Characterization of T cell hybridomas raised against a glycopeptide containing the tumor-associated T antigen, (betaGal (1-3) alphaGalNAc-O/Ser).
T cell hybridomas were raised against the glycopeptide S(72) (Core-1) containing the tumor-associated disaccharide betaGal (1-3) alphaGalNAc (Core-1) O-linked to serine at position 72 in the mouse hemoglobin derived decapeptide Hb (67-76). All hybridomas recognized the glycopeptide S(72) (Core-1). Two of the selected hybridomas responded, however, much better to the S(72) (Tn) glycopeptide containing the monosaccharide alphaGalNAc O-linked to serine. In addition, one hybridoma cross-responded to the glycopeptide T(72) (Core-1) having a threonine at position 72 instead of a serine. No cross-responses were found to other glycopeptides consisting of the same hemoglobin peptide with different glycans attached or to the unglycosylated peptides. The T cell receptor Valpha and Vbeta usage was clearly diverse. The CDR3alpha regions demonstrated moreover a predominance of small polar amino acid side chains, and three hybridomas contained a common sequence motif. All the sequenced CDR3beta regions contained furthermore a conserved proline-glycine motif. In conclusion, immunization with the disaccharide containing glycopeptides S(72) (Core-1) created a heterogeneous population of glycopeptide specific T cells with the ability of cross-responding toward related glycopeptides.
Gad
Monika
M
Institute for Medical Microbiology and Immunology, University of Copenhagen, Denmark.
Werdelin
Ole
O
Meldal
Morten
M
Komba
Shiro
S
Jensen
Teis
T
D000595:Amino Acid Sequence; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000737:chemistry / Q000276:immunology*; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D011948:Receptors, Antigen, T-Cell / Q000235:genetics
D000957:Antigens, Viral, Tumor; D022801:Complementarity Determining Regions; D006020:Glycopeptides; D011948:Receptors, Antigen, T-Cell