Lectin Array Analysis for Wild-Type and α-Gal-knockout Pig Islets Versus Healthy Human Islets

Surg Today. 2013 Dec;43(12):1439-47. doi: 10.1007/s00595-013-0569-6. Epub 2013 Apr 3.

Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylgalactosamine / analysis*
  • Animals
  • Antigens
  • Cells, Cultured
  • Disaccharides / analysis*
  • Galactans / analysis*
  • Galactosyltransferases / genetics*
  • Gene Knockout Techniques
  • Humans
  • Islets of Langerhans / immunology
  • Islets of Langerhans / metabolism*
  • Lectins*
  • Mannose
  • Polysaccharides / immunology
  • Protein Array Analysis / methods*
  • Swine
  • Transplantation, Heterologous

Substances

  • Antigens
  • Disaccharides
  • Galactans
  • Lectins
  • Polysaccharides
  • galactosyll(1,3)-N-acetylgalactosamine
  • Galactosyltransferases
  • Acetylgalactosamine
  • Mannose