TLR1/2通路活化对脐带间充质干细胞免疫状态影响的研究  

The Study of Altered Immune Status of UCMSC upon the Activation of TLR1/2 Pathway

作　　者：马宇[1] 张立[2] 江婷[3] 刘志强 李全生[3] 

机构地区：[1]四川大学华西医院甲状腺乳腺外科,成都610041 [2]四川大学华西医院病理研究室,成都610041 [3]四川大学华西医院胆道外科,成都610041 [4]德阳市第六人民医院外二科,德阳618000

出　　处：《四川大学学报（医学版）》2015年第5期679-683,701,共6页Journal of Sichuan University(Medical Sciences)

摘　　要：目的研究Toll样受体1/2(Toll like receptor 1/2,TLR1/2)活化后是否改变脐带间充质干细胞(umbilical cord mesenchymal stem cell,UCMSC)的免疫状态。方法UCMSC与外周血单个核细胞(peripheral blood mononuclear cell,PBMC)共培养体系中加入TLR1/2激动剂,用流式细胞术检测PBMC的增殖,并检测PBMC对UCMSC的杀伤作用;检测TLR1/2通路活化后UCMSC表面共刺激分子和干细胞标志物的表达变化;荧光定量PCR检测UCMSC多个免疫相关因子表达变化;对UCMSC进行定向诱导分化,检测TLR1/2活化对UCMSC分化能力的影响。结果 TLR1/2通路活化可促进PBMC增殖,并增强PBMC对UCMSC的免疫杀伤效应,但对UCMSC表面共刺激分子〔CD80/CD86/人类白细胞抗原(HLA)-E〕和干细胞标志物(CD29/CD59/CD90)无影响;荧光定量PCR结果表明UCMSC中多个免疫相关因子被TLR1/2明显诱导〔白细胞介素(IL-2,IL-6,IL-10,IL-12),干扰素-γ(IFN-γ),核因子-κB(NF-κB),肿瘤坏死因子-α(TNF-α)〕;TLR1/2不影响UCMSC的定向分化能力。结论 TLR1/2可改变UCMSC的免疫状态,在一定程度诱导针对UCMSC的免疫攻击。Objective To study whether the activation of Toll like receptor 1/2 （TLR1/2） has influence on the immune status of umbilical cord mesenchymal stem cell （UCMSC）. Methods TLR1/2 agonist Pam3CSK4 was conducted in UCMSC-peripheral blood mononuclear cell （PBMC） co-culture system. Flow cytometry and lactate dehydrogenase （LDH） detection were used to measure the proliferation of PBMC and immune attack of PBMC to UCMSC, respectively. The detection of expressions of co-stimulator and stem cell markers of UCMSC upon TLR1/ 2 activation was completed by flow cytometry. Real-time PCR was introduced to assay the expression of many immune-related molecules in UCMSC. Cell differentiation staining was conducted to study the change of differentiation ability of UCMSC. Results Activation of TLR1/2 pathway in UCMSC increased the proliferation of PBMC and attack of PBMC to UCMSC, but without influenced the expressions of co-stimulator and stem cell markers. The results of real-time PCR showed that many immune-related molecules were dramatically induced in UCMSC upon TLR1/2 activation, such as Cinterleukin （IL） -2, IL-6, IL-10, IL-12, interferon-γ （IFN-γ）, nuclear factor-κB （NF-κB）, tumor necrosis factor-α （TNF-α）]. However, the activation of TLR1/2 had no influence on the differentiation ability of UCMSC. Conclusion Activation of TLR1/2 altered the immune status of UCMSC, including increase the immune attack to UCMSC.

关 键 词：脐带间充质干细胞 Toll样受体1/2 免疫相关因子 免疫状态 

分 类 号：R392.1[医药卫生—免疫学]