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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
Author(s) -
Gao Hanchao,
Liu Lu,
Zhao Yanli,
Hara Hidetaka,
Chen Pengfei,
Xu Jia,
Tang Jia,
Wei Ling,
Li Zesong,
Cooper David K.C.,
Cai Zhiming,
Mou Lisha
Publication year - 2017
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12291
Subject(s) - tissue factor , xenotransplantation , proinflammatory cytokine , chemokine , biology , microbiology and biotechnology , allotransplantation , tumor necrosis factor alpha , immunology , cytokine , interleukin 10 , transplantation , inflammation , medicine , coagulation
Background 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. Methods 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. Results We found that NF ‐κB and mitogen‐activated protein kinases ( MAPK s) were activated by recombinant human IL ‐17 (rh IL ‐17), rh IL ‐1β, and rh TNF ‐α, while rh IL ‐6 activated signal transducer and activator of transcription 3 ( STAT 3) in PAEC s. 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 rh IL ‐17, rh IL ‐1β, and rh TNF ‐α, while rh IL ‐6 only increased the expression of MCP ‐1 and tissue factor. Using flow cytometry analysis, SLA class I was upregulated in PAEC s after exposure to rh IL ‐1β and rh TNF ‐α, but not rh IL ‐6 or rh IL ‐17, whereas SLA class II DR could not be induced by rh IL ‐6, rh IL ‐17, rh IL ‐1β, or rh TNF ‐α, although it could by recombinant porcine IFN ‐γ (rp IFN ‐γ). Although activation of PAEC s by rh IL ‐17 alone was not strong, rh IL ‐17 combined with rh TNF ‐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 rh IL ‐6‐mediated STAT 3 activation in PAEC s. Human IFN ‐γ could not activate STAT 1 or induce the downstream gene expression in PAEC s, which was consistent with a previous report. Conclusion In conclusion, our data suggest that human IL ‐6, IL ‐17, IL ‐1β, and TNF ‐α significantly activate PAEC s and are likely to promote inflammation and coagulation reaction in response to xenograft.