title: Antigen binding to GM1 ganglioside results in delayed presentation: minimal effects of GM1 on presentation of antigens internalized via other pathways.
abstract: Plasma membrane rafts are sphingolipid- and cholesterol-rich patches that function as membrane trafficking and surface signalling regions. Ganglioside GM1 is an integral component of these microdomains, and Escherichia coli enterotoxin B subunit (EtxB) is a pentamer that binds with high affinity to GM1 resulting in GM1 cross-linking. We previously demonstrated that antigen coupled directly to EtxB resulted in enhanced presentation relative to antigen taken up by fluid-phase endocytosis. Here we demonstrate a new role for GM1 in antigen presentation by examining the effects of cross-linking GM1 on the kinetics of presentation and processing of antigen by the B-cell receptor (BCR), fluid-phase endocytosis and GM1-targeted antigen. EtxB bound to B cells does not augment the subsequent kinetics or magnitude of presentation of either BCR-internalized antigen or soluble antigen. Moreover, presentation of GM1-bound antigen is significantly slower than antigen presentation following BCR-mediated uptake. In contrast to the rapid internalization of BCR-bound antigen (which has a half life of 60 min), the majority of EtxB-bound antigen forms a plasma membrane depot detectable for many hours after initial incubation (and with a half life of 12 hr). We conclude that cross-linking of GM1 by EtxB minimally affects the processing and presentation of antigens internalized via other pathways. Nevertheless, binding of antigens to GM1 results in delayed presentation that has important implications for in vivo immunization using GM1-targeted adjuvants.
authors.affiliation: Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Bristol, UK. t.o.nashar@bristol.ac.uk
authors.forename: Toufic O
authors.initials: TO
authors.lastname: Nashar
authors.forename: Zoe E
authors.initials: ZE
authors.lastname: Betteridge
authors.forename: Richard N
authors.initials: RN
authors.lastname: Mitchell
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000187:drug effects / Q000276:immunology*; D001427:Bacterial Toxins / Q000276:immunology / Q000378:metabolism; D002462:Cell Membrane / Q000276:immunology; D004768:Enterotoxins / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000276:immunology; D029968:Escherichia coli Proteins; D005677:G(M1) Ganglioside / Q000378:metabolism*; D051379:Mice; D010047:Ovalbumin / Q000276:immunology; D010873:Pinocytosis / Q000276:immunology; D011947:Receptors, Antigen, B-Cell / Q000276:immunology; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011947:Receptors, Antigen, B-Cell; D005677:G(M1) Ganglioside; D010047:Ovalbumin; C027538:heat-labile enterotoxin, E coli