Abstract

Granulocyte reactive antibodies have been found to cause clinical disorders such as transfusion related acute lung injury (TRALI), febrile transfusion reactions, alloimmune neonatal neutropenia, immune neutropenia after bone marrow transplantation, refractoriness to granulocyte transfusion, drug-induced neutropenia and autoimmune neutropenia. Using the granulocyte immunofluorescence test (GIFT) by microscopic and flow cytometric analysis, the frequencies of the neutrophil antigens HNA−1a, −1b, −2a, −3a and −4a were determined among 100 random Brazilian blood donors from the Blood Center of Universidade Federal de Sao Paulo, SP, Brazil. Granulocytes were separated from mononuclear cells and red cells by sedimentation with 5% dextran, followed by centrifugation on Ficoll-paque (d = 1.077), and then incubated with anti-sera (anti-HNA−1a, −1b, −2a, −3a, and −4a obtained from American Red Cross, North Central Blood Services, St. Paul, MN) conjugated with fluorescein isothiocyanate (FITC) labeled F(ab’)2 fragments of anti-human IgG. Only cell suspensions containing ≥95% neutrophils with viability ≥90% according to the trypan-blue staining were analyzed. The frequencies of HNA−1a, −1b and −2a were 65%, 83% and 94%, respectively, and for such alloantigens exact same results were observed using either the GIFT performed by microscopy or by flow cytometry. The frequency of HNA-3a was 86% by the microscopic GIFT, and 95% by the flow cytometry analysis; while the frequency of HNA−4a was 93% by the microscopic GIFT, and 94% by the flow cytometric technique. These results indicate that:

  1. GIFT by flow cytometry is more sensitive than the GIFT by microscopy to detect HNA−3a;

  2. the phenotypic frequencies found for neutrophil antigens HNA−1a and −1b among Brazilian blood donors are quite similar to those reported among African Americans, but different from those reported for Japanese and Chinese individuals;

  3. the phenotype frequencies of the neutrophil antigens HNA−2a, −3a, and −4a in Brazilians are quite similar to those found among Caucasians (Table). (These studies were funded by FAPESP, SP, Brazil - 05/55237–9).

AsiansBrazilians
AntigenAfrican AmericansChinesesHindusJapanesesCaucasiansMFC
M, microscopy; FC, flow cytometry; nd, not done 
HNA-1a 46 – 68 90 44 88 52 – 54 65 65 
HNA-1b 78 – 84 52 83 51 – 64 87 – 89 83 83 
HNA-2a nd 99 nd 89 87 – 97 94 94 
HNA-3a nd nd nd nd 99 86 95 
HNA-4a nd nd nd nd 96 93 94 
AsiansBrazilians
AntigenAfrican AmericansChinesesHindusJapanesesCaucasiansMFC
M, microscopy; FC, flow cytometry; nd, not done 
HNA-1a 46 – 68 90 44 88 52 – 54 65 65 
HNA-1b 78 – 84 52 83 51 – 64 87 – 89 83 83 
HNA-2a nd 99 nd 89 87 – 97 94 94 
HNA-3a nd nd nd nd 99 86 95 
HNA-4a nd nd nd nd 96 93 94 

Disclosure: No relevant conflicts of interest to declare.

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