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

Cross-Najafi, Arthur; Farag, Kristine; Isidan, Abdulkadir; Li, Wei; Zhang, Wenjun; Lin, Zhan; Walsh, Julia; Lopez, Kevin; Park, Yujin; Higgins, Nasra; Cooper, D. K. C.; Ekser, Burçin; Li, Ping

doi:10.3389/fimmu.2023.1217809

Cited by 6 publications

(4 citation statements)

References 49 publications

(64 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several studies have shown that porcine cells expressing non-classical HLA class I molecules, including HLA-E and/or HLA-G, can protect porcine cells from human NK cell cytotoxicity [35][36][37]. Further, our recent study indicated that co-expressing HLA-E and HLA-G on porcine cells effectively inhibits human NK cell activation [29].…”

Section: Discussionmentioning

confidence: 76%

See 1 more Smart Citation

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

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 76%

“…NK cell activation was assessed by CD107a degranulation assays as previously reported [29]. Buffy coats were purchased from Versiti Indiana Blood Center.…”

Section: Human Nk Cell Activation In Response To Porcine Endothelial ...mentioning

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…The use of genetic engineering to express on pig cells inhibitory ligands such as nonclassical human leukocyte antigens HLA-E and -G can be a strategy to overcome NK responses in xenotransplantation. Indeed, at least in vitro, co-expression of both these HLA molecules on pig cells is associated with a significant reduction of human NK cell degranulation [ 19 ].…”

Section: Immunological Challengesmentioning

confidence: 99%

Current challenges in xenotransplantation

Vadori,

Cozzi

2024

Current Opinion in Organ Transplantation

View full text Add to dashboard Cite

Purpose of review In recent years, the xenotransplantation science has advanced tremendously, with significant strides in both preclinical and clinical research. This review intends to describe the latest cutting-edge progress in knowledge and methodologies developed to overcome potential obstacles that may preclude the translation and successful application of clinical xenotransplantation. Recent findings Preclinical studies have demonstrated that it is now possible to extend beyond two years survival of primate recipients of life saving xenografts. This has been accomplished thanks to the utilization of genetic engineering methodologies that have allowed the generation of specifically designed gene-edited pigs, a careful donor and recipient selection, and appropriate immunosuppressive strategies. In this light, the compassionate use of genetically modified pig hearts has been authorized in two human recipients and xenotransplants have also been achieved in human decedents. Although encouraging the preliminary results suggest that several challenges have yet to be fully addressed for a successful clinical translation of xenotransplantation. These challenges include immunologic, physiologic and biosafety aspects. Summary Recent progress has paved the way for the initial compassionate use of pig organs in humans and sets the scene for a wider application of clinical xenotransplantation.

show abstract

“…Considerable discussion surrounds the reactivity of antibodies to human leukocyte antigens (HLAs) and swine leukocyte antigens (SLAs) in the immunomodulatory pathway post‐xenotransplantation. The co‐expression of HLA‐E and HLA‐G on genetically modified porcine endothelial cells was observed to attenuate human natural killer (NK) cell‐mediated degranulation 30 …”

Section: Modifications To the Xenograftmentioning

confidence: 99%