Anti‐Granulocyte Antibody Screening with Extracted Granulocyte Antigens by a Micro‐Mixed Passive Hemagglutination Method

Abstract

This new method is preferable for screening for granulocyte antibodies. In addition, it has the advantage of requiring only 5 microl of serum for each test.

Granulocyte antibody detection – the role of MPHA

“…We and others [2] are applying this method for the testing of granulocyte antibodies, but presently, it is not fully developed nor largely applied even in Japan. Modifications should be introduced to improve the detection ability of antibodies against HNA-3.…”

“…Modifications should be introduced to improve the detection ability of antibodies against HNA-3. Since Araki et al [2] were able to detect the antibody against 5b (presently, HNA-3a), and they have used extracted granulocyte antigens, without fixatives, it is possible that the fixative agent affects HNA-3 antigenicity. Also, some antigenicities, such as HNA-4, can be lost by the treatment of granulocytes with chloroquine to remove HLA antigenicity.…”

“…The extracted granulocyte antigen panels can be stored frozen for at least 1 year at )80°C. (2) Use of Terasaki plates instead of 96-well plates: it was also previously reported by Araki et al [2] Granulocyte specific antibodies could be determined in the MPHA assay using Terasaki plate. The advantage of this plate is the need of small volumes of the samples for testing.…”

“…For example: (1) Use of extracted granulocyte antigens instead of intact cells: it was previously reported by Araki et al [2]. By this procedure, human antibodies specific for NA1 (now HNA-1a), NA2 (now HNA-1b), NB1 (now HNA-2a), 5b (now HNA-3a) and Sar-a could be determined, as well as monoclonal antibodies against CD11a, CD11b, CD13, CD16, CD18 and HLA class I.…”

Detection of Auto- And Alloantibodies Against Granulocytes by Means of Gel Agglutination Technique

“…Присутствие HLA антигенов на тест-гранулоцитах может затруднить определе-ние специфичности антител в сыворотках [1]. Для уточнения специфичности антител к антиге-нам гранулоцитов целесообразно в дальнейшем проводить генотипирование гранулоцитов до-норов, что позволит охарактеризовать антигены и получить антигранулоцитарные сыворотки, ко-торые в дальнейшем можно использовать в каче-стве контроля при оценке чувствительности ме-тодов исследования.…”