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.

Autor: Kamada H; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Takahashi D; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Shimizu M; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Uchida M; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Watanabe Y; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Nakajima F; GenoDive Pharma Inc., Atsugi, Kanagawa, Japan., Miyata S; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan., Satake M; Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
Jazyk: angličtina
Zdroj: Transfusion [Transfusion] 2024 May; Vol. 64 (5), pp. 906-918. Date of Electronic Publication: 2024 Mar 26.
DOI: 10.1111/trf.17813
Abstrakt: Background: To 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.
Methods: We 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).
Results: ICFA-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.
Conclusion: The novel ICFA is highly sensitive and exhibits specificity similar to MAIGA and LABM for detecting HNA-1 antibodies.
(© 2024 AABB.)
Databáze: MEDLINE
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