Antigen modulation as a potential mechanism of anti-KEL immunoprophylaxis in mice.
Red blood cell (RBC) alloimmunization is a serious complication of transfusion or pregnancy. Despite the widespread use of Rh immune globulin to prevent pregnancy associated anti-D alloimmunization, its mechanism of action remains elusive. We have previously described a murine model in which immunoprophylaxis with polyclonal anti-KEL sera prevents alloimmunization in wild-type recipients transfused with transgenic murine RBCs expressing the human KEL glycoprotein. To investigate the mechanism of action, we have now evaluated the outcome of immunoprophylaxis treatment in mice lacking Fcγ receptors (FcγRs), complement (C3), both, or none. Whereas polyclonal anti-KEL sera completely prevented alloimmunization in wild-type and single-knockout (KO) mice lacking FcγRs or C3, double-KO mice lacking both FcγRs and C3 became alloimmunized despite immunoprophylaxis. Rapid clearance of essentially all transfused RBCs with detectable KEL glycoprotein antigen occurred within 24 hours in wild-type and single-KO recipients treated with immunoprophylaxis, with the transfused RBCs remaining in circulation having minimal KEL glycoprotein antigen detectable by flow cytometry or western blot. In contrast, transfused RBCs with the KEL glycoprotein antigen fully intact continued to circulate for days in double-KO mice despite treatment with immunoprophylaxis. Further, in vitro phagocytosis assays showed no consumption of opsonized murine RBCs by double-KO splenocytes. Taken in combination, our data suggest that modulation of the KEL antigen (and potentially RBC clearance) by redundant recipient pathways involving both FcγRs and C3 may be critical to the mechanism of action of polyclonal anti-KEL immunoprophylaxis. These findings could have implications for the development of immunoprophylaxis programs in humans.
Department of Laboratory Medicine and.
Jingchun
J
Liu
Department of Laboratory Medicine and.
Manjula
M
Santhanakrishnan
Department of Laboratory Medicine and.
Prabitha
P
Natarajan
Department of Laboratory Medicine and.
David R
DR
Gibb
Department of Laboratory Medicine and.|Department of Immunobiology, Yale University School of Medicine, New Haven, CT.
Stephanie C
SC
Eisenbarth
Department of Laboratory Medicine and.|Pathology and Laboratory Medicine Service, VA Connecticut Healthcare System, West Haven, CT.
Christopher A
CA
Tormey
Department of Laboratory Medicine and.|Department of Pathology, Yale University School of Medicine, New Haven, CT.
Alexa J
AJ
Siddon
Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA.
Sean R
SR
Stowell
Centre for Innovation, Canadian Blood Services, Toronto, ON, Canada; and.
Donald R
DR
Branch
Department of Laboratory Medicine and.|Department of Pediatrics, Yale University School of Medicine, New Haven, CT.
Jeanne E
JE
Hendrickson
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism; D000941:Antigens / Q000378:metabolism*; D001803:Blood Transfusion; D015153:Blotting, Western; D003176:Complement C3 / Q000378:metabolism; D003433:Crosses, Genetic; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D007114:Immunization; D008562:Membrane Glycoproteins / Q000276:immunology*; D008666:Metalloendopeptidases / Q000276:immunology*; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009895:Opsonin Proteins / Q000378:metabolism; D010587:Phagocytosis; D011971:Receptors, Immunologic / Q000378:metabolism; D013154:Spleen / Q000166:cytology
D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D003176:Complement C3; D008562:Membrane Glycoproteins; D009895:Opsonin Proteins; D011971:Receptors, Immunologic; C585978:KEL protein, human; D008666:Metalloendopeptidases
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