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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...

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Summary A case of acquired hemolytic anemia has been presented in a patient who had multiple exposures to insecticides and whose serum displayed a positive indirect Coombs test which was dependent on insecticide preparations. Evidence has been presented which shows differences between the insecticide dependent serum factor and the factors coating the patient's erythrocytes. The clinical syndrome in this case was similar to that of two other cases demonstrating a positive indirect Coombs test which was drug dependent. Résumé Présentation d'un cas d'anémie hémolytique acquise chez un malade qui avait été exposé à plusieurs reprises à des insecticides. Le test de Coombs indirect avec le sérum du malade était positif, mais seulement en présence de préparations d'insecticide. Le facteur sérique responsable de ce phénomène et le facteur adhérent aux érythrocytes du malade et causant un test de Coombs direct fortement positif ne semblaient pas être entièrement identiques. Le syndrome clinique dans le cas présenté ressemblait à celui de deux autres cas dans lesquels le test de Coombs indirect était également positif en présence de médicaments. Zusammenfassung Es wird über eine erworbene hämolytische Anämie bei einem Patienten, welcher wiederholt mit Insektiziden in Berührung kam, berichtet. Das Patientenserum zeigte bei Zugabe von Insektiziden einen positiven indirekten Coombstest. Derjenige Serumfaktor, welcher mit den Insektiziden reagierte, envies sich als nicht identisch mit denjenigen Faktoren, welche die Patientenerythrozyten blockierten. Das klinische Bild dieses Falles deckte sich weitgehend mit demjenigen von zwei andern Fällen, bei welchen ein an die Anwesenheit von Arzneimitteln gebundener positiver indirekter Coombstest festgestellt worden war.

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