Human DAF on pig cells protects against human and non-human primate sera cytotoxicity mediated by exogenous or endogenous complement, as determined by flow cytometry

Díaz-Román, Tomás M.; Máñez, Rafael; Lopez-Pelaez, Eduardo; Centeno, Alberto; Moscoso, Isabel; Pértegaz, Sonia; Doménech, Nieves

doi:10.1016/j.trim.2006.03.008

Cited by 9 publications

(5 citation statements)

References 27 publications

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“…While it is not clear what role NK cells play in direct rejection of xenograft organs, they clearly could be important in rejection of cellular transplants, such as those involved in the induction of tolerance through mixed chimerism [40]. Also using in vitro assays, Diaz-Roman et al [48] have provided evidence that some of the lack of protection from complement-mediated lysis by hDAF in vivo in non-human primates could be due to a difference between the complement of humans versus that of non-human primates. Manzi et al [49] have shown that transfection of a pig endothelial cell line with a truncated form of human SCR1 is capable of protecting these cells from complement-mediated lysis by human natural antibodies.…”

Section: Xenotransplantation Using Transgenic Animalsmentioning

confidence: 99%

Genetic manipulation in pigs

Sachs¹,

Galli²

2009

Current Opinion in Organ Transplantation

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Purpose of Review Recent developments in the field of genetic engineering have made it possible to add, delete or exchange genes from one species to another. This technology has special relevance to the field of xenotransplantation, in which the elimination of a species-specific disparity could make the difference between success or failure of an organ transplant. This review focuses on developments in both the techniques and applications of genetically modified animals. Recent Findings Advances have been made using existing techniques for genetic modifications of swine and in the development of new, emerging technologies, including enzymatic engineering and the use of siRNA. Applications of the modified animals have provided evidence that genetically modified swine have the potential to overcome both physiologic and immunologic barriers that have previously impeded this field. Use of GalT-KO animals as donors have shown marked improvements in xenograft survivals. Summary Techniques for genetic engineering of swine have been directed toward avoiding naturally existing cellular and antibody responses to species-specific antigens. Organs from genetically engineered animals have enjoyed markedly improved survivals in non-human primates, especially in protocols directed toward the induction of tolerance, presumably by avoiding immunization to new antigens.

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Section: Xenotransplantation Using Transgenic Animalsmentioning

confidence: 99%

Genetic manipulation in pigs

Sachs¹,

Galli²

2009

Current Opinion in Organ Transplantation

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“…However, it must be highlighted that data collected independently for each transgenic specimen revealed clear differences in performance for the hemolysis assay. In that sense, it has been indicated that the level of protection decreases when low hDAF levels occur [ 34 ]. Taking this into account, our observations about performance differences at the hemolysis assay existing between specimens would be an attestation of the functional limits imposed by random integration transgenesis techniques, and so the need for methodologies, as our proposal, helping in discriminating highly profitable specimens from the rest.…”

Section: Discussionmentioning

confidence: 99%

Profiling Human CD55 Transgene Performance Assist in Selecting Best Suited Specimens and Tissues for Swine Organ Xenotransplantation

Martı́nez-Alarcón

Liarte

Quereda

et al. 2021

Biology

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Xenotransplantation of pig organs receives substantial attention for being comparable to human’s. However, compatibility constraints involving hyper-acute rejection (HAR) still block clinical applications. Transgenesis of human complement regulatory proteins has been proposed to overcome xenorejection. Pigs expressing human-CD55 have been widely tested in experimental surgery. Still, no standardized method has been developed to determine tissue expression of human decay-accelerating factor (DAF), hCD55’s product, or to predict the ability to overpass HAR. Here we describe objective procedures addressing this need. Organs and tissues from five hCD55 transgenic pigs were collected and classified according to their xenotransplantation value. The ability to overcome HAR was assessed by classical complement pathway hemolysis assays. Quantitative PCR mRNA expression and Western blot protein level studies were performed. Real-time cytotoxicity assays (RTCA) on fibroblast cultures exposed to baboon and human sera informed on longer-term rejection dynamics. While greater hCD55/DAF expression correlated with better performance, the results obtained varied among specimens. Interestingly, the individual with highest mRNA and protein levels showed positive feedback for hCD55 transcript after challenge with human and baboon sera. Moreover, hCD55 expression correlated to DAF levels in the liver, lung and intestine, but not in the heart. Moreover, we found significant correlations among valuable and non-valuable tissues. In sum, the methodology proposed allows us to characterize the hCD55 transgene functioning and performance. Moreover, the correlations found could allow us to predict hCD55/DAF expression in surrogate tissues, thus eliminating the need for direct biopsies, resulting in preservation of organ integrity before xenotransplantation.

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“…Although immunosuppressive agents are also effective for transplantation therapies in cats, anti-pig antibodies frequently trigger hyperacute rejection before the immunosuppressive agent starts to have its effects. In xenotransplantation procedures between humans and pigs, human DAF-expressing swine cells have been successfully used to prevent hyperacute rejection [ 36 ]. Therefore, the same strategy could be utilized for transplantation between pigs and cats.…”

Section: Discussionmentioning

confidence: 99%

Generation of a Felinized Swine Endothelial Cell Line by Expression of Feline Decay-Accelerating Factor

et al. 2015

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Embryonic stem cell research has facilitated the generation of many cell types for the production of tissues and organs for both humans and companion animals. Because ≥30% of pet cats suffer from chronic kidney disease (CKD), xenotransplantation between pigs and cats has been studied. For a successful pig to cat xenotransplant, the immune reaction must be overcome, especially hyperacute rejection. In this study, we isolated the gene for feline decay-accelerating factor (fDAF), an inhibitor of complement proteins, and transfected a swine endothelial cell line with fDAF to “felinize” the pig cells. These fDAF-expressing cells were resistant to feline serum containing anti-pig antibodies, suggesting that felinized pig cells were resistant to hyperacute rejection. Our results suggest that a “felinized” pig kidney can be generated for the treatment of CKD in cats in the future.

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Human DAF on pig cells protects against human and non-human primate sera cytotoxicity mediated by exogenous or endogenous complement, as determined by flow cytometry

Cited by 9 publications

References 27 publications

Genetic manipulation in pigs

Genetic manipulation in pigs

Profiling Human CD55 Transgene Performance Assist in Selecting Best Suited Specimens and Tissues for Swine Organ Xenotransplantation

Generation of a Felinized Swine Endothelial Cell Line by Expression of Feline Decay-Accelerating Factor

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