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Human galectin‐9 on the porcine cells affects the cytotoxic activity of M1‐differentiated THP ‐1 cells through inducing a shift in M2‐differentiated THP ‐1 cells
Author(s) -
Jung Sung Han,
Hwang Jeong Ho,
Kim Sang Eun,
Kim Young Kyu,
Park Hyo Chang,
Lee Hoon Taek
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.12305
Subject(s) - cytotoxic t cell , thp1 cell line , microbiology and biotechnology , immune system , biology , flow cytometry , cell culture , immunology , chemistry , in vitro , biochemistry , genetics
Background In xenotransplantation, immune rejection by macrophages occurs rapidly and remains a major obstacle. Studies to control immune rejection in macrophages have been continuing to date. Recent studies have reported that human galectin‐9 ( hG al‐9) can regulate the function of regulatory T cells (Treg), as well as cytotoxicity T cells ( CTL ) and natural killer cells ( NK ). Although the effect of hG al‐9 on lymphocytes has been well studied, the relationship between hG al‐9 and myeloid cells has been scarcely studied. Methods To confirm the decreased cytotoxic activity effect by hG al‐9 in M1‐differentiated THP ‐1 cells, we established the hG al‐9 expressing transgenic porcine cell line. hG al‐9 si RNA was transfected to transgenic cells and recombinant hG al‐9 (rhGal‐9) was treated to co‐culturing condition, and then, flow cytometry assay was conducted for analyzing the cytotoxic activity of M1‐differentiated THP ‐1 cells. Related inflammatory cytokines ( IL ‐1β, IL ‐10, TNF ‐α, IL ‐6, IL ‐12, IL ‐23, and TGF ‐β) and related enzymes ( iNOS and Arginase 1) were analyzed by qPCR and Western blot assay. To identify the shift in M1/M2‐differentiated THP ‐1 cells, expression levels of CCR 7, CD 163, iNOS , and Arginase 1 and population of M2 marker positive cells were analyzed. Results The expression levels of pro‐inflammatory cytokines in M1‐differentiated THP ‐1 cells co‐cultured with hG al‐9‐expressing porcine kidney epithelial cells were decreased, but not in co‐cultured THP ‐1 cells. However, the expression levels of anti‐inflammatory cytokines were also increased in co‐cultured M1‐differentiated THP ‐1 cells. The cytotoxicity effect of M1‐differentiated THP ‐1 cells on transgenic cells was decreased while the expression levels of anti‐inflammatory cytokines and M2 macrophages‐related molecules were increased. M2 differentiation program was turned on while M1 program was turned down by enhancing the phosphorylation levels of Akt and PI 3K and the expression level of PPAR ‐γ. Due to these changes, differentiation of M2 program was enhanced in cells co‐cultured with hG al‐9. Conclusions These data suggested that hG al‐9 has a reduction in M1‐differentiated THP ‐1 cell cytotoxic activity‐related acute immune rejection in pig‐to‐human xenotransplantation in addition to its role in lymphoid lineage immune cell regulation.