Drug-Dependent Coombs (Antiglobulin) Test and Anemia

Abstract

The Coombs (antiglobulin) test as applied to erythrocytes has been especially of value in the identification and study of isoimmune states1,2 and in acquired hemolytic anemia3,4 in which an autoimmune state has been proposed.5,6 Recently Harris7 described an additional application of the Coombs test to erythrocytes which was dependent on humoral factors in the patient's serum plus the trivalent antimony compound stibophen (Fuadin). This phenomenon, associated with a hemolytic state due to multiple contacts by the patient with the drug stibophen, may be compared with the drug-dependent agglutination of platelets described by Ackroyd 8,9 in the thrombocytopenia induced by allylisopropylacetylcarbamide (Sedormid) and varified for quinidine 10-12 and quinine.13 It is also of interest that the agglutination of leukocytes dependent on serum factors and a drug, similar to Ackroyd's phenomenon for platelets, has been described.14 In the present report we wish to record the encounter of a positive Coombs test for erythrocytes which appears dependent on serum factors plus the drugs quinine and acetophenetidin ( phenacetin ).The study began with observations on a patient who developed a fulminant hemo¬ lytic anemia upon the second contact with quinine. It was subsequently extended to the serum from 112 normal subjects. More recently studies of a second patient with an anemia associated with the intake of acetophenetidin yielded similar information. MethodThe Coombs (antiglobulin) test1,2 was per¬ formed as follows: 0.1 ml. of serum, 0.1 ml. of a 2% suspension of erythrocytes, and 0.1 ml. of a solution of the drug to be tested were incubated for one hour in a water bath at 37 C. After this the erythrocytes were washed three times with abundant saline (buffered to pH 6.8-7.0). Follow¬ ing the third wash the test tube was turned upside down and was allowed to drain. The erythrocytes were allowed to settle into the residual saline, and to this was added 0.1 ml. of the antiglobulin serum.The mixture stood for 15 minutes at room tem¬ perature and then was spun at 1000 rpm for one minute. The button of erythrocytes was shaken gently, and the test was read grossly ; following this the contents were poured onto a glass slide and observed microscopically for stability or lack of stability of the clumps. A solid clump without free erythrocytes was read as 4+; a few major stable clumps with scattered intervening erythro¬ cytes were read as 3+ ; smaller stable clumps were read as 2-\-; scattered fine clumps which were stable for five minutes microscopically were read as 1+; other findings were considered negative ( Figs. 1 and 2). The Coombs (antiglobulin) serum used was pre¬ pared locally. The serum was prepared in the rabbit, with use of pooled whole human serum as described previously for the preparation of canine antiglobulin serum (whole-serum rabbit serum).15 This serum gave good results when tested with Rh+ (D-f-) erythrocytes lightly and heavily coated with incomplete anti-D antibody.The trypsin-treated erythrocytes were prepare...

Acquired Hemolytic Anemia, Exposures to Insecticides and Positive Coombs Test Dependent on Insecticide Preparations

“…The type of case reported in this discussion has features similar to those of two other cases reported from this laboratory in which a drug dependent indirect Coombs test was noted [7]. In the latter cases the drugs quinine and acetophenetidin were involved.…”

“…The laboratory methods used were the same as referred to in two former reports [5,7]. There are a few special features in the present communication.…”

A Case of Haemolytic Anaemia due to Phenacetin Allergy

“…In the case reported by Muirhead et al [15] the patient was a coloured woman, the sensitising dose was not indicated, and the crisis was not accompanied by renal compli cations. The antibody was of the incomplete type, active only in the presence of complement.…”

Hemolytic effects of drugs and chemical agents

“…Occasionally cells from an affected patient will agglutinate directly with antiglobulin serum, but usually, for some inexplicable reason, the presence of the antibody can only be demonstrated in the patient's plasma and not on his cells. 98 The following drugs have been incriminated in causing immune hemolysis:…”

“…Stibophen (Fuadin) 58 Quinine 98 Quinidine 48 Acetophenetidin 84 Para-aminosalicylic acid"o, 84 Antazoline (Antistine) 86 Salicylazosulfapyridine (Azulfidine) 115 The mechanism leading to the production of antibody by these drugs in certain individuals only is not clear. Two principal possibilities present themselves.…”