Leukocyte-Reactive Antibodies in Female Blood Donors: The Austrian Experience

Schönbacher, Marlies; Aichinger, Nicole; Weidner, Lisa; Jungbauer, Christof; Grabmer, Christoph; Schuha, Beate; Rohde, Eva; Mayr, Wolfgang R.; Körmöczi, Günther F.

doi:10.1159/000509946

Cited by 5 publications

(6 citation statements)

References 38 publications

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“…6,10,11 However, falsepositive LABM reactions are common due to denaturation of bead-bound HNA antigens, and some antibodies cannot be detected. 4,6,12 Therefore, the ISBT and IGIW recommend the MAIGA assay, which detects antibodies in a molecule-specific manner, as the gold standard for determining HNA antibody specificity. 1,12 However, the MAIGA assay is susceptible to falsenegative results when epitopes of the alloantibodies against the molecule of interest overlap with those of the molecule-specific capture mAbs.…”

Section: Discussionmentioning

confidence: 99%

“…Our data suggest that the novel ICFA using CD16 mAb‐conjugated Luminex beads and HNA‐1‐expressing cells is highly sensitive and exhibits specificity similar to that of the MAIGA and LABM assays for detecting antibodies against HNA‐1a and HNA‐1b. The recently developed fluorescent bead method, LABM, is widely used for HNA antibody testing 6,10,11 . However, false‐positive LABM reactions are common due to denaturation of bead‐bound HNA antigens, and some antibodies cannot be detected 4,6,12 .…”

Section: Discussionmentioning

confidence: 99%

“…The recently developed fluorescent bead method, LABM, is widely used for HNA antibody testing 6,10,11 . However, false‐positive LABM reactions are common due to denaturation of bead‐bound HNA antigens, and some antibodies cannot be detected 4,6,12 . Therefore, the ISBT and IGIW recommend the MAIGA assay, which detects antibodies in a molecule‐specific manner, as the gold standard for determining HNA antibody specificity 1,12 .…”

Section: Discussionmentioning

confidence: 99%

“…4 Furthermore, LABM may misclassify the specificity of some antibodies; its use is thus recommended in combination with assays utilizing intact neutrophils. [4][5][6] In this study, we developed a novel immunocomplex capture fluorescence assay (ICFA) using Luminex beads and HNA-1-transfected cells and compared it with the MAIGA and LABM assays for identifying anti-HNA-1 antibodies.…”

Section: Introductionmentioning

confidence: 99%

“…However, LABM may exhibit high false‐positive rates due to denaturation of the three‐dimensional structure of purified antigens coating the beads 4 . Furthermore, LABM may misclassify the specificity of some antibodies; its use is thus recommended in combination with assays utilizing intact neutrophils 4–6 …”

Section: Introductionmentioning

confidence: 99%

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A novel immunocomplex capture fluorescence assay (ICFA) using fluorescent beads and transfected cells for specific identification of human neutrophil antigen (HNA)‐1a and ‐1b antibodies

Kamada,

Takahashi,

Shimizu

et al. 2024

Transfusion

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BackgroundTo identify specific human neutrophil antigen (HNA) antibodies, assays using neutrophils such as monoclonal antibody‐specific immobilization of granulocyte antigens (MAIGA) are recommended. However, these assays are limited by labor‐intensive neutrophil preparation and varying antigen expression levels.MethodsWe evaluated a newly developed immunocomplex capture fluorescence assay (ICFA) for identifying HNA‐1 antibodies and compared it to MAIGA and LABScreen Multi (LABM), which utilizes recombinant HNA‐coated Luminex beads. For ICFA, HNA‐1a or HNA‐1b transfected cells replaced neutrophils. Cells incubated with serum were lysed, and immune complexes were captured using five CD16 monoclonal antibody‐conjugated Luminex beads. Nine antisera with known specificity and 26 samples suspected of containing HNA antibodies were analyzed by ICFA and MAIGA using neutrophils or transfected cells (ICFA‐N or ICFA‐T, and MAIGA‐N or MAIGA‐T, respectively).ResultsICFA‐T and MAIGA‐N accurately determined the specificity of all antibodies in the nine antiserum samples. The ICFA‐T detection limit was 2048‐fold for anti–HNA‐1a and 256‐fold for anti‐HNA‐1b; the limits of MAIGA‐T, MAIGA‐N, and LABM were 32‐, 4 ~ 64‐, and 128‐fold for anti‐HNA‐1a and 64‐, 16 ~ 64‐, and 32‐fold for anti–HNA‐1b, respectively. Twelve and 7 of the remaining 26 samples tested negative and positive, respectively, in both ICFA‐T and MAIGA‐N. Antibody specificity against HNA‐1a or HNA‐1b determined using ICFA‐T agreed with that determined using MAIGA‐N and LABM. Another seven samples tested positive in ICFA‐T but negative in MAIGA‐N.ConclusionThe novel ICFA is highly sensitive and exhibits specificity similar to MAIGA and LABM for detecting HNA‐1 antibodies.

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