title: Human neutrophil antigen-3a antibodies induce neutrophil aggregation in a plasma-free medium.
abstract: BACKGROUND
  Antibodies against the human neutrophil alloantigen-3a (HNA-3a) are involved in severe cases of transfusion-related acute lung injury (TRALI), but the susceptibility of patients towards HNA-3a antibody differs largely. HNA-3a antibodies induce granulocyte aggregation. However, it is unresolved whether plasma proteins are required for granulocyte aggregation.
  
  
  METHODS
  We investigated whether HNA-3a-antibody-induced aggregation of polymorphonuclear cells is dependent on plasma factors by using and modifying the granulocyte agglutination test (GAT).
  
  
  RESULTS
  Polymorphonuclear cells homozygous for HNA-3a did not aggregate when incubated with HNA-3a antibodies in a plasma-protein-free GAT setup. When the GAT was performed using polymorphonuclear cells re-suspended in phosphate-buffered saline containing proteins, HNA-3-mediated aggregation was observed. Moreover, using Tween® 20 for blocking the plates, reconstituted the granulocyte aggregation in a protein-free medium. This indicates that granulocyte aggregation probably occurs by direct granulocyte-granulocyte interaction(s) or is mediated by substances released by neutrophils after activation.
  
  
  CONCLUSIONS
  Granulocyte aggregation induced by HNA-3a antibodies does not require human plasma proteins. Interindividual variability in the response to HNA-3a antibodies does not depend on differences in patient's plasma proteins.
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Schubert
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Tom
authors.initials: T
authors.lastname: Berthold
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Muschter
authors.affiliation: ZIK HIKE, Innovation Center - Humoral Immune Responses in Cardiovascular Disorders, Cell Response Group, Greifswald, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Wesche
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Birgitt
authors.initials: B
authors.lastname: Fürll
authors.affiliation: Red Cross Blood Donation Service West, Hagen, Germany.
authors.forename: Angelika
authors.initials: A
authors.lastname: Reil
authors.affiliation: Red Cross Blood Donation Service West, Hagen, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bux
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Tamam
authors.initials: T
authors.lastname: Bakchoul
authors.affiliation: Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University Greifswald, Greifswald, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Greinacher
mesh_terms: D055371:Acute Lung Injury / Q000209:etiology / Q000276:immunology; D000371:Agglutination / Q000187:drug effects / Q000276:immunology; D001323:Autoantibodies / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D005260:Female; D006801:Humans; D007519:Isoantigens / Q000276:immunology*; D008297:Male; D009504:Neutrophils / Q000276:immunology*; D065227:Transfusion Reaction
chemical_list: D001323:Autoantibodies; D016895:Culture Media, Serum-Free; C518134:HNA-3a antigen, human; D007519:Isoantigens