Double Transgenic Pigs with Combined Expression of Human α1,2-Fucosyltransferase and α-Galactosidase Designed to Avoid Hyperacute Xenograft Rejection View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2014-02-20

AUTHORS

Joanna Zeyland, Anna Woźniak, Barbara Gawrońska, Wojciech Juzwa, Jacek Jura, Agnieszka Nowak, Ryszard Słomski, Zdzisław Smorąg, Marlena Szalata, Urszula Mazurek, Daniel Lipiński

ABSTRACT

Hyperacute rejection (HAR) depends on the response of xenoreactive antibodies principally against porcine α-Gal epitope. Methods eliminating HAR include GGTA1 inactivation, regulation of the complement system and modification of the oligosaccharide structure of surface proteins in donor's cells. Transgenic animals designed for the purpose of xenotransplantation with single modification do not display full reduction of the α-Gal epitope level, which means that a accumulation of several modifications in one transgenic individual is needed. The aim of the study was to create a molecular and cytogenetic profile of a double transgenic animal with α1,2-fucosyltransferase and α-galactosidase expression. As a result of interbreeding of an individual with α1,2-fucosyltransferase expression with an individual with α-galactosidase expression 12 living piglets were obtained. PCR revealed the pCMVFUT gene construct was present in four individuals and pGAL-GFPBsd in three, including one with a confirmed integration of both the gene constructs. Fluorescence in situ hybridization confirmed the site of transgene integration, which corresponded to the mapping site of the transgenes which occurred in the parental generations. Karyotype analysis did not show any changes in the structure or the number of chromosomes (2n = 38, XX). As for the results pertaining to the single transgenic individuals, expression analysis demonstrated a high extent of α-Gal epitope level reduction on the surface of cells, whereas human serum cytotoxicity tests revealed the smallest decrease in longevity of cells in the obtained double transgenic individual (4.35 %). The tests suggest that the co-expression of both the transgenes leads to a considerable reduction of the α-Gal antigen level on the surface of cells and a decrease of xenotransplant immunogenicity. More... »

PAGES

411-422

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00005-014-0280-3

DOI

http://dx.doi.org/10.1007/s00005-014-0280-3

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1001161754

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24554032


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64 fluorescence
65 fucosyltransferase
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69 galactosidase
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71 gene constructs
72 generation
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75 humans
76 hybridization
77 hyperacute rejection
78 hyperacute xenograft rejection
79 immunogenicity
80 inactivation
81 individuals
82 integration
83 karyotype analysis
84 level reduction
85 levels
86 longevity
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88 mapping sites
89 method
90 modification
91 number
92 number of chromosomes
93 oligosaccharide structures
94 pCMVFUT gene construct
95 pGAL-GFPBsd
96 parental generation
97 piglets
98 pigs
99 porcine α
100 profile
101 protein
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103 purpose of xenotransplantation
104 reduction
105 regulation
106 rejection
107 response
108 results
109 serum cytotoxicity tests
110 single modification
111 single transgenic individuals
112 sites
113 situ hybridization
114 small decrease
115 structure
116 study
117 surface
118 surface of cells
119 surface proteins
120 system
121 test
122 transgene
123 transgene integration
124 transgenic animals
125 transgenic individuals
126 transgenic pigs
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