Strategies to induce natural killer cell tolerance in xenotransplantation

Lopez, Kevin; Cross-Najafi, Arthur; Farag, Kristine; Obando, Benjamin; Thadasina, Deepthi; Isidan, Abdulkadir; Park, Yujin; Zhang, Wenjun; Ekser, Burçin; Li, Ping

doi:10.3389/fimmu.2022.941880

Cited by 5 publications

(4 citation statements)

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“…Expressing inhibitory ligands on porcine cells to induce human NK cell tolerance is a practical approach to protect xenografts from human NK cell-mediated destruction ( 43 ). We showed that co-expressing HLA-E and HLA-G on a genetically modified 5GKO cell line synergistically reduced human NK cell activation as compared to cells expressing either HLA-E or HLA-G alone as well as 5GKO parental cells.…”

Section: Discussionmentioning

confidence: 99%

Co-expression of HLA-E and HLA-G on genetically modified porcine endothelial cells attenuates human NK cell-mediated degranulation

et al. 2023

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Natural killer (NK) cells play an important role in immune rejection in solid organ transplantation. To mitigate human NK cell activation in xenotransplantation, introducing inhibitory ligands on xenografts via genetic engineering of pigs may protect the graft from human NK cell-mediated cytotoxicity and ultimately improve xenograft survival. In this study, non-classical HLA class I molecules HLA-E and HLA-G were introduced in an immortalized porcine liver endothelial cell line with disruption of five genes (GGTA1, CMAH, β4galNT2, SLA-I α chain, and β-2 microglobulin) encoding three major carbohydrate xenoantigens (αGal, Neu5Gc, and Sda) and swine leukocyte antigen class I (SLA-I) molecules. Expression of HLA-E and/or HLA-G on pig cells were confirmed by flow cytometry. Endogenous HLA-G molecules as well as exogenous HLA-G VL9 peptide could dramatically enhance HLA-E expression on transfected pig cells. We found that co-expression of HLA-E and HLA-G on porcine cells led to a significant reduction in human NK cell activation compared to the cells expressing HLA-E or HLA-G alone and the parental cell line. NK cell activation was assessed by analysis of CD107a expression in CD3-CD56+ population gated from human peripheral blood mononuclear cells. CD107a is a sensitive marker of NK cell activation and correlates with NK cell degranulation and cytotoxicity. HLA-E and/or HLA-G on pig cells did not show reactivity to human sera IgG and IgM antibodies. This in vitro study demonstrated that co-expression of HLA-E and HLA-G on genetically modified porcine endothelial cells provided a superior inhibition in human xenoreactive NK cells, which may guide further genetic engineering of pigs to prevent human NK cell mediated rejection.

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Section: Discussionmentioning

confidence: 99%

Co-expression of HLA-E and HLA-G on genetically modified porcine endothelial cells attenuates human NK cell-mediated degranulation

et al. 2023

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“…Future studies will focus on identifying and eliminating porcine ligands for human NKG2D receptor from porcine cells, as well as introducing HLA-E and HLA-G inhibitory ligands into porcine cells through genetic engineering. These strategies have great potential to block human NK cell activation and NK cell-mediated rejection [38]. Additionally, investigating the interplay between NK cells and other immune cell populations, such as macrophages and T cells, could provide a more comprehensive understanding of the immune response to xenogeneic cells.…”

Section: Discussionmentioning

confidence: 99%

Porcine UL-16 Binding Protein 1 Is Not a Functional Ligand for the Human Natural Killer Cell Activating Receptor NKG2D

Lopez,

Spence,

et al. 2023

Cells

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Natural killer (NK) cells play a vital role in xenotransplantation rejection. One approach to induce NK cell immune tolerance is to prevent the NK cell-mediated direct killing of porcine cells by targeting the interaction of the activating receptor NKG2D and its ligands. However, the identity of porcine ligands for the human NKG2D receptor has remained elusive. Previous studies on porcine UL-16 binding protein 1 (pULBP-1) as a ligand for human NKG2D have yielded contradictory results. The goal of the present study was to clarify the role of pULBP-1 in the immune response and its interaction with human NKG2D receptor. To accomplish this, the CRISPR/Cas9 gene editing tool was employed to disrupt the porcine ULBP-1 gene in a 5-gene knockout porcine endothelial cell line (GGTA1, CMAH, β4galNT2, SLA-I α chain, and β-2 microglobulin, 5GKO). A colony with two allele mutations in pULBP-1 was established as a 6-gene knockout pig cell line (6GKO). We found that pULBP-1-deficient pig cells exhibited a reduced binding capacity to human NKG2D-Fc, a recombinant chimera protein. However, the removal of ULBP-1 from porcine endothelial cells did not significantly impact human NK cell degranulation or cytotoxicity upon stimulation with the pig cells. These findings conclusively demonstrate that pULBP-1 is not a crucial ligand for initiating xenogeneic human NK cell activation.

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“…Nevertheless, we are just scratching the surface of the iceberg about the function and mechanisms of each type of cells. For instance, NK cells may play an effector role by releasing cytotoxicity granules against xenogeneic cells, or an affector role on other immune cells through cytokine secretion ( 117 ), and much work need to be carried out to promote xenograft acceptance by driving NK cells ( 118 ).…”

Section: Research Interestsmentioning

confidence: 99%

Developments in kidney xenotransplantation

Xu,

2024

Front. Immunol.

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The search for kidney xenografts that are appropriate for patients with end-stage renal disease has been ongoing since the beginning of the last century. The major cause of xenograft loss is hyperacute and acute rejection, and this has almost been overcome via scientific progress. The success of two pre-clinical trials of α1,3-galactosyltransferase gene-knockout porcine kidneys in brain-dead patients in 2021 triggered research enthusiasm for kidney xenotransplantation. This minireview summarizes key issues from an immunological perspective: the discovery of key xenoantigens, investigations into key co-stimulatory signal inhibition, gene-editing technology, and immune tolerance induction. Further developments in immunology, particularly immunometabolism, might help promote the long-term outcomes of kidney xenografts.

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Strategies to induce natural killer cell tolerance in xenotransplantation

Cited by 5 publications

References 96 publications

Co-expression of HLA-E and HLA-G on genetically modified porcine endothelial cells attenuates human NK cell-mediated degranulation

Co-expression of HLA-E and HLA-G on genetically modified porcine endothelial cells attenuates human NK cell-mediated degranulation

Porcine UL-16 Binding Protein 1 Is Not a Functional Ligand for the Human Natural Killer Cell Activating Receptor NKG2D

Developments in kidney xenotransplantation

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