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[Association of autoimmune thrombopenic purpura and leiomyoma. Recovery after excision of the tumor].

A case of auto-immune thrombocytopenic purpura in a woman with benign cystic leiomyoma in Retzius' space is reported. The platelet Coombs' test was positive on platelets; the indirect Coombs' test was negative on serum but strongly positive on cyst fluid. Following excision of the leiomyoma thrombocytopenia rapidly subsided and the serological findings became negative, which strongly suggests that the antibody was produced by the tumour.

Abdominal Neoplasms↗

Hyperthyroidism associated with autoimmune hemolytic anemia and periodic paralysis: a report of a case in which antihyperthyroid therapy alone was effective against hemolysis.

We describe a case of a 29-year-old man with hyperthyroidism associated with autoimmune hemolytic anemia and periodic paralysis. Euthyroidism, which was achieved by propylthiouracil, brought inhibition of hemolysis and amelioration of anemia in spite of continuously positive direct and indirect Coombs' tests. Neither adrenocortical steroid nor blood transfusion was administered. Since indirect monospecific Coombs' test was negative against anti-human complements serum, the membrane of red blood cells may be less fragile. This is one reason why hemolysis was inhibited by anti-hyperthyroid therapy only. This may indicate that the hyperdynamic circulatory state secondary to hyperthyroidism plays an important role in the destruction of red blood cells which were coated by anti-red blood cell antibody.

Adult↗

Management of fetal hemolytic disease by cordocentesis. I. Prediction of fetal anemia.

Between January 1985 and November 1990, 128 pregnancies complicated by maternal red blood cell alloimmunization were referred to our Fetal Diagnosis and Treatment Unit. We examined the premise that an evaluation of fetal blood would accurately identify fetuses at risk of requiring antenatal transfusion therapy. Two hundred seventy-two diagnostic cordocenteses were performed. Criteria for the timing of repeat cordocenteses were developed retrospectively on the basis of the fetal hematocrit values, reticulocyte counts, and direct Coombs' test results of the first 84 pregnancies. These criteria were tested and confirmed prospectively on the next 44 pregnancies. On the basis of the first blood sample, four hematologic patterns (and their distributions) were identified in the 98 antigen-positive fetuses. Pattern 1: fetuses at low risk of having significant antenatal anemia (hematocrit less than 30%) (n = 11, 11%). These fetuses had normal hematocrit values and reticulocyte counts coupled with negative or trace-positive direct Coombs' test. No fetus in this group had significant antenatal anemia. Pattern 2: fetuses at intermediate risk of having anemia (n = 29, 31%). Pattern 2 fetuses had normal hematocrit values and either direct Coombs' titers of more than trace less than or equal to 2+ and normal reticulocyte counts or low reticulocyte counts (less than 2.5th percentile for gestation). Twenty-one percent (n = 6) of fetuses in pattern 2 had significant antenatal anemia. Patterns 3 and 4: fetuses at greatest risk of having severe anemia. These fetuses had normal hematocrit values associated with either reticulocyte counts greater than 97.5th percentile for gestation or a direct Coombs' test greater than or equal to 3+ (pattern 3, n = 49, 50%) or both, or a mild anemia (greater than 30% but less than 2.5th percentile for gestation) (pattern 4, n = 9, 10%). Eighty percent (n = 39) of fetuses with pattern 3 and 90% (n = 8) with pattern 4 developed a hematocrit value less than 30%. We conclude that evaluation of fetal hemolytic disease with a fetal blood specimen permits the identification of fetuses at high risk of having antenatal anemia.

Amniocentesis↗

Application of a high sensitivity aggregate-haemagglutination test for the diagnosis of autoimmune haemolytic anaemia with a negative direct antiglobulin test.

An aggregate-haemagglutination test has been used for determining antierythrocyte autoantibodies. The first antiglobulin variant of the test allows us to establish the presence of antibodies in 33 cases with a negative direct Coombs' test. The test II (antiantiglobulin variant) proved to be positive in 88 cases of auto-immune haemolytic anaemia (AIHA) that showed a negative direct Coombs' test and a negative antiglobulin variant. Immunoglobulin G has been revealed in the majority of AIHA patients. IgM has been determined in 1 case of a symptomatic form associated with chronic lymphocytic leukaemia. IgA has been recorded in 3 cases with an idiopathic AIHA form and in 6 cases of chronic lymphocytic leukaemia. Both types of light chains were found on the surface of erythrocytes in all cases of AIHA.

Anemia, Hemolytic, Autoimmune↗

A manual semi-micro method for quantitation of haemagglutination.

A manual semi-micro method for quantitation of haemagglutination is described. It appears to be of wide use for most agglutination reactions usually performed in test tube, including Coombs-test. It is easy to handle and requires the usual equipment of an immunohaematological laboratory. The accuracy obtained for quantitation of antibody concentrations is similar to that of the auto-analyser.

Autoanalysis↗

Anti-erythrocyte natural antibody activity in the unconventional 'heavy chain' immunoglobulins of Indian desert camel (Camelus dromedarius).

Members of the family Camelidae contain unconventional 'heavy chain' antibodies (HCAbs) that are devoid of light chains (LCs) in their structure and occur under physiological conditions. The spectrum of antigenic specificities in HCAbs and hence their biological significance is not known at present. Recent studies have however indicated that they contribute significantly towards the immunoglobulin (Ig) receptor diversity. The present study was planned to investigate the natural antibody (NAb) activity in camel HCAbs as further indication of the wide spectrum of their antigenic specificities. Detection of NAbs in the sera and isolated HCAbs of Indian Thar desert camels was undertaken against erythrocyte antigens (E-Ags) from eleven animal species including nine mammals, chicken and frog by using direct haemagglutination (HA) and indirect Coombs' test. HCAbs were found to behave as 'incomplete antibodies' and agglutinated erythrocytes of different animal species in indirect Coombs' test using rabbit anti-camel IgG3 (HCAbs) antiserum. Variations were noticed in the Coombs' titres against erythrocytes from different species. HCAbs also reacted against E-Ags in immunoblots. These findings provide further evidence that camelid HCAbs are produced against diverse antigens (Ags) under natural conditions, thereby contributing to camelid Ig receptor diversity.

Animals↗

Application of the enzyme-linked immunosorbent assay to detect canine erythrocyte antibodies.

A double-antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies bound to canine RBC. The method used a horseradish peroxidase conjugate, o-dianisidine, as a water-soluble chromogen and osmotically lysed RBC on which antibody was detected. The ELISA method was compared with the traditional antiglobulin (Coombs') test on positive controls, using sheep RBC sensitized with dilutions of canine anti-sheep RBC serum and on RBC from canine patients tested for autoantibodies to RBC. The ELISA method appeared to be more sensitive than the Coombs' test and gave an estimate of the amount of antibody present without the need for serial antiserum dilutions to determine the antibody titer.

Anemia, Hemolytic, Autoimmune↗

Specific antibodies detected during relapse of human brucellosis.

We studied 10 patients who had a relapse of brucellosis for significant serological changes during relapse. By the Coombs test, the pre-relapse and post-relapse median (range) titers of antibody to Brucella were 1:1120 (1:40-1:10,240) and 1:10,240 (1:40-1:81,920), respectively (P = .0069); by enzyme-linked immunosorbent assay (ELISA) for IgG, these values were 1:2240 (1:60-1:10,240) and 1:10,240 (1:160-1:81,920; P = .0069); by ELISA for IgA, the median titer increased 3.36 times but did not reach statistical significance (P = .0929); by tube agglutination, dithiothreitol (DTT) agglutination, and ELISA for IgM, median titers did not change. Nine patients had a significant titer increase by Coombs test and IgG ELISA, three had a significant increase by tube and DTT agglutination, none had significant increases by IgM ELISA, and one had no significant increase after relapse. Our findings show that for most patients with a relapse of brucellosis, there is an increase in IgG titers, as detected by ELISA and Coombs test, but no change in IgM titers.

Adolescent↗

[IgA-lambda type myeloma complicating with autoimmune hemolytic anemia].

A 66-year-old female was admitted to our hospital with lumbago. On admission, laboratory examination revealed hemolytic anemia. Direct Coombs' test was positive, and also direct monospecific-Coombs' test by anti-C3d serum was positive. Immunoelectrophoresis showed IgA-lambda type M proteins in serum. Bone marrow aspiration disclosed increased atypical plasma cells. X-ray of skull showed punched-out lesion. From these findings, she was diagnosed as IgA (lambda) myeloma complicating with autoimmune hemolytic anemia. Hemolysis was improved by chemotherapy. It was thought that IgA and C3 were related well to hemolysis in this case.

Aged↗

[A case of autoimmune hemolytic anemia and bullous pemphigoid-like skin lesion combined with idiopathic thrombocytopenic purpura].

A case of autoimmune hemolytic anemia (AIHA) and bullous pemphigoid (BP)-like skin lesion combined with Idiopathic thrombocytopenic purpura (ITP) is reported. A 25-years-old male, who had been diagnosed as ITP and treated at another hospital, was admitted in this hospital recently complaining of disseminated bullous-vesicular eruptions on the whole body and autoimmune hemolytic anemia. Examinations, disclosed that RBC was 364 X 10(4)/microliters, reticulocyte 40, platelet 3000/microliters, direct and indirect Coombs test positive, and platelet Coombs consumption test was positive leading to the diagnosis of AIHA and ITP, known as "Evans syndrome." Vesicular biopsy-findings and immunofluorescence study showed suspicion of BP, but clinical course and blister was not improved though the administration of prednisolone was performed. Reports of cases of BP complicated by Evans syndrome are very few. AIHA, ITP and BP are considered to have autoimmune disorders and their pathogenetic mechanism are discussed. This patient consulted another hospital one year later, when we heard that skin eruptions already had disappeared.

Adult↗

[Intravenous administration of anti-D hyperimmunoglobulin after mismatched transfusion (author's transl)].

620 ml of 0 Rh-positive blood (310 ml CCD.ee + 310 ml CcD.EE) were administered to a 27-year-old 0 Rh-negative (ccddee) woman in connection with an emergency curettage. 16 hours after this mismatched transfusion an intravenous infusion of anti-D hyperimmunoglobulin was started: 8000 mug (13 mug/ml blood) were given within 14 hours (570 mug/h) under strict supervision. The 10% immunoglobulin preparation had been diluted 20 times. There were only slight variations in heart rate and blood pressure, slight fever and mild increase in serum bilirubin after the infusion. There was slight haemoglobinuria in one urine sample, but no signs of kidney damage. The direct Coombs test was slightly positive after two days, negative after four and six days. The indirect Coombs test was negative after six months. The patient has not become pregnant since. Slow intravenous infusion of highly diluted anti-D hyperimmunoglobulin, under strict clinical supervision, is probably appropriate for Rh prophylaxis after transfusion mismatch. The dose of 13 mug/ml could probably be considerably reduced. 250 mug anti-D per hour should not be exceeded.

Adult↗

Erythrocyte autoantibodies and expression of CD59 on the surface of red blood cells of polytransfused patients with beta-thalassaemia major.

Repeated transfusions for the treatment of thalassaemia major cause an insult to the patient's immune system and provoke post-transfusion purpura and haemolytic reactions that can be severe and life threatening. This study aims to investigate the presence of erythrocyte autoantibodies and CD59 expression on the surface of red blood cells (RBCs) in patients with beta-thalassaemia major, and any relationship to frequency of blood transfusion. The study looks at a total of 49 patients (both children and adults) with beta-thalassaemia major, divided into four groups according to the number of blood transfusions received and the presence or absence of the spleen. Glycosylated haemoglobin, Coombs' test (direct and indirect) and CD59 level on the RBC surface (by flow cytometry) are estimated in all patients studied. Glycosylated haemoglobin level was significantly lower in those who had received less than 10 units of blood (group III) than in those who had received more than 25 units of blood and had undergone splenectomy (group Ib), and was significantly lower in those who had received 10-25 units of blood (group II) than in those that comprised group Ib (F=3.598, P=0.0205). Considering CD59 expression, there was a marked difference between the groups. Expression was highest in group III and diminished progressively through groups II, Ia (polytransfused, non-splenectomised) and Ib (F=19.83, P=0.0000). No relationship was observed between CD59 expression and either blood group or gender. A significant negative correlation between CD59 expression and reticulocyte percentage (r=-0.538, P=0.000) and normoblast count (r=-0.5455, P=0.000) was found. A negative correlation between lymphocytosis and CD59 expression was also noted in groups III (r=-0.745, P=0.013), Ia (r=-0.5849, P=0.022) and Ib (r=-0.6711, P=0.009). Direct Coombs' test was positive in only one patient in group Ib, who also showed the lowest haemoglobin level. Thalassaemia patients exposed to multiple antigens through repeated blood transfusions showed lower CD59 expression than did those who had received fewer transfused units, which is a good method of detecting potential autoantibodies. Furthermore, a negative Coombs' test does not exclude autoimmunisation in such patients.

Adolescent↗

[Comparative study of different serological tests for the diagnosis of brucellosis (author's transl)].

The diagnosis of brucellosis is frequently a difficult one, and it is founded basically on the results of blood cultures and serological tests. Wright's classical agglutination test is commonly utilized in the serological diagnosis of brucellosis and its value is well established in the acute forms of the disease; for the diagnosis of subacute or chronic forms, however, a Coombs test must be performed to show incomplete antibodies. The agglutination carried out with serum treated with 2-mercaptoethanol allows the determination of the type of immunoglobulins present. Indirect immunofluorescence has been infrequently utilized in the diagnosis of brucellosis. The first three tests have been compared with indirect immunofluorescence using sera from 100 patients presenting with fever of unknown origin. All tests were negative in 40 sera, and of the 60 remaining sera only 32 were positive to all four tests. The highest titers were found with the Coombs test, while immunofluorescence yielded titers which were never above 1/320. Significant correlation coefficients were obtained between the tests. The best correlation was obtained between immunofluorescence and 2-mercaptoethanol, followed by immunofluorescence and Coombs, and by immunofluorescence and agglutination. Only immunofluorescence titers above 1/80 corresponded to the same of superior titers in the remaining tests; thus immunofluorescence titers below 1/80 can not be considered as diagnostic of the disease. The authors believe that immunofluorescence to 1/80 can be utilized as a quick routine test for the diagnosis of brucellosis, its disadvantages being a high cost and the requirement for an immunofluorescence microscope which may not be available in many laboratories.

Agglutination Tests↗

[Cold agglutinins, clinical presentation and significance; retrospective analysis of 58 patients].

PURPOSE: To describe clinical, biological characteristics and associated diseases of cold agglutinins in adults. METHODS: Retrospective study in a single department of internal medicine from 1997 to 2002. The inclusion criteria were a positive direct Coombs test and a positive research for cold-reactive autoantibodies. We recorded for each patient: clinical presentation at onset and during follow-up, biological parameters of haemolysis, biological characteristics of the cold agglutinin and associated diseases. RESULTS: Fifty-eight patients (34 females, 24 males), with medium age of 58.8 were included in the study. Clinical presentation was highly variable between acute life-threatening haemolysis and absence of symptoms. Results of direct antiglobulin test were C3 (74%), IgG + C3 (22.4%), IgG (3.4%). Titer, thermal amplitude, strength and specificity of Coombs test were correlated, in all cases except 6, with cold agglutinin haemolytic activity. In 77.6% of cases cold agglutinin was secondary; related to: autoimmune disorders (n = 19), lymphoproliferative disorders (n = 11) and infections (n = 10). CONCLUSION: Clinical presentation of cold agglutinin is highly variable and not always related to the biological characteristics of the bound antibody (titer, thermal amplitude, specificity). In our single center study, diseases associated with cold agglutinin were various with the highest frequency of auto-immune disorders. Our study underlined also the high frequency of lymphoproliferative disorders and justifies a close follow-up of these patients. Finally, we reported a high frequency of hepatitis C virus infection among the infectious aetiologies.

Adult↗

[Immunological properties of S-1090, cefmatilen hydrochloride hydrate].

S-1090, a cefmatilen hydrochloride hydrate, is being developed as a cephalosporin antibiotic for oral use. Immunogenicity, hypersensitivity-eliciting antigenicity and immunological cross-reactivity with other antibiotics were evaluated by active systemic anaphylaxis (ASA) test, passive cutaneous anaphylaxis (PCA) test and enzyme-linked immunosorbent assay (ELISA) using guinea pigs and mice/rats. In addition, in vitro direct Coombs' test was also performed to examine the possibility of hemolytic anemia in clinical use. Immunogenicity of S-1090 was not observed in guinea pigs after repeated immunization with S-1090 by ASA or PCA tests. Even in ELISA, only weak antibody production against S-1090 was found in some guinea pigs from the intraperitoneal groups showing the antibody titers only 10(1) to 10(2). When the sera collected from C3H/He mice and C57BL/6J mice immunized with S-1090 were tested for immunogenicity, rat PCA was elicited in a C3H/He mouse serum by S-1090 and antibodies against S-1090 were detected in a C57BL/6J mouse serum by ELISA. When adjuvant was used in mice and guinea pigs, the production of antibody against S-1090 was less frequent in comparison with other antibiotics such as cefmetazole (CMZ) and cefotiam (CTM). When hypersensitivity-eliciting antigenicity of S-1090 was examined using S-1090 as an eliciting antigen in ASA and PCA tests, positive ASA and PCA were observed in guinea pigs and positive PCA in a C3H/He mouse. Hypersensitivity-eliciting antigenicity was also observed in other reference antibiotics, i.e. cephalothin (CET), CMZ and CTM. Immunological cross-reactivity among S-1090, penicillin G (PCG), CET, CMZ and CTM was tested by ASA and PCA tests. S-1090 was found to immunologically cross-react only with CET in guinea pigs. In the present study, immunological cross-reactivities were also noted between PCG and CET, PCG and CMZ, PCG and CTM, and between CET and CMZ. In in vitro direct Coombs' test using human red blood cells, S-1090.Na, PCG and CET gave positive reactions at the final concentrations of 40 mg/mL, 20 to 40 mg/mL and 2.5 to 10 mg/mL, respectively.

Animals↗

[A case of pure red cell aplasia associated with sodium valproate therapy].

We reported a 14-year-old female with petit mal epilepsy who suffered from pure red cell aplasia 12 weeks after sodium valproate (VPA) therapy started. On admission she presented anemia but no jaundice. Her hemoglobin was 5.4g/dl and bone-marrow biopsy revealed hypocellularity with erythroid hypoplasia. The Direct Coombs' test and anti-nucleotide antibody were positive. Reticulocytosis occurred 10 days after discontinuation of VPA. The Direct Coombs' test became negative one year after discontinuation of VPA. In general, drug-induced PRCA is caused by toxic or allergic agents, but this case suggests the immune mechanism of VPA-induced PRCA.

Adolescent↗

[Elderly dyserythropoietic anemia first diagnosed after presentation of hemorrhagic gastric ulcer].

A 71-year-old man presented with epigastralgia and tarry stool. Laboratory examination showed anemia (Hb 7.1 g/dl) due to hemorrhagic gastric ulcer and positive Coombs' test without features of hemolysis. Initial bone marrow smears disclosed normal granulocytes and megakaryocytes, but only erythroid hyperplasia with multinuclearity and megaloblastosis was identified. Cytogenetic studies revealed normal karyotype. Congenital dyserythropoietic anemia (CDA) type II was initially suspected. Serologically, however, acid hemolysis and anti-I * i tests were negative as were the results of another Coombs' test. The second bone marrow aspiration disclosed a marked decrease in multinucleated erythroblasts together with an increase in those that had internuclear chromatin bridges. Electron microscopy of bone marrow specimens demonstrated morphological features of CDA types I and II, thus yielding a final diagnosis of elderly dyserythropoietic anemia, which is similar to both CDA types. It was suggested that the reactive secretion of erythropoietin due to bleeding played a role in these pathologic changes.

Aged↗

[Detection and side effects of isoantibodies in intravenously administered immunoglobulin preparations].

BACKGROUND: Intravenous immunoglobulins (IvIg) contain not only the declared antibodies against pathogenic microorganisms, but also all the other antibodies of the blood donors, e.g. against erythrocytic antigens. MATERIALS AND METHODS: We tested 14 IvIg from 7 manufacturers (a total of 40 charges) for isoantibodies and irregular antibodies. To improve the reading of our tests we used the gel centrifugation method (ID-Microtyping-System, Fa. Diamed, Bensheim, Germany). RESULTS: The highest isoantibody titers were (in 8 charges) 1:32 or 1:64 in the Liss-Coombs test. Irregular antibodies were found in 5 IvIg (maximal titer 1:8). CONCLUSIONS: Isoantibodies in the IvIg can influence blood group serologic tests. With an example of a newborn who had received IvIg we point to the potential danger of misinterpretation of a positive direct antiglobulin test after administration of IvIg. Therefore we recommend to carry out the direct antiglobulin test before administration of IvIg and to examine all eluates after a positive direct Coombs test not only with 0 RBCs but also with A or B RBCs of the AB0 blood group of the patient.

ABO Blood-Group System↗