Neutrophil Testing
Neutrophil Testing
The Red Cross National Neutrophil laboratory areas of expertise include granulocyte antibody screening, extended antibody identification (MAINA), HLA antibody screening and drug antibody testing. The detection of antibodies to human neutrophil alloantigens (HNA) remains very technically difficult due to the labile nature of granulocytes and the frequent presence of HLA Class I antibodies which also react with neutrophils. Consequently, expertise in this area has been confined to a small number of laboratories. One of the founding members of the International Society of Blood Transfusion (ISBT) Working Party on Granulocyte Immunobiology, our laboratory has participated in its quality assurance exercises, the International Granulocyte Immunology Workshop (IGIW), since its inception in 1989.
Neutrophil testing can now be ordered through Connect. See below for additional ordering information.
American Red Cross Neutrophil Immunology Laboratory
North Central Blood Services
100 South Robert Street
St. Paul, MN 55107-1489
Monday-Friday: 8:00am – 4:30pm Central
Email: BRSTPaulMNNeutrophil@redcross.org
Phone: (855) 216-9202
Benefits of selecting the ARC National Neutrophil Laboratory
- We are the only laboratory in the United States that routinely utilizes a combination of the granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) for detecting HNA antibodies.
- The ISBT Working Party on Granulocyte Immunobiology has recommended that a combination of GIFT and GAT remains the best means of HNA antibody detection, and has been the standard approach in most laboratories participating in the IGIW1,2,3,4.
- The Red Cross laboratory has achieved a 100% score when detecting and identifying HNA antibodies in the 2008-2017 IGIW surveys.
- The laboratory is one of four that are responsible for organizing, preparing and distributing IGIW survey samples to participating laboratories.
- The only neutrophil laboratory in the United States that routinely offers a monoclonal antibody immobilization of neutrophil antigen (MAINA) assay. This test is used to distinguish neutrophil antibodies from HLA Class I antibodies when both are present in the same specimen. MAINA is highly sensitive and is suggested when testing for neutrophil antibodies in children with a presumptive autoimmune neutropenia diagnosis.
- The laboratory includes extremely rare donor cells in our neutrophil panel that assists in the detection of HNA-2, HNA-3a, HNA-3b, HNA-4a, HNA-4b, HNA-5a and FcγRIIIb (CD16) antibodies.
- The laboratory provides phenotyping for HNA-2. Other neutrophil laboratories that provide molecular typing for HNA alleles do not provide HNA-2 typing results.
Clinical Indications
The presence of antibodies directed toward neutrophils results in their destruction and leads to a clinical condition known as neutropenia. Neutropenia is defined as an abnormally low concentration of neutrophils. Neutrophils are the most abundant white blood cells found in your blood stream and are essential in fighting off bacterial and fungal infections. Neutrophil antibodies can be found in the blood of women following pregnancy, multi-transfused individuals, patients with autoimmune disease and donors causing transfusion reactions. Our tests can aid in the clinical diagnosis of the following disorders:
Specialized Methods
The International Society of Blood Transfusion (ISBT) - Granulocyte Immunobiology Working Party (GIWP) has strongly recommended the use of both the granulocyte immunofluorescence test (GIFT) AND the granulocyte agglutination test (GAT) techniques in the detection of granulocyte (neutrophil) antibodies. It is of vital importance to use both the GAT and GIFT methods when detecting HNA antibodies. International Granulocyte Immunology Workshop (IGIW) external quality assessments have demonstrated that HNA antibodies detected by GAT have NOT BEEN DECTECTED by GIFT. Two IGIW assessments have included HNA-3a antibodies where laboratories using GIFT alone failed to detect antibody to HNA-3a samples involved in TRALI fatalities.
Both of these laboratory assays require viable intact granulocytes for detection and identification of human neutrophil antigen (HNA) antibodies. Granulocytes are intrinsically designed to respond to physiologic priming signals, so they must be handled very carefully. Once activated it is impossible to interpret any serologic test result involving these cells, as false positive results abound. Granulocytes are also extremely labile and must be used within 24 hours following collection. This necessitates ready access to panel cell donors with known HNA types. Finally, the presence of HLA antibodies in the test sera can make identifying HNA antibodies difficult as granulocytes also express HLA class I antigens on their cellular membrane. This obstacle can be overcome with the use of the monoclonal antibody immobilization of neutrophil antigens (MAINA) technique.
The isolation of a pure granulocyte suspension from peripheral blood is accomplished using the Ficoll-Hypaque discontinuous gradient technique. The double-density gradient centrifugation of leukocyte rich plasma yields a granulocyte suspension with a purity of 90 - 99%. Preparing a panel of donors to include all known HNA currently defined by ISBT using a suspension of pure granulocytes will aid in the detection and identification of HNA antibodies.
HNA Nomenclature
How to Order Testing
For customers using Connect, please select Request for Neutrophil Services form and complete the electronic form. Print a copy of the form and include with the sample.
To submit a sample using a paper request, please select the Order Form option below.
Orders placed without an ordering physician or test type selected will not be accepted.