원문정보
초록
영어
To avoid hyperacute rejection of xenografts, α1,3-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of α1,3-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that α2,3-sialyltransferase (α2,3ST) and α2,6-sialyltransferase (α2,6ST) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of α-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of α2,6-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.
목차
INTRODUCTION
MATERIALS AND METHODS
Construction of α1,3-galactosyltransferase Gene KO Vector
PCR Analysis of Targeted Cells
Production of Heterozygote GalT KO Pigs
Sample Preparation and Protein Determination
RNA Isolation and Real-time RT-PCR
Assay of Glycosyltransferase Activity
Lectin Blotting
Analysis of Sialic Acid Contents
Statistical Analysis
RESULTS
Production of Heterozygote GalT KO Gnotobiotic Miniature Pigs
Heterozygote GalT KO Pig-Derived Liver Displays Higher Non-Gal Xenoantigen
DISCUSSION
REFERENCES
