Granulocyte immunofluorescence and granulocyte agglutination tests are standard methods for detecting human neutrophil antigen (HNA) antibodies (Abs); however, these require a typed panel of neutrophils, which can be time-consuming to develop, and it remains difficult to determine antibody specificity in some cases. We established and evaluated four detection systems for HNA-1a Abs based on an HNA-1a-expressing cell line (KY cells) and antigen capture. We additionally evaluated a commercial solid-phase system. Eleven HNA-1a antibody-positive samples, including the World Health Organization Reference Reagent, and 40 serum samples derived from male blood donors were used as positive and negative control samples, respectively. Although specificity was >0.90 in all systems evaluated, the sensitivity varied among the systems. The KY cell-based monoclonal antibody specific immobilisation of granulocyte antigens (KY-MAIGA) system using certain, but not all, monoclonal Abs, and the solid-phase system revealed higher sensitivity than other systems. In conclusion, the KY-MAIGA and commercial solid-phase systems were superior in terms of specific and sensitive detection of HNA-1a Abs.
Background and Objectives The Fcgamma receptor IIIb (FccRIIIb) carries the human neutrophil antigen (HNA)-1 system. The three FCGR3B alleles encode four antigens: one allele encodes HNA-1a, and two alleles encode two antigens each, HNA-1b and HNA-1d and HNA-1b and HNA-1c, respectively. Several other nonsynonymous substitutions have also been reported. Little is known about the properties of HNA-1d antibodies because HNA-1d was recently discovered, and HNA-1d antibodies have been reported only in a couple of cases. These findings raise the possibility that the polymorphisms in the HNA-1 system and spectrum of the specificity of resulting antibodies are more complex than previously thought. We address this possibility in this work. Materials and MethodsSeveral HNA-1 antibodies, which had been classified as HNA-1a or HNA-1b before HNA-1d was discovered, were re-evaluated for their reactivity to different recombinant variants of FccRIIIb. ResultsThe HNA-1a antibody, but not the reference HNA-1a antibody, crossreacted to HNA-1d. Further, nucleotide replacement in FCGR3B alleles *02 at 193G?A (AA position 54 E?K) resulted in the disappearance of HNA-1d, but not HNA-1b, antigenicity. ConclusionThe HNA-1 system may be more complex than previously understood.
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