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Blood, Vol. 93 No. 2 (January 15), 1999:
pp. 713-720
Extreme Leukoreduction of Major Histocompatibility Complex Class II
Positive B Cells Enhances Allogeneic Platelet Immunity
John W. Semple,
Edwin R. Speck,
Donna Cosgrave,
Alan H. Lazarus,
Victor S. Blanchette, and
John Freedman
From the Divisions of Hematology, St. Michael's Hospital and The
Hospital for Sick Children, The University of Toronto and The Toronto
Platelet Immunobiology Group, Toronto, Ontario, Canada.
In a murine model of platelet alloimmunization, we examined the
definitive role that mononuclear cells (MC) have in modulating platelet
immunity by using platelets from severe combined immunodeficient (SCID)
mice. CB.17 (H-2d) SCID or BALB/c (H-2d) mouse
platelets were transfused weekly into fully allogeneic CBA
(H-2k) mice and antidonor antibodies measured by flow
cytometry. MC levels in BALB/c platelets were 1.1 ± 0.6/µL and SCID
mouse platelets could be prepared to have significantly lower
(<0.05/µL) MC numbers. Transfusions with 108 BALB/c
platelets (containing  100 MC/transfusion) stimulated IgG antidonor
antibodies in 100% of the recipients by the fifth transfusion, whereas
108 SCID mouse platelets (containing 5 MC/transfusion)
stimulated higher-titered IgG alloantibodies by the second
transfusion. When titrations of BALB/c peripheral blood MC were added
to the SCID mouse platelets, levels approaching 1 MC/µL reduced SCID
platelet immunity to levels similar to BALB/c platelets.
Characterization of the alloantibodies showed that the low levels of MC
significantly influenced the isotype of the antidonor IgG; the presence
of 1 MC/µL was associated with induction of noncomplement fixing IgG1 antidonor antibodies, whereas platelet transfusions, devoid of MC
(<0.05/µL), were responsible for complement-fixing IgG2a
production. When magnetically sorted defined subpopulations of MC were
added to the SCID platelets, major histocompatability complex (MHC) class II positive populations, particularly B cells, were found to be
primarily responsible for the reduced SCID mouse platelet immunity. The
presence of low numbers of MC within the platelets was also associated
with an age-dependent reduction in platelet immunogenicity; this
relationship however, was not observed with SCID mouse platelets devoid
of MC. The results suggest that a residual number of MHC class II
positive B cells within allogeneic platelets are required for maximally
reducing alloimmunization.
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