Toll样受体7活化调节脐带间充质干细胞免疫原性的实验研究  

The Study of Increased Immunogenicity of UCMSC Stimulated by TLR7 Agonist CL264

作　　者：杨海青[1] 张盛[2] 况小串 杨银忠[3] 邱发麒[1] 

机构地区：[1]四川省医学科学院.四川省人民医院城东病区内一科,成都610041 [2]四川省医学科学院.四川省人民医院城东病区质管科,成都610041 [3]四川省医学科学院.四川省人民医院城东病区检验科,成都610041

出　　处：《四川大学学报（医学版）》2015年第2期191-196,共6页Journal of Sichuan University(Medical Sciences)

摘　　要：目的研究脐带间充质干细胞(umbilical cord mesenchymal stem cell,UCMSC)的Toll样受体7(toll like receptor 7,TLR7)通路活化后,是否会增加UCMSC的免疫原性。方法用TLR7的激动剂CL264刺激UCMSC后,用流式细胞术检测UCMSC表面共刺激抗原〔人白细胞抗原-E(HLA-E),CD80,CD86〕和干细胞标志物(CD29,CD59,CD90)的表达变化;定量PCR检测CL264刺激UCMSC后多个免疫相关因子的表达变化;细胞分化实验检测TLR7活化后对UCMSC分化能力的影响;分离健康人外周血单个核细胞(peripheral blood mononuclear cells,PBMC)并与UCMSC共培养,用细胞杀伤实验检测免疫细胞对UCMSC的免疫杀伤效应。结果 CL264活化TLR7通路后,可明显刺激UCMSC表面共刺激分子CD86和HLA-E的表达;定量PCR结果表明CL264诱导多个促炎症相关因子〔白细胞介素(IL)-1β,IL-6,IL-8,IL-10,干扰素(IFN)-β,IFN-γ,核因子-κB(NF-κB),转化生长因子-β(TGF-β)〕表达,并抑制多个干细胞标志物〔Kruppel样因子4(Klf4),巢蛋白(Nestin),胚胎干细胞关键蛋白质(Sox2),RNA结合蛋白质(Lin28)〕表达;TLR7通路活化不影响UCMSC的分化潜能;CL264活化TLR7通路后增加免疫细胞对UCMSC的免疫攻击。结论 TLR7激动剂CL264可增加UCMSC的免疫原性。Objective To study the change of immune status of umbilical cord mesenchymal stem cells（UCMSC）stimulated by toll like receptor 7（TLR7）agonist CL264.Methods TLR7 specific ligand CL264 was used to stimulate the UCMSC.Flow cytometry was conducted to assay the expression of co-stimulators 〔human leukocyte antigen（HLA）-E,CD80 and CD86〕and surface markers of stem cells（CD29,CD59 and CD90）.Quantitative PCR was applied to measure the expression variation of immune-related molecules and stem cell markers.Cell differentiation experiment was used to study the change of differentiation ability of UCMSC upon CL264 stimulation.Peripheral blood mononuclear cells（PBMC）were isolated from healthy human and then cocultured with UCMSC in the presence of CL264.Cytotoxicity assay was used to measure the attack of PBMC to UCMSC.Results Expression of co-stimulatory molecules CD86 and HLA-E were enhanced in UCMSC upon CL264 stimulation.Real-time PCR indicated that many pro-inflammatory molecules 〔interleukin（IL）-1β,IL-6,IL-8,IL-10,interferon（IFN）-β,IFN-γ,nuclear factor-κB（NF-κB）,transforming growth factor-β（TGF-β）〕were induced in the presence of CL264 while the expression of stem cells markers were inhibited 〔Kruppel-like factor-4（Klf4）,Nestin,SRY-related high-mobility-group-box protein-2（Sox2）,Lin28〕.Activation of TLR7 also increased the immune attack of PBMC on UCMSC.Our study also indicated that the treatment of CL264 did not influence the differentiation ability of UCMSC.Conclusion TLR7 agonist CL264could increase the immunogenicity of UCMSC.

关 键 词：脐带间充质干细胞 TOLL样受体7 CL264 免疫原性 

分 类 号：R392.12[医药卫生—免疫学]