IMMUNOHEMATOLOGY: Rapid screening of granulocyte antibodies with a novel assay: flow cytometric granulocyte immunofluorescence test

Abstract

The Flow-GIFT method permits rapid detection of granulocyte antibodies requiring fewer donor test cells. This method is ideal for automation and will potentially open the way for screening of granulocyte antibodies in a large donor population.

A Novel Flow Cytometric Method for the Simultaneous Detection of Antibodies Against Platelet, Lymphocyte, And Neutrophil

“…In particular, HNA-3a is present on lymphocytes and platelets (11,19). For example, HNA-3, HNA-4, and HNA-5 are present on WBCs other than neutrophils.…”

“…While the anti-platelet Ab1 control sera reacted with platelets alone, anti-HLA Ab1 control sera reacted with all three cell types (Figs. This may be because some human neutrophil antigens (HNAs) (e.g., HNA-3a) are also expressed on platelets or on lymphocytes (11,19). Although we expected that anti-neutrophil Ab1 control sera might react with neutrophils alone, two cases from one infant with autoimmune neutropenia reacted strongly with platelets also (Fig.…”

Granulocyte antibodies in male blood donors: can they trigger transfusion‐related acute lung injury?

“…Granulocyte test cells were obtained from freshly drawn Na 2 ethylenediaminetetraacetate (EDTA) whole blood of healthy blood donors previously typed for neutrophil antigens HNA‐1a, ‐1b, ‐1c, ‐2, ‐3a, ‐4a, and –5a . Granulocytes were isolated as described . Sera from healthy blood donors without WBC‐specific antibodies were used as controls.…”

“…All pipetting steps were performed using an automated workstation (Biomek NX p Multichannel Laboratory Automation Workstation, Beckman‐Coulter). Flow cytometric analysis was performed on a flow cytometer (FC 500 MPL, Beckman‐Coulter) . The median fluorescence intensity (MFI) was used for sample analyses.…”

“…Recently, we established a high‐throughput automated flow‐cytometry–based granulocyte immunofluorescence test (Flow‐GIFT) for routine detection of HNA antibodies . The test has since also been referred to as the WBC immunofluorescence test (WIFT) .…”