Glycosphingolipid expression in pig aorta: Identification of possible target antigens for human natural antibodies

Abstract

Total non-acid glycosphingolipids were isolated from the aortas of more than 80 pigs. The glycolipids were separated by HPLC, analysed by thin-layer chromatography, and tested for reactivity with monoclonal anti-blood group antibodies. The fractions were structurally characterized by NMR spectroscopy and mass spectrometry. Reactivity with both antiblood group A and H antibodies was seen. The major glycosphingolipid constituents were globotriand globotetraosylceramides and blood group H pentaglycosylceramides based on type 1 and type 2 core saccharide chains. Globopentaosylceramides, blood group H hexaglycosylceramides based on type 4 chain, and blood group A hexaglycosylceramides based on type 1 core chain were also present. Two structures, that may be important targets for human antibodies initiating hyperacute rejection following pig to human xenotransplantation, were present as minor constituents compared to the blood group components. These were Galα1,3neolactotetraosylceramide and a Galα1,3Le x structure. A Le b /Y hexaglycosylceramide was also present.

Survey of glycoantigens in cells from α1-3galactosyltransferase knockout pig using a lectin microarray

“…Concerning glyco-antigens on the pig cell, before the a-Gal was knocked out, Bouhors et al, in their analysis of glycosphingolipids from the pig kidney in 1997, reported that the Gala1-3Lew x is a new epitope capable of being recognized by the human natural antibody (NA) [18]. Using a similar method of analysis, Hallberg et al also indicated that Gal a1-3Lew x and Gala1-3nLc4(neolactotetraosylceramid) were important targets for human NA [19]. In addition, quite recently, Kim et al reported that NeuGc-Gal-GlcNAc and Gala1-3Lew x were novel antigens, as evidenced by a structural analysis of N-glycans from the miniature pig kidney, using MALDI-TOF/MS and MS/MS [20].…”

Complement regulation in the GalT KO era

“…However, concerning Gala1-3Lew x , in 1997, Bouhors et al already reported it as a new epitope for the human natural antibody in their analysis of glycosphingolipids from the pig kidney [146]. Hallberg et al also indicated that Gal a1-3Lew x and Gala1-3nLc4 were important targets for the human natural antibody [147].…”

Structural analysis of α‐Gal and new non‐Gal carbohydrate epitopes from specific pathogen‐free miniature pig kidney

“…Since permethylation also improves the stability and sensitivity of the charged glycans, especially sialic acid, neutral and sialylated glycans were observed in positive mode as [M 1 Na] 1 ions. The identified sialylated Nglycan structures are listed in Table 1 [26,27] analyzed glycosphingolipid expressed in pig kidney and aorta and firstly reported the expression of Gala1,3Lewis x as well as a-Gal-terminated GSL in pig organ. However, the Gala1,3Lewis x epitopes have not been reported in N-glycans from pig organs.…”