Analysis of granulocyte‐reactive antibodies using an immunoassay based upon monoclonal‐antibody‐specific immobilization of granulocyte antigens

Abstract

To detect human granulocyte-reactive antibodies, a glycoprotein-specific enzyme immunoassay for platelet antibodies was adapted for the use of granulocytes as target cells. Peripheral blood granulocytes were simultaneously incubated with a monoclonal antibody (mAb) and the serum to be investigated. After solubilization, aliquots of the cell lysate were transferred to plastic tubes coated with goat anti-mouse antibodies. Following immobilization of the trimolecular (mAb-glycoprotein-human antibody) complex it was detected by addition of enzyme-labelled goat anti-human antibodies using a luminescence technique. This assay allowed identification of different granulocyte-reactive antibodies present in the same sample without the need for complicated absorption studies. Alloantibodies against HLA and the granulocyte-specific NA antigens as well as isoantibodies against the Fc-gamma-receptor III (FcRIII) were detectable using mAb-specific immobilization of granulocyte antigens (MAIGA). Binding of autoantibodies to the FcRIII and to the CD 11b/CD18 complex could be shown.

The incidence, risk factors, and outcome of transfusion-related acute lung injury in a cohort of cardiac surgery patients: a prospective nested case-control study

“…Finally, the detected antibodies were confirmed in the Monoclonal Antibody Immobilization of Granulocyte Antigens assay, as previously described. 38 …”

Human alloantibody anti-Mart interferes with Mac-1–dependent leukocyte adhesion

“…The specificity of all sera and IgG preparations against the CD11b subunit was confirmed by mAb immobilization of the granulocyte antigen (MAIGA) assay using a panel of CD11b-specific mAbs as capture antibodies and granulocytes from donors who underwent Mart phenotyping. 22 …”

Transfusion Related Acute Lung Injury (TRALI) Caused by Red Blood Cell Transfusion Involving Residual Plasma Anti‐HLA Antibodies: A report on two Cases and General Considerations

“…The serum IgG antibody reacted strongly and remained granulo-agglutinating at a titer of 1 -64. Furthermore, the presence of additional antigranulocyte antibodies was suspected because the donor's serum proved capable of reacting, both in flow cytometry and in the Lazezari's microagglutination technique (GAT), with HNA1a, HNA1b, HNA1c, and HNA2a granulocytes (Lazezari et al 1976;Veys et al 1989;Bux et al 1993). Monoclonal antibody (mAb)-specific immobilization of granulocyte antigens (MAIGA) test positivity indicated the presence of specific anti-granulocyte antibody because the donor serum gave a weak positive reaction with mAb DJ-130c (Dakocytomation, Trappes, France), but not mAb 3G8 (Beckton Dickinson, Le Pont-de-Claix, France).…”