HLA-G1, but Not HLA-G3, Suppresses Human Monocyte/Macrophage-mediated Swine Endothelial Cell Lysis
HLA-G1, but Not HLA-G3, Suppresses Human Monocyte/Macrophage-mediated Swine Endothelial Cell Lysis
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DOI：
10.1016/j.transproceed.2015.10.070
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发表时间：
2016-05-01
影响因子：
0.9
通讯作者：
Miyagawa, S.
中科院分区：
文献类型：
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作者：
Eguchi, H.;Maeda, A.;Miyagawa, S.
检测Ib类分子HLA-G1对单核/巨噬细胞介导的细胞毒作用的抑制作用。用流式细胞术检测体外培养的巨噬细胞和佛波酯(PMA)激活的THP-1细胞上与人类白细胞抗原G、免疫球蛋白样转录物2(ILT2)、ILT4和KIR2DL4(CD158d)相互作用的抑制受体的表达。制备人类白细胞抗原-G1、人类白细胞抗原-G3、人类白细胞抗原-E和人β2-微球蛋白的基因片段，将其导入猪内皮细胞和巨噬细胞，检测THP-1细胞对猪内皮细胞的杀伤作用。体外培养的巨噬细胞不仅表达ILT2和ILT4，还表达CD158d。PEC上的转基因HLA-G1对巨噬细胞介导的细胞毒作用有明显的抑制作用，与转基因的HL A-E相当。HLAG1在PEC细胞表面有明显的表达，而HLAG3的表达水平较低，主要分布在细胞间隙。另一方面，PMA激活的THP-1细胞比体外产生的巨噬细胞更少表达这些抑制分子。因此，与体外产生的巨噬细胞相比，PECS表面的HLA-G1对THP-1细胞介导的细胞毒作用具有显著的抑制作用，但相对较小。这些结果表明，转基因猪通过产生人类白细胞抗原-G1，而不是人类白细胞抗原-G3，可以保护移植物免受单核/巨噬细胞介导的细胞毒性。
The inhibitory function of HLA-G1, a class Ib molecule, on monocyte/macrophage mediated cytotoxicity was examined. The expression of inhibitory receptors that interact with HLA-G, immunoglobulin-like transcript 2 (ILT2), ILT4, and KIR2DL4 (CD158d) on in vitro-generated macrophages obtained from peripheral blood mononuclear cells and the phorbol 12-myristate 13-acetate (PMA)-activated THP-1 cells were examined by flow cytometry. cDNAs of HLA-G1, HLA-G3, HLA-E, and human beta 2-microglobulin were prepared, transfected into pig endothelial cells (PECs), and macrophage- and the THP-1 cell mediated PEC cytolysis was then assessed. In vitro generated macrophages expressed not only ILT2 and ILT4 but CD158d as well. The transgenic HLA-G1 on PEC indicated a significant suppression in macrophage-mediated cytotoxicity, which was equivalent to that of transgenic HLA-E. HLA-G1 was clearly expressed on the cell surface of PEC, whereas the levels of HLA-G3 were much lower and remained in the intracellular space. On the other hand, the PMA-activated THP-1 cell was less expressed these inhibitory molecules than in vitro generated macrophages. Therefore, the HLA-G1 on PECs showed a significant but relatively smaller suppression to THP-1 cell mediated cytotoxicity compared to in vitro generated macrophages. These results indicate that by generating HLA-G1, but not HLA-G3, transgenic pigs can protect porcine grafts from monocyte/macrophage-mediated cytotoxicity.