title: The production of red blood cell alloantibodies in mice transfused with blood from transgenic Fyb-expressing mice.
abstract: BACKGROUND
  A murine model would be useful to identify which immune mechanisms could be manipulated to treat or prevent red blood cell (RBC) alloimmunization in patients who become sensitized to multiple or widely expressed antigens.
  
  
  METHODS
  Transgenic mice (B6CBAF1/J-Tg-Fy(b)) expressing the human Fy(b) antigen of the Duffy (Fy) blood group were donors. Recipient B6CBA-F1 mice received four weekly intravenous (IV) transfusions: either 0.3 mL of washed buffy coat-depleted RBCs or 0.3 mL of RBCs with spleen cells. Titers of immunoglobulin M (IgM) and immunoglobulin G (IgG) were measured in recipient serum samples by flow cytometry with RBCs from donor mice as target cells. Recipient serum samples were also tested against human RBCs of various Fy phenotypes. Additionally, RBC survival studies were performed in alloimmunized mice utilizing biotin-labeled Fy(b) transgenic mouse RBCs.
  
  
  RESULTS
  B6CBA-F1 mice receiving washed buffy coat-depleted RBCs first made IgM, followed by IgG alloantibodies to transgenic mouse Fy(b)-positive RBCs. Recipients of Fy(b)-positive RBCs mixed with spleen cells also produced IgM and IgG alloantibodies, but at a slower rate than recipients of washed buffy coat-depleted RBCs. Serum samples showed specificity for Fy3, Fy(b), and Fy6. Decreased survival of transfused RBCs was evident at 24 hours after transfusion.
  
  
  CONCLUSIONS
  It is possible to elicit the formation of anti-Fy alloantibodies by IV transfusion in mice that lack Fy antigens. The transfusion of RBCs alone was adequate to stimulate alloantibody production in B6CBA-F1 recipient mice. The survival of transfused Fy(b)-positive RBCs is diminished in sensitized mice. This model will be useful in further studies of RBC alloimmunization.
authors.affiliation: Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, Maryland 21287, USA. scampbe1@jhmi.edu
authors.forename: Sally A
authors.initials: SA
authors.lastname: Campbell-Lee
authors.forename: Jinhuan
authors.initials: J
authors.lastname: Liu
authors.forename: Randall W
authors.initials: RW
authors.lastname: Velliquette
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Halverson
authors.forename: R Sue
authors.initials: RS
authors.lastname: Shirey
authors.forename: Asok
authors.initials: A
authors.lastname: Chaudhuri
authors.forename: Marion E
authors.initials: ME
authors.lastname: Reid
authors.forename: Paul M
authors.initials: PM
authors.lastname: Ness
authors.forename: William M
authors.initials: WM
authors.lastname: Baldwin
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology; D001803:Blood Transfusion / Q000379:methods; D004375:Duffy Blood-Group System / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology*; D007518:Isoantibodies / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D065227:Transfusion Reaction
chemical_list: D004375:Duffy Blood-Group System; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007518:Isoantibodies