A lectin array analysis for wild-type and α-Gal-knockout pig islets versus healthy human islets
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We performed lectin microarray analyses of islets from wild-type (WT) pigs and α1-3galactosyltransferase gene knockout (GKO) pigs and compared the results with the corresponding values for islets from healthy humans.
Islets were isolated from the pancreas. After sonication and centrifugation, the proteins in the supernatant from each islet were labeled with Cy3 and applied to a lectin array.
Despite negligible expression of the Gal antigen on the adult pig islets (APIs), GKO-islets showed weaker signals, not only for GS-I-B4 but also for PNA, WFA, PTL-I, and GS-I-A4, than the WT islets, indicating reduced contents of α-linked GalNAc and Galβ1-3GalNAc. In comparing the islets of pigs vs. humans, human islets showed stronger signals for UEA-I, AAL, TJA-II, EEL, WFA, HPA, DBA, SBA and PTL-I, indicating that besides ABO blood type antigens, high levels of fucose and α-linked GalNAc are present. On the other hand, the high mannose form was very rich in the APIs.
GKO reduced alpha-linked GalNAc, despite negligible expression of the Gal antigen on WT-API. On the other hand, the high-mannose form was richer in both APIs than in healthy human islets. These results provide useful information for future studies.
KeywordsXenotransplantation Glycoantigen Lectin array Islets
The α-Gal knockout
Adult pig islets
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