Frontiers in Veterinary Science (Apr 2022)
Production of Triple-Gene (GGTA1, B2M and CIITA)-Modified Donor Pigs for Xenotransplantation
- Kaixiang Xu,
- Kaixiang Xu,
- Kaixiang Xu,
- Honghao Yu,
- Shuhan Chen,
- Shuhan Chen,
- Shuhan Chen,
- Yaxuan Zhang,
- Yaxuan Zhang,
- Yaxuan Zhang,
- Jianxiong Guo,
- Jianxiong Guo,
- Chang Yang,
- Chang Yang,
- Deling Jiao,
- Deling Jiao,
- Deling Jiao,
- Tien Dat Nguyen,
- Tien Dat Nguyen,
- Tien Dat Nguyen,
- Heng Zhao,
- Heng Zhao,
- Heng Zhao,
- Jiaoxiang Wang,
- Jiaoxiang Wang,
- Jiaoxiang Wang,
- Taiyun Wei,
- Taiyun Wei,
- Honghui Li,
- Honghui Li,
- Honghui Li,
- Baoyu Jia,
- Baoyu Jia,
- Baoyu Jia,
- Muhammad Ameen Jamal,
- Muhammad Ameen Jamal,
- Muhammad Ameen Jamal,
- Hong-Ye Zhao,
- Hong-Ye Zhao,
- Xingxu Huang,
- Hong-Jiang Wei,
- Hong-Jiang Wei,
- Hong-Jiang Wei,
- Hong-Jiang Wei
Affiliations
- Kaixiang Xu
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Kaixiang Xu
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Kaixiang Xu
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Honghao Yu
- College of Biotechnology, Guilin Medical University, Guilin, China
- Shuhan Chen
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Shuhan Chen
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Shuhan Chen
- College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China
- Yaxuan Zhang
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Yaxuan Zhang
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Yaxuan Zhang
- College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China
- Jianxiong Guo
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Jianxiong Guo
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Chang Yang
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Chang Yang
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Deling Jiao
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Deling Jiao
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Deling Jiao
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Tien Dat Nguyen
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Tien Dat Nguyen
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Tien Dat Nguyen
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Heng Zhao
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Heng Zhao
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Heng Zhao
- College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China
- Jiaoxiang Wang
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Jiaoxiang Wang
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Jiaoxiang Wang
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Taiyun Wei
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Taiyun Wei
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Honghui Li
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Honghui Li
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Honghui Li
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Baoyu Jia
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Baoyu Jia
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Baoyu Jia
- College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China
- Muhammad Ameen Jamal
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Muhammad Ameen Jamal
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Muhammad Ameen Jamal
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Hong-Ye Zhao
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Hong-Ye Zhao
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Xingxu Huang
- School of Life Science and Technology, ShanghaiTech University, Shanghai, China
- Hong-Jiang Wei
- Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China
- Hong-Jiang Wei
- Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University, Kunming, China
- Hong-Jiang Wei
- Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China
- Hong-Jiang Wei
- College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China
- DOI
- https://doi.org/10.3389/fvets.2022.848833
- Journal volume & issue
-
Vol. 9
Abstract
Activation of human immune T-cells by swine leukocyte antigens class I (SLA-I) and class II (SLA-II) leads to xenograft destruction. Here, we generated the GGTA1, B2M, and CIITA (GBC) triple-gene-modified Diannan miniature pigs, analyzed the transcriptome of GBC-modified peripheral blood mononuclear cells (PBMCs) in the pig's spleen, and investigated their effectiveness in anti-immunological rejection. A total of six cloned piglets were successfully generated using somatic cell nuclear transfer, one of them carrying the heterozygous mutations in triple genes and the other five piglets carrying the homozygous mutations in GGTA1 and CIITA genes, but have the heterozygous mutation in the B2M gene. The autopsy of GBC-modified pigs revealed that a lot of spot bleeding in the kidney, severe suppuration and necrosis in the lungs, enlarged peripulmonary lymph nodes, and adhesion between the lungs and chest wall were found. Phenotyping data showed that the mRNA expressions of triple genes and protein expressions of B2M and CIITA genes were still detectable and comparable with wild-type (WT) pigs in multiple tissues, but α1,3-galactosyltransferase was eliminated, SLA-I was significantly decreased, and four subtypes of SLA-II were absent in GBC-modified pigs. In addition, even in swine umbilical vein endothelial cells (SUVEC) induced by recombinant porcine interferon gamma (IFN-γ), the expression of SLA-I in GBC-modified pig was lower than that in WT pigs. Similarly, the expression of SLA-II DR and DQ also cannot be induced by recombinant porcine IFN-γ. Through RNA sequencing (RNA-seq), 150 differentially expressed genes were identified in the PBMCs of the pig's spleen, and most of them were involved in immune- and infection-relevant pathways that include antigen processing and presentation and viral myocarditis, resulting in the pigs with GBC modification being susceptible to pathogenic microorganism. Furthermore, the numbers of human IgM binding to the fibroblast cells of GBC-modified pigs were obviously reduced. The GBC-modified porcine PBMCs triggered the weaker proliferation of human PBMCs than WT PBMCs. These findings indicated that the absence of the expression of α1,3-galactosyltransferase and SLA-II and the downregulation of SLA-I enhanced the ability of immunological tolerance in pig-to-human xenotransplantation.
Keywords