title: Molecular immunology profiles of monkeys following xenografting with the islets and heart of α-1,3-galactosyltransferase knockout pigs.
abstract: Effective immunosuppression strategies and genetically modified animals have been used to prevent hyperacute and acute xenograft rejection; however, the underlying mechanisms remain unknown. In this study, we evaluated the expression of a comprehensive set of immune system-related genes (89 genes, including five housekeeping genes) in the blood of cynomolgus monkeys (~5 yr old) used as graft recipients, before and after the xenografting of the islets and heart from single and double α-1,3-galactosyltransferase (GalT) knockout (KO) pigs (<6 weeks old). The immunosuppressive regimen included administration of cobra venom factor, anti-thymocyte globulin, rituximab, and anti-CD154 monoclonal antibodies to recipients before and after grafting. Islets were xenografted into the portal vein in type 1 diabetic monkeys, and the heart was xenografted by heterotopic abdominal heart transplantation. Genes from recipient blood were analyzed using RT(2) profiler PCR arrays and the web-based RT(2) profiler PCR array software v.3.5. Recipients treated with immunosuppressive agents without grafting showed significant downregulation of CCL5, CCR4, CCR6, CD4, CD40LG, CXCR3, FASLG, CXCR3, FOXP3, GATA3, IGNG, L10, IL23A, TRAF6, MAPK8, MIF, STAT4, TBX21, TLR3, TLR7, and TYK2 and upregulation of IFNGR1; thus, genes involved in protection against viral and bacterial infection were downregulated, confirming the risk of infection. Notably, C3-level control resulted in xenograft failure within 2 days because of a 7- to 11-fold increase in all xenotransplanted models. Islet grafting using single GalT-KO pigs resulted in upregulation of CXCL10 and MX1, early inflammation, and acute rejection-associated signals at 2 days after xenografting. We observed at least 5-fold upregulation in recipients transplanted with islets grafts from single (MX1) or double (C3, CCR8, IL6, IL13, IRF6, CXCL10, and MX1) GalT-KO pigs after 77 days; single GalT-KO incurred early losses owing to immune attacks. Our results suggest that this novel, simple, non-invasive, and time-efficient procedure (requiring only 1.5 ml blood) for evaluating graft success, minimizing immune rejection, and blocking infection.
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea. ouksuna@hotmail.com, ocksa@korea.kr.
authors: Sun A
authors: SA
authors: Ock
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: Jungkyu
authors: J
authors: Lee
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: Keon Bong
authors: KB
authors: Oh
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: Seongsoo
authors: S
authors: Hwang
authors: Department of Surgery, Konkuk University School of Medicine, Seoul, Korea.
authors: Ik Jin
authors: IJ
authors: Yun
authors: Transplantation Center, Seoul National University Hospital, Seoul, Korea.|Designed Animal & Transplantation Research Institute, Institute of Green BioScience & Technology, Seoul National University, Pyeongchang, Gangwon-do, Korea.
authors: Curie
authors: C
authors: Ahn
authors: Department of Cardiothoracic Surgery, Konkuk University School of Medicine, Seoul, Korea.
authors: Hyun Keun
authors: HK
authors: Chee
authors: Transplantation Center, Seoul National University Hospital, Seoul, Korea.
authors: Hwajung
authors: H
authors: Kim
authors: Department of Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors: Jae Berm
authors: JB
authors: Park
authors: Department of Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors: Sung Joo
authors: SJ
authors: Kim
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: Youngim
authors: Y
authors: Kim
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: Gi-Sun
authors: GS
authors: Im
authors: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun, Jeollabuk-do, Korea.
authors: EungWoo
authors: E
authors: Park
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D005700:Galactosyltransferases / Q000172:deficiency / Q000276:immunology*; D006084:Graft Rejection / Q000188:drug therapy / Q000276:immunology; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D016027:Heart Transplantation; D064593:Heterografts / Q000276:immunology*; D007165:Immunosuppression Therapy / Q000379:methods; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016381:Islets of Langerhans Transplantation; D008252:Macaca fascicularis; D013552:Swine; D014183:Transplantation, Heterologous / Q000379:methods
chemical_list: D007166:Immunosuppressive Agents; D005700:Galactosyltransferases
keywords: immune response; monkey; pig; xenotransplantation; α-1,3-galactosyltransferase