Structural and immunological characterization of the N-glycans from the major yellow jacket allergen Ves v 2: the N-glycan structures are needed for the human antibody recognition.
Yellow jacket (Vespula vulgaris) hyaluronidase (Ves v 2) is a glycoprotein and a mixture of two isoallergens, Ves v 2.01 and Ves v 2.02. Wasp and bee sensitized individuals frequently show IgE antibodies that in vitro recognize common carbohydrate structures across the hymenoptera species. The aim of the study was to characterize the glycosylation patterns in Ves v 2 isoallergens and to assess their immunological properties regarding antibody binding and T cell activation. The glycosylation sites and the carbohydrate structures were verified by use of tandem mass spectrometry (MS/MS). The immunological characterization of the N-glycan structures was assessed by antibody binding, T cell proliferation and T cell epitope assays comparing native (n) and non-glycosylated recombinant (r) Ves v 2. Analyses of the Ves v 2 glycopeptides revealed that glycan attachments were found for residues 79, 99 and 127 of Ves v 2.01, and residues 66 and 81 of Ves v 2.02. Structural analysis of the glycopeptides showed that the majority of the N-glycans contained at least one alpha1,3-fucose and/or alpha1,6-fucose residues in a structure. Interestingly, serum IgE antibodies from vespid allergic patients recognized nVes v 2 but not rVes v 2. Non-glycosylated rVes v 2, however, induced T cell and cytokine responses comparable to glycosylated nVes v 2. The present study shows that N-glycan structures are needed for the antibody recognition but not for the T cell reactivity of Ves v 2 in vitro. The occurrences of carbohydrate-specific antibodies against nVes v 2, however, suggest that non-mammalian glycan structures as in nVes v 2 may provide a link between T cells and other effector cells in allergic responses.
Vaccine Research & Discovery, ALK-Abelló, Boge Allé 6-8, 2970 Horsholm, Denmark. usedk@alk-abello.com
Ulla
U
Seppälä
David
D
Selby
Rafael
R
Monsalve
Te Piao
TP
King
Christof
C
Ebner
Peter
P
Roepstorff
Barbara
B
Bohle
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D049109:Cell Proliferation; D016207:Cytokines / Q000096:biosynthesis; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000737:chemistry* / Q000276:immunology*; D007122:Immunoelectrophoresis; D007073:Immunoglobulin E / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry* / Q000276:immunology*; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D014862:Wasp Venoms / Q000737:chemistry* / Q000276:immunology*; D014863:Wasps / Q000737:chemistry* / Q000276:immunology*
D000906:Antibodies; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D011134:Polysaccharides; D014862:Wasp Venoms; D007073:Immunoglobulin E; C101836:allergen Ves v 2; D006821:Hyaluronoglucosaminidase