胡文宝, 潘登科, David K.C.Cooper, 等. hCD47诱导人巨噬细胞对猪内皮细胞的免疫耐受[J]. 器官移植, 2019, 10(2): 165-170. DOI: 10.3969/j.issn.1674-7445.2019.02.008
引用本文: 胡文宝, 潘登科, David K.C.Cooper, 等. hCD47诱导人巨噬细胞对猪内皮细胞的免疫耐受[J]. 器官移植, 2019, 10(2): 165-170. DOI: 10.3969/j.issn.1674-7445.2019.02.008
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Hu Wenbao, Pan Dengke, David K.C.Cooper, et al. hCD47 induces immune tolerance of human macrophages to porcine endothelial cells[J]. ORGAN TRANSPLANTATION, 2019, 10(2): 165-170. DOI: 10.3969/j.issn.1674-7445.2019.02.008
Citation: Hu Wenbao, Pan Dengke, David K.C.Cooper, et al. hCD47 induces immune tolerance of human macrophages to porcine endothelial cells[J]. ORGAN TRANSPLANTATION, 2019, 10(2): 165-170. DOI: 10.3969/j.issn.1674-7445.2019.02.008
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hCD47诱导人巨噬细胞对猪内皮细胞的免疫耐受

hCD47 induces immune tolerance of human macrophages to porcine endothelial cells

    摘要
  • 摘要:
      目的  探讨人CD47（hCD47）在诱导人巨噬细胞对猪内皮细胞免疫耐受中的作用。
      方法  将转染了pCDH-hCD47-FLAG质粒的猪髂总动脉内皮细胞（PIEC）设为pCDH-hCD47组；将转染了pCDH-FLAG空载体质粒的PIEC为pCDH组；将转染了hCD47-dN的PIEC设为pCDH-hCD47-dN组；将人脐静脉内皮细胞（HUVEC）设为阳性对照组。将细胞分别与人巨噬细胞共培养，检测信号调节蛋白α（SIRPα）的磷酸化情况以及人巨噬细胞对PIEC的杀伤作用。进一步从GT－/－及GT－/－/hCD47基因编辑猪中分离了猪主动脉血管内皮细胞（PAEC）, 并分析SIRPα的磷酸化情况及人巨噬细胞对PAEC的杀伤作用。
      结果  pCDH组细胞不能诱导SIRPα的磷酸化，而pCDH-hCD47组细胞与人巨噬细胞共培养10 min后便能够激活SIRPα的磷酸化，且随着共培养时间的延长，SIRPα的磷酸化程度也随之增强。pCDH-hCD47-dN组细胞不能激活SIRPα的磷酸化。人巨噬细胞对pCDH组细胞产生了明显的杀伤作用，pCDH-hCD47组细胞可明显抑制人巨噬细胞的杀伤作用（P < 0.05），而pCDH-hCD47-dN组细胞则不能抑制人巨噬细胞的杀伤作用。GT－/－-PAEC与人巨噬细胞共培养后，不能激活SIRPα的磷酸化；而GT－/－/hCD47-PAEC与人巨噬细胞共培养后则明显激活了SIRPα的磷酸化。人巨噬细胞对GT－/－-PAEC产生了明显的杀伤作用，GT－/－/hCD47-PAEC可明显抑制人巨噬细胞的杀伤作用（P < 0.05）。
      结论  在猪内皮细胞中表达hCD47可以通过激活SIRPα的磷酸化抑制人巨噬细胞对内皮细胞的杀伤作用。

     

    Abstract:
      Objective  To investigate the effect of human CD47 (hCD47) in inducing the immune tolerance of human macrophages to porcine endothelial cells.
      Methods  The porcine iliac endothelial cell (PIEC) transfected with pCDH-hCD47-FLAG plasmid was assigned into the pCDH-hCD47 group, PIEC transfected with pCDH-FLAG empty vector plasmid was assigned into the pCDH group, PIEC transfected with hCD47-dN was assigned into the pCDH-hCD47-dN group and human umbilical vein endothelial cell (HUVEC) was assigned into the positive control group. The cells were co-cultured with human macrophages to detect and analyze the phosphorylation of signal regulatory protein α (SIRPα) and the killing effect of human macrophages on PIEC. Furthermore, porcine arteriae endothelial cell (PAEC) was isolated from GT－/－ and GT－/－/ hCD 47 gene editing pigs to analyze the phosphorylation of SIRPα and the killing effect of human macrophages on PAEC.
      Results  The pCDH group cells could not induce the phosphorylation of SIRPα, whereas the pCDH-hCD47 group cells could activate the phosphorylation of SIRPα after 10 min co-culture with human macrophages, and the degree of phosphorylation of SIRPα was increased with the prolongation of the co-culture time. The pCDH-hCD47-dN group cells failed to activate the phosphorylation of SIRPα. Human macrophages exerted significant effect on killing the pCDH group cells. The pCDH-hCD47 group cells could evidently inhibit the killing effect of human macrophages (P < 0.05), whereas the pCDH-hCD47-dN cells failed to suppress the killing effect of human macrophages. GT－/－-PAEC could not activate the phosphorylation of SIRPα after co-culture with human macrophages. However, GT－/－/hCD47-PAEC significantly activated the phosphorylation of SIRPα after co-culture with human macrophages. Human macrophages exerted significant killing effect on GT－/－-PAEC, and GT－/－/hCD47-PAEC could obviously inhibit the killing effect of human macrophages (P < 0.05).
      Conclusions  The expression of hCD47 in the porcine endothelial cells can inhibit the killing effect of human macrophages on endothelial cells by activating the phosphorylation of SIRPα.

     

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