Human IgG/Fc gamma R interactions are modulated by streptococcal IgG glycan hydrolysis.
BACKGROUND
The human pathogen Streptococcus pyogenes produces an endoglycosidase, EndoS that hydrolyzes the chitobiose core of the asparagine-linked glycan on the heavy chain of human IgG. IgG-binding to Fc gamma receptors (Fc gamma R) on leukocytes triggers effector functions including phagocytosis, oxidative burst and the release of inflammatory mediators. The interactions between Fc gamma R and the Fc domain of IgG depend on the IgG glycosylation state.


RESULTS
Here we show for the first time that EndoS hydrolyzes the heavy chain glycan of all four human IgG subclasses (IgG1-4), in purified form and in a plasma environment. An inactive form of EndoS, obtained by site-directed mutagenesis, binds IgG with high affinity, in contrast to wild type EndoS that only transiently interacts with IgG, as shown by Slot-blotting and surface plasmon resonance technology. Furthermore, EndoS hydrolysis of the IgG glycan influences the binding of IgG to immobilized soluble Fc gamma R and to an erythroleukemic cell line, K562, expressing Fc gamma RIIa. Incubation of whole blood with EndoS results in a dramatic decrease of IgG binding to activated monocytes as analyzed by flow cytometry. Moreover, the IgG bound to K562 cells dissociates when cells are treated with EndoS. Likewise, IgG bound to immobilized Fc gamma RIIa and subsequently treated with EndoS, dissociates from the receptor as analyzed by surface plasmon resonance and Western blot.


CONCLUSIONS
We provide novel information about bacterial enzymatic modulation of the IgG/Fc gamma R interaction that emphasizes the importance of glycosylation for antibody effector functions. Moreover, EndoS could be used as a biochemical tool for specific IgG N-glycan hydrolysis and IgG purification/detection, or as a potential immunosuppressing agent for treatment of antibody-mediated pathological processes.
Allhorn
Maria
M
Division of Infection Medicine, Department of Clinical Sciences, Lund University, Lund, Sweden. maria.allhorn@med.lu.se
Olin
Anders I
AI
Nimmerjahn
Falk
F
Collin
Mattias
M
D015153:Blotting, Western; D002460:Cell Line; D006801:Humans; D006868:Hydrolysis; D007074:Immunoglobulin G / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011134:Polysaccharides / Q000378:metabolism*; D017452:Receptors, IgG / Q000378:metabolism*; D013291:Streptococcus / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D007074:Immunoglobulin G; D011134:Polysaccharides; D017452:Receptors, IgG