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An enzyme-linked antiglobulin test for the detection of erythrocyte-bound antibodies in canine autoimmune haemolytic anaemia.

A method has been developed which allows identification and quantitation of red cell-bound immunoglobulins and complement in canine blood. The technique utilizes ELISA methodology and the assay identifies cases of autoimmune haemolytic anaemia which are negative by the Direct Coombs test. Further, the amount of antibody present on the red cells shows a close correlation to the haemoglobin level; suggesting that the degree of sensitization of the RBCs influences their rate of destruction. Favourable response to treatment correlates with a decrease in the levels of bound antibody and complement.

Anemia, Hemolytic, Autoimmune↗

Idiopathic pure red cell aplasia and nonregenerative immune-mediated anemia in dogs: 43 cases (1988-1999).

OBJECTIVE: To examine clinical features, laboratory test results, treatment, and outcome of dogs with pure red cell aplasia (PRCA) and idiopathic nonregenerative immune-mediated anemia (NRIMA). DESIGN: Retrospective study. ANIMALS: 43 dogs with severe nonregenerative anemia. PROCEDURE: Medical records of dogs determined to have PRCA, NRIMA, or ineffective erythropoiesis on the basis of bone marrow analysis between 1988 and 1999 were reviewed. Criteria for inclusion were > or = 5-day history of severe nonregenerative anemia (Hct < 20%; < 60.0 x 10(3) reticulocytes/microliter) with no underlying diseases. Information was retrieved on signalment, clinical signs, laboratory test results, treatment, and outcome. RESULTS: Median age of the dogs was 6.5 years. Spayed females and Labrador Retrievers were significantly overrepresented. Median Hct was 11% with no evidence of regeneration (median, 1.5 x 10(3) reticulocytes/microliter). Direct Coombs' test results were positive in 57% of dogs. Biochemical abnormalities included hyperferremia and high percentage saturation of transferrin. Bone marrow findings ranged from PRCA (5%) to erythroid hyperplasia (55%). Myelofibrosis was common. Dogs were treated with immunosuppressive drugs and the response was complete, partial, and poor in 55, 18, and 27% of the dogs, respectively. Mortality rate was 28%. CONCLUSIONS AND CLINICAL RELEVANCE: An immune-mediated pathogenesis should be considered in dogs with severe, nonregenerative anemia, normal WBC and platelet counts, hyperferremia, mild clinical signs, and no evidence of underlying disease. Bone marrow findings range from the rare PRCA to erythroid hyperplasia. Myelofibrosis is often detected in affected dogs and may prevent bone marrow aspiration.

Anemia, Hemolytic, Autoimmune↗

In vivo binding of circulating immune complexes by C3b receptors (CR1) of transfused erythrocytes.

The effects of packed erythrocyte transfusion with high CR1 activity on circulating immune complex concentrations were studied in 14 transfusion experiments involving 12 patients with immune complex related diseases. Before erythrocyte transfusion circulating immune complex concentrations ranged from 8 to 128 micrograms/ml. After transfusion (2-3 units) immune complex concentrations decreased depending on the levels of CH50 titres in the recipients. In 11 experiments, in which the patients' CH50 titres ranged from 21 to 44, immune complex concentrations decreased by 75-100% within five days. The CH50 titres were also decreased after erythrocyte transfusion but subsequently increased to initial ranges within 6-35 days. In three patients with low CH50 titres (1.0-10.0) decreases in immune complexes were not observed. Direct Coombs' tests for IgG and C3 were performed before and after erythrocyte transfusion to determine potential in vivo binding of circulating immune complexes. Thus in eight of 14 experiments, in which erythrocytes carried no IgG before packed erythrocyte transfusion, seven became Coombs' positive for IgG after the transfusion. In seven of 14 experiments, in which erythrocytes were negative for complement before transfusion, five became positive afterwards. Moreover, in 12 instances slight increases of CR1 activity of patients' erythrocytes were observed within eight days, which improved further within 35 days after erythrocyte transfusion. These studies suggest that transfusion of erythrocytes with high CR1 activity results in the removal of circulating immune complexes and that this process is dependent on complement consumption. These experiments support the hypothesis that erythrocyte-CR1 has a functional role in the removal of circulating immune complexes and may thereby inhibit the deposition of immune complexes within body tissue constituents.

Antigen-Antibody Complex↗

Red-cell sensitization in myelofibrosis.

Using an enzyme-linked immunosorbent assay (ELISA), elevated RBC-Ig of the IgG and IgM class were found in 8 of 14 patients with idiopathic myelofibrosis. In 2 patients with high levels of RBC-Ig the direct Coombs' test was positive. It is supposed that immune haemolysis may contribute to the anaemia in some patients with idiopathic myelofibrosis.

Aged↗

Severe immune hemolytic anemia in disseminated tuberculosis with response to antituberculosis therapy.

Severe hemolytic anemia in patients with disseminated tuberculosis is exceedingly rare. We report an episode of Coombs'-positive hemolytic anemia in a previously healthy young man with miliary tuberculosis, resulting in a hemoglobin level of 5 g/dL and an undetectable haptoglobin level. The patient responded well to treatment with antituberculosis drugs, and the results of the direct Coombs' test became negative without the need of blood transfusion or steroid therapy.

Adult↗

Transmission of auto-immune haemolytic anaemia and murine leukaemia virus in NZB-BALB/c hybrid mice.

When mated to normal BALB/c partners, male and female NZB mice transmitted auto-immune haemolytic anaemia to three generations of their hybrid progeny. Red cell auto-antibodies (positive Coombs tests) were detected, on average, 11 months later in the F1 hybrids than in the parental strain, and the course of the disease was protracted. In explicably, the auto-immune reactions then appeared progressively earlier in successive generations of both croses. The Coombs reactions of the F1 and F2 hybrids were often weak and inconsistent, while those of F3 offspring showed the strong and stable picture typical of NZB mice. The incidence of auto-immune disease in each generation, although similar in the reciprocal crosses, indicated that the pattern of inheritance was very complex. The hybrids did not develop the lymphoid type B reticulum cell neoplasia which characterizes auto-immune NZB mice. Instead, and irrespective of Coombs status, they had lymphocytic leukaemias, lung adenomas, hepatomas and type A reticulum cell neoplasms of the liver. Murine leukaemia virus was identified electronmicroscopically in F1 embryos, and in all the adults examined. It was also isolated from leukaemic spleen cells passaged briefly in vivo, and from malignant hepatic (reticulum) cells maintained in vitro. These isolated were leukaemogenic in newborn BALB/c, NZB, and F1 hybrid recipients, but did not induce or accelerate positive Coombs reactions. Only a small proportion of the hybrids had significant glomerulonephritis, and overt kidney disease was minimal. The lesions were not confined to Coombs-positive mice. Possible links between auto-immunity, malignancy, and virus infection in NZB mice are discussed in the light of these results.

Anemia, Hemolytic, Autoimmune↗

[Typing and detection of antibodies in the PLA system (platelet). Application to the study of neonatal thrombopenia by feto-maternal PLA allo-immunisation].

A platelet indirect radio-active Coombs test has been described. The technique for purification and labelling the antiglobulin has been precised. This test allows the typing of platelets in the PLA system and the study of sera from mothers of thrombocytopenic child. As examples, four families of neonatal thrombocytopenia are reported, with PLA1 negative mother. In the serum of three of these mothers, we could demonstrate anti-PLA antibodies in spite of a negative platelet complement fixation. This test has many advantages compared to other tests such as platelet complement fixation, assay for blocking antibodies or antiglobulin consumption: it gives objective and quantitative results and is highly reproducible, anticomplementary serum may be tested. It has enabled us to select PLA1 negative donors for exsanguino-transfusions of thrombocytopenic children born from PLA1 negative mothers.

Blood Platelets↗

Quantitation of erythrocyte-bound IgG by competitive solid-phase enzyme immunoassay.

A competitive solid-phase enzyme immunoassay technique for the quantitation of erythrocyte (RBC)-bound IgG was devised. This method is highly sensitive, reproducible and can be performed more simply. Anemic dogs with a positive Coombs' test result were observed to have higher RBC-bound IgG values (IgG concentration, ng/ml) than healthy dogs.

Anemia↗

Probenecid induced immune hemolytic anemia.

We report a patient who, while receiving probenecid and colchicine for acute gouty arthritis, developed severe hemolytic anemia in association with a generalized rash. The hemolysis was immune mediated as shown by a positive direct Coombs' test. In vitro hematologic studies showed that a probenecid dependent antibody was present.

Anemia, Hemolytic, Autoimmune↗

Neurobrucellosis in childhood: six new cases and a review of the literature.

Neurobrucellosis accounts for <1% of cases of brucellosis in children. Six new cases of neurobrucellosis are presented and data from 39 previously published cases are analysed. The incidence is equal in males and females, and the source of infection is likely to be unpasteurised milk. Clinical presentation varies from severe meningoencephalitis or peripheral neuropathy/radiculopathy to behavioural disturbance. Diagnostic certainty requires isolation of the organism from the CSF, but as this is rarely possible serological diagnosis can be performed with the Coombs test on the CSF. Treatment requires combination antibiotic therapy and should continue for at least 8 weeks.

Animals↗

Significance of anti-A and anti-B isohemagglutinins in cord blood of ABO incompatible newborn infants: correlation with hyperbilirubinemia.

Transcutaneous bilirubin index measurements (TcBI) were determined in ABO-incompatible term white newborn infants who had an indirect Coombs' test for detection of ABO incompatibility (ABO test) performed on cord blood. Serial transcutaneous measurements in the first 72 hours of life in 63 ABO test positive (group I) and 31 ABO test negative (group II) infants were compared with a control group of 109 term ABO compatible infants (group III). During the first phase of this study, the accuracy of TcBI was confirmed in 33 infants who had serum bilirubin determinations demonstrating a correlation coefficient of r = 0.93 between the jaundice meter index reading and total serum bilirubin. Determination of total serum bilirubin using TcBI did not show any statistically significant differences when groups I, II, and III were compared for maximum serum bilirubin values. These results reveal that a positive ABO test does not increase the predictive value of hyperbilirubinemia in ABO-incompatible white newborn infants.

ABO Blood-Group System↗

Developing self-instructional learning materials--a case study.

A case study in the development of a self-instructional learning unit is presented. A seven-step systems approach "Design for Learning Programs" was followed in the development of a self-instructional unit on "The Direct Coombs' Test." Each step of the process as well as the problems encountered in producing a self-instructional package is described. This model is adaptable to other learning design situations.

Coombs Test↗

A case of thymoma-associated autoimmune haemolytic anaemia.

A 50-year-old female presented with severe Coombs-positive haemolytic anaemia. Chest roentgenogram revealed a right-sided paracardial mass in the anterior mediastinum that was proven to be a thymoma. The patient was treated with oral prednisone (1 mg/kg/day) and subsequent thymectomy. Haemolysis did not return although the direct Coombs test remained weakly positive. In reviewing the literature 15 additional cases of thymoma-associated autoimmune haemolytic anaemia were identified.

Administration, Oral↗

[Quantitative determination of EAIgG by ABC-ELISA and its clinical significance].

OBJECTIVE: To explore a method for diagnosis of autoimmune hemolytic anemia (AIHA) and evaluate its clinical significance. METHODS: Erythrocyte associated IgG (EAIgG) was quantified in 60 healthy people, 44 patients with warm antibody AIHA (WAIHA), 108 non-immune anemia and 44 other immune diseases (total 256 cases) by using ABC-ELISA were estimated. RESULTS: EAIgG in WAIHA patients were significantly higher than those in other groups (P < 0.01). In the 44 WAIHA patients, 38 were Coombs positive and 6 Coombs negative. EAIgG in Coombs negative was significantly high. The dynamic changes of EAIgG in 12 WAIHA patients were observed during treatment. When patient's condition got better, Hb and RBC increased and EAIgG decreased. CONCLUSION: Quantitative detection of EAIgG is an index in the diagnosis of WAIHA, especially in AIHA with negative Coombs' test. It is also a useful factor for evaluating therapeutic effect and prognosis in WAIHA.

Adolescent↗

A long-term study of diclophenac sodium in the treatment of rheumatoid arthritis and osteo-arthrosis.

One hundred and two patients with either rheumatoid arthritis or osteo-arthrosis were treated for prolonged periods with diclophenac sodium (Voltaren; Geigy) to evaluate the efficacy and tolerability of the drug. Fifty-seven patients completed a trial of 12 months. A total of 70% showed an improvement in functional class, and 40% of the total had complete functional capacity by the end of the trial. The drug was well tolerated. The side-effects (heartburn, abdominal cramps, headache and dizziness) were mild and in most cases did not require cessation of treatment. In 9 patients the Coombs test became positive during the trial, but this did not require cessation of therapy.

Adult↗

[Haemolytic anaemia and acute renal failure associated with cephalothin and furosemide therapy (author's transl)].

A case report is presented of a 79 year old patient with a bilateral brochopneumonia. Four days after beginning the therapy with cephalothin and furosemide an acute renal failure appeared and six days later a haemolytic anaemia was observed. The modificated Coombs test was positive. The post mortem histiological examination of the kidneys revealed cloudy swelling, necrosis of tubuli, containing protein cylinders ("chromoproteid kidney").

Acute Kidney Injury↗

Imipenem/cilastatin in the treatment of serious bacterial infections.

Imipenem (N-formimidoyl thienamycin) is a new carbapenem beta-lactam antibiotic with a broad antibacterial spectrum. Forty-five patients were treated with either 500 or 1,000 mg of imipenem/cilastatin four times daily, the duration varying according to clinical response. The diagnoses were urinary tract infection, 10 patients; septicemia, six; intraabdominal sepsis, six; pneumonia, six (two cases of Legionnaires' disease); skin and soft tissue infection, four; and other diagnoses, 13. Of the 32 clinically assessable patients, 17 were cured, nine improved, three died, and three were withdrawn from the trial. Of 21 patients who were microbiologically assessable, 13 were cured. In six cases of complicated urinary tract infection, the organism--which had been eradicated from the urine during treatment--reappeared after completion of antibiotic therapy. Two patients developed adverse clinical reactions that were thought to be drug-related (drug-induced fever and nausea plus vomiting, respectively). Both patients had mildly abnormal results in liver function tests, and one developed a positive direct Coombs' test. Fifty-seven percent of the patients developed some degree of phlebitis, which was moderate to severe in 19%. In this study imipenem/cilastatin proved to be a highly effective agent for the treatment of a variety of serious bacterial infections.

Adolescent↗