any circulating Tc(a+) red cells. The antibody was strongly reactive in a monocyte monolayer assay. CONCLUSION: Although Cromer system antibodies have generally not been proven to be clinically significant in transfusion therapy, the destruction of red cells from six units of transfused Tc(a+) red cells in this patient indicates that anti-Tc(a) may have destructive potential in some patients.     

Lower hemoglobin levels in human immunodeficiency virus-infected patients with a positive direct antiglobulin test (DAT): relationship with DAT strength and clinical stages

BACKGROUND: There are conflicting opinions regarding the effect of positive direct antiglobulin test (DAT) on hemoglobin (Hb) levels in human immunodeficiency virus-infected (HIV+) patients. STUDY DESIGN AND METHODS: A total of 166 samples from HIV+ outpatients were studied. The DAT was performed with the tube test and column agglutination technology (CAT). RESULTS: The DAT was positive in 18.67 percent with the tube method and 33.73 percent with the CAT. Patients with DAT-positive results showed lower Hb levels than DAT-negative patients, 12.3 g per dL versus 14.3 g per dL (p = 0.0002). The univariate logistic regression enabled us to study the phenomenon better and fit the probability of having a DAT-positive result on the basis of the Hb levels. The relationship between the CAT and the tube test when washing the red blood cells (RBC) at 4 degrees C was stronger than when washing these at room temperature (phi = 0.8156; p = 0.000). The Hb levels were significantly lower in the positive DATs of Stage C (acquired immune deficiency syndrome [AIDS]) and Stage B (symptomatic non-AIDS patients), which showed decreasing Hb values for increasing agglutination strengths (p = 0.000). Anemia was related with the DAT results (odds ratio [OR], 8.005; p = 0.000) but not to the AIDS condition (OR, 1.741; p = 0.221). DISCUSSION: Our study indicates that the DAT-positive results may be specifically related to lower Hb levels in HIV+ patients. The immunologic RBC clearance could be part of the anemic multifactorial condition in HIV+ patients.     

Haemolytic anaemia and renal failure associated with antibodies to trimethoprim and sulfamethoxazole

Aim: The aim of this study was to support a clinical diagnosis of drug‐induced immune haemolytic anaemia (DIIHA). Background: DIIHA is rare and has only been described twice with the antibiotic combination of trimethoprim (TMP) and sulfamethoxazole (SMX). Methods/Materials: Serologic tests for drug antibodies were performed using methods previously published by our laboratory. Results: A 44‐year‐old woman experienced body aches, chills, chest pressure, nausea and a rash while receiving TMP–SMX; a week later her haemoglobin was low and she was in renal failure. At the hospital, the direct antiglobulin test (DAT) was positive (C3 only) and the serum reacted with all red blood cells (RBCs) by the gel method only (TMP–SMX is present in the RBC diluent used for the gel method). At the Red Cross immunohaematology laboratory, the patient's serum was reactive in the presence of TMP–SMX (haemolysis and positive antiglobulin test), pure TMP (positive antiglobulin test using anti‐IgG only) and pure SMX (haemolysis and positive antiglobulin test using both anti‐IgG and anti‐C3). The patient was treated with transfusions and haemodialysis and was discharged after a week in stable condition. Conclusion: We describe a patient who appeared to have haemolytic anaemia and renal failure associated with antibodies to both TMP and SMX.     

Anti-Jka, -C, and -E in a single patient, initially demonstrable only by the manual hexadimethrine bromide (Polybrene) test, with incompatibilities confirmed by 51Cr-labeled red cell studies

Published reports have confirmed the superior sensitivity of the manual hexadimethrine bromide (Polybrene) test (MPT) for demonstrating many alloantibodies in vitro; however, the clinical significance of alloantibodies demonstrable exclusively by MPT has not been shown conclusively. A patient with macroglobulinemia experienced chills, fever, hemoglobinemia, and hemoglobinuria following the transfusion of 1 unit of red cells (RBCs) shown to be compatible by the low-ionic-strength antiglobulin (LIS-AG) method. Serologic investigation was negative. Intravascular hemolysis occurred with a second "compatible" unit. Serologic studies were again negative by LIS-AG and ficin-AG methods, but revealed anti-Jka by MPT. Both donors were Jk(a+b-), and 51Cr studies of the second donor's RBCs revealed a t1/2 of less than 30 minutes, with marked intravascular hemolysis. A LIS-AG-compatible Jk(a-) unit was transfused uneventfully, but with no rise in hematocrit. MPT next revealed anti-C; subsequent 51Cr studies with the Jk(a-), Cc donor's RBCs showed a 51Cr t1/2 of 100 minutes with slight intravascular lysis. Four transfusions of Jk(a-), C- blood were uneventful, but 5 days later the patient's hemoglobin declined. The following day, anti-E was demonstrable exclusively by MPT. 51Cr-labeled Jk(a-), C-, E- RBCs had normal 24-hour survival. The patient's hemoglobin rose to 11 g per dl following transfusions of Jk(a-), C-, E- RBCs, and he was discharged. In vitro studies employing the patient's purified IgM paraprotein revealed no interference with alloantibody binding or detection.     

Removing IgG antibodies from intact red cells: comparison of acid and EDTA, heat, and chloroquine elution methods

BACKGROUND: To accurately phenotype red cell from patients with a positive direct antiglobulin test (DAT), nonlytic elution procedures were assessed for their ability to dissociate IgG from antibody-coated red cells without altering red cell antigen expression. STUDY DESIGN AND METHODS: Antibodies coating red cells that were sensitized in vivo (warm-reactive autoantibodies: 8 patients) or in vitro (42 alloantibodies) were eluted by using glycine-HCl and EDTA (acid/ EDTA), heat (56 degrees C, 10 min), or chloroquine method. RESULTS: Acid/EDTA elution gave the best results, reducing DAT positivity to microscopic levels or rendering the DAT negative in 48 of 50 instances, whereas 4 samples remained resistant to heat elution and 24 to chloroquine. Standard DAT agglutination scores demonstrated that both acid/EDTA and heat elution were superior to the chloroquine method (p < 0.0001). With the gel low-ionic-strength saline indirect antiglobulin test, acid/ EDTA was superior to heat (p < 0.001). Overall, acid/ EDTA elution dissociated more antibodies than heat (p < 0.0001), especially for Kell system (K, k, Kpa, Kpb) alloantibodies. Common red cell antigens, other than Kell system antigens, were unaffected by acid/EDTA elution. In contrast, the expression of most blood group antigens was diminished after heat elution. However, it was possible to type red cell antigens by using gel low-ionic-strength saline indirect antiglobulin tests or tube agglutination methods. CONCLUSION: Although heat elution may be used on a limited basis, the acid/EDTA method appears to be the procedure of choice for typing red cell coated with warm-reactive IgG alloantibodies or autoantibodies.     

Membrane-bound immunoglobulins and complement components on young and old red cells

In order to characterize changes in membrane-bound immunoglobulins and complement components, red cells (RBCs) were separated into young and old populations by simple centrifugation. Old RBCs had reduced mean corpuscular hemoglobin volume, increased mean corpuscular hemoglobin concentration, and reduced sialic acid. Using radioactive anti- antiglobulin techniques, old RBCs were shown to have more IgG, IgM, IgA, and C3d on their surfaces than did young RBCs; there was no increase on old RBCs of C3b, factor B, C4b, or C5. Similar results were observed with RBCs strongly coated with C3d in vivo from a patient with cold agglutinin disease. RBCs taken into ethylenediamine tetraacetate, washed thoroughly in saline, and then stored for prolonged periods in Alsever's solution or kept in autologous ethylenediamine tetraacetate plasma, at 4 degrees C, showed no increase in RBC-bound C3d with increased storage time. If, however, blood was taken into citrate- phosphate-dextrose and maintained at 4 degrees C in autologous plasma, a significant increase in RBC-bound C3d was observed in the mixed-cell population with prolonged storage time. Order donor blood units, taken into citrate-phosphate-dextrose and stored at 4 degrees C as packed red cells, showed higher levels of RBC-bound C3d in the mixed-cell population than did units stored for a shorter time. In no case did donor unit RBCs give a positive direct antiglobulin test on serologic testing with anti-C3d. The findings complement data already collected on membrane and cytoplasmic changes in aging RBCs and may contribute to an understanding of RBC senescence.     

Coombs' negative immune hemolytic anemia with anti-E occurring in the red blood cell eluate of an E-negative patient

A previously untransfused 20-year-old man presented with a seven day history of malaise, fatigue, jaundice, dark urine and splenomegaly. Hemolytic anemia was indicated by a hemoglobin of 8.7 mg/dl, reticulocyte count 8 per cent, Lactic dehydrogenase 389 iu/L, bilirubin 4.3 mg/dl (direct 0.1 mg/dl), and undetectable haptoglobins. Tests for nonimmunologic mediated hemolytic anemia were negative. The direct antiglobulin test (DAT) was repeatedly negative with polyspecific, anti-IgG, -IgA, -IgM and anti-C3 antisera. The patient's serum contained a weak anti-I, anti-E strongly reactive by indirect antiglobulin test (IAT) and an antibody reactive against all cells tested. The latter antibody reacted weakly by the IAT but strongly against enzyme-treated cells (Titer 160). Eluates from the patient's red blood cells only reacted with E+ red blood cells. The patient typed E negative. He was treated for warm autoimmune hemolytic anemia (AIHA) with high doses of prednisone. By the twelfth day his response allowed the medication to be tapered and by one month from the onset of treatment laboratory studies had returned to normal. The DAT remained negative, however, following recovery, anti-E could not be eluted from the red blood cells. Anti-E remained in his serum and the titer of the enzyme reactive antibody had decreased to 16. It is suggested that the anti-“E-like” antibody may represent auto anti-Hr preferentially reacting with E+ red blood cells. A unique feature in the case is the presence of a specific antibody eluted from cells that appear to lack that antigen in a DAT-negative patient with AIHA.     

An acute hemolytic transfusion reaction caused by an anti-P1 that reacted at 37 degrees C

BACKGROUND: Hemolytic transfusion reactions (HTRs) due to anti-P1 have rarely been reported. There is only one report (from 1945) of an acute HTR due to anti-P1. CASE REPORT: A 74-year-old woman with anti-P1 was given blood that had been found to be compatible by the use of prewarmed serum and saline-suspended red cells (RBCs) and of an antiglobulin test with anti-IgG. The test mixtures were not centrifuged or inspected for agglutination after the 37 degrees C incubation phase. After transfusion of 50 mL of P1 + blood, the patient had an acute HTR (hemoglobinemia, hemoglobinuria, and increased blood pressure, temperature, and respiration). RESULTS: When studied by a reference laboratory, the anti-P1 was shown to be easily detectable (3+ agglutination) by a prewarming technique (saline or low-ionic-strength saline [LISS]), which included centrifugation at 37 degrees C, but only weak reactions were observed when centrifugation after 37 degrees C incubation was omitted. The indirect antiglobulin test was weakly positive (1+) with anti-IgG, but polyspecific anti-human globulin reacted 2+. The anti-P1 agglutinin was IgM, and its titer was 16 at 37 degrees C (prewarmed) and 256 at 23 degrees C; it caused hemolysis of RBCs at 37 degrees C under conditions known to enhance hemolysis. An indirect monocyte monolayer assay gave results of 11.2 and 22 percent in testing of P1 + RBCs incubated with the patient's serum alone and with patient's serum plus fresh normal serum (as a source of complement), respectively (normal < or = 3%). CONCLUSION: An acute HTR was caused by a hemolytic anti-P1 that reacted at 37 degrees C. This antibody was not detected by the hospital in a prewarmed crossmatch that omitted 1) the addition of LISS, 2) the reading for agglutination after the 37 degrees C incubation, and 3) the use of antiglobulin sera containing anti-complement.     

An immediate hemolytic reaction induced by repeated administration of oxaliplatin

BACKGROUND: Platinum-based chemotherapy agents have been associated with potentially fatal acute immune-mediated hemolytic anemia. The target antigen, cause of the positive direct antiglobulin test (DAT) and mechanism of hemolysis have been the subject of controversy. CASE REPORT: We report a patient who developed a DAT-positive hemolytic episode after a red cell (RBC) transfusion was delivered during the infusion of her 17th cycle of oxaliplatin. Standard pretransfusion testing was uncomplicated; however, after infusion, the serum was no longer compatible with the transfused units and a strong (4+) panreactive IgG antibody was detected. RESULTS: The patient's serum from 10 days after the episode, only when therapeutic concentrations of oxaliplatin were added, reacted with all RBCs tested using the indirect antiglobulin test (IAT) (3+). The effect was retained with a purified IgG fraction and almost eliminated with IgG-depleted serum. Immunoprecipitation analysis revealed a band with the molecular weight of the Band 3 anion channel only in the presence of the patient's serum and oxaliplatin. CONCLUSION: Our investigations indicated that oxaliplatin interacted with both an IgG antibody and a RBC membrane epitope probably located on the Band 3 anion channel.     

A potent cold autoagglutinin that recognizes type 2H determinant on red cells

An example of an IgM (lambda) cold autoagglutinin against a Type 2H determinant of the ABO blood group system is described. The direct antiglobulin test was positive due to C3d on the patient's red cells. The antibody in the patient's serum was active at 37 degrees C, and the degree of agglutination was almost the same at 37 and 4 degrees C. It agglutinated group O red cells preferentially, but not Bombay (Oh) red cells. The specificity was determined by adsorption with synthetic oligosaccharide immunoadsorbent.