A novel immunosuppressive pathway involving peroxynitrite-mediated [corrected] nitration of platelet antigens within antigen-presenting cells.
BACKGROUND
Studies have demonstrated that immunity against platelet (PLT) transfusions is dependent on recipient antigen-presenting cells (APCs) and their ability to produce nitric oxide (NO). To further analyze this, we focused on NO's major metabolite peroxynitrite (ONOO(-)) and its ability to affect PLT immunity.


METHODS
To address how NO and its major metabolite may mediate PLT immunity, GP91(PHOX) knockout (KO) mice that lack the ability to produce the ONOO(-) were transfused weekly with allogeneic BALB/c PLTs, and donor antibody development was analyzed.


RESULTS
Compared with controls, GP91(PHOX) KO mice developed significantly (p < 0.0001) higher-titered immunoglobulin G (IgG) donor antibodies by two transfusions, and this immune response could be inhibited by treating the recipient mice with aminoguanidine, a relatively selective inhibitor of inducible nitric oxide synthase. In vitro nitration of PLTs did not alter PLT antibody binding but significantly inhibited the transfused PLT's ability to stimulate IgG immunity in either wild-type or KO mice. The lack of nitrated PLT immunity correlated with an inability of APCs to mediate phagocytosis of nitrated PLTs. The lack of nitrated PLT immunity could only be restored when normal PLTs were mixed with the nitrated PLTs and transfused.


CONCLUSIONS
The results identify a dual role for NO metabolism within APCs that significantly modulates PLT immunity; nitration of PLT antigens leads to lack of immunity due to an inability of APCs to move PLT antigens intracellularly whereas there exists an NO-dependent pathway that stimulates anti-PLT immunity.
Semple
John W
JW
Keenan Research Centre, Li Ka Shing Knowledge Institute, St Michael's Hospital, Toronto, Ontario, Canada. Semplej@smh.toronto.on.ca
Speck
Edwin R
ER
Fabron
Antonio
A
Kim
Rukhsana Aslam Michael
RA
Freedman
John
J
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D016824:Antigens, Human Platelet / Q000276:immunology / Q000378:metabolism*; D006146:Guanidines / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics; D009566:Nitrates / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors; D017713:Platelet Transfusion / Q000009:adverse effects; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D016824:Antigens, Human Platelet; D006146:Guanidines; D008562:Membrane Glycoproteins; D009566:Nitrates; C048844:anti-IgG; C039642:peroxynitric acid; D052247:Nitric Oxide Synthase Type II; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; C004479:pimagedine