Analysis of granulocyte-reactive antibodies using an immunoassay based upon monoclonal-antibody-specific immobilization of granulocyte antigens.
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.
Bux
J
J
Institute for Clinical Immunology and Transfusion Medicine, Justus-Liebig-University Giessen, Germany.
Kober
B
B
Kiefel
V
V
Mueller-Eckhardt
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000097:blood; D000961:Antilymphocyte Serum / Q000032:analysis*; D003124:Colorimetry; D005258:Felty Syndrome / Q000097:blood / Q000276:immunology; D005260:Female; D006098:Granulocytes / Q000276:immunology*; D006801:Humans; D007112:Immunity, Maternally-Acquired; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D007223:Infant; D007231:Infant, Newborn; D007518:Isoantibodies / Q000032:analysis*; D007519:Isoantigens / Q000276:immunology*; D008163:Luminescent Measurements; D051379:Mice; D009503:Neutropenia / Q000097:blood / Q000151:congenital* / Q000276:immunology; D011247:Pregnancy / Q000097:blood / Q000276:immunology*; D011961:Receptors, Fc / Q000276:immunology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000961:Antilymphocyte Serum; D007518:Isoantibodies; D007519:Isoantigens; D011961:Receptors, Fc; C067704:neutrophil-specific antigen NA1, human; C072753:neutrophil-specific antigen NA2