title: Human IL-6, IL-17, IL-1β, and TNF-α differently regulate the expression of pro-inflammatory related genes, tissue factor, and swine leukocyte antigen class I in porcine aortic endothelial cells.
abstract: Pro-inflammatory cytokines play important pathological effects in various diseases and allotransplantation; however, their pathological role in xenotransplantation remains elusive. In pig-to-human cell or organ transplantation, whether porcine cells or organs are activated by human cytokines or not as an important question needs to be investigated.
  
  
  
  We investigated the effect of human IL-6, IFN-γ, IL-17, IL-1β, and TNF-α in xenotransplantation using several in vitro models and porcine aortic endothelial cells (PAECs) as target cells. The downstream signaling pathways activated by these cytokines were studied with Western blotting, the regulation of the pro-inflammatory related genes and pro-coagulation factor were assessed using real-time PCR or enzyme-linked immunosorbent assay, and swine leukocyte antigen (SLA) class I and SLA class II DR were analyzed by flow cytometry.
  
  
  
  We found that NF-κB and mitogen-activated protein kinases (MAPKs) were activated by recombinant human IL-17 (rhIL-17), rhIL-1β, and rhTNF-α, while rhIL-6 activated signal transducer and activator of transcription 3 (STAT3) in PAECs. The adhesion molecules (E-selectin, VCAM-1, and ICAM-1), pro-inflammatory gene (IL-6), chemokines (IL-8 and MCP-1), and the pro-coagulation factor (tissue factor) were induced by rhIL-17, rhIL-1β, and rhTNF-α, while rhIL-6 only increased the expression of MCP-1 and tissue factor. Using flow cytometry analysis, SLA class I was upregulated in PAECs after exposure to rhIL-1β and rhTNF-α, but not rhIL-6 or rhIL-17, whereas SLA class II DR could not be induced by rhIL-6, rhIL-17, rhIL-1β, or rhTNF-α, although it could by recombinant porcine IFN-γ (rpIFN-γ). Although activation of PAECs by rhIL-17 alone was not strong, rhIL-17 combined with rhTNF-α amplified the expression of E-selectin, IL-6, and IL-8. Unexpectedly, we found that tocilizumab, a humanized anti-human IL-6 receptor antibody, could not block rhIL-6-mediated STAT3 activation in PAECs. Human IFN-γ could not activate STAT1 or induce the downstream gene expression in PAECs, which was consistent with a previous report.
  
  
  
  In conclusion, our data suggest that human IL-6, IL-17, IL-1β, and TNF-α significantly activate PAECs and are likely to promote inflammation and coagulation reaction in response to xenograft.
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.|Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Hanchao
authors.identifier: 0000-0002-3225-801X
authors.initials: H
authors.lastname: Gao
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.|Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Lu
authors.identifier: 0000-0002-4173-9886
authors.initials: L
authors.lastname: Liu
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Xenotransplantation Program/Department of Surgery, The University of Alabama at Birmingham, Birmingham, AL, USA.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Hara
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.|Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Chen
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Xu
authors.affiliation: Medical Genetics Center, Jiangmen Maternity and Child health Care Hospital, Jiangmen, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Tang
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Wei
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Zesong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Xenotransplantation Program/Department of Surgery, The University of Alabama at Birmingham, Birmingham, AL, USA.
authors.forename: David K C
authors.initials: DKC
authors.lastname: Cooper
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Cai
authors.affiliation: Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
authors.forename: Lisha
authors.initials: L
authors.lastname: Mou
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000276:immunology; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects* / Q000276:immunology; D015870:Gene Expression / Q000187:drug effects*; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000276:immunology; D020381:Interleukin-17 / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000494:pharmacology*; D015850:Interleukin-6 / Q000494:pharmacology*; D013552:Swine; D013925:Thromboplastin / Q000235:genetics; D014183:Transplantation, Heterologous / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; C508585:IL1B protein, human; C508600:IL6 protein, human; D020381:Interleukin-17; D053583:Interleukin-1beta; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; C438668:swine leukocyte antigen; D013925:Thromboplastin
keywords: IFN-γ; IL-17; IL-1β; IL-6; TNF-α; porcine aortic endothelial cells; xenotransplantation