Carbohydrate residues downstream of the terminal Galα(1,3)Gal epitope modulate the specificity of xenoreactive antibodies

• Published in: Immunology and cell biology Vol. 85; no. 8; pp. 623 - 632
• Main Authors: Milland, Julie, Yuriev, Elizabeth, Xing, Pei‐Xiang, McKenzie, Ian F C, Ramsland, Paul A, Sandrin, Mauro S
• Format: Journal Article
• Language: English
• Published: Nature Publishing Group 01.11.2007
• Subjects: antibody specificity; antibody structure and function; carbohydrate antigens; natural antibodies; xenotransplantation
• ISSN: 0818-9641; 1440-1711
• DOI: 10.1038/sj.icb.7100111

Carbohydrates are involved in many immunological responses including the rejection of incompatible blood, tissues and organs. Carbohydrate antigens with Galα(1,3)Gal epitopes are recognized by natural antibodies in humans and pose a major barrier for pig‐to‐human xenotransplantation. Genetically modified pigs have been established that have no functional α1,3‐galactosyltransferase (α1,3GT), which transfers αGal to N‐acetyllactosamine (LacNAc) type oligosaccharides. However, a low level of Galα(1,3)Gal is still expressed in α1,3GT knockout animals in the form of a lipid, isoglobotrihexosylceramide (iGb3), which is produced by iGb3 synthase on lactose (Lac) type core structures. Here, we define the reactivity of a series of monoclonal antibodies (mAb) generated in α1,3GT−/− mice immunized with rabbit red blood cells (RbRBC), as a rich source of lipid‐linked antigens. Interestingly, one mAb (15.101) binds weakly to synthetic and cell surface‐expressed Galα(1,3)Gal on LacNAc, but strongly to versions of the antigen on Lac cores, including iGb3. Three‐dimensional models suggest that the terminal α‐linked Gal binds tightly into the antibody‐binding cavity. Furthermore, antibody interactions were predicted with the second and third monosaccharide units. Collectively, our findings suggest that although the terminal carbohydrate residues confer most of the binding affinity, the fine specificity is determined by subsequent residues in the oligosaccharide.